Baculovirus synergism for improved management of false codling moth Thaumatotibia leucotreta Meyr. (Lepidoptera: Tortricidae)
- Authors: Taylor, David Graham
- Date: 2021-04
- Subjects: Baculoviruses , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Biological control , Biological pest control agents , Citrus -- Diseases and pests , Codling moth , Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV)
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/176942 , vital:42774
- Description: Baculoviruses are an environmentally friendly and effective agent for managing lepidopteran pests. This includes the management of Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), a serious pest of citrus in Southern Africa and a major threat to the South African citrus export industry. For more than 15 years, CrleGV-SA- based biopesticides have been used as part of an integrated pest management strategy for the control of T. leucotreta in citrus orchards in South Africa, under the names Cryptogran™ and Cryptex®. While these biopesticides have been effective during this period, there are some areas in which baculovirus use could potentially be improved. Baculoviruses are notoriously slow to kill in comparison to chemical-based pesticides, and lately, pest resistance to baculoviruses has become a major concern with the development of resistance by Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) to its granulovirus occurring in the field in Europe. The consistent use of CrleGV-SA for more than 15 years in the field has raised concern that T. leucotreta could develop resistance to this virus, and has made it necessary to alter baculovirus-based management strategies to prevent this from occurring. A second baculovirus, Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV), has recently been isolated and was shown to be effective against T. leucotreta. However, the interactions between CrleGV-SA and CrpeNPV are not yet understood and so it is important to test these interactions before both viruses are applied on the same orchards. Not only is it important to know whether these viruses could negatively impact each other, but it is also important to test whether they could interact synergistically. A synergistic interaction could not only provide a potential tool for the management of resistance, but it could also be exploited to improve baculovirus-based management of T. leucotreta. In this study, a stock of CrleGV-SA was purified by glycerol gradient centrifugation from T. leucotreta cadavers, while a stock of CrpeNPV purified from Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae) cadavers was provided by River Bioscience (Pty) Ltd. These stocks were screened for purity by a multiplex polymerase chain reaction (mPCR) protocol designed to detect CrleGV-SA and CrpeNPV. The occlusion body (OB) density was then calculated using darkfield microscopy and a counting chamber. Both stocks were shown to be pure within the limits of the mPCR protocol, and the CrleGV-SA and CrpeNPV stocks were calculated to contain 3.08 × 1011 OBs/mL and 1.92 × 1011 OBs/mL respectively The first aspect of the interaction between CrleGV-SA and CrpeNPV that was investigated was the dose mortality, in terms of lethal concentration. This was calculated using 7-day surface-dose biological assays for each virus and a 1:1 mixture of OBs of the two against T. leucotreta neonates. The lethal concentrations of each treatment required to kill 50 % of larvae (LC50) and 90 % of larvae (LC90) for each treatment were then calculated and compared using a probit regression. The mixed infection performed significantly better than either virus by itself, while each virus by itself did not differ significantly from the other. The LC50 for CrleGV-SA, CrpeNPV and the mixed infection were 1.53 × 104 OBs/mL, 1.15 × 104 OBs/mL and 4.38 × 103 OBs/mL respectively. The LC90 of CrleGV-SA, CrpeNPV and the mixed infection were calculated to be 4.10 × 105 OBs/mL, 1.05 × 105 OBs/mL, and 4.09 × 104 OBs/mL respectively. The second aspect of the interaction between CrleGV-SA and CrpeNPV that was investigated was the speed of kill. A time-response biological assay protocol was created that allowed for effective observation of the larvae. This was then used to generate time-mortality data that were analysed by a logit regression function to calculate and compare the treatments at the time of 50 % larval mortality (LT50) and the time of 90 % mortality (LT90). Each virus by itself did not differ significantly from the other, while the mixed infection took significantly longer to kill 50 % and 90 % of the larvae, suggesting that there is competition for resources between viruses during the secondary, systemic phase of infection. The LT50 for CrleGV-SA, CrpeNPV and the mixed infection were 117.5 hours, 113.5 hours and 139.0 hours respectively. The LT90 for CrleGV-SA, CrpeNPV and the mixed infection were 153.2 hours, 159.3, and 193.4 hours respectively. Finally, the composition of OBs recovered from the cadavers produced by the time-response biological assays were investigated by mPCR. A method for extracting gDNA from OBs in neonate-sized T. leucotreta larvae is described. The presence of CrpeNPV along with CrleGV-SA was noted in 4 out of 9 larvae inoculated with only CrleGV-SA. The presence of CrleGV-SA as well as CrpeNPV was noted in all but one larva inoculated with only CrpeNPV, and both CrleGV-SA and CrpeNPV were noted in all but one larva inoculated with a 1:1 mixture of the two, with one larva only being positive for CrleGV-SA. This suggests either stock contamination or the presence of covert infections of CrleGV-SA and CrpeNPV in the T. leucotreta population used in this study. This is the second study to report an improved lethal concentration of a mixed infection of CrleGV-SA and CrpeNPV against T. leucotreta neonates, and the first study to report the slower speed of kill of a mixed infection of CrleGV-SA and CrpeNPV against T. leucotreta neonates. While the improved lethal concentration of the mixed infection is a promising step in the future improvement of baculovirus-based biopesticides, it is at the cost of a slower speed of kill. , Thesis (MSc) -- Faculty of Science, Department of Zoology and Entomology, 2021
- Full Text:
- Date Issued: 2021-04
- Authors: Taylor, David Graham
- Date: 2021-04
- Subjects: Baculoviruses , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Biological control , Biological pest control agents , Citrus -- Diseases and pests , Codling moth , Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV)
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/176942 , vital:42774
- Description: Baculoviruses are an environmentally friendly and effective agent for managing lepidopteran pests. This includes the management of Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), a serious pest of citrus in Southern Africa and a major threat to the South African citrus export industry. For more than 15 years, CrleGV-SA- based biopesticides have been used as part of an integrated pest management strategy for the control of T. leucotreta in citrus orchards in South Africa, under the names Cryptogran™ and Cryptex®. While these biopesticides have been effective during this period, there are some areas in which baculovirus use could potentially be improved. Baculoviruses are notoriously slow to kill in comparison to chemical-based pesticides, and lately, pest resistance to baculoviruses has become a major concern with the development of resistance by Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) to its granulovirus occurring in the field in Europe. The consistent use of CrleGV-SA for more than 15 years in the field has raised concern that T. leucotreta could develop resistance to this virus, and has made it necessary to alter baculovirus-based management strategies to prevent this from occurring. A second baculovirus, Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV), has recently been isolated and was shown to be effective against T. leucotreta. However, the interactions between CrleGV-SA and CrpeNPV are not yet understood and so it is important to test these interactions before both viruses are applied on the same orchards. Not only is it important to know whether these viruses could negatively impact each other, but it is also important to test whether they could interact synergistically. A synergistic interaction could not only provide a potential tool for the management of resistance, but it could also be exploited to improve baculovirus-based management of T. leucotreta. In this study, a stock of CrleGV-SA was purified by glycerol gradient centrifugation from T. leucotreta cadavers, while a stock of CrpeNPV purified from Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae) cadavers was provided by River Bioscience (Pty) Ltd. These stocks were screened for purity by a multiplex polymerase chain reaction (mPCR) protocol designed to detect CrleGV-SA and CrpeNPV. The occlusion body (OB) density was then calculated using darkfield microscopy and a counting chamber. Both stocks were shown to be pure within the limits of the mPCR protocol, and the CrleGV-SA and CrpeNPV stocks were calculated to contain 3.08 × 1011 OBs/mL and 1.92 × 1011 OBs/mL respectively The first aspect of the interaction between CrleGV-SA and CrpeNPV that was investigated was the dose mortality, in terms of lethal concentration. This was calculated using 7-day surface-dose biological assays for each virus and a 1:1 mixture of OBs of the two against T. leucotreta neonates. The lethal concentrations of each treatment required to kill 50 % of larvae (LC50) and 90 % of larvae (LC90) for each treatment were then calculated and compared using a probit regression. The mixed infection performed significantly better than either virus by itself, while each virus by itself did not differ significantly from the other. The LC50 for CrleGV-SA, CrpeNPV and the mixed infection were 1.53 × 104 OBs/mL, 1.15 × 104 OBs/mL and 4.38 × 103 OBs/mL respectively. The LC90 of CrleGV-SA, CrpeNPV and the mixed infection were calculated to be 4.10 × 105 OBs/mL, 1.05 × 105 OBs/mL, and 4.09 × 104 OBs/mL respectively. The second aspect of the interaction between CrleGV-SA and CrpeNPV that was investigated was the speed of kill. A time-response biological assay protocol was created that allowed for effective observation of the larvae. This was then used to generate time-mortality data that were analysed by a logit regression function to calculate and compare the treatments at the time of 50 % larval mortality (LT50) and the time of 90 % mortality (LT90). Each virus by itself did not differ significantly from the other, while the mixed infection took significantly longer to kill 50 % and 90 % of the larvae, suggesting that there is competition for resources between viruses during the secondary, systemic phase of infection. The LT50 for CrleGV-SA, CrpeNPV and the mixed infection were 117.5 hours, 113.5 hours and 139.0 hours respectively. The LT90 for CrleGV-SA, CrpeNPV and the mixed infection were 153.2 hours, 159.3, and 193.4 hours respectively. Finally, the composition of OBs recovered from the cadavers produced by the time-response biological assays were investigated by mPCR. A method for extracting gDNA from OBs in neonate-sized T. leucotreta larvae is described. The presence of CrpeNPV along with CrleGV-SA was noted in 4 out of 9 larvae inoculated with only CrleGV-SA. The presence of CrleGV-SA as well as CrpeNPV was noted in all but one larva inoculated with only CrpeNPV, and both CrleGV-SA and CrpeNPV were noted in all but one larva inoculated with a 1:1 mixture of the two, with one larva only being positive for CrleGV-SA. This suggests either stock contamination or the presence of covert infections of CrleGV-SA and CrpeNPV in the T. leucotreta population used in this study. This is the second study to report an improved lethal concentration of a mixed infection of CrleGV-SA and CrpeNPV against T. leucotreta neonates, and the first study to report the slower speed of kill of a mixed infection of CrleGV-SA and CrpeNPV against T. leucotreta neonates. While the improved lethal concentration of the mixed infection is a promising step in the future improvement of baculovirus-based biopesticides, it is at the cost of a slower speed of kill. , Thesis (MSc) -- Faculty of Science, Department of Zoology and Entomology, 2021
- Full Text:
- Date Issued: 2021-04
Biology, ecology and management of the Keurboom moth, Leto venus Cramer and the leafhopper Molopopterus sp. Jacobi in cultivated Honeybush (Cyclopia spp.)
- Authors: Mushore, Tapiwa Gift
- Date: 2021-04
- Subjects: Legumes , Legumes -- Diseases and pests , Hepialidae , Leafhoppers , Pests -- Biological control , Entomopathogenic fungi , Leafhoppers -- Biological control , Hepialidae -- Biological control , Keurboom moth (Leto venus Cramer) , Molopopterus sp. Jacobi , Honeybush (Cyclopia spp.)
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/177125 , vital:42792
- Description: Honeybush, Cyclopia spp. Vent (Fabaceae), farmers have raised pest concerns following commercial cultivation. The Keurboom moth Leto venus Cramer (Lepidoptera: Hepialidae) and the leafhopper Molopopterus sp. Jacobi (Hemiptera: Cicadellidae), are two of the major pests identified in cultivated Honeybush. Laboratory and field studies were conducted to gain an understanding of the biology of these two pests to inform future pest management solutions. Additionally, entomopathogenic fungi were isolated from Honeybush farms and screened for virulence against Molopopterus sp. as a possible management strategy. This study showed that the L. venus infestation on Honeybush was a product of four fixed effects; stem diameter, species of Cyclopia, Farm location and age of the plants. Cyclopia subternata, had the highest likelihood of infestation. Increase in age of the plants resulted in an increase in the stem diameter and therefore a higher probability of infestation. Stem diameter was also shown to be a significant predictor of infestation likelihood. Infestation severity, determined by the number of larvae per plant, was shown to be influenced by three fixed effects; stem diameter, plant species and Farm location. The results also showed that L. venus prefers to initiate penetration at, or just aboveground level. Laboratory studies showed that the leafhopper Molopopterus sp. undergoes five nymphal instars with an average egg incubation time of 20 days, development time from 1st instar to adult of 26 days and average generation time of 47 days. Laboratory experiments revealed variations in host preference by the leafhopper over a period of 15 days. Cyclopia longifolia was identified to be the most preferred species for feeding compared to the two other commonly cultivated species, C. subternata and C. maculata. The results were consistent with those obtained from the field survey which showed that leafhopper density was influenced by four fixed effects; plant species, age of the plant, Farm location and harvesting practices. There were significant differences in leafhopper density in different species with C. longifolia having the highest number of leafhoppers per plant. There were differences in leafhopper density in different farms as 57% of the sampled farms had leafhopper infestations, of these farms, Lodestone and Kurland had the highest leafhopper densities. Harvested plants were shown to have significantly higher leafhopper density than non-harvested plants. Age was also shown to influence leafhopper density, which reduced with an increase in the age of the plants. A total of 20 fungal isolates were recovered from 98 soil samples of which 70% were from Honeybush fields and 30% were from surrounding refugia. Fusarium oxysporum isolates comprised 20% of the recovered isolates, with Metarhizium anisopliae isolates making up the remainder. Laboratory bioassays against adults and nymphs of the leafhopper, Molopopterus sp., showed that F. oxysporum isolates induced 10 – 45% mortality and M. anisopliae isolates induce 30 – 80% mortality. Metarhizium anisopliae isolates J S1, KF S3, KF S11, KF S13, LS1 and LS2 were the most virulent and induced over 60% mortality in both Molopopterus sp. nymphs and adults. The results of this study showed pest preference towards different Cyclopia species. As such, they should be managed differently. Furthermore, L. venus was observed to occur in low densities, hence, it cannot be considered a major pest. However, Molopopterus sp. recorded high population densities making it a major pest in Honeybush production. Positive results indicated that some of the isolated fungal isolates have potential for control, an avenue worth investigating further. , Thesis (MSc) -- Faculty of Science, Department of Zoology and Entomology, 2021
- Full Text:
- Date Issued: 2021-04
- Authors: Mushore, Tapiwa Gift
- Date: 2021-04
- Subjects: Legumes , Legumes -- Diseases and pests , Hepialidae , Leafhoppers , Pests -- Biological control , Entomopathogenic fungi , Leafhoppers -- Biological control , Hepialidae -- Biological control , Keurboom moth (Leto venus Cramer) , Molopopterus sp. Jacobi , Honeybush (Cyclopia spp.)
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/177125 , vital:42792
- Description: Honeybush, Cyclopia spp. Vent (Fabaceae), farmers have raised pest concerns following commercial cultivation. The Keurboom moth Leto venus Cramer (Lepidoptera: Hepialidae) and the leafhopper Molopopterus sp. Jacobi (Hemiptera: Cicadellidae), are two of the major pests identified in cultivated Honeybush. Laboratory and field studies were conducted to gain an understanding of the biology of these two pests to inform future pest management solutions. Additionally, entomopathogenic fungi were isolated from Honeybush farms and screened for virulence against Molopopterus sp. as a possible management strategy. This study showed that the L. venus infestation on Honeybush was a product of four fixed effects; stem diameter, species of Cyclopia, Farm location and age of the plants. Cyclopia subternata, had the highest likelihood of infestation. Increase in age of the plants resulted in an increase in the stem diameter and therefore a higher probability of infestation. Stem diameter was also shown to be a significant predictor of infestation likelihood. Infestation severity, determined by the number of larvae per plant, was shown to be influenced by three fixed effects; stem diameter, plant species and Farm location. The results also showed that L. venus prefers to initiate penetration at, or just aboveground level. Laboratory studies showed that the leafhopper Molopopterus sp. undergoes five nymphal instars with an average egg incubation time of 20 days, development time from 1st instar to adult of 26 days and average generation time of 47 days. Laboratory experiments revealed variations in host preference by the leafhopper over a period of 15 days. Cyclopia longifolia was identified to be the most preferred species for feeding compared to the two other commonly cultivated species, C. subternata and C. maculata. The results were consistent with those obtained from the field survey which showed that leafhopper density was influenced by four fixed effects; plant species, age of the plant, Farm location and harvesting practices. There were significant differences in leafhopper density in different species with C. longifolia having the highest number of leafhoppers per plant. There were differences in leafhopper density in different farms as 57% of the sampled farms had leafhopper infestations, of these farms, Lodestone and Kurland had the highest leafhopper densities. Harvested plants were shown to have significantly higher leafhopper density than non-harvested plants. Age was also shown to influence leafhopper density, which reduced with an increase in the age of the plants. A total of 20 fungal isolates were recovered from 98 soil samples of which 70% were from Honeybush fields and 30% were from surrounding refugia. Fusarium oxysporum isolates comprised 20% of the recovered isolates, with Metarhizium anisopliae isolates making up the remainder. Laboratory bioassays against adults and nymphs of the leafhopper, Molopopterus sp., showed that F. oxysporum isolates induced 10 – 45% mortality and M. anisopliae isolates induce 30 – 80% mortality. Metarhizium anisopliae isolates J S1, KF S3, KF S11, KF S13, LS1 and LS2 were the most virulent and induced over 60% mortality in both Molopopterus sp. nymphs and adults. The results of this study showed pest preference towards different Cyclopia species. As such, they should be managed differently. Furthermore, L. venus was observed to occur in low densities, hence, it cannot be considered a major pest. However, Molopopterus sp. recorded high population densities making it a major pest in Honeybush production. Positive results indicated that some of the isolated fungal isolates have potential for control, an avenue worth investigating further. , Thesis (MSc) -- Faculty of Science, Department of Zoology and Entomology, 2021
- Full Text:
- Date Issued: 2021-04
Selection for improved virulence of Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV) to False Codling Moth, Thaumatotibia leucotreta, by serial passage through a heterologous host
- Authors: Iita, Petrus Paulus
- Date: 2021-04
- Subjects: Cryptophlebia leucotreta -- Biological control , Biological pest control agents , Citrus -- Diseases and pests , Baculoviruses , Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV)
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/178180 , vital:42918
- Description: The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is endemic to southern Africa, and strongly associated with citrus. As South African citrus production is mainly for export to foreign markets, the market access risk due to the phytosanitary status of this pest is considerable and its control is therefore imperative. Various control measures as part of a rigorous integrated pest management (IPM) programme targeted against T. leucotreta have been effective at suppressing the pest in citrus, but there is still a growing need for continued improvement of the programme and augmentation of the available control options. Of these control options, biological control, particularly the use of Cryptophlebia leucotreta granulovirus (CrleGV-SA), is a key component of IPM in citrus orchards and it has been very successful at reducing T. leucotreta populations in the field for almost two decades. There is however, a growing need for more baculovirus variants with an improved virulence against T. leucotreta for a more efficient pest management system. The newly identified insect virus, Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV) offers a unique opportunity for an additional biopesticide in IPM for control of T. leucotreta in the field. This study aimed to conduct serial passaging of CrpeNPV through a heterologous host, T. leucotreta, in order to determine the potential for improved virulence or speed of kill against it. In order to select for a variant of CrpeNPV with improved virulence against T. leucotreta, a high dose (LC90) of the virus OBs was used to perform 12 serial passages through T. leucotreta larvae in surface-dose bioassays. Whole genome sequencing and analysis of the passaged virus, along with restriction endonuclease profiling in silico was performed to determine if the genetic identity of the virus had changed during serial passage, in relation to the original virus. These analyses indicated that the dominant genotype of CrpeNPV was maintained following 12 serial passages through the heterologous host. The biological activity of the passaged virus, along with the original virus was evaluated against neonate T. leucotreta in surface-dose bioassays and compared. Results from dose-response bioassays showed that the virulence of CrpeNPV did not improve after 12 serial passages. The LC50 values of the passaged virus and the original virus were estimated at 1.96 × 104 and 1.58 × 104 OBs/ml, respectively, whereas the LC90 values were estimated at 3.46 × 104 OBs/ml for the passaged virus and 3.68 × 104 for the original virus. Similarly, the results from time-response bioassays showed that the speed of kill of CrpeNPV did not improve after 12 serial passages. The LT50 values of the passaged virus and the original virus were 88.44 hours (3 days and 16 hours) and 83.74 hours (3 days and 12 hours), respectively, whereas the LT90 values were 115 hours (4 days 19 hours) for the passaged virus and 102 hours (4 days 6 hours) for the original virus. The virulence and speed of kill of the passaged virus decreased significantly, in relation to the original virus. When the full genome of the passaged virus was sequenced and analysed, only a few SNPs were detected in the viral genome, in comparison to the original virus. No detectable difference in REN digestion patterns were observed following REN analysis of gDNA of the passaged virus with several restriction enzymes in silico. The results for this study suggest that CrpeNPV may already be optimally suited to the heterologous host as it persists under these conditions without significant changes to the genome. These results have positive implications for the genetic integrity of CrpeNPV as a potential biocontrol agent in the field. This study is the first to report the virulence selection of CrpeNPV by serial passage through a heterologous host, and also the first to record bioassay data in terms of dose response (or lethal concentration) against T. leucotreta second instars. The data obtained have added to the knowledge about interactions between CrpeNPV and its heterologous host, and may be fundamental to continued investigation into the effect of serial passage on pathogenicity and genetic diversity of CrpeNPV. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2021
- Full Text:
- Date Issued: 2021-04
- Authors: Iita, Petrus Paulus
- Date: 2021-04
- Subjects: Cryptophlebia leucotreta -- Biological control , Biological pest control agents , Citrus -- Diseases and pests , Baculoviruses , Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV)
- Language: English
- Type: thesis , text , Masters , MSc
- Identifier: http://hdl.handle.net/10962/178180 , vital:42918
- Description: The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is endemic to southern Africa, and strongly associated with citrus. As South African citrus production is mainly for export to foreign markets, the market access risk due to the phytosanitary status of this pest is considerable and its control is therefore imperative. Various control measures as part of a rigorous integrated pest management (IPM) programme targeted against T. leucotreta have been effective at suppressing the pest in citrus, but there is still a growing need for continued improvement of the programme and augmentation of the available control options. Of these control options, biological control, particularly the use of Cryptophlebia leucotreta granulovirus (CrleGV-SA), is a key component of IPM in citrus orchards and it has been very successful at reducing T. leucotreta populations in the field for almost two decades. There is however, a growing need for more baculovirus variants with an improved virulence against T. leucotreta for a more efficient pest management system. The newly identified insect virus, Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV) offers a unique opportunity for an additional biopesticide in IPM for control of T. leucotreta in the field. This study aimed to conduct serial passaging of CrpeNPV through a heterologous host, T. leucotreta, in order to determine the potential for improved virulence or speed of kill against it. In order to select for a variant of CrpeNPV with improved virulence against T. leucotreta, a high dose (LC90) of the virus OBs was used to perform 12 serial passages through T. leucotreta larvae in surface-dose bioassays. Whole genome sequencing and analysis of the passaged virus, along with restriction endonuclease profiling in silico was performed to determine if the genetic identity of the virus had changed during serial passage, in relation to the original virus. These analyses indicated that the dominant genotype of CrpeNPV was maintained following 12 serial passages through the heterologous host. The biological activity of the passaged virus, along with the original virus was evaluated against neonate T. leucotreta in surface-dose bioassays and compared. Results from dose-response bioassays showed that the virulence of CrpeNPV did not improve after 12 serial passages. The LC50 values of the passaged virus and the original virus were estimated at 1.96 × 104 and 1.58 × 104 OBs/ml, respectively, whereas the LC90 values were estimated at 3.46 × 104 OBs/ml for the passaged virus and 3.68 × 104 for the original virus. Similarly, the results from time-response bioassays showed that the speed of kill of CrpeNPV did not improve after 12 serial passages. The LT50 values of the passaged virus and the original virus were 88.44 hours (3 days and 16 hours) and 83.74 hours (3 days and 12 hours), respectively, whereas the LT90 values were 115 hours (4 days 19 hours) for the passaged virus and 102 hours (4 days 6 hours) for the original virus. The virulence and speed of kill of the passaged virus decreased significantly, in relation to the original virus. When the full genome of the passaged virus was sequenced and analysed, only a few SNPs were detected in the viral genome, in comparison to the original virus. No detectable difference in REN digestion patterns were observed following REN analysis of gDNA of the passaged virus with several restriction enzymes in silico. The results for this study suggest that CrpeNPV may already be optimally suited to the heterologous host as it persists under these conditions without significant changes to the genome. These results have positive implications for the genetic integrity of CrpeNPV as a potential biocontrol agent in the field. This study is the first to report the virulence selection of CrpeNPV by serial passage through a heterologous host, and also the first to record bioassay data in terms of dose response (or lethal concentration) against T. leucotreta second instars. The data obtained have added to the knowledge about interactions between CrpeNPV and its heterologous host, and may be fundamental to continued investigation into the effect of serial passage on pathogenicity and genetic diversity of CrpeNPV. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2021
- Full Text:
- Date Issued: 2021-04
An Integrated Management System to reduce False Codling Moth, Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) infested citrus fruit from being packed for export
- Authors: Mac Aleer, Clint
- Date: 2019
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Biological control -- South Africa , Citrus -- Diseases and pests -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Insecticides , Citrus fruit industry -- South Africa , South Africa -- Commerce -- European Economic Community Countries
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92219 , vital:30691
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is indigenous to southern Africa and is an important pest of citrus in this region. As a result of its endemism to sub-Saharan Africa, several countries to which South Africa exports citrus, regulate it as a phytosanitary pest. Consequently, it is necessary to ship fruit to these markets under cold-disinfestation protocols. This has been possible, as until recently, all of these markets could be considered relatively small niche markets. The South African citrus industry exports approximately 130 million cartons of fruit (15 kg equivalent) annually. During the 2017 season, a total of 48 million cartons were exported to the European Union (EU), which is the equivalent of 41% of South Africa’s total export volume, thus making the EU South Africa’s most important export market. In 2013 the European and Mediterranean Plant Protection Organisation (EPPO) conducted a pest risk analysis (PRA) on FCM, leading to the EU declaring it an officially regulated pest for this region, effective of 1 January 2018. Citrus is regarded as a preferred non-native host of FCM and South African citrus was identified as a primary focus due to large volumes being exported to Europe. Shipping under cold disinfestation is not possible with such large volumes of fruit. Additionally, several cultivars would suffer high levels of chilling injury under such conditions. In this study, an Integrated Management System was tested with pre- and postharvest controls to test the hypothesis that pre-harvest interventions resulted in lower post-harvest infection. Thirty orchards ranging from soft citrus cultivars such as Nule and Nova Mandarins, to Navel orange cultivars such as Newhall, Palmer and Late Navel and ending with Valencia cultivars such as Midknight and Delta, were identified for this study. This system relies on pre-harvest inspections such as FCM trap counts and fruit infestation on data trees in every orchard, with associated thresholds for action or continued compliance. Inspections were conducted on a weekly basis. There was a significant relationship between the moth catches and FCM infestation for the full monitoring period, using a two-week lag period for infestation. Inspections of harvested fruit were conducted at the packhouse to determine FCM infestation. This included inspection of the fruit on delivery to the packhouse, on the packing line, and a final fruit sample taken from the packed product and inspected for FCM. The highest levels of infestation were recorded on the Navel cultivars, thus confirming that Navels cultivars are a preferred host for FCM. Significant positive relationships were recorded between FCM infestation during the last 4 weeks before harvest and the level of infestation in the fruit delivered to the packhouse and between the fruit delivered to the packhouse and in the fruit packed in a carton for export. There was a substantial reduction in infestation between the fruit delivered to the packhouse and the fruit packed in a carton for export, with certain orchards recording as much as a 93% reduction in the fruit packed in a carton, which indicated that the packhouse could effectively identify and remove FCM infested fruit. The outcome of the study is that a holistic management approach minimizes the risk of FCM in citrus fruit destined for export and therefore mitigate the risk associated with FCM.
- Full Text:
- Date Issued: 2019
- Authors: Mac Aleer, Clint
- Date: 2019
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Biological control -- South Africa , Citrus -- Diseases and pests -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Insecticides , Citrus fruit industry -- South Africa , South Africa -- Commerce -- European Economic Community Countries
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92219 , vital:30691
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is indigenous to southern Africa and is an important pest of citrus in this region. As a result of its endemism to sub-Saharan Africa, several countries to which South Africa exports citrus, regulate it as a phytosanitary pest. Consequently, it is necessary to ship fruit to these markets under cold-disinfestation protocols. This has been possible, as until recently, all of these markets could be considered relatively small niche markets. The South African citrus industry exports approximately 130 million cartons of fruit (15 kg equivalent) annually. During the 2017 season, a total of 48 million cartons were exported to the European Union (EU), which is the equivalent of 41% of South Africa’s total export volume, thus making the EU South Africa’s most important export market. In 2013 the European and Mediterranean Plant Protection Organisation (EPPO) conducted a pest risk analysis (PRA) on FCM, leading to the EU declaring it an officially regulated pest for this region, effective of 1 January 2018. Citrus is regarded as a preferred non-native host of FCM and South African citrus was identified as a primary focus due to large volumes being exported to Europe. Shipping under cold disinfestation is not possible with such large volumes of fruit. Additionally, several cultivars would suffer high levels of chilling injury under such conditions. In this study, an Integrated Management System was tested with pre- and postharvest controls to test the hypothesis that pre-harvest interventions resulted in lower post-harvest infection. Thirty orchards ranging from soft citrus cultivars such as Nule and Nova Mandarins, to Navel orange cultivars such as Newhall, Palmer and Late Navel and ending with Valencia cultivars such as Midknight and Delta, were identified for this study. This system relies on pre-harvest inspections such as FCM trap counts and fruit infestation on data trees in every orchard, with associated thresholds for action or continued compliance. Inspections were conducted on a weekly basis. There was a significant relationship between the moth catches and FCM infestation for the full monitoring period, using a two-week lag period for infestation. Inspections of harvested fruit were conducted at the packhouse to determine FCM infestation. This included inspection of the fruit on delivery to the packhouse, on the packing line, and a final fruit sample taken from the packed product and inspected for FCM. The highest levels of infestation were recorded on the Navel cultivars, thus confirming that Navels cultivars are a preferred host for FCM. Significant positive relationships were recorded between FCM infestation during the last 4 weeks before harvest and the level of infestation in the fruit delivered to the packhouse and between the fruit delivered to the packhouse and in the fruit packed in a carton for export. There was a substantial reduction in infestation between the fruit delivered to the packhouse and the fruit packed in a carton for export, with certain orchards recording as much as a 93% reduction in the fruit packed in a carton, which indicated that the packhouse could effectively identify and remove FCM infested fruit. The outcome of the study is that a holistic management approach minimizes the risk of FCM in citrus fruit destined for export and therefore mitigate the risk associated with FCM.
- Full Text:
- Date Issued: 2019
Augmentative releases of Dactylopius austrinus De Lotto (Dactylopiidae; Hemiptera) for biological control of Opuntia aurantiaca Lindley (Cactaceae), in South Africa
- Authors: Mulateli, Thifhelimbilu
- Date: 2019
- Subjects: Opuntia aurantiaca -- Biolotical control -- South Africa , Invasive plants -- Biolotical control -- South Africa , Dactylopius austrinus De Lotto -- South Africa , Dactylopius -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92931 , vital:30765
- Description: Opuntia aurantiaca Lindely (Cactaceae) is an invasive alien cactus which has detrimental effects on agroecosystems and indigenous biodiversity in South Africa. Dense infestations over large areas reduce grazing capacity and indigenous biodiversity. Despite the release of a biological control agent, the cochineal insect Dactylopius austrinus De Lotto (Dactylopiidae), the weed is still considered a major problem in many parts of the country. Biological control has relied heavily on classical biological control, with little augmentative biological control implemented. This study investigated the outcome of mass-rearing and augmentative releases of D. austrinus for the control of O. aurantiaca. Augmentative releases are thought to improve the level of control by increasing agent densities in the field and thus increasing the level of damage inflicted to the plants. All data were collected with the intention to optimize release strategies so that the maximum benefit from the biological control agent could be achieved. An impact study was conducted using potted plants in a greenhouse to quantify the efficacy of multiple releases of the agent on the target weed. All three of the release treatments showed consistently higher proportion of cochineal than the controls, as well as the insect exclusion treatments, and these differences were statistically significant. The number of cladodes per plant increased significantly for the insect exclusion and control treatment over the period of the study, whilst all three release treatments decreased steadily over the same period. This study indicated that the agent is damaging to O. aurantiaca and that a single release event was beneficial but that multiple releases did not result in greater levels of control. A post-release evaluation was carried out to quantify the impact of releases of D. austrinus on O. aurantiaca in the field. Plots where the agent was excluded were compared with those where the agent was left at natural field densities and three treatments where agent populations were augmented to varying degrees through releases. The percentage of cochineal infested cladodes for all treatments decreased over time from the initiation of the experiment in October 2017 until the end of the experiment in October 2018. Opuntia aurantiaca densities also decreased over time for all treatments. The insect exclusion treatment had the greatest number of plants for the duration of the study, but this was not significantly different from other treatments. Dactylopius austrinus was damaging to O. aurantiaca, but climatic conditions in the field limited the efficacy of releases. Although O. aurantiaca density decreased during the experiment, it was evident that the reduced number of plants was not due to augmentation of the cochineal populations from the releases that were conducted. The experiment was conducted over a very dry period, when cochineal was particularly effective, so although augmentative releases did not improve the level of control, the natural population of cochineal was high and very damaging to O. aurantiaca over the course of the experiment. Releasing during wet periods, when the agent is less effective, could augment agent populations at a time when natural populations would be low, and hence improve levels of control further. Although this study was limited to a short period of two years, the results of this study suggest that the number of releases is less important than the timing of releases. Releasing immediately after periods of high rainfall is likely to be beneficial, while releasing during dry periods, or during winter when temperatures are low, is less effective. Dactylopius austrinus populations should be constantly monitored so that releases can be conducted when cochineal populations are low and the climatic conditions are correct. If the timing of release events is appropriate, then the over level of control of O. aurantiaca using D. austrinus could be improved.
- Full Text:
- Date Issued: 2019
- Authors: Mulateli, Thifhelimbilu
- Date: 2019
- Subjects: Opuntia aurantiaca -- Biolotical control -- South Africa , Invasive plants -- Biolotical control -- South Africa , Dactylopius austrinus De Lotto -- South Africa , Dactylopius -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92931 , vital:30765
- Description: Opuntia aurantiaca Lindely (Cactaceae) is an invasive alien cactus which has detrimental effects on agroecosystems and indigenous biodiversity in South Africa. Dense infestations over large areas reduce grazing capacity and indigenous biodiversity. Despite the release of a biological control agent, the cochineal insect Dactylopius austrinus De Lotto (Dactylopiidae), the weed is still considered a major problem in many parts of the country. Biological control has relied heavily on classical biological control, with little augmentative biological control implemented. This study investigated the outcome of mass-rearing and augmentative releases of D. austrinus for the control of O. aurantiaca. Augmentative releases are thought to improve the level of control by increasing agent densities in the field and thus increasing the level of damage inflicted to the plants. All data were collected with the intention to optimize release strategies so that the maximum benefit from the biological control agent could be achieved. An impact study was conducted using potted plants in a greenhouse to quantify the efficacy of multiple releases of the agent on the target weed. All three of the release treatments showed consistently higher proportion of cochineal than the controls, as well as the insect exclusion treatments, and these differences were statistically significant. The number of cladodes per plant increased significantly for the insect exclusion and control treatment over the period of the study, whilst all three release treatments decreased steadily over the same period. This study indicated that the agent is damaging to O. aurantiaca and that a single release event was beneficial but that multiple releases did not result in greater levels of control. A post-release evaluation was carried out to quantify the impact of releases of D. austrinus on O. aurantiaca in the field. Plots where the agent was excluded were compared with those where the agent was left at natural field densities and three treatments where agent populations were augmented to varying degrees through releases. The percentage of cochineal infested cladodes for all treatments decreased over time from the initiation of the experiment in October 2017 until the end of the experiment in October 2018. Opuntia aurantiaca densities also decreased over time for all treatments. The insect exclusion treatment had the greatest number of plants for the duration of the study, but this was not significantly different from other treatments. Dactylopius austrinus was damaging to O. aurantiaca, but climatic conditions in the field limited the efficacy of releases. Although O. aurantiaca density decreased during the experiment, it was evident that the reduced number of plants was not due to augmentation of the cochineal populations from the releases that were conducted. The experiment was conducted over a very dry period, when cochineal was particularly effective, so although augmentative releases did not improve the level of control, the natural population of cochineal was high and very damaging to O. aurantiaca over the course of the experiment. Releasing during wet periods, when the agent is less effective, could augment agent populations at a time when natural populations would be low, and hence improve levels of control further. Although this study was limited to a short period of two years, the results of this study suggest that the number of releases is less important than the timing of releases. Releasing immediately after periods of high rainfall is likely to be beneficial, while releasing during dry periods, or during winter when temperatures are low, is less effective. Dactylopius austrinus populations should be constantly monitored so that releases can be conducted when cochineal populations are low and the climatic conditions are correct. If the timing of release events is appropriate, then the over level of control of O. aurantiaca using D. austrinus could be improved.
- Full Text:
- Date Issued: 2019
Climatic suitability of Dichrorampha odorata Brown and Zachariades (Lepidoptera: Tortricidae), a shoot-boring moth for the biological control of Chromolaena odorata (L.) R.M. King and H. Robinson (Asteraceae) in South Africa
- Authors: Nqayi, Slindile Brightness
- Date: 2019
- Subjects: CLIMEX , Chromolaena odorata -- Biological control -- South Africa , Tortricidae -- South Africa , Bioclimatology -- Software
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92208 , vital:30681
- Description: Biological control using natural enemies introduced from the native range is an integral component of the management of Chromolaena odorata, a serious invader in the eastern regions of South Africa. A number of biological control agents for C. odorata have been released in South Africa, and one of them, Dichrorampha odorata, has failed to establish. To understand if D. odorata failed to establish due to climate incompatibility, its thermal physiology was investigated. Thermal tolerance data were used to determine the developmental thresholds and number of generations that D. odorata is capable of going through in South Africa per year. These predictions were generated using CLIMEX temperature data and the degree-day parameters K and t0. Developmental time decreased with increasing temperatures ranging from 20 °C to 30°C, with immature stages not able to complete development at 18°C and 32°C. The developmental threshold, to, was determined as 8.45 °C with 872.4 degree-days required to complete development (K), indicating that D. odorata is capable of producing a maximum number of 6.5 generations per year in South Africa. The CLIMEX data indicated that the east coast regions of South Africa, which are the heaviest invaded areas by C. odorata in South Africa, were climatically most suitable for D. odorata to. D. odorata lower (LLT50) and upper (ULT50) lethal temperatures were -4.5°C and 39.64°C for larvae and 1.83 and 41.02°C for adults, and D. odorata adults were able to maintain locomotory functioning at 4.4 to 43.7°C, respectively. Acclimation at low and high temperatures indicate that when D. odorata was kept at a lower temperature of 20°C for 7 days, it became tolerant to warmer and cooler temperatures (1.95 and 44.41°C) when compared to D. odorata reared at 25°C (3.36 and 43.67°C) and 30°C (5.92 and 42.93°C). Dichrorampha odorata is therefore climatically suitable for release and should establish in South Africa to control C. odorata. The establishment and persistence of D. odorata will not be limited by climatic conditions but rather the distribution of its host weed, C. odorata in South Africa. Also, this study presents a decision-making protocol for the release of D. odorata to allow better performance in the field.
- Full Text:
- Date Issued: 2019
- Authors: Nqayi, Slindile Brightness
- Date: 2019
- Subjects: CLIMEX , Chromolaena odorata -- Biological control -- South Africa , Tortricidae -- South Africa , Bioclimatology -- Software
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92208 , vital:30681
- Description: Biological control using natural enemies introduced from the native range is an integral component of the management of Chromolaena odorata, a serious invader in the eastern regions of South Africa. A number of biological control agents for C. odorata have been released in South Africa, and one of them, Dichrorampha odorata, has failed to establish. To understand if D. odorata failed to establish due to climate incompatibility, its thermal physiology was investigated. Thermal tolerance data were used to determine the developmental thresholds and number of generations that D. odorata is capable of going through in South Africa per year. These predictions were generated using CLIMEX temperature data and the degree-day parameters K and t0. Developmental time decreased with increasing temperatures ranging from 20 °C to 30°C, with immature stages not able to complete development at 18°C and 32°C. The developmental threshold, to, was determined as 8.45 °C with 872.4 degree-days required to complete development (K), indicating that D. odorata is capable of producing a maximum number of 6.5 generations per year in South Africa. The CLIMEX data indicated that the east coast regions of South Africa, which are the heaviest invaded areas by C. odorata in South Africa, were climatically most suitable for D. odorata to. D. odorata lower (LLT50) and upper (ULT50) lethal temperatures were -4.5°C and 39.64°C for larvae and 1.83 and 41.02°C for adults, and D. odorata adults were able to maintain locomotory functioning at 4.4 to 43.7°C, respectively. Acclimation at low and high temperatures indicate that when D. odorata was kept at a lower temperature of 20°C for 7 days, it became tolerant to warmer and cooler temperatures (1.95 and 44.41°C) when compared to D. odorata reared at 25°C (3.36 and 43.67°C) and 30°C (5.92 and 42.93°C). Dichrorampha odorata is therefore climatically suitable for release and should establish in South Africa to control C. odorata. The establishment and persistence of D. odorata will not be limited by climatic conditions but rather the distribution of its host weed, C. odorata in South Africa. Also, this study presents a decision-making protocol for the release of D. odorata to allow better performance in the field.
- Full Text:
- Date Issued: 2019
Genetic characterisation of a range of geographically distinct Helicoverpa armigera nucleopolyhedrovirus (HearNPV) isolates and evaluation of biological activity against South African populations of the African bollworm, Helicoverpa armigera (Hu bner) (Lepidoptera: Noctuidae)
- Mtambanengwe, Kudzai Tapiwanashe Esau
- Authors: Mtambanengwe, Kudzai Tapiwanashe Esau
- Date: 2019
- Subjects: Helicoverpa armigera -- Biological control -- South Africa , Baculoviruses -- Genetics , Agricultural pests -- Biological control -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/97334 , vital:31426
- Description: The African bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a pest of economic and agricultural importance globally. It is a polyphagous pest that feeds on a wide range of host plants including economically important crops. The impact it has on agricultural systems makes its control a priority. The most common method of control is using chemical pesticides; however, continuous application of the pesticides has resulted in the development of resistance. The use of biological control has been investigated and established as an effective method of control as a standalone or part of an integrated pest management (IPM) system. The use of the baculovirus Helicoverpa armigera nucleopolyhedrovirus (HearNPV), has shown promise in the control of H. armigera. Commercial formulations based on the virus are available in many global markets. However, the identification of novel HearNPV isolates will aid in the control of H. armigera as well as provide alternative isolates that may have better virulence. Three new HearNPV isolates were purified and identified from three distinct geographical South African locations H. armigera cadavers and named HearNPV-Albany, HearNPV-KZN and HearNPV-Haygrove. The genomes of two of the HearNPV isolates, namely HearNPV-Albany and HearNPV-KZN were genetically characterised and compared to other geographically distinct HearNPV isolates. Virulence studies were performed comparing the new HearNPV isolates against established commercial HearNPV formulations, Helicovir™ and Helicovex® and other geographically distinct isolated HearNPV, HearNPV-G4 and HearNPV-SP1. Two laboratory colonies were established using H. armigera collected from South African fields in the Belmont Valley near Grahamstown labelled as Albany colony and a colony provided from Haygrove Eden farm near George labelled as Haygrove colony. Biological studies were carried out using the Albany H. armigera colony comparing the rate of development, survival and fertility on bell green peppers, cabbage leaves and on artificial diet. From the biological studies, it was recorded that development and survivorship was best on artificial diet. Regular quality control was required for the maintenance of the colony and continuous generations of healthy larvae were eventually established. Diseased cadavers with signs of baculovirus infection were collected after bioprospecting from the Kwa-Zulu Natal Province in South Africa and were labelled KZN isolate; Belmont Valley near Grahamstown and were labelled Albany isolate; and Haygrove Eden farm near George and were labelled Haygrove isolate for the study. A fourth isolate made up of a crude extract of occlusion bodies (OBs) first described by Whitlock was also analysed and labelled Whitlock isolate. Occlusion bodies were extracted, purified and morphologically identified from the KZN, Albany, Haygrove and Whitlock isolates using TEM. Genomic DNA, which was extracted from the purified OBs. Using PCR, the identity of the OBs as HearNPV was confirmed. Genomic analyses were performed on HearNPV-Albany and HearNPV-KZN through genetic characterisation and comparison with other geographically distinct HearNPV genomes to confirm novelty and establish potential genetic relationships between the isolates through evolutionary distances. Full genomic sequencing of the isolated HearNPV and comparison with other geographically distinct HearNPV isolates identified genomic differences that showed that the HearNPV isolates were novel. HearNPV-Albany and HearNPV-KZN were successfully sequenced and identified as novel isolates with unique fragment patterns and unique gene sequences through deletions or insertions when compared to other geographically distinct HearNPV. This raised the potential for differences in biological activity against H. armigera larvae when tested through biological assays. HearNPV-Whit genome assembly had low quality data which resulted in many gaps and failed assembly. The biological activity of HearNPV isolates from Spain, China, South Africa and two commercial formulations were studied against the laboratory established H. armigera South African colony. The LC50 values of the different South African HearNPV isolates were established to be between 7.7 × 101 OBs.ml-1 for the most effective and 3.2 × 102 OBs.ml-1 for the least effective. The Spanish and Chinese HearNPV isolates resulted in LC50 values of 2.0 × 102 OBs.ml-1 and 1.2 × 101 OBs.ml-1 respectively. The commercial formulations resulted in the least virulence observed with an LC50 of 5.84× 102 OBs.ml-1 and 9.0 × 102 OBs.ml-1 for Helicovex® and Helicovir™ respectively. In this study, novel South African HearNPV isolates were isolated and identified. Through characterisation and bioassays against South African H. armigera populations the HearNPV isolates were shown to have different virulence in comparison to geographically distinct isolates. From this research, there is potential for development of new H. armigera biopesticides based on the novel isolates after field trial testing.
- Full Text:
- Date Issued: 2019
- Authors: Mtambanengwe, Kudzai Tapiwanashe Esau
- Date: 2019
- Subjects: Helicoverpa armigera -- Biological control -- South Africa , Baculoviruses -- Genetics , Agricultural pests -- Biological control -- South Africa
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/97334 , vital:31426
- Description: The African bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a pest of economic and agricultural importance globally. It is a polyphagous pest that feeds on a wide range of host plants including economically important crops. The impact it has on agricultural systems makes its control a priority. The most common method of control is using chemical pesticides; however, continuous application of the pesticides has resulted in the development of resistance. The use of biological control has been investigated and established as an effective method of control as a standalone or part of an integrated pest management (IPM) system. The use of the baculovirus Helicoverpa armigera nucleopolyhedrovirus (HearNPV), has shown promise in the control of H. armigera. Commercial formulations based on the virus are available in many global markets. However, the identification of novel HearNPV isolates will aid in the control of H. armigera as well as provide alternative isolates that may have better virulence. Three new HearNPV isolates were purified and identified from three distinct geographical South African locations H. armigera cadavers and named HearNPV-Albany, HearNPV-KZN and HearNPV-Haygrove. The genomes of two of the HearNPV isolates, namely HearNPV-Albany and HearNPV-KZN were genetically characterised and compared to other geographically distinct HearNPV isolates. Virulence studies were performed comparing the new HearNPV isolates against established commercial HearNPV formulations, Helicovir™ and Helicovex® and other geographically distinct isolated HearNPV, HearNPV-G4 and HearNPV-SP1. Two laboratory colonies were established using H. armigera collected from South African fields in the Belmont Valley near Grahamstown labelled as Albany colony and a colony provided from Haygrove Eden farm near George labelled as Haygrove colony. Biological studies were carried out using the Albany H. armigera colony comparing the rate of development, survival and fertility on bell green peppers, cabbage leaves and on artificial diet. From the biological studies, it was recorded that development and survivorship was best on artificial diet. Regular quality control was required for the maintenance of the colony and continuous generations of healthy larvae were eventually established. Diseased cadavers with signs of baculovirus infection were collected after bioprospecting from the Kwa-Zulu Natal Province in South Africa and were labelled KZN isolate; Belmont Valley near Grahamstown and were labelled Albany isolate; and Haygrove Eden farm near George and were labelled Haygrove isolate for the study. A fourth isolate made up of a crude extract of occlusion bodies (OBs) first described by Whitlock was also analysed and labelled Whitlock isolate. Occlusion bodies were extracted, purified and morphologically identified from the KZN, Albany, Haygrove and Whitlock isolates using TEM. Genomic DNA, which was extracted from the purified OBs. Using PCR, the identity of the OBs as HearNPV was confirmed. Genomic analyses were performed on HearNPV-Albany and HearNPV-KZN through genetic characterisation and comparison with other geographically distinct HearNPV genomes to confirm novelty and establish potential genetic relationships between the isolates through evolutionary distances. Full genomic sequencing of the isolated HearNPV and comparison with other geographically distinct HearNPV isolates identified genomic differences that showed that the HearNPV isolates were novel. HearNPV-Albany and HearNPV-KZN were successfully sequenced and identified as novel isolates with unique fragment patterns and unique gene sequences through deletions or insertions when compared to other geographically distinct HearNPV. This raised the potential for differences in biological activity against H. armigera larvae when tested through biological assays. HearNPV-Whit genome assembly had low quality data which resulted in many gaps and failed assembly. The biological activity of HearNPV isolates from Spain, China, South Africa and two commercial formulations were studied against the laboratory established H. armigera South African colony. The LC50 values of the different South African HearNPV isolates were established to be between 7.7 × 101 OBs.ml-1 for the most effective and 3.2 × 102 OBs.ml-1 for the least effective. The Spanish and Chinese HearNPV isolates resulted in LC50 values of 2.0 × 102 OBs.ml-1 and 1.2 × 101 OBs.ml-1 respectively. The commercial formulations resulted in the least virulence observed with an LC50 of 5.84× 102 OBs.ml-1 and 9.0 × 102 OBs.ml-1 for Helicovex® and Helicovir™ respectively. In this study, novel South African HearNPV isolates were isolated and identified. Through characterisation and bioassays against South African H. armigera populations the HearNPV isolates were shown to have different virulence in comparison to geographically distinct isolates. From this research, there is potential for development of new H. armigera biopesticides based on the novel isolates after field trial testing.
- Full Text:
- Date Issued: 2019
Interaction between the root-feeding beetle, Longitarsus bethae (Coleoptera: Chrysomelidae) and the root-knot nematode, Meloidogyne javanica (Nematoda: Heteroderidae): Implications for the biological control of Lantana camara L. (Verbenaceae) in South Africa
- Authors: Musedeli, Jufter
- Date: 2019
- Subjects: Insect-plant relationships , Insects -- Host plants , Flea beetles , Symbiosis , Longitarsus , Chrysomelidae , Lantana camara -- Biological control -- South Africa , Heteroderidae , Root-knot nematodes , Weeds -- Biological control -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/71130 , vital:29788
- Description: Plants often are simultaneously attacked by several herbivores that can affect each other’s performance, and their interaction may affect their host plant fitness. The current study was conducted to determine the interaction between the root-feeding beetle, Longitarsus bethae Savini & Escalona (Coleoptera: Chrysomelidae) and a root-knot nematode, Meloidogyne javanica (Treub) Chitwood (Tylenchida: Heteroderidae), with implications for the biological control of Lantana camara L. (Verbenaceae) in South Africa. The studies were conducted under quarantine conditions at the Agricultural Research Council-PHP, Roodeplaat, Pretoria, South Africa. Specifically, the study determined; (i) whether root damage by the flea beetle enhanced infection by M. javanica, (ii) whether L. camara roots infected with the nematode enhanced the performance of the beetle, (iii) whether single or combined effect of the two organisms (i.e. L. bethae and M. javanica) had an overall effect on the growth and biomass of their shared host, L. camara, and (iv) the susceptibility of 10 L. camara varieties that are commonly found in South Africa to M. javanica. The study found that galling on the roots of L. camara by the nematode occurs at the highest inoculation of 300 eggs of L. bethae per plant, and no galling occurred at inoculation of 200 eggs per plant and below. The findings also showed that L. bethae performed better on M. javanica-infected than on healthy L. camara roots, and that more L. bethae adult progeny with slightly bigger body size emerged from M. javanica-infected, than from healthy plants. Fresh weight (galls) of plant roots from treatments where both species (i.e., L. bethae and M. javanica) were combined was significantly higher than that from plants infected with the nematode only, suggesting that the combination of both species induces more galling than the nematode does alone. The above-ground dry biomass was significantly lower both in combined and M. javanica only treatments, than in L. bethae only treatment. The study also found that selected L. camara varieties were infected with M. javanica, albeit at varying degrees of infection. Among the 10 L. camara varieties, Orange Red OR 015 was the most susceptible. Other susceptible varieties included Light Pink 009 LP, Total Pink 021 TP and Dark Pink 018 DP, and these, together with variety Orange Red OR 015, constituted 40% of the L. camara varieties evaluated in the current study. Fifty percent of the varieties displayed slight to moderate susceptibility to M. javanica, while 10% displayed lack of susceptibility. The study concluded that the symbiotic relationship between L. bethae and M. javanica was mutual, resulting in increase in the fitness of the beetle. The combined herbivory by L. bethae and M. javanica was also found to be additive on one of the most common varieties of L. camara in South Africa, and therefore co-infestation by both species might enhance the biological control of this weed in South Africa. The study further concluded that the suitability of some invasive L. camara cultivars such as Light Pink 009 LP and Orange Red 015 OR for M. javanica, might also contribute towards biological control of this weed in South Africa, particularly in areas where the two herbivores species co-exist.
- Full Text:
- Date Issued: 2019
- Authors: Musedeli, Jufter
- Date: 2019
- Subjects: Insect-plant relationships , Insects -- Host plants , Flea beetles , Symbiosis , Longitarsus , Chrysomelidae , Lantana camara -- Biological control -- South Africa , Heteroderidae , Root-knot nematodes , Weeds -- Biological control -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/71130 , vital:29788
- Description: Plants often are simultaneously attacked by several herbivores that can affect each other’s performance, and their interaction may affect their host plant fitness. The current study was conducted to determine the interaction between the root-feeding beetle, Longitarsus bethae Savini & Escalona (Coleoptera: Chrysomelidae) and a root-knot nematode, Meloidogyne javanica (Treub) Chitwood (Tylenchida: Heteroderidae), with implications for the biological control of Lantana camara L. (Verbenaceae) in South Africa. The studies were conducted under quarantine conditions at the Agricultural Research Council-PHP, Roodeplaat, Pretoria, South Africa. Specifically, the study determined; (i) whether root damage by the flea beetle enhanced infection by M. javanica, (ii) whether L. camara roots infected with the nematode enhanced the performance of the beetle, (iii) whether single or combined effect of the two organisms (i.e. L. bethae and M. javanica) had an overall effect on the growth and biomass of their shared host, L. camara, and (iv) the susceptibility of 10 L. camara varieties that are commonly found in South Africa to M. javanica. The study found that galling on the roots of L. camara by the nematode occurs at the highest inoculation of 300 eggs of L. bethae per plant, and no galling occurred at inoculation of 200 eggs per plant and below. The findings also showed that L. bethae performed better on M. javanica-infected than on healthy L. camara roots, and that more L. bethae adult progeny with slightly bigger body size emerged from M. javanica-infected, than from healthy plants. Fresh weight (galls) of plant roots from treatments where both species (i.e., L. bethae and M. javanica) were combined was significantly higher than that from plants infected with the nematode only, suggesting that the combination of both species induces more galling than the nematode does alone. The above-ground dry biomass was significantly lower both in combined and M. javanica only treatments, than in L. bethae only treatment. The study also found that selected L. camara varieties were infected with M. javanica, albeit at varying degrees of infection. Among the 10 L. camara varieties, Orange Red OR 015 was the most susceptible. Other susceptible varieties included Light Pink 009 LP, Total Pink 021 TP and Dark Pink 018 DP, and these, together with variety Orange Red OR 015, constituted 40% of the L. camara varieties evaluated in the current study. Fifty percent of the varieties displayed slight to moderate susceptibility to M. javanica, while 10% displayed lack of susceptibility. The study concluded that the symbiotic relationship between L. bethae and M. javanica was mutual, resulting in increase in the fitness of the beetle. The combined herbivory by L. bethae and M. javanica was also found to be additive on one of the most common varieties of L. camara in South Africa, and therefore co-infestation by both species might enhance the biological control of this weed in South Africa. The study further concluded that the suitability of some invasive L. camara cultivars such as Light Pink 009 LP and Orange Red 015 OR for M. javanica, might also contribute towards biological control of this weed in South Africa, particularly in areas where the two herbivores species co-exist.
- Full Text:
- Date Issued: 2019
Post-release evaluation of Megamelus scutellaris Berg. (hemiptera: delphacidae): a biological control agent of water hyacinth Eichhornia crassipes (Mart.) Solms-Laub (Pontederiaceae) in South Africa
- Authors: Miller, Benjamin Erich
- Date: 2019
- Subjects: Megamelus scutellaris Berg. , Delphacidae , Noxious weeds -- Biological control -- South Africa , Aquatic weeds -- Biological control -- South Africa , Water hyacinth -- Biological control -- South Africa , Biological pest control agents
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92330 , vital:30710
- Description: Water hyacinth, Eichhornia crassipes (Mart.) Solms-Laub. (Pontederiaceae) is a free-floating aquatic macrophyte from South America that was introduced to South Africa in the 1900s for its attractive ornamental flowers. The plant was classified as a serious invader in the country in the 1970s, eventually becoming the worst invasive aquatic plant in South Africa. Biological control is widely regarded as the most effective method of managing water hyacinth, as it is ecologically safe, cost-effective, and self-sustaining. To date, nine biological control agents have been released in South Africa against water hyacinth, including eight arthropods and a pathogen. Due to the cumulative effects of highly eutrophic waterbodies, which mitigate the damage caused by biological control, and the cold winters which inhibit the rate of biological control agent population build up, South Africa currently has more biological control agents released on water hyacinth than anywhere else in the world. The need for a cold-tolerant agent that can reproduce and develop quickly, while still being damaging to water hyacinth in eutrophic systems, led to the introduction of the most recently released water hyacinth biological control agent, the planthopper Megamelus scutellaris Berg (Hemiptera: Delphacidae), which was initially collected from Argentina. This thesis formed the first post-release evaluation of M. scutellaris since its release in South Africa in 2013. It included a greenhouse experiment to measure the agent’s feeding damage in relation to different nutrient levels and stocking rates, as well as a field component to evaluate both the post-winter recovery of M. scutellaris, and a nationwide survey to measure the establishment of the agent around the country in relation to climate, water quality, and plant health. In the greenhouse experiment, the feeding damage was quantified using measurements of plant growth parameters and chlorophyll fluorometry. It was found that, like other biological control agents of water hyacinth, M. scutellaris was most damaging when released in high numbers on plants grown at medium nutrient levels, and less effective on plants grown at elevated nutrient levels. A water hyacinth infestation on the Kubusi River was selected for the evaluation of the post-winter recovery of M. scutellaris. The Kubusi River is both the first site where M. scutellaris was released, and the coldest site where water hyacinth biological control agents have established successfully in South Africa. Monthly visits tracking seasonal plant health characteristics and agent population densities indicated that the populations of M. scutellaris were impacted most significantly by the season. Low temperatures led to the water hyacinth plants being of poor quality during the winter, which had a subsequent negative effect on the agent populations. The agents could only fully recover by late summer, which meant that the plants were without any significant biological control through the initial phases of the growing season, when they were most vulnerable, and a significant lag-phase occurred between the recovery of the plants and the recovery of the agent population after the winter bottleneck. A survey of all sites where M. scutellaris had been released in South Africa yielded 16 sites where the agents had successfully established, having survived at least one full winter. Among these sites were four sites where the agents were found without them having been released, indicating that they can disperse unaided to new sites. The temperature was a major factor responsible for the success or failure of establishment, with very few agents surviving in the hot areas of South Africa or in areas with a high frost incidence. The density of M. scutellaris was higher in nutrient-rich water, and on plants with more leaves, suggesting that the quality of the plants also contributed to establishment. The results of this thesis showed that M. scutellaris is able to establish successfully in South Africa, and that the agents are capable of causing significant damage to water hyacinth, making it a promising addition to the biological control programme. Novel methods of measuring subtle insect feeding damage in plants and quantifying agent populations are also discussed, along with suggestions for the future implementation of M. scutellaris in South Africa.
- Full Text:
- Date Issued: 2019
- Authors: Miller, Benjamin Erich
- Date: 2019
- Subjects: Megamelus scutellaris Berg. , Delphacidae , Noxious weeds -- Biological control -- South Africa , Aquatic weeds -- Biological control -- South Africa , Water hyacinth -- Biological control -- South Africa , Biological pest control agents
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/92330 , vital:30710
- Description: Water hyacinth, Eichhornia crassipes (Mart.) Solms-Laub. (Pontederiaceae) is a free-floating aquatic macrophyte from South America that was introduced to South Africa in the 1900s for its attractive ornamental flowers. The plant was classified as a serious invader in the country in the 1970s, eventually becoming the worst invasive aquatic plant in South Africa. Biological control is widely regarded as the most effective method of managing water hyacinth, as it is ecologically safe, cost-effective, and self-sustaining. To date, nine biological control agents have been released in South Africa against water hyacinth, including eight arthropods and a pathogen. Due to the cumulative effects of highly eutrophic waterbodies, which mitigate the damage caused by biological control, and the cold winters which inhibit the rate of biological control agent population build up, South Africa currently has more biological control agents released on water hyacinth than anywhere else in the world. The need for a cold-tolerant agent that can reproduce and develop quickly, while still being damaging to water hyacinth in eutrophic systems, led to the introduction of the most recently released water hyacinth biological control agent, the planthopper Megamelus scutellaris Berg (Hemiptera: Delphacidae), which was initially collected from Argentina. This thesis formed the first post-release evaluation of M. scutellaris since its release in South Africa in 2013. It included a greenhouse experiment to measure the agent’s feeding damage in relation to different nutrient levels and stocking rates, as well as a field component to evaluate both the post-winter recovery of M. scutellaris, and a nationwide survey to measure the establishment of the agent around the country in relation to climate, water quality, and plant health. In the greenhouse experiment, the feeding damage was quantified using measurements of plant growth parameters and chlorophyll fluorometry. It was found that, like other biological control agents of water hyacinth, M. scutellaris was most damaging when released in high numbers on plants grown at medium nutrient levels, and less effective on plants grown at elevated nutrient levels. A water hyacinth infestation on the Kubusi River was selected for the evaluation of the post-winter recovery of M. scutellaris. The Kubusi River is both the first site where M. scutellaris was released, and the coldest site where water hyacinth biological control agents have established successfully in South Africa. Monthly visits tracking seasonal plant health characteristics and agent population densities indicated that the populations of M. scutellaris were impacted most significantly by the season. Low temperatures led to the water hyacinth plants being of poor quality during the winter, which had a subsequent negative effect on the agent populations. The agents could only fully recover by late summer, which meant that the plants were without any significant biological control through the initial phases of the growing season, when they were most vulnerable, and a significant lag-phase occurred between the recovery of the plants and the recovery of the agent population after the winter bottleneck. A survey of all sites where M. scutellaris had been released in South Africa yielded 16 sites where the agents had successfully established, having survived at least one full winter. Among these sites were four sites where the agents were found without them having been released, indicating that they can disperse unaided to new sites. The temperature was a major factor responsible for the success or failure of establishment, with very few agents surviving in the hot areas of South Africa or in areas with a high frost incidence. The density of M. scutellaris was higher in nutrient-rich water, and on plants with more leaves, suggesting that the quality of the plants also contributed to establishment. The results of this thesis showed that M. scutellaris is able to establish successfully in South Africa, and that the agents are capable of causing significant damage to water hyacinth, making it a promising addition to the biological control programme. Novel methods of measuring subtle insect feeding damage in plants and quantifying agent populations are also discussed, along with suggestions for the future implementation of M. scutellaris in South Africa.
- Full Text:
- Date Issued: 2019
Assessment of pheromone specificity in Thaumatotibia leucotreta (Meyrick) populations with focus on pest monitoring and the regional rollout of the sterile insect technique in citrus
- Authors: Joubert, Francois D
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Pheromone traps , Citrus -- Diseases and pests -- South Africa , Cryptophlebia leucotreta -- Contol , Cryptophlebia leucotreta -- Biological control
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/60665 , vital:27812
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. It is recognized by several markets as a phytosanitary organism and the efficient control of this pest is now more important than ever. The pheromone communication between the male and female moths has been exploited in order to control FCM through the sterile insect technique (SIT). The sterilized males used for all SIT programmes across South Africa come from a colony that originates from wild material collected from the Citrusdal area of the Western Cape Province. The aim of this study was to determine if any differences in attractiveness of females to males exist between different geographical populations of FCM and if so what impact this would have on the male’s ability to locate females from other populations via the volatile sex pheromone released by the female. Laboratory trials with Y-tube olfactometers and flight tunnels tested the attraction of male moths to virgin females, but did not yield any consistent results. Field experiments were conducted with sterile male Citrusdal moths released and recaptured in yellow delta traps in two separate trials. For one trial, the traps were baited with live virgin females from five different geographical populations including Addo, Nelspruit, Marble Hall, Citrusdal and the Old colony, which is a mixture of several populations. For the other trial traps were baited with various synthetic pheromone blends including three regional blends which included South Africa, Ivory Coast and Malawi and three commercial blends including Pherolure, Isomate and Checkmate. For the virgin female trial the Citrusdal males showed a significant preference for females from their own population. There was also a significant difference in the recaptures from the different synthetic pheromones. The South African blend was the most attractive of all the regional and commercial blends. A cross-mating trial was also conducted under laboratory conditions in petri dishes with five different FCM populations including Citrusdal, Addo, Marble Hall, Nelspruit and Old (mixed origin). Females produced more eggs when mated with males from the same population for the Addo, Marble Hall, Nelspruit and Old (mixed origin) populations. The only case in which this was statistically significant was for the Marble Hall population. All the crosses produced viable eggs and the origin of the male or female did not influence egg hatch. The results from this study may lead to improvements in both the control and monitoring of FCM populations. The control methods include mating disruption, attract-and-kill and SIT. Tailoring these methods for a specific growing area with a pheromone blend originating from the area or releasing sterile moths from a colony that originates from the area may optimize the available monitoring and control options.
- Full Text:
- Date Issued: 2018
- Authors: Joubert, Francois D
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Pheromone traps , Citrus -- Diseases and pests -- South Africa , Cryptophlebia leucotreta -- Contol , Cryptophlebia leucotreta -- Biological control
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/60665 , vital:27812
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. It is recognized by several markets as a phytosanitary organism and the efficient control of this pest is now more important than ever. The pheromone communication between the male and female moths has been exploited in order to control FCM through the sterile insect technique (SIT). The sterilized males used for all SIT programmes across South Africa come from a colony that originates from wild material collected from the Citrusdal area of the Western Cape Province. The aim of this study was to determine if any differences in attractiveness of females to males exist between different geographical populations of FCM and if so what impact this would have on the male’s ability to locate females from other populations via the volatile sex pheromone released by the female. Laboratory trials with Y-tube olfactometers and flight tunnels tested the attraction of male moths to virgin females, but did not yield any consistent results. Field experiments were conducted with sterile male Citrusdal moths released and recaptured in yellow delta traps in two separate trials. For one trial, the traps were baited with live virgin females from five different geographical populations including Addo, Nelspruit, Marble Hall, Citrusdal and the Old colony, which is a mixture of several populations. For the other trial traps were baited with various synthetic pheromone blends including three regional blends which included South Africa, Ivory Coast and Malawi and three commercial blends including Pherolure, Isomate and Checkmate. For the virgin female trial the Citrusdal males showed a significant preference for females from their own population. There was also a significant difference in the recaptures from the different synthetic pheromones. The South African blend was the most attractive of all the regional and commercial blends. A cross-mating trial was also conducted under laboratory conditions in petri dishes with five different FCM populations including Citrusdal, Addo, Marble Hall, Nelspruit and Old (mixed origin). Females produced more eggs when mated with males from the same population for the Addo, Marble Hall, Nelspruit and Old (mixed origin) populations. The only case in which this was statistically significant was for the Marble Hall population. All the crosses produced viable eggs and the origin of the male or female did not influence egg hatch. The results from this study may lead to improvements in both the control and monitoring of FCM populations. The control methods include mating disruption, attract-and-kill and SIT. Tailoring these methods for a specific growing area with a pheromone blend originating from the area or releasing sterile moths from a colony that originates from the area may optimize the available monitoring and control options.
- Full Text:
- Date Issued: 2018
Baculovirus synergism: investigating mixed alphabaculovirus and betabaculovirus infections in the false codling moth, thaumatotibia leucotreta, for improved pest control
- Authors: Jukes, Michael David
- Date: 2018
- Subjects: Baculoviruses , Cryptophlebia leucotreta -- Biological control , Citrus -- Diseases and pests -- South Africa , Pests -- Integrated control , Nucleopolyhedroviruses , Natural pesticides , Cryptophlebia leucotreta granulovirus (CrleGV)
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/61797 , vital:28061
- Description: Baculovirus based biopesticides are an effective and environmentally friendly approach for the control of agriculturally important insect pests. The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is indigenous to southern Africa and is a major pest of citrus crops. This moth poses a serious risk to export of fruit to foreign markets and the control of this pest is therefore imperative. The Cryptophlebia leucotreta granulovirus (CrleGV) has been commercially formulated into the products Cryptogran™ and Cryptex®. These products have been used successfully for over a decade as part of a rigorous integrated pest management (IPM) programme to control T. leucotreta in South Africa. There is however, a continuous need to improve this programme while also addressing new challenges as they arise. An example of a rising concern is the possibility of resistance developing towards CrleGV. This was seen in Europe with field populations of the codling moth, Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae), which developed resistance to the Mexican isolate of the Cydia pomonella granulovirus (CpGV-M). To prevent such a scenario occurring in South Africa, there is a need to improve existing methods of control. For example, additional baculovirus variants can be isolated and characterised for determining virulence, which can then be developed as new biopesticides. Additionally, the potential for synergistic effects between different baculoviruses infecting the same host can be explored for improved virulence. A novel nucleopolyhedrovirus was recently identified in T. leucotreta larval homogenates which were also infected with CrleGV. This provided unique opportunities for continued research and development. In this study, a method using C. pomonella larvae, which can be infected by the NPV but not by CrleGV, was developed to separate the NPV from GV-NPV mixtures in an in vivo system. Examination of NPV OBs by transmission electron microscopy showed purified occlusion bodies with a single nucleopolyhedrovirus morphology (SNPV). Genetic characterisation identified the novel NPV as Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV), which was recently isolated from the litchi moth, Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae). To begin examining the potential for synergism between the two viruses, a multiplex PCR assay was developed to accurately detect CrleGV and/or CrpeNPV in mixed infections. This assay was applied to various samples to screen for the presence of CrpeNPV and CrleGV. Additionally, a validation experiment was performed using different combinations of CrpeNPV and/or CrleGV to evaluate the effectiveness of the mPCR assay. The results obtained indicated a high degree of specificity with the correct amplicons generated for each test sample. The biological activity of CrpeNPV and CrleGV were evaluated using surface dose bioassays, both individually and in various combinations, against T. leucotreta neonate larvae in a laboratory setting. A synergistic effect was recorded in the combination treatments, showing improved virulence when compared against each virus in isolation. The LC90 for CrpeNPV and CrleGV when applied alone against T. leucotreta was calculated to be 2.75*106 and 3.00*106 OBs.ml"1 respectively. These values decreased to 1.07*106 and 7.18*105 OBs.ml"1 when combinations of CrleGV and CrpeNPV were applied at ratios of 3:1 and 1:3 respectively. These results indicate a potential for developing improved biopesticides for the control of T. leucotreta in the field. To better understand the interactions between CrleGV and CrpeNPV, experiments involving the serial passage of these viruses through T. leucotreta larvae were performed. This was done using each virus in isolation as well as both viruses in different combinations. Genomic DNA was extracted from recovered occlusion bodies after each passage and examined by multiplex and quantitative PCR. This analysis enabled the detection of each virus present throughout this assay, as well as recording shifts in the ratio of CrleGV and CrpeNPV at each passage. CrleGV rapidly became the dominant virus in all treatments, indicating a potentially antagonistic interaction during serial passage. Additionally, CrpeNPV and CrleGV were detected in treatments which were not originally inoculated with one or either virus, indicating potential covert infections in T. leucotreta. Occlusion bodies recovered from the final passage were used to inoculate C. pomonella larvae to isolate CrpeNPV from CrleGV. Genomic DNA was extracted from these CrpeNPV OBs and examined by restriction endonuclease assays and next generation sequencing. This enabled the identification of potential recombination events which may have occurred during the dual GV and NPV infections throughout the passage assay. No recombination events were identified in the CrpeNPV genome sequences assembled from virus collected at the end of the passage assay. Lastly, the efficacy of CrpeNPV and CrleGV, both alone and in various combinations, was evaluated in the field. In two separate trials conducted on citrus, unfavorable field conditions resulted in no significant reduction in fruit infestation for both the virus and chemical treatments. While not statistically significant, virus treatments were recorded to have the lowest levels of fruit infestation with a measured reduction of up to 64 %. This study is the first to report a synergistic effect between CrleGV and CrpeNPV in T. leucotreta. The discovery of beneficial interactions creates an opportunity for the development of novel biopesticides for improved control of this pest in South Africa.
- Full Text:
- Date Issued: 2018
- Authors: Jukes, Michael David
- Date: 2018
- Subjects: Baculoviruses , Cryptophlebia leucotreta -- Biological control , Citrus -- Diseases and pests -- South Africa , Pests -- Integrated control , Nucleopolyhedroviruses , Natural pesticides , Cryptophlebia leucotreta granulovirus (CrleGV)
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/61797 , vital:28061
- Description: Baculovirus based biopesticides are an effective and environmentally friendly approach for the control of agriculturally important insect pests. The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is indigenous to southern Africa and is a major pest of citrus crops. This moth poses a serious risk to export of fruit to foreign markets and the control of this pest is therefore imperative. The Cryptophlebia leucotreta granulovirus (CrleGV) has been commercially formulated into the products Cryptogran™ and Cryptex®. These products have been used successfully for over a decade as part of a rigorous integrated pest management (IPM) programme to control T. leucotreta in South Africa. There is however, a continuous need to improve this programme while also addressing new challenges as they arise. An example of a rising concern is the possibility of resistance developing towards CrleGV. This was seen in Europe with field populations of the codling moth, Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae), which developed resistance to the Mexican isolate of the Cydia pomonella granulovirus (CpGV-M). To prevent such a scenario occurring in South Africa, there is a need to improve existing methods of control. For example, additional baculovirus variants can be isolated and characterised for determining virulence, which can then be developed as new biopesticides. Additionally, the potential for synergistic effects between different baculoviruses infecting the same host can be explored for improved virulence. A novel nucleopolyhedrovirus was recently identified in T. leucotreta larval homogenates which were also infected with CrleGV. This provided unique opportunities for continued research and development. In this study, a method using C. pomonella larvae, which can be infected by the NPV but not by CrleGV, was developed to separate the NPV from GV-NPV mixtures in an in vivo system. Examination of NPV OBs by transmission electron microscopy showed purified occlusion bodies with a single nucleopolyhedrovirus morphology (SNPV). Genetic characterisation identified the novel NPV as Cryptophlebia peltastica nucleopolyhedrovirus (CrpeNPV), which was recently isolated from the litchi moth, Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae). To begin examining the potential for synergism between the two viruses, a multiplex PCR assay was developed to accurately detect CrleGV and/or CrpeNPV in mixed infections. This assay was applied to various samples to screen for the presence of CrpeNPV and CrleGV. Additionally, a validation experiment was performed using different combinations of CrpeNPV and/or CrleGV to evaluate the effectiveness of the mPCR assay. The results obtained indicated a high degree of specificity with the correct amplicons generated for each test sample. The biological activity of CrpeNPV and CrleGV were evaluated using surface dose bioassays, both individually and in various combinations, against T. leucotreta neonate larvae in a laboratory setting. A synergistic effect was recorded in the combination treatments, showing improved virulence when compared against each virus in isolation. The LC90 for CrpeNPV and CrleGV when applied alone against T. leucotreta was calculated to be 2.75*106 and 3.00*106 OBs.ml"1 respectively. These values decreased to 1.07*106 and 7.18*105 OBs.ml"1 when combinations of CrleGV and CrpeNPV were applied at ratios of 3:1 and 1:3 respectively. These results indicate a potential for developing improved biopesticides for the control of T. leucotreta in the field. To better understand the interactions between CrleGV and CrpeNPV, experiments involving the serial passage of these viruses through T. leucotreta larvae were performed. This was done using each virus in isolation as well as both viruses in different combinations. Genomic DNA was extracted from recovered occlusion bodies after each passage and examined by multiplex and quantitative PCR. This analysis enabled the detection of each virus present throughout this assay, as well as recording shifts in the ratio of CrleGV and CrpeNPV at each passage. CrleGV rapidly became the dominant virus in all treatments, indicating a potentially antagonistic interaction during serial passage. Additionally, CrpeNPV and CrleGV were detected in treatments which were not originally inoculated with one or either virus, indicating potential covert infections in T. leucotreta. Occlusion bodies recovered from the final passage were used to inoculate C. pomonella larvae to isolate CrpeNPV from CrleGV. Genomic DNA was extracted from these CrpeNPV OBs and examined by restriction endonuclease assays and next generation sequencing. This enabled the identification of potential recombination events which may have occurred during the dual GV and NPV infections throughout the passage assay. No recombination events were identified in the CrpeNPV genome sequences assembled from virus collected at the end of the passage assay. Lastly, the efficacy of CrpeNPV and CrleGV, both alone and in various combinations, was evaluated in the field. In two separate trials conducted on citrus, unfavorable field conditions resulted in no significant reduction in fruit infestation for both the virus and chemical treatments. While not statistically significant, virus treatments were recorded to have the lowest levels of fruit infestation with a measured reduction of up to 64 %. This study is the first to report a synergistic effect between CrleGV and CrpeNPV in T. leucotreta. The discovery of beneficial interactions creates an opportunity for the development of novel biopesticides for improved control of this pest in South Africa.
- Full Text:
- Date Issued: 2018
Elevated CO2 determines cell damage and nitrogen allocation in barley subjected to aphid herbivory
- Authors: Gallagher, Sean
- Date: 2018
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64337 , vital:28535
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2018
- Authors: Gallagher, Sean
- Date: 2018
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/64337 , vital:28535
- Description: Expected release date-May 2019
- Full Text:
- Date Issued: 2018
Isolation, identification and genetic characterisation of a microsporidium isolated from the carob moth, Ectomyelois ceratoniae (Lepidoptera: Pyralidae)
- Authors: Lloyd, Melissa
- Date: 2018
- Subjects: Pyralidae , Pyralidae -- Genetics , Pyralidae -- Phylogeny , Pyralidae -- Pathogens , Cladistic analysis , Transmission electron microscopy , Carob moth (Ectomyelois ceratoniae)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/61894 , vital:28075
- Description: Carob moth, Ectomyelois ceratoniae (Zeller) (Lepidoptera: Pyralidae) is an economically important pest, yet its biology and pest status on citrus in South Africa was, until recently, poorly understood. A study was initiated to determine the cause of collapse of a laboratory carob moth colony that was established to investigate the biology of carob moth on citrus and to develop integrated management strategies for the pest. An organism was isolated from deceased larvae and was morphologically identified as a microsporidium, based on transmission electron microscopy. Microsporidia are obligate intracellular parasites that have been found to infect almost all eukaryotes. Several Nosema species have been isolated from economically important insect pests, yet little genetic information is available from online databases for identification. Mature spores were recovered and measured using transmission electron microscopy. Spores were ovocylindrical with a wrinkled exospore, and had a length of 2.8 ± 0.02 pm and a width of 1.6 ± 0.04 pm. The identity of the microsporidium was confirmed by PCR amplification, sequencing and analysis of the regions encoding the ribosomal RNA. BLAST analysis of the different rRNA regions amplified showed that the microsporidium shared a 96 - 99 % identity with Nosema sp. M-Pr, Nosema carpocapsae, Nosema oulemae, Nosema sp. CO1, Microsporidium 57864, and Nosema bombi. Phylogenetic analysis of the SSU and LSU rRNA genes showed that the microsporidium clustered with the Nosema / Vairimorpha clade, supported by a bootstrap value of 100. The organisation of the RNA cistron was determined by PCR amplification using the primer set 18f and L1328r to be 5’-SSU-ITS-LSU-IGS-5S-3’, which confirms the placement of the microsporidium within the Nosema / Vairimorpha clade. Because the BLAST results showed a close relationship with Nosema carpocapsae, a microsporidium infecting codling moth, the pathogenicity of the microsporidium was tested against codling moth by inoculating artificial diet with a high spore concentration of 1.1 x 107 spores/ml and a low spore concentration of 1.1 x 104 spores/ml. DNA was extracted from deceased larvae inoculated with the high concentration, and PCR of the SSU rRNA gene and bacterial 16S region was performed. Mortality in the high concentration experiment was significant (p = 0.05), but the cause of infection was determined to be a bacterium, through sequencing and BLAST analysis of the bacterial 16S rDNA. The bacterium shared a 99 % identity with Bacillus cereus. Percentage mortality (p = 0.09), larval mass (p = 0.09) and instar (p = 0.24) did not differ significantly between treatments in the low concentration experiment. DNA was extracted from the larvae and PCR amplification of the SSU rRNA gene was performed to determine whether microsporidia were present. No SSU bands were observed in any of the treatments and percentage mortality was not significant, thus it was determined that no infection occurred. This is the first study to report the genetic characterisation of a microsporidium isolated from carob moth and provides important genetic information for classification of microsporidia within the Nosema / Vairimorpha clade. It is also one of few studies in which the complete rRNA cistron of a species within the Nosema / Vairimorpha clade has been sequenced. The identification of a microsporidium from a laboratory colony of carob moth is important as it provides information about pathogens infecting the carob moth and constraints to carob moth rearing, which is useful for further studies on rearing carob moth and for establishment of a clean colony for research purposes.
- Full Text:
- Date Issued: 2018
- Authors: Lloyd, Melissa
- Date: 2018
- Subjects: Pyralidae , Pyralidae -- Genetics , Pyralidae -- Phylogeny , Pyralidae -- Pathogens , Cladistic analysis , Transmission electron microscopy , Carob moth (Ectomyelois ceratoniae)
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/61894 , vital:28075
- Description: Carob moth, Ectomyelois ceratoniae (Zeller) (Lepidoptera: Pyralidae) is an economically important pest, yet its biology and pest status on citrus in South Africa was, until recently, poorly understood. A study was initiated to determine the cause of collapse of a laboratory carob moth colony that was established to investigate the biology of carob moth on citrus and to develop integrated management strategies for the pest. An organism was isolated from deceased larvae and was morphologically identified as a microsporidium, based on transmission electron microscopy. Microsporidia are obligate intracellular parasites that have been found to infect almost all eukaryotes. Several Nosema species have been isolated from economically important insect pests, yet little genetic information is available from online databases for identification. Mature spores were recovered and measured using transmission electron microscopy. Spores were ovocylindrical with a wrinkled exospore, and had a length of 2.8 ± 0.02 pm and a width of 1.6 ± 0.04 pm. The identity of the microsporidium was confirmed by PCR amplification, sequencing and analysis of the regions encoding the ribosomal RNA. BLAST analysis of the different rRNA regions amplified showed that the microsporidium shared a 96 - 99 % identity with Nosema sp. M-Pr, Nosema carpocapsae, Nosema oulemae, Nosema sp. CO1, Microsporidium 57864, and Nosema bombi. Phylogenetic analysis of the SSU and LSU rRNA genes showed that the microsporidium clustered with the Nosema / Vairimorpha clade, supported by a bootstrap value of 100. The organisation of the RNA cistron was determined by PCR amplification using the primer set 18f and L1328r to be 5’-SSU-ITS-LSU-IGS-5S-3’, which confirms the placement of the microsporidium within the Nosema / Vairimorpha clade. Because the BLAST results showed a close relationship with Nosema carpocapsae, a microsporidium infecting codling moth, the pathogenicity of the microsporidium was tested against codling moth by inoculating artificial diet with a high spore concentration of 1.1 x 107 spores/ml and a low spore concentration of 1.1 x 104 spores/ml. DNA was extracted from deceased larvae inoculated with the high concentration, and PCR of the SSU rRNA gene and bacterial 16S region was performed. Mortality in the high concentration experiment was significant (p = 0.05), but the cause of infection was determined to be a bacterium, through sequencing and BLAST analysis of the bacterial 16S rDNA. The bacterium shared a 99 % identity with Bacillus cereus. Percentage mortality (p = 0.09), larval mass (p = 0.09) and instar (p = 0.24) did not differ significantly between treatments in the low concentration experiment. DNA was extracted from the larvae and PCR amplification of the SSU rRNA gene was performed to determine whether microsporidia were present. No SSU bands were observed in any of the treatments and percentage mortality was not significant, thus it was determined that no infection occurred. This is the first study to report the genetic characterisation of a microsporidium isolated from carob moth and provides important genetic information for classification of microsporidia within the Nosema / Vairimorpha clade. It is also one of few studies in which the complete rRNA cistron of a species within the Nosema / Vairimorpha clade has been sequenced. The identification of a microsporidium from a laboratory colony of carob moth is important as it provides information about pathogens infecting the carob moth and constraints to carob moth rearing, which is useful for further studies on rearing carob moth and for establishment of a clean colony for research purposes.
- Full Text:
- Date Issued: 2018
The implementation of a push-pull programme for the control of Eldana saccharina (Lepidoptera: Pyralidae) in sugarcane in the coastal regions of Kwazulu-Natal, South Africa
- Authors: Mulcahy, Megan Marie
- Date: 2018
- Subjects: Pyralidae -- South Africa -- KwaZulu-Natal , Pests -- Integrated control , Sugarcane -- Diseases and pests -- South Africa -- KwaZulu-Natal , Stem borers -- Effect of habitat modification on -- South Africa -- KwaZulu-Natal , Insect-plant relationships -- South Africa -- KwaZulu-Natal
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/63290 , vital:28390
- Description: Eldana saccharina, an indigenous lepidopteran stemborer, is considered the most damaging pest of sugarcane in South Africa. Researchers have advocated the use of an area-wide integrated pest management (AW-IPM) programme as a means of improving the control of this pest. A push-pull strategy was developed as a component of this AW-IPM approach. The push-pull strategy in sugarcane is a habitat management method of pest control that uses plants that are both repellent (Melinis Minutiflora) and attractive (Cyperus dives, Cyperus papyrus and Bt-maize) to E. saccharina. Previous research into push-pull has shown that this strategy is an effective tool for the control of E. saccharina. Push-pull has been implemented successfully in the Midlands North sugarcane growing region of KwaZulu-Natal (KZN), South Africa. Despite the proven efficacy of push-pull, very little push-pull research has been conducted in the coastal sugarcane growing regions of KZN, and adoption of this technology has been poor in these regions. Therefore, the aim of this research was to facilitate the implementation of push-pull for the management of E. saccharina in sugarcane in the coastal regions of KZN. This was done by focussing on on-farm field trials and farmer participatory research. On-farm push-pull field trials were conducted on five model farms in the North and South Coast sugarcane growing regions of KZN. High levels of E. saccharina were recorded during this study. The push-pull treatment sites showed a significant reduction of E. saccharina damage on four of the five farms used in the study. Mean percentage of stalks damaged decreased by up to 50 % in the presence of the repellent grass species, M. minutiflora. The number of E. saccharina found per 100 stalks also decreased significantly at these farms. The farm which did not show a significant reduction in E. saccharina populations or damage had low numbers of this pest in the sugarcane throughout the experiment. This demonstrates that push-pull is more effective in areas that have high levels of E. saccharina. Stem borer surveys in wetlands on sugarcane farms revealed that high numbers of E. saccharina were found within the pull plants, C. papyrus and C. dives, in comparison to the push-pull sites. This verifies that the pull plants do work efficiently to attract E. saccharina away from sugarcane. Additionally, eight parasitoids emerged from E. saccharina larvae collected in wetland sedges. The beneficial roles that push-pull plants play in attracting and maintaining natural enemies in the agroecosystem are discussed, and these findings further demonstrate the important ecosystem, and pest management services that wetlands provide on sugarcane farms. The success of the push-pull trials in this study show that this technology can be an effective tool for controlling E. saccharina in the coastal sugarcane growing regions. The timing of the planting of push-pull plants was shown to play a role in the efficacy of this technology. The study also confirmed that push-pull should be used as a component of AW-IPM in conjunction with good crop management practices. Surveys were undertaken to determine large-scale sugarcane growers' (LSGs) knowledge and perceptions of E. saccharina and other pests. Research regarding the farmers' perceptions of push- pull was also conducted to better understand the drivers and barriers to adoption of push-pull, and other new technologies. The surveys found that large-scale farmers in the coastal regions suffer from high infestations of E. saccharina. As such there is scope for the introduction of new pest management practices such as push-pull in this area. Farmers also demonstrated a good basic knowledge of E. saccharina and IPM. However, LSGs had a poor understanding of push-pull and how it works, as well as the plants that make up the push-pull system that is being implemented against E. saccharina in South Africa. A dearth in practical knowledge regarding the implementation of push-pull was seen as a major barrier to the adoption of this strategy, as was financial instability, farmer attitudes and poor institutional support. Farmers recommended collaboration amongst stakeholders, improved education, proof of the efficacy of push-pull and incentives as tools to improve the implementation of this strategy in the coastal sugarcane growing regions of KZN. Farmers preferred direct contact with extension personnel and experiential learning opportunities when acquiring information about push- pull and other new pest management practices. If opportunities for push-pull education are increased through direct contact with extension personnel, and through on-farm demonstrations, and if inputs are provided in the form of push-pull plants, it is likely that push-pull will succeed amongst coastal LSGs, especially since farmers had an overall positive attitude towards the technology. Surveys amongst small-scale sugarcane growers (SSGs) showed that sugarcane is important in the lives of these farmers. The SSGs perceive pests to be a major constraint to their farming systems, and they identified E. saccharina as a major pest of sugarcane. The farmers also demonstrated good knowledge of sugarcane pests and vegetable pests. However, SSGs lacked knowledge regarding pest management practices and beneficial insects. Extension and advisory services should to continue concentrating on pest management practices to educate SSGs on the variety and application of pest control strategies. SSGs were found to employ complex, diverse and integrated agricultural systems that are well-suited to the implementation of IPM technologies such as push-pull. Since insect pests act were found to be a major constraint to SSG sugarcane production, push-pull was deemed a feasible pest management strategy for coastal farmers and its implementation by SSGs should be further explored. SSGs in this study were also concerned about vegetable pests, therefore if push-pull can be adapted to help protect additional crops, adoption of this technology by small-scale growers will improve.
- Full Text:
- Date Issued: 2018
- Authors: Mulcahy, Megan Marie
- Date: 2018
- Subjects: Pyralidae -- South Africa -- KwaZulu-Natal , Pests -- Integrated control , Sugarcane -- Diseases and pests -- South Africa -- KwaZulu-Natal , Stem borers -- Effect of habitat modification on -- South Africa -- KwaZulu-Natal , Insect-plant relationships -- South Africa -- KwaZulu-Natal
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/63290 , vital:28390
- Description: Eldana saccharina, an indigenous lepidopteran stemborer, is considered the most damaging pest of sugarcane in South Africa. Researchers have advocated the use of an area-wide integrated pest management (AW-IPM) programme as a means of improving the control of this pest. A push-pull strategy was developed as a component of this AW-IPM approach. The push-pull strategy in sugarcane is a habitat management method of pest control that uses plants that are both repellent (Melinis Minutiflora) and attractive (Cyperus dives, Cyperus papyrus and Bt-maize) to E. saccharina. Previous research into push-pull has shown that this strategy is an effective tool for the control of E. saccharina. Push-pull has been implemented successfully in the Midlands North sugarcane growing region of KwaZulu-Natal (KZN), South Africa. Despite the proven efficacy of push-pull, very little push-pull research has been conducted in the coastal sugarcane growing regions of KZN, and adoption of this technology has been poor in these regions. Therefore, the aim of this research was to facilitate the implementation of push-pull for the management of E. saccharina in sugarcane in the coastal regions of KZN. This was done by focussing on on-farm field trials and farmer participatory research. On-farm push-pull field trials were conducted on five model farms in the North and South Coast sugarcane growing regions of KZN. High levels of E. saccharina were recorded during this study. The push-pull treatment sites showed a significant reduction of E. saccharina damage on four of the five farms used in the study. Mean percentage of stalks damaged decreased by up to 50 % in the presence of the repellent grass species, M. minutiflora. The number of E. saccharina found per 100 stalks also decreased significantly at these farms. The farm which did not show a significant reduction in E. saccharina populations or damage had low numbers of this pest in the sugarcane throughout the experiment. This demonstrates that push-pull is more effective in areas that have high levels of E. saccharina. Stem borer surveys in wetlands on sugarcane farms revealed that high numbers of E. saccharina were found within the pull plants, C. papyrus and C. dives, in comparison to the push-pull sites. This verifies that the pull plants do work efficiently to attract E. saccharina away from sugarcane. Additionally, eight parasitoids emerged from E. saccharina larvae collected in wetland sedges. The beneficial roles that push-pull plants play in attracting and maintaining natural enemies in the agroecosystem are discussed, and these findings further demonstrate the important ecosystem, and pest management services that wetlands provide on sugarcane farms. The success of the push-pull trials in this study show that this technology can be an effective tool for controlling E. saccharina in the coastal sugarcane growing regions. The timing of the planting of push-pull plants was shown to play a role in the efficacy of this technology. The study also confirmed that push-pull should be used as a component of AW-IPM in conjunction with good crop management practices. Surveys were undertaken to determine large-scale sugarcane growers' (LSGs) knowledge and perceptions of E. saccharina and other pests. Research regarding the farmers' perceptions of push- pull was also conducted to better understand the drivers and barriers to adoption of push-pull, and other new technologies. The surveys found that large-scale farmers in the coastal regions suffer from high infestations of E. saccharina. As such there is scope for the introduction of new pest management practices such as push-pull in this area. Farmers also demonstrated a good basic knowledge of E. saccharina and IPM. However, LSGs had a poor understanding of push-pull and how it works, as well as the plants that make up the push-pull system that is being implemented against E. saccharina in South Africa. A dearth in practical knowledge regarding the implementation of push-pull was seen as a major barrier to the adoption of this strategy, as was financial instability, farmer attitudes and poor institutional support. Farmers recommended collaboration amongst stakeholders, improved education, proof of the efficacy of push-pull and incentives as tools to improve the implementation of this strategy in the coastal sugarcane growing regions of KZN. Farmers preferred direct contact with extension personnel and experiential learning opportunities when acquiring information about push- pull and other new pest management practices. If opportunities for push-pull education are increased through direct contact with extension personnel, and through on-farm demonstrations, and if inputs are provided in the form of push-pull plants, it is likely that push-pull will succeed amongst coastal LSGs, especially since farmers had an overall positive attitude towards the technology. Surveys amongst small-scale sugarcane growers (SSGs) showed that sugarcane is important in the lives of these farmers. The SSGs perceive pests to be a major constraint to their farming systems, and they identified E. saccharina as a major pest of sugarcane. The farmers also demonstrated good knowledge of sugarcane pests and vegetable pests. However, SSGs lacked knowledge regarding pest management practices and beneficial insects. Extension and advisory services should to continue concentrating on pest management practices to educate SSGs on the variety and application of pest control strategies. SSGs were found to employ complex, diverse and integrated agricultural systems that are well-suited to the implementation of IPM technologies such as push-pull. Since insect pests act were found to be a major constraint to SSG sugarcane production, push-pull was deemed a feasible pest management strategy for coastal farmers and its implementation by SSGs should be further explored. SSGs in this study were also concerned about vegetable pests, therefore if push-pull can be adapted to help protect additional crops, adoption of this technology by small-scale growers will improve.
- Full Text:
- Date Issued: 2018
The thermal physiology of Stenopelmus rufinasus and Neohydronomus affinis (Coleoptera: Curculionidae), biological control agents for the invasive alien aquatic weeds Azolla filiculoides and Pistia stratiotes respectively
- Authors: Mvandaba, Sisanda F
- Date: 2018
- Subjects: Beetles -- South Africa , Curculionidae -- South Africa , Azolla filiculoides -- South Africa , Water lettuce -- South Africa , Aquatic weeds -- Biological control -- South Africa , Stenopelmus rufinasus , Neohydronomus affinis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/62362 , vital:28158
- Description: Water lettuce, Pistia stratiotes L. (Araceae), and red water fern, Azolla filiculoides Lam. (Azollaceae), are floating aquatic macrophytes that have become problematic invaders in numerous South African waterbodies. Two weevils, Neohydronomus affinis Hustache 1926 (Coleoptera: Curculionidae) and Stenopelmus rufinasus Gyllenhal 1936 (Coleoptera: Curculionidae), are successful biological control agents of these two species, respectively, in South Africa. However, nothing is known about the thermal physiology of these two species Therefore, the aim of this study was to investigate the thermal physiologies of these two species to explain their establishment, distribution and impact in the field. Laboratory based thermal physiology trials showed that both weevils were widely tolerant of cold and warm temperatures. The CTmin of N. affinis was determined to be 5.5 ± 0.312°C and the CTmax was 44 ± 0.697°C, while the CTmin of S. rufinasus was 5.4 ± 0.333°C and the CTmax was 44.5 ± 0.168°C. In addition, the lower lethal temperatures were -9.8 ± 0.053°C and -7.2 ± 0.19°C, and the upper lethal temperatures were 42.8 ± 0.053°C and 41.9 ± 0.19°C respectively. These results suggest that both species should not be limited by cold winter temperatures, as previously thought. This is evident in the field, where S. rufinasus has established widely on A. filiculoides, despite local cold climates in some areas of the plant’s distribution. Even though N. affinis has a similar thermal range, and should therefore theoretically reflect a similar distribution to S. rufinasus throughout South Africa, its distribution is limited by the range of its host, which is restricted to the warmer regions of the country, as is its biocontrol agent. Using the reduced major axis regression method, the development for N. affinis was described using the formulay=12.976x+435.24, while the development of S. rufinasus was described by y=13.6x+222.45. These results showed that S. rufinasus develops much faster, in fact almost twice as quickly, than N. affinis. Using these formulae and temperature data obtained from the South African Weather Service, N. affinis was predicted to complete between 4 and 9 generations per year in South Africa, while S. rufinasus was predicted to complete between 5 and 14 generations per year around the country. This study showed that although the native range of these two species is warm temperate to tropical, they possess sufficient thermal plasticity to not only establish, but also damage their respective host plants in far cooler climates. Thus, in South Africa N. affinis and S. rufinasus are limited by the distribution of their target weeds and not climate.
- Full Text:
- Date Issued: 2018
- Authors: Mvandaba, Sisanda F
- Date: 2018
- Subjects: Beetles -- South Africa , Curculionidae -- South Africa , Azolla filiculoides -- South Africa , Water lettuce -- South Africa , Aquatic weeds -- Biological control -- South Africa , Stenopelmus rufinasus , Neohydronomus affinis
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/62362 , vital:28158
- Description: Water lettuce, Pistia stratiotes L. (Araceae), and red water fern, Azolla filiculoides Lam. (Azollaceae), are floating aquatic macrophytes that have become problematic invaders in numerous South African waterbodies. Two weevils, Neohydronomus affinis Hustache 1926 (Coleoptera: Curculionidae) and Stenopelmus rufinasus Gyllenhal 1936 (Coleoptera: Curculionidae), are successful biological control agents of these two species, respectively, in South Africa. However, nothing is known about the thermal physiology of these two species Therefore, the aim of this study was to investigate the thermal physiologies of these two species to explain their establishment, distribution and impact in the field. Laboratory based thermal physiology trials showed that both weevils were widely tolerant of cold and warm temperatures. The CTmin of N. affinis was determined to be 5.5 ± 0.312°C and the CTmax was 44 ± 0.697°C, while the CTmin of S. rufinasus was 5.4 ± 0.333°C and the CTmax was 44.5 ± 0.168°C. In addition, the lower lethal temperatures were -9.8 ± 0.053°C and -7.2 ± 0.19°C, and the upper lethal temperatures were 42.8 ± 0.053°C and 41.9 ± 0.19°C respectively. These results suggest that both species should not be limited by cold winter temperatures, as previously thought. This is evident in the field, where S. rufinasus has established widely on A. filiculoides, despite local cold climates in some areas of the plant’s distribution. Even though N. affinis has a similar thermal range, and should therefore theoretically reflect a similar distribution to S. rufinasus throughout South Africa, its distribution is limited by the range of its host, which is restricted to the warmer regions of the country, as is its biocontrol agent. Using the reduced major axis regression method, the development for N. affinis was described using the formulay=12.976x+435.24, while the development of S. rufinasus was described by y=13.6x+222.45. These results showed that S. rufinasus develops much faster, in fact almost twice as quickly, than N. affinis. Using these formulae and temperature data obtained from the South African Weather Service, N. affinis was predicted to complete between 4 and 9 generations per year in South Africa, while S. rufinasus was predicted to complete between 5 and 14 generations per year around the country. This study showed that although the native range of these two species is warm temperate to tropical, they possess sufficient thermal plasticity to not only establish, but also damage their respective host plants in far cooler climates. Thus, in South Africa N. affinis and S. rufinasus are limited by the distribution of their target weeds and not climate.
- Full Text:
- Date Issued: 2018
Biotic and abiotic factors promoting the development and proliferation of water hyacinth (eichhornia crassipes (Mart.) Solms-Laub.) in the Wouri Basin (Douala-Cameroon) and environs, with implications for its control
- Voukeng, Sonia Nadege Kenfack
- Authors: Voukeng, Sonia Nadege Kenfack
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/7644 , vital:21281
- Description: The Wouri River, situated in the Wouri Basin, is one of the main rivers of the Littoral Region in the city of Douala in Cameroon. It is a source of income and food for the population living around these areas. Since the 1990s, the fishing, transportation, irrigation and sand extraction activities have been impeded by the invasion of aquatic plants, specifically water hyacinth (Eichhornia crassipes [Mart.] Solms-Laubach: Pontederiaceae). Introduced in 1997 to the shore of Lake Chad, water hyacinth has invaded almost 114 ha of the Wouri Basin. Furthermore, Douala, the economic capital of the Cameroon and location for more than 70% of the country’s industries, uses the Wouri River and its tributaries to deposit its effluent and waste, which has worsened the problem of water hyacinth. This thesis examined the ecological and socio-economic impacts of water hyacinth in the Wouri Basin and its possible control. An increase in the nutrients in the water has provided water hyacinth with appropriate conditions for its fast growth during both the rainy and dry seasons. The availability of nutrients in these areas is enhanced by the constant, daily tidal fluctuation of water, providing enough water to the plant for easy nutrient uptake. A survey of the impacts of water hyacinth on aquatic plant communities in the Wouri Basin showed that this plant is able to out-compete native species. Assessment of the impact of water hyacinth on the abundance and diversity of plant communities indicated that at some invaded sites, 65% of the vegetation consisted of water hyacinth. Species found in association with water hyacinth with a high level of abundance-dominance were Pistia stratiotes L. (Araceae) (another invader), Commelina benghalensis L. (Commelinaceae) and Echinochloa pyramidalis (Lam.) Hitchc. & Chase (Poaceae). This component of the study also showed that habitats rich in water hyacinth were poor in diversity, while habitats without water hyacinth were rich in diversity, thus raising awareness of the importance of monitoring invasive aquatic weeds along the Wouri Basin, and of implementing correct control management of all invasive aquatic weeds. Communities living along the invaded rivers are well aware of the range of problems caused by the weed; because as the rivers and water bodies used for fishing, transportation, and sand extraction are progressively invaded by the weed, the riparian population is the first to feel the impact. The impact on people has been noticeable, with an increase in diseases, such as malaria, cholera, diarrhoea, typhoid, filariasis, schistosomiasis, scabies and yellow fever increasing the need for a medicine and hospitalization. Economic losses due to the management of invasive aquatic weeds were recorded, and the Ministry of Environment spent an estimated US$1 200 000 between 2010 and 2015 to manage this scourge. In 2016, an amount of US$160 000 was transferred to these regions to manage invasive aquatic weeds, especially water hyacinth, although manual clearing is still the only method used to control this weed. Isolation of fungi from diseased water hyacinth plants in the Wouri Basin revealed several fungal species, most of which have been isolated from water hyacinth species in water bodies elsewhere, which showed a higher diversity during the dry season than during the rainy season. These fungi included Acremonium zonatum (Sawada). W. Gams (Hypocreaceae), Alternaria eichhorniae Nag Raj & Ponnappa (Pleosporaceae), Chaetomium sp., Colletotrichum sp., Curvularia pallescens Boedjin (Pleosporaceae), Curvalaria sp., Epicoccum nigrum Link (Pleosporaceae), Fusarium sp., Pithomyces chartarum fBerk. & M. A. Curtis) M. B. Ellis (Montagnulaceae), to a lesser extent Myrothecium roridum Tode ex Fr. (Incertae sedis) and Nigrospora sp. Although never released in Cameroon, arthropod biological control agents (Neochetina eichhorniae Warner (Coleoptera, Curculionidae) and N. bruchi Hustache (Coleoptera, Curculionidae)) were present, but their populations were relatively low. The slow spread of the insect population was explained by several factors, among them the tidal fluctuation of water, which has an impact on the population growth of the weevils. Whilst adults may be able to survive tidal fluctuations, larvae are severely impacted by them, contributing to the slow success of biological control. In this study, a significant increase in pathogen-induced disease severity and incidence was noted when Neochetina eichhorniae weevils were present, possibly because larvae tunnelling on the petiole created openings for the penetration of fungal spores. This study highlights the negative impacts of water hyacinth, on the environment, people, and thus economy of Cameroon. The presence of biological control agents and pathogens offers Cameroon the possibility of initiating and properly implementing the biological control option, or an integrated management solution, to manage water hyacinth in the Wouri Basin, and in the rest of Cameroon.
- Full Text:
- Date Issued: 2017
- Authors: Voukeng, Sonia Nadege Kenfack
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/7644 , vital:21281
- Description: The Wouri River, situated in the Wouri Basin, is one of the main rivers of the Littoral Region in the city of Douala in Cameroon. It is a source of income and food for the population living around these areas. Since the 1990s, the fishing, transportation, irrigation and sand extraction activities have been impeded by the invasion of aquatic plants, specifically water hyacinth (Eichhornia crassipes [Mart.] Solms-Laubach: Pontederiaceae). Introduced in 1997 to the shore of Lake Chad, water hyacinth has invaded almost 114 ha of the Wouri Basin. Furthermore, Douala, the economic capital of the Cameroon and location for more than 70% of the country’s industries, uses the Wouri River and its tributaries to deposit its effluent and waste, which has worsened the problem of water hyacinth. This thesis examined the ecological and socio-economic impacts of water hyacinth in the Wouri Basin and its possible control. An increase in the nutrients in the water has provided water hyacinth with appropriate conditions for its fast growth during both the rainy and dry seasons. The availability of nutrients in these areas is enhanced by the constant, daily tidal fluctuation of water, providing enough water to the plant for easy nutrient uptake. A survey of the impacts of water hyacinth on aquatic plant communities in the Wouri Basin showed that this plant is able to out-compete native species. Assessment of the impact of water hyacinth on the abundance and diversity of plant communities indicated that at some invaded sites, 65% of the vegetation consisted of water hyacinth. Species found in association with water hyacinth with a high level of abundance-dominance were Pistia stratiotes L. (Araceae) (another invader), Commelina benghalensis L. (Commelinaceae) and Echinochloa pyramidalis (Lam.) Hitchc. & Chase (Poaceae). This component of the study also showed that habitats rich in water hyacinth were poor in diversity, while habitats without water hyacinth were rich in diversity, thus raising awareness of the importance of monitoring invasive aquatic weeds along the Wouri Basin, and of implementing correct control management of all invasive aquatic weeds. Communities living along the invaded rivers are well aware of the range of problems caused by the weed; because as the rivers and water bodies used for fishing, transportation, and sand extraction are progressively invaded by the weed, the riparian population is the first to feel the impact. The impact on people has been noticeable, with an increase in diseases, such as malaria, cholera, diarrhoea, typhoid, filariasis, schistosomiasis, scabies and yellow fever increasing the need for a medicine and hospitalization. Economic losses due to the management of invasive aquatic weeds were recorded, and the Ministry of Environment spent an estimated US$1 200 000 between 2010 and 2015 to manage this scourge. In 2016, an amount of US$160 000 was transferred to these regions to manage invasive aquatic weeds, especially water hyacinth, although manual clearing is still the only method used to control this weed. Isolation of fungi from diseased water hyacinth plants in the Wouri Basin revealed several fungal species, most of which have been isolated from water hyacinth species in water bodies elsewhere, which showed a higher diversity during the dry season than during the rainy season. These fungi included Acremonium zonatum (Sawada). W. Gams (Hypocreaceae), Alternaria eichhorniae Nag Raj & Ponnappa (Pleosporaceae), Chaetomium sp., Colletotrichum sp., Curvularia pallescens Boedjin (Pleosporaceae), Curvalaria sp., Epicoccum nigrum Link (Pleosporaceae), Fusarium sp., Pithomyces chartarum fBerk. & M. A. Curtis) M. B. Ellis (Montagnulaceae), to a lesser extent Myrothecium roridum Tode ex Fr. (Incertae sedis) and Nigrospora sp. Although never released in Cameroon, arthropod biological control agents (Neochetina eichhorniae Warner (Coleoptera, Curculionidae) and N. bruchi Hustache (Coleoptera, Curculionidae)) were present, but their populations were relatively low. The slow spread of the insect population was explained by several factors, among them the tidal fluctuation of water, which has an impact on the population growth of the weevils. Whilst adults may be able to survive tidal fluctuations, larvae are severely impacted by them, contributing to the slow success of biological control. In this study, a significant increase in pathogen-induced disease severity and incidence was noted when Neochetina eichhorniae weevils were present, possibly because larvae tunnelling on the petiole created openings for the penetration of fungal spores. This study highlights the negative impacts of water hyacinth, on the environment, people, and thus economy of Cameroon. The presence of biological control agents and pathogens offers Cameroon the possibility of initiating and properly implementing the biological control option, or an integrated management solution, to manage water hyacinth in the Wouri Basin, and in the rest of Cameroon.
- Full Text:
- Date Issued: 2017
Chemical composition of leaf essential oils of Lantana camara varieties in South Africa and their effect on the behavioural preference of Falconia intermedia
- Authors: Ngxande-Koza, Samella W
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/4877 , vital:20739
- Description: Lantana camara L. (Verbenaceae) is one the most problematic invaders in South Africa invading forest edges, sand dunes, and shorelines by forming impenetrable thickets. Lantana camara invasions degrade natural biodiversity, reduce the value of land and consequently it has been a target for biological control, over the last 50 years in South Africa. Studies that have reported on chemical profile of Lantana camara have been conducted around the world but not in South Africa. Hence, the first aim of the current study was to identify the chemical baseline of L. camara varieties in the Eastern Cape, South Africa. Recent studies have shown that feeding by one of the agents released against L. camara, Falconia intermedia (Distant) (Hemiptera: Miridae), induces anti-herbivory response through increased leaf toughness and trichome density. A preliminary study conducted also reported the production of volatile chemicals by one variety, Whitney Farm, due to feeding by the mirids. Therefore, the second aim was to determine the induced changes in chemical compounds of L. camara varieties after feeding by F. intermedia. A third aim was to determine the effect these chemical compounds have on the behaviour of F. intermedia. To identify the chemical baseline of L. camara varieties, the essential oils of four L. camara varieties (East London, Port Alfred, Whitney Farm and Heather Glen) were analysed using gas chromatography mass spectrometry (GC-MS) and that resulted to the identification of 163 constitutive and 75 induced chemicals across the varieties tested. Lantana camara varieties showed different chemical classes but were highly dominated by terpenes. A great variation in the number of constitutive chemical compounds was found in all the varieties. There were 56 constitutive chemical compounds in the Whitney Farm variety, 41 in the East London variety, 36 in the Heather Glen variety and 30 in the Port Alfred variety. The Whitney Farm variety had the highest number (22) of unique constitutive chemicals identified when compared with other varieties. This indicates the chemical distinctiveness of the Whitney Farm variety from the other varieties. In the varieties tested, there were common chemical compounds identified in constitutive and induced (discussed below) states of the plants such as caryophyllene, hexane, naphthalene, copaene and a-caryophyllene. Besides naphthalene, the majority of chemical compounds in South African L. camara varieties were similar to compounds that have been identified across the world, suggesting that they are closely related. The expression of naphthalene in these varieties may be due to changes in the chemicals expressed over evolutionary time as predicted by the Novel Weapons Hypothesis. Amongst the varieties, a great variation in chemical compounds and their concentrations was shown in the induced states of the plants. The concentration of constitutive caryophyllene ranged from (3.13 - 15.7) %, to (4.02 - 11.10) % after feeding. The concentration of constitutive hexane ranged from (6.13 - 71.19) %, to (33.3 - 75.8) % after feeding. The concentration of constitutive naphthalene ranged from (0.21 - 4.79) %, to (0.92 - 2.11) % after feeding. The concentration of constitutive copaene ranged from (0.57 - 1.57) %, to (1.20 - 2.72) %. Lastly, the concentration of constitutive a-caryophyllene ranged from (1.18 - 9.03) %, to (0.78 - 5.48) % after feeding. The changes in chemical concentrations in lantana varieties indicated that feeding by the mirid on L. camara varieties causes an induction by either reducing or increasing the chemical concentrations. To determine the effect of the identified compounds on the behaviour of F. intermedia adults, olfactometer bioassays were conducted using a Y-tube technique. A significantly higher proportion of F. intermedia were attracted to undamaged leaves over damaged leaves and purified air. Undamaged leaves attracted 52 % of F. intermedia from the East London variety, 62.5 % from the Port Alfred variety, 56 % from the Whitney Farm variety, 58 % from the Lyndhurst variety and 54.5 % from the Heather Glen variety in dual choice trials versus damaged leaves. Furthermore, a significantly higher proportion of F. intermedia were attracted to damaged leaves over purified air. Damaged leaves attracted 67 % of F. intermedia from the East London variety, 67 % from the Port Alfred variety, 65.9 % from the Whitney Farm variety, 65.3 % from the Heather Glen variety and 64.5 % from the Lyndhurst variety. Olfactometer bioassays were also conducted using purified standard compounds of four chemical compounds identified from essential oils, hexane was used as a positive control as it is reported to be an insect attractant in literature. Hexane was highly attractive to the mirids compared to three standard compounds caryophyllene, caryophyllene oxide and naphthalene at the rate of 80 %, 73 % and 80 %, respectively. The standard compounds tested against F. intermedia are major compounds contained by L. camara varieties and they have proven to have a repellent effect. This may indicate that after feeding by F. intermedia, the major compounds expressed by the plant varieties repel F. intermedia contributing to the invasiveness of this weed. The increased expression of hexane and caryophyllene after feeding may also indicate increased attraction to some insects, opening up the potential for third trophic level interactions in varieties where this is the case. This is the first study on the chemical composition of essential oils of L. camara in South Africa. Therefore, we recommend that where appropriate chemical profile studies of the invasive alien plants should be considered during host specificity testing, and the vital role of chemical compounds on agent-weed interactions must be taken into consideration with other factors before and after the biological control agents are released.
- Full Text:
- Date Issued: 2017
- Authors: Ngxande-Koza, Samella W
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/4877 , vital:20739
- Description: Lantana camara L. (Verbenaceae) is one the most problematic invaders in South Africa invading forest edges, sand dunes, and shorelines by forming impenetrable thickets. Lantana camara invasions degrade natural biodiversity, reduce the value of land and consequently it has been a target for biological control, over the last 50 years in South Africa. Studies that have reported on chemical profile of Lantana camara have been conducted around the world but not in South Africa. Hence, the first aim of the current study was to identify the chemical baseline of L. camara varieties in the Eastern Cape, South Africa. Recent studies have shown that feeding by one of the agents released against L. camara, Falconia intermedia (Distant) (Hemiptera: Miridae), induces anti-herbivory response through increased leaf toughness and trichome density. A preliminary study conducted also reported the production of volatile chemicals by one variety, Whitney Farm, due to feeding by the mirids. Therefore, the second aim was to determine the induced changes in chemical compounds of L. camara varieties after feeding by F. intermedia. A third aim was to determine the effect these chemical compounds have on the behaviour of F. intermedia. To identify the chemical baseline of L. camara varieties, the essential oils of four L. camara varieties (East London, Port Alfred, Whitney Farm and Heather Glen) were analysed using gas chromatography mass spectrometry (GC-MS) and that resulted to the identification of 163 constitutive and 75 induced chemicals across the varieties tested. Lantana camara varieties showed different chemical classes but were highly dominated by terpenes. A great variation in the number of constitutive chemical compounds was found in all the varieties. There were 56 constitutive chemical compounds in the Whitney Farm variety, 41 in the East London variety, 36 in the Heather Glen variety and 30 in the Port Alfred variety. The Whitney Farm variety had the highest number (22) of unique constitutive chemicals identified when compared with other varieties. This indicates the chemical distinctiveness of the Whitney Farm variety from the other varieties. In the varieties tested, there were common chemical compounds identified in constitutive and induced (discussed below) states of the plants such as caryophyllene, hexane, naphthalene, copaene and a-caryophyllene. Besides naphthalene, the majority of chemical compounds in South African L. camara varieties were similar to compounds that have been identified across the world, suggesting that they are closely related. The expression of naphthalene in these varieties may be due to changes in the chemicals expressed over evolutionary time as predicted by the Novel Weapons Hypothesis. Amongst the varieties, a great variation in chemical compounds and their concentrations was shown in the induced states of the plants. The concentration of constitutive caryophyllene ranged from (3.13 - 15.7) %, to (4.02 - 11.10) % after feeding. The concentration of constitutive hexane ranged from (6.13 - 71.19) %, to (33.3 - 75.8) % after feeding. The concentration of constitutive naphthalene ranged from (0.21 - 4.79) %, to (0.92 - 2.11) % after feeding. The concentration of constitutive copaene ranged from (0.57 - 1.57) %, to (1.20 - 2.72) %. Lastly, the concentration of constitutive a-caryophyllene ranged from (1.18 - 9.03) %, to (0.78 - 5.48) % after feeding. The changes in chemical concentrations in lantana varieties indicated that feeding by the mirid on L. camara varieties causes an induction by either reducing or increasing the chemical concentrations. To determine the effect of the identified compounds on the behaviour of F. intermedia adults, olfactometer bioassays were conducted using a Y-tube technique. A significantly higher proportion of F. intermedia were attracted to undamaged leaves over damaged leaves and purified air. Undamaged leaves attracted 52 % of F. intermedia from the East London variety, 62.5 % from the Port Alfred variety, 56 % from the Whitney Farm variety, 58 % from the Lyndhurst variety and 54.5 % from the Heather Glen variety in dual choice trials versus damaged leaves. Furthermore, a significantly higher proportion of F. intermedia were attracted to damaged leaves over purified air. Damaged leaves attracted 67 % of F. intermedia from the East London variety, 67 % from the Port Alfred variety, 65.9 % from the Whitney Farm variety, 65.3 % from the Heather Glen variety and 64.5 % from the Lyndhurst variety. Olfactometer bioassays were also conducted using purified standard compounds of four chemical compounds identified from essential oils, hexane was used as a positive control as it is reported to be an insect attractant in literature. Hexane was highly attractive to the mirids compared to three standard compounds caryophyllene, caryophyllene oxide and naphthalene at the rate of 80 %, 73 % and 80 %, respectively. The standard compounds tested against F. intermedia are major compounds contained by L. camara varieties and they have proven to have a repellent effect. This may indicate that after feeding by F. intermedia, the major compounds expressed by the plant varieties repel F. intermedia contributing to the invasiveness of this weed. The increased expression of hexane and caryophyllene after feeding may also indicate increased attraction to some insects, opening up the potential for third trophic level interactions in varieties where this is the case. This is the first study on the chemical composition of essential oils of L. camara in South Africa. Therefore, we recommend that where appropriate chemical profile studies of the invasive alien plants should be considered during host specificity testing, and the vital role of chemical compounds on agent-weed interactions must be taken into consideration with other factors before and after the biological control agents are released.
- Full Text:
- Date Issued: 2017
Improving the cold tolerance of false codling moth, thaumatotibia leucotreta, for better performance in a sterile insect release programme
- Authors: Daniel, Claire Ashleigh
- Date: 2017
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Reproduction -- Effect of temperature on , Cryptophlebia leucotreta -- Biological control -- South Africa , Citrus -- Diseases and pests -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Insecticides , Citrus fruit industry -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/5271 , vital:20803
- Description: The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a major pest of citrus and other important crops in Sub-Saharan Africa. The introduction of a sterile insect technique (SIT) programme for FCM in South Africa has proven to be very effective in the control of FCM. The objective was to flood citrus orchards with large numbers of sterile males resulting in a ratio of at least 10 sterile to 1 wild moth, increasing the probability of a female moth mating with a sterile male. This target is often achieved and the programme is generally successful, however there are some challenges regarding this programme. The mass rearing environment, artificial diet, handling methods and irradiation have an impact on sterile insect quality as environmental differences between the rearing facility and field, influences the insect phenotype and competitiveness. This is evident as wild male moths can theoretically actively fly at a temperature of 12°C and laboratory-reared sterilized moths, due to the radiation treatment, appear unable to fly below 20°C. As a consequence, sterile males are out-competed by wild males during the cooler months of the year. This is detrimental to the SIT programme as FCM do not undergo diapause, meaning they are active during winter and will still reproduce. Therefore, to maximize the effect of the SIT programme, it is vital to increase the flight ability of mass reared sterile males at lower temperatures. Various studies have shown that by adding cryoprotectants to the basic laboratory diets increases the cold tolerance of certain insects and thus may allow them to be mobile at lower temperatures, however it imperative that any chemical used to augment the commercial diet of the insect has no negative effects on the insect physiology and development. To investigate this detail for FCM, five generations of FCM were reared on diets augmented with various known insect cryoprotectants. These augmented FCM were subsequently used in experiments designed to determine firstly, if the cryoprotectants had a positive result on the cold tolerance of the FCM, and secondly, if they had any adverse effects on other physiological aspects such as duration of development. Laboratory trials indicated that the flight ability of male FCM was improved when larvae were reared on diets augmented with trehalose and cholesterol (with an average of 40 % of cholesterol and trehalose augmented males that flew at 15 °C where 0 % of the control flew). Results obtained during the field trials support the laboratory results as there was a significant increase in the number of trehalose augmented moths caught in the field during March and July (winter). Results also showed potential for cholesterol to be used as an additive. Other important findings show that both cholesterol and trehalose have no negative impacts on developmental rate, pupal size, and egg production and viability. Trehalose was found to increase the pupal mass of male and female FCM, as well as the number of eggs laid per female. Cholesterol was found to increase developmental rate and the number of eggs laid. The main findings of this study were that diet additives could improve the massrearing of FCM for SIT and the competitiveness of the males, especially at lower temperatures. However, the additives were expensive and cost could well be a constraint to the wide scale implementation of the new technology.
- Full Text:
- Date Issued: 2017
- Authors: Daniel, Claire Ashleigh
- Date: 2017
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Reproduction -- Effect of temperature on , Cryptophlebia leucotreta -- Biological control -- South Africa , Citrus -- Diseases and pests -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Insecticides , Citrus fruit industry -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/5271 , vital:20803
- Description: The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a major pest of citrus and other important crops in Sub-Saharan Africa. The introduction of a sterile insect technique (SIT) programme for FCM in South Africa has proven to be very effective in the control of FCM. The objective was to flood citrus orchards with large numbers of sterile males resulting in a ratio of at least 10 sterile to 1 wild moth, increasing the probability of a female moth mating with a sterile male. This target is often achieved and the programme is generally successful, however there are some challenges regarding this programme. The mass rearing environment, artificial diet, handling methods and irradiation have an impact on sterile insect quality as environmental differences between the rearing facility and field, influences the insect phenotype and competitiveness. This is evident as wild male moths can theoretically actively fly at a temperature of 12°C and laboratory-reared sterilized moths, due to the radiation treatment, appear unable to fly below 20°C. As a consequence, sterile males are out-competed by wild males during the cooler months of the year. This is detrimental to the SIT programme as FCM do not undergo diapause, meaning they are active during winter and will still reproduce. Therefore, to maximize the effect of the SIT programme, it is vital to increase the flight ability of mass reared sterile males at lower temperatures. Various studies have shown that by adding cryoprotectants to the basic laboratory diets increases the cold tolerance of certain insects and thus may allow them to be mobile at lower temperatures, however it imperative that any chemical used to augment the commercial diet of the insect has no negative effects on the insect physiology and development. To investigate this detail for FCM, five generations of FCM were reared on diets augmented with various known insect cryoprotectants. These augmented FCM were subsequently used in experiments designed to determine firstly, if the cryoprotectants had a positive result on the cold tolerance of the FCM, and secondly, if they had any adverse effects on other physiological aspects such as duration of development. Laboratory trials indicated that the flight ability of male FCM was improved when larvae were reared on diets augmented with trehalose and cholesterol (with an average of 40 % of cholesterol and trehalose augmented males that flew at 15 °C where 0 % of the control flew). Results obtained during the field trials support the laboratory results as there was a significant increase in the number of trehalose augmented moths caught in the field during March and July (winter). Results also showed potential for cholesterol to be used as an additive. Other important findings show that both cholesterol and trehalose have no negative impacts on developmental rate, pupal size, and egg production and viability. Trehalose was found to increase the pupal mass of male and female FCM, as well as the number of eggs laid per female. Cholesterol was found to increase developmental rate and the number of eggs laid. The main findings of this study were that diet additives could improve the massrearing of FCM for SIT and the competitiveness of the males, especially at lower temperatures. However, the additives were expensive and cost could well be a constraint to the wide scale implementation of the new technology.
- Full Text:
- Date Issued: 2017
The effect of shade on the biological control of Salvinia molesta D.S. Mitchell [Salviniaceae] by the weevil, Cyrtobagous salviniae Calder and Sands [Curculionidae]
- Authors: Maseko, Zolile
- Date: 2017
- Subjects: Salvinia molesta , Aquatic weeds -- Biological control , Beetles , Insects as biological pest control agents
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/4808 , vital:20726
- Description: Salvinia molesta, a floating aquatic weed, is a global menace in many water bodies and waterways. The weed disrupts the ecological balance wherever it invades and also has wide ranging economic and health impacts. Its impact has resulted in the need to control it, and while chemical and mechanical control are often ineffective, biological control by the weevil Cyrtobagous salviniae is largely successful. However, in many parts of the world, including South Africa, biological control of S. molesta has been less effective where the weed grows as an understory species. Shallow and shaded waters characteristically found at the margins of water bodies provide a refuge for S. molesta. Therefore, the aim of the study was to determine the effect of shade on the efficacy of biological control of S. molesta. Investigations into the problems associated with control in the shade were carried out in a greenhouse and in the field. In controlled greenhouse experiments, plants were grown at high and low nutrient levels in individual mesocosms, at three varying levels of shade, where half the mesocosms were inoculated with C. salviniae. Plants in high nutrient conditions were significantly more productive in terms of biomass accumulation, compared to those grown at low nutrient levels, in both the absence and presence of herbivory at all levels of shade tested. Plants grown in the shade and in high nutrient conditions had significantly higher quality compared to the ones exposed to full sun. Higher plant quality in the shade consequently resulted in accelerated fecundity for C. salviniae resulting in significantly higher weevil populations. Furthermore, less damage was recorded on plants in the shade, possibly due to the high nitrogen concentrations which may have deterred grazing by C. salviniae. However, in the absence of shading, plants were of lower quality and consequently sustained more damage from herbivores despite lower weevil populations. In contrast to the greenhouse, there were no differences in biomass recorded in the field at two nutrient levels and in the presence of herbivory for both shade and open sites. Nutrients added to high nutrient treatment quadrats diffused evenly across the water body resulting in uniform nutrient distribution, hence uniform plant biomass and carbon-nitrogen ratio. Despite the lack of statistical differences, more weevils were found in the full sun plots, while modest populations were recorded in the shade. Higher weevil populations consequently led to more damage in the sun, a situation that has been observed in most field sites in South Africa. The results therefore suggest that nutrients were pivotal in plant growth compared to light regimes (amount of PAR). Plant quality significantly influenced weevil populations and plant damage in both studies, demonstrating that plant nitrogen plays a crucial role in the plant- herbivore system. Furthermore, in both the greenhouse and field, despite lack of statistical differences, plants exposed to full sunlight suffered more damage than shade plants, pointing towards better control in the sun compared to the shade, suggesting that there is greater preference of sun-exposed plants for grazing by the weevils. The results from this study add to the growing body of literature that plant quality is a major factor in determining the success of aquatic weed biological control programmes.
- Full Text:
- Date Issued: 2017
- Authors: Maseko, Zolile
- Date: 2017
- Subjects: Salvinia molesta , Aquatic weeds -- Biological control , Beetles , Insects as biological pest control agents
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/4808 , vital:20726
- Description: Salvinia molesta, a floating aquatic weed, is a global menace in many water bodies and waterways. The weed disrupts the ecological balance wherever it invades and also has wide ranging economic and health impacts. Its impact has resulted in the need to control it, and while chemical and mechanical control are often ineffective, biological control by the weevil Cyrtobagous salviniae is largely successful. However, in many parts of the world, including South Africa, biological control of S. molesta has been less effective where the weed grows as an understory species. Shallow and shaded waters characteristically found at the margins of water bodies provide a refuge for S. molesta. Therefore, the aim of the study was to determine the effect of shade on the efficacy of biological control of S. molesta. Investigations into the problems associated with control in the shade were carried out in a greenhouse and in the field. In controlled greenhouse experiments, plants were grown at high and low nutrient levels in individual mesocosms, at three varying levels of shade, where half the mesocosms were inoculated with C. salviniae. Plants in high nutrient conditions were significantly more productive in terms of biomass accumulation, compared to those grown at low nutrient levels, in both the absence and presence of herbivory at all levels of shade tested. Plants grown in the shade and in high nutrient conditions had significantly higher quality compared to the ones exposed to full sun. Higher plant quality in the shade consequently resulted in accelerated fecundity for C. salviniae resulting in significantly higher weevil populations. Furthermore, less damage was recorded on plants in the shade, possibly due to the high nitrogen concentrations which may have deterred grazing by C. salviniae. However, in the absence of shading, plants were of lower quality and consequently sustained more damage from herbivores despite lower weevil populations. In contrast to the greenhouse, there were no differences in biomass recorded in the field at two nutrient levels and in the presence of herbivory for both shade and open sites. Nutrients added to high nutrient treatment quadrats diffused evenly across the water body resulting in uniform nutrient distribution, hence uniform plant biomass and carbon-nitrogen ratio. Despite the lack of statistical differences, more weevils were found in the full sun plots, while modest populations were recorded in the shade. Higher weevil populations consequently led to more damage in the sun, a situation that has been observed in most field sites in South Africa. The results therefore suggest that nutrients were pivotal in plant growth compared to light regimes (amount of PAR). Plant quality significantly influenced weevil populations and plant damage in both studies, demonstrating that plant nitrogen plays a crucial role in the plant- herbivore system. Furthermore, in both the greenhouse and field, despite lack of statistical differences, plants exposed to full sunlight suffered more damage than shade plants, pointing towards better control in the sun compared to the shade, suggesting that there is greater preference of sun-exposed plants for grazing by the weevils. The results from this study add to the growing body of literature that plant quality is a major factor in determining the success of aquatic weed biological control programmes.
- Full Text:
- Date Issued: 2017
The isolation and genetic characterisation of a novel alphabaculovirus for the microbial control of Cryptophlebia peltastica and closely related tortricid pests
- Authors: Marsberg, Tamryn
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59292 , vital:27543
- Description: Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae) is an economically damaging pest of litchis and macadamias in South Africa. Cryptophlebia peltastica causes both pre- and post-harvest damage to litchis, reducing overall yields and thus classifying the pest as a phytosanitary risk. Various control methods have been implemented against C. peltastica in an integrated pest management programme. These control methods include chemical control, cultural control and biological control. However, these methods have not yet provided satisfactory control as of yet. As a result, an alternative control option needs to be identified and implemented into the IPM programme. An alternative method of control that has proved successful in other agricultural sectors and not yet implemented in the control of C. peltastica is that of microbial control, specifically the use of baculovirus biopesticides. This study aimed to isolate and characterise a novel baculovirus from a laboratory culture of C. peltastica that could be used as a commercially available baculovirus biopesticide. In order to isolate a baculovirus a laboratory culture of C. peltastica was successfully established at Rhodes University, Grahamstown, South Africa. This is the first time a laboratory culture of C. peltastica has been established. This allowed for various biological aspects of the pest to be determined, which included: length of the life cycle, fecundity and time to oviposition, egg and larval development and percentage hatch. The results obtained from these studies found that the biology of C. peltastica was similar to that of Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). Once the laboratory culture had reached high densities, larvae showing symptoms of baculovirus infection were observed. Symptomatic larvae were collected and examined for the presence of a baculovirus. An alphabaculovirus (NPV) was successfully isolated and morphologically identified using purified OBs that were sectioned and observed by transmission electron microscopy. This was then confirmed by amplifying the polyhedrin gene region using degenerate primers. A BLAST analysis found a 93% similarity to a partial polyhedrin gene sequence to be that of Epinotia granitalis (Butler) (Lepidoptera: Tortricidae). The alphabaculovirus was then genetically characterised by generating restriction profiles and sequencing the whole genome. Due to the novelty of the virus, no comparison could be made. The biological activity of the alphabaculovirus was then tested against C. peltastica and two closely related Tortricidae pests: T. leucotreta and Cydiapomonella (Linnaeous) (Lepidoptera: Tortricidae). The alphabaculovirus was highly virulent against all three species. The lethal concentrations (LC50 and LC90) for the virus against C. peltastica was 8.19 x 103 and 3.33 x 105 OBs/ml. The LC50 and LC90 for T. leucotreta was 2.29 x 103 and 9.97 x 104 OBs/ml, respectively and C. pomonella had a LC50 of 1.43 x 103 OBs/ml and LC90 1.26 x 104 OBs/ml. The virus was particularly virulent against T. leucotreta and C. pomonella as compared to C. peltastica. The biological activity of the alphabaculovirus was also tested against CpGV resistant European C. pomonella. From the results it was observed that the virus had the ability to overcome the resistance in C. pomonella and could potentially be used in the resistance management of C. pomonella. With the successful biological activity results obtained from this study, preliminary investigation were made into the mass production of the alphabaculovirus using both the in vivo and in vitro production methods. For in vivo production both the homologous host (C. peltastica) and a heterologous host (T leucotreta) were investigated. Preliminary studies focused on determining the biological activity in fifth instars of both hosts. Fifth instar LC50 and LC90 values for C. peltastica were 3.43 x 103 and 1.11 x 107 OBs/ml and for T. leucotreta the LC50 and LC90 values were 2.53 x 103 and 8.82 x 106 OBs/ml, respectively. The average yield of virus produced in each species was also determined. Cryptophlebia peltastica had the highest viral yield of 5.37 x 1010 OBs/larva and 2.93 x 1010 OBs/larva for T. leucotreta. The results obtained, from the preliminary investigation concluded that the virus could be produced in vivo in both C. peltastica and T. leucotreta, however further research is required into the mass production in both hosts. The in vitro production of the virus was also considered and the susceptibility of the virus was tested against the C. pomonella cell line, Cp14R. After infection of the Cp14R cells with budded virus collected from fifth instar C. peltastica larvae, OBs could be observed after three days. Thus, the alphabaculovirus is susceptible to the Cp14R cell line, thus has the potential to be produced in vitro and further characterised. This study is the first to report of the identification and characterisation of a novel alphabaculovirus isolated from a laboratory reared culture of C. peltastica and the potential for it to be commercially developed into a bipoesticide and used against Tortricidae pests.
- Full Text:
- Date Issued: 2017
- Authors: Marsberg, Tamryn
- Date: 2017
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/59292 , vital:27543
- Description: Cryptophlebia peltastica (Meyrick) (Lepidoptera: Tortricidae) is an economically damaging pest of litchis and macadamias in South Africa. Cryptophlebia peltastica causes both pre- and post-harvest damage to litchis, reducing overall yields and thus classifying the pest as a phytosanitary risk. Various control methods have been implemented against C. peltastica in an integrated pest management programme. These control methods include chemical control, cultural control and biological control. However, these methods have not yet provided satisfactory control as of yet. As a result, an alternative control option needs to be identified and implemented into the IPM programme. An alternative method of control that has proved successful in other agricultural sectors and not yet implemented in the control of C. peltastica is that of microbial control, specifically the use of baculovirus biopesticides. This study aimed to isolate and characterise a novel baculovirus from a laboratory culture of C. peltastica that could be used as a commercially available baculovirus biopesticide. In order to isolate a baculovirus a laboratory culture of C. peltastica was successfully established at Rhodes University, Grahamstown, South Africa. This is the first time a laboratory culture of C. peltastica has been established. This allowed for various biological aspects of the pest to be determined, which included: length of the life cycle, fecundity and time to oviposition, egg and larval development and percentage hatch. The results obtained from these studies found that the biology of C. peltastica was similar to that of Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae). Once the laboratory culture had reached high densities, larvae showing symptoms of baculovirus infection were observed. Symptomatic larvae were collected and examined for the presence of a baculovirus. An alphabaculovirus (NPV) was successfully isolated and morphologically identified using purified OBs that were sectioned and observed by transmission electron microscopy. This was then confirmed by amplifying the polyhedrin gene region using degenerate primers. A BLAST analysis found a 93% similarity to a partial polyhedrin gene sequence to be that of Epinotia granitalis (Butler) (Lepidoptera: Tortricidae). The alphabaculovirus was then genetically characterised by generating restriction profiles and sequencing the whole genome. Due to the novelty of the virus, no comparison could be made. The biological activity of the alphabaculovirus was then tested against C. peltastica and two closely related Tortricidae pests: T. leucotreta and Cydiapomonella (Linnaeous) (Lepidoptera: Tortricidae). The alphabaculovirus was highly virulent against all three species. The lethal concentrations (LC50 and LC90) for the virus against C. peltastica was 8.19 x 103 and 3.33 x 105 OBs/ml. The LC50 and LC90 for T. leucotreta was 2.29 x 103 and 9.97 x 104 OBs/ml, respectively and C. pomonella had a LC50 of 1.43 x 103 OBs/ml and LC90 1.26 x 104 OBs/ml. The virus was particularly virulent against T. leucotreta and C. pomonella as compared to C. peltastica. The biological activity of the alphabaculovirus was also tested against CpGV resistant European C. pomonella. From the results it was observed that the virus had the ability to overcome the resistance in C. pomonella and could potentially be used in the resistance management of C. pomonella. With the successful biological activity results obtained from this study, preliminary investigation were made into the mass production of the alphabaculovirus using both the in vivo and in vitro production methods. For in vivo production both the homologous host (C. peltastica) and a heterologous host (T leucotreta) were investigated. Preliminary studies focused on determining the biological activity in fifth instars of both hosts. Fifth instar LC50 and LC90 values for C. peltastica were 3.43 x 103 and 1.11 x 107 OBs/ml and for T. leucotreta the LC50 and LC90 values were 2.53 x 103 and 8.82 x 106 OBs/ml, respectively. The average yield of virus produced in each species was also determined. Cryptophlebia peltastica had the highest viral yield of 5.37 x 1010 OBs/larva and 2.93 x 1010 OBs/larva for T. leucotreta. The results obtained, from the preliminary investigation concluded that the virus could be produced in vivo in both C. peltastica and T. leucotreta, however further research is required into the mass production in both hosts. The in vitro production of the virus was also considered and the susceptibility of the virus was tested against the C. pomonella cell line, Cp14R. After infection of the Cp14R cells with budded virus collected from fifth instar C. peltastica larvae, OBs could be observed after three days. Thus, the alphabaculovirus is susceptible to the Cp14R cell line, thus has the potential to be produced in vitro and further characterised. This study is the first to report of the identification and characterisation of a novel alphabaculovirus isolated from a laboratory reared culture of C. peltastica and the potential for it to be commercially developed into a bipoesticide and used against Tortricidae pests.
- Full Text:
- Date Issued: 2017