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
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
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 pest status and integrated management programme of carob moth, Ectomyelois ceratoniae Zeller, attacking citrus in South Africa
- Authors: Thackeray, Sean Robin
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7758 , vital:21293
- Description: The carob moth, Ectomyelois ceratoniae Zeller, is a pest of agricultural commodities and stored products around the world. Carob moth is known to infest citrus in the Mediterranean region and in southern Africa. In grapefruit cultivars, carob moth infestations are associated with high levels of mealybug. However, although this relationship has been observed in other citrus types such as Navel oranges, this has never been quantified. A recent survey of infested fruit from various production areas in South Africa indicated that the pest status of carob moth on Navel oranges may have been underestimated. As a result of the incidental pest status of carob moth on citrus in South Africa in the past, a species specific integrated pest management (IPM) programme does not exist. Therefore, the overriding aim of this theses was to evaluate the pest status of carob moth in citrus and establish a species specific IPM programme by determine the autecology of carob moth in citrus. Reliable methods for monitoring carob moth in citrus orchards both for producers and for research purposes were developed. A user-friendly monitoring method for determining weekly carob moth infestation through dropped fruit was suitable for producers. A timed scouting method was also developed; although the accuracy of this method varied with the experience of the scout. The pest status of carob moth was highest in the Loskop Valley, Nelspruit and the Vaalharts production areas and economic injury to growers ranged from R512.35 to R3 719.80 per hectare as a direct result of infestation. No infestation was recorded in the Sundays River Valley and Citrusdal production areas over both the 2014-15 and 201516 growing seasons. A laboratory study showed the survival of carob moth larvae infesting citrus is less than 10% in the absence of mealybug. However, this increases to almost 40% in the presence of mealybug residues and sooty mould. There was a significant relationship between carob infestation at harvest and mealybug infestation in the middle months of the growing season. The relationship between carob moth and mealybug indicates that current production guidelines for the management of mealybug in citrus may need to be amended. Consequently, it is proposed that an orchard with a history of carob moth infestation and a high mealybug infestation in the previous season should be subjected to an early season preventative application of a registered control product. Also, if mealybug infestation in December is higher than a 5% of fruit per tree, then a corrective application of a registered product is recommended. The application of 2,4-D at petal drop reduced the size of the navel-end opening, decreasing the proportion of mealybug found in the navel-end, subsequently reducing carob moth infestation, resulting in a direct benefit for producers. Products registered for the control of false codling moth (FCM), Thaumatotibia leucotreta Meyrick, were effective in reducing carob moth infestation. In a spray trial conducted over two seasons, Delegate® and Runner® reduced infestation significantly in the 2014-15 season (over 80%), while only Delegate® was effective in the 2015-16 season (over 80%). If a late season corrective chemical application is targeted at both FCM and carob moth, this application should take place between 6-7 weeks prior to harvest. The mating disruption product, SPLAT® EC, reduced carob moth infestation by 70% compared to the untreated control. A laboratory culture was established and head-capsule size categories were determined for all five carob moth instars. A parasitoid survey indicated that parasitism of carob moth larvae is generally less than 5% in citrus orchards and a new species of Braconidae was described as Phanterotoma carobivora van Achterberg and Thackeray. Carob moth fifth instar were found to be the most cold-tolerant larval stage, and were shown to be more cold susceptible than the most cold-tolerant FCM instars at -0.55ºC for eighteen days. This cold treatment resulted in a mortality of 94.6% fifth instar carob moth compared to a combined fourth and fifth instar mortality of 87.8% for FCM after eighteen days. These results indicate that post-harvest cold treatments targeting FCM will be as, if not more, effective against carob moth, suggesting that current phytosanitary legislation for carob moth should be amended to incorporate this study’s findings.
- Full Text:
- Date Issued: 2017
- Authors: Thackeray, Sean Robin
- Date: 2017
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/7758 , vital:21293
- Description: The carob moth, Ectomyelois ceratoniae Zeller, is a pest of agricultural commodities and stored products around the world. Carob moth is known to infest citrus in the Mediterranean region and in southern Africa. In grapefruit cultivars, carob moth infestations are associated with high levels of mealybug. However, although this relationship has been observed in other citrus types such as Navel oranges, this has never been quantified. A recent survey of infested fruit from various production areas in South Africa indicated that the pest status of carob moth on Navel oranges may have been underestimated. As a result of the incidental pest status of carob moth on citrus in South Africa in the past, a species specific integrated pest management (IPM) programme does not exist. Therefore, the overriding aim of this theses was to evaluate the pest status of carob moth in citrus and establish a species specific IPM programme by determine the autecology of carob moth in citrus. Reliable methods for monitoring carob moth in citrus orchards both for producers and for research purposes were developed. A user-friendly monitoring method for determining weekly carob moth infestation through dropped fruit was suitable for producers. A timed scouting method was also developed; although the accuracy of this method varied with the experience of the scout. The pest status of carob moth was highest in the Loskop Valley, Nelspruit and the Vaalharts production areas and economic injury to growers ranged from R512.35 to R3 719.80 per hectare as a direct result of infestation. No infestation was recorded in the Sundays River Valley and Citrusdal production areas over both the 2014-15 and 201516 growing seasons. A laboratory study showed the survival of carob moth larvae infesting citrus is less than 10% in the absence of mealybug. However, this increases to almost 40% in the presence of mealybug residues and sooty mould. There was a significant relationship between carob infestation at harvest and mealybug infestation in the middle months of the growing season. The relationship between carob moth and mealybug indicates that current production guidelines for the management of mealybug in citrus may need to be amended. Consequently, it is proposed that an orchard with a history of carob moth infestation and a high mealybug infestation in the previous season should be subjected to an early season preventative application of a registered control product. Also, if mealybug infestation in December is higher than a 5% of fruit per tree, then a corrective application of a registered product is recommended. The application of 2,4-D at petal drop reduced the size of the navel-end opening, decreasing the proportion of mealybug found in the navel-end, subsequently reducing carob moth infestation, resulting in a direct benefit for producers. Products registered for the control of false codling moth (FCM), Thaumatotibia leucotreta Meyrick, were effective in reducing carob moth infestation. In a spray trial conducted over two seasons, Delegate® and Runner® reduced infestation significantly in the 2014-15 season (over 80%), while only Delegate® was effective in the 2015-16 season (over 80%). If a late season corrective chemical application is targeted at both FCM and carob moth, this application should take place between 6-7 weeks prior to harvest. The mating disruption product, SPLAT® EC, reduced carob moth infestation by 70% compared to the untreated control. A laboratory culture was established and head-capsule size categories were determined for all five carob moth instars. A parasitoid survey indicated that parasitism of carob moth larvae is generally less than 5% in citrus orchards and a new species of Braconidae was described as Phanterotoma carobivora van Achterberg and Thackeray. Carob moth fifth instar were found to be the most cold-tolerant larval stage, and were shown to be more cold susceptible than the most cold-tolerant FCM instars at -0.55ºC for eighteen days. This cold treatment resulted in a mortality of 94.6% fifth instar carob moth compared to a combined fourth and fifth instar mortality of 87.8% for FCM after eighteen days. These results indicate that post-harvest cold treatments targeting FCM will be as, if not more, effective against carob moth, suggesting that current phytosanitary legislation for carob moth should be amended to incorporate this study’s findings.
- Full Text:
- Date Issued: 2017
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