Sexual attraction and mating compatibility between Thaumatotibia leucotreta populations and implications for semiochemical dependent technologies
- Authors: Upfold, Jennifer Kate
- Date: 2020
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Reproduction , Citrus -- Disease and pests -- Control -- South Africa , Insect sterilization -- South Africa , Pheromones , Pheromone traps
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/148526 , vital:38747
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick), is the most important pest for the cultivation of citrus in South Africa. False codling moth is indigenous to southern Africa and is a regulated pest of many international markets for phytosanitary concerns. Considerable research efforts have been invested in the past decades to develop semiochemcial technologies, such as monitoring with sex pheromones, attract-and-kill, mating disruption and the sterile insect technique. One of the potential obstacles identified with semiochemical control is the differences in the ratio of the compounds comprising the sex pheromone at different geographical locations, resulting in what is known as regional attraction. This has been identified in FCM populations from three different countries, however, regional attraction within South African FCM populations was unknown. Therefore, the study assessed the genetic integrity of five laboratory-reared FCM populations originating from geographically isolated populations in South Africa using the AFLP technique in order to assess regional attractiveness within the country. The results found isolated populations from Addo, Citrusdal, Marble Hall, Nelspruit and a fifth group found to be closely related to Addo and Citrusdal called the ‘Old’ colony. These five genetically isolated populations as well as a population from Xsit (Pty) Ltd, used for the sterile insect technique (SIT), were used in regional attractiveness trials. Males were significantly (P = <0.05) more attracted to females originating from the same population. No significant attraction could be determined from the sterile males, as the recapture rates in the trap were too low. Furthermore, regional attractiveness was assessed through choice/ no-choice mating compatibility trials. Significant sexual isolation (ISI) occurred between mating combinations Addo × Nelspruit (ISI = 0,13; t2 = 6.23; p = 0.02), Addo × Marble Hall (ISI = 0,11; t2 = 4.72; p = 0.04), Citrusdal × Nelspruit (ISI = 0,11; t2 = 4.95; p = 0.04), and Citrusdal × Marble Hall (ISI = 0,12; t2 = 4.31; p = 0.04). In these combinations, Addo and Citrusdal males were found to have outcompeted Nelspruit and Marble Hall males for more mating events. Significant sexual isolation was also recorded for Sterile × Marble Hall (ISI = 0.12; t2 = 4.98; p =0.01) and Sterile × Citrusdal (ISI = 0.13; t2 = 3.96; p = 0.01) populations. The male relative performance index was significant in both combinations, indicating that non-sterile laboratory males outcompeted the sterile males in these two combinations. When given no choice, evaluated as spermatophore transfer/ female/ 48h, all males (including sterile) were successful in transferring spermatophores to all FCM populations, with no significant differences. These results indicate that there may be incipient pre-isolation mechanisms affected by local natural selection, resulting in localised sexual attraction via differences in the sex pheromone ratios. These findings provide important information for semiochemical technologies and the implication of these results with regard to monitoring with sex pheromones, attract-and-kill, mating disruption and sterile insect technique are discussed.
- Full Text:
- Date Issued: 2020
- Authors: Upfold, Jennifer Kate
- Date: 2020
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Reproduction , Citrus -- Disease and pests -- Control -- South Africa , Insect sterilization -- South Africa , Pheromones , Pheromone traps
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/148526 , vital:38747
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick), is the most important pest for the cultivation of citrus in South Africa. False codling moth is indigenous to southern Africa and is a regulated pest of many international markets for phytosanitary concerns. Considerable research efforts have been invested in the past decades to develop semiochemcial technologies, such as monitoring with sex pheromones, attract-and-kill, mating disruption and the sterile insect technique. One of the potential obstacles identified with semiochemical control is the differences in the ratio of the compounds comprising the sex pheromone at different geographical locations, resulting in what is known as regional attraction. This has been identified in FCM populations from three different countries, however, regional attraction within South African FCM populations was unknown. Therefore, the study assessed the genetic integrity of five laboratory-reared FCM populations originating from geographically isolated populations in South Africa using the AFLP technique in order to assess regional attractiveness within the country. The results found isolated populations from Addo, Citrusdal, Marble Hall, Nelspruit and a fifth group found to be closely related to Addo and Citrusdal called the ‘Old’ colony. These five genetically isolated populations as well as a population from Xsit (Pty) Ltd, used for the sterile insect technique (SIT), were used in regional attractiveness trials. Males were significantly (P = <0.05) more attracted to females originating from the same population. No significant attraction could be determined from the sterile males, as the recapture rates in the trap were too low. Furthermore, regional attractiveness was assessed through choice/ no-choice mating compatibility trials. Significant sexual isolation (ISI) occurred between mating combinations Addo × Nelspruit (ISI = 0,13; t2 = 6.23; p = 0.02), Addo × Marble Hall (ISI = 0,11; t2 = 4.72; p = 0.04), Citrusdal × Nelspruit (ISI = 0,11; t2 = 4.95; p = 0.04), and Citrusdal × Marble Hall (ISI = 0,12; t2 = 4.31; p = 0.04). In these combinations, Addo and Citrusdal males were found to have outcompeted Nelspruit and Marble Hall males for more mating events. Significant sexual isolation was also recorded for Sterile × Marble Hall (ISI = 0.12; t2 = 4.98; p =0.01) and Sterile × Citrusdal (ISI = 0.13; t2 = 3.96; p = 0.01) populations. The male relative performance index was significant in both combinations, indicating that non-sterile laboratory males outcompeted the sterile males in these two combinations. When given no choice, evaluated as spermatophore transfer/ female/ 48h, all males (including sterile) were successful in transferring spermatophores to all FCM populations, with no significant differences. These results indicate that there may be incipient pre-isolation mechanisms affected by local natural selection, resulting in localised sexual attraction via differences in the sex pheromone ratios. These findings provide important information for semiochemical technologies and the implication of these results with regard to monitoring with sex pheromones, attract-and-kill, mating disruption and sterile insect technique are discussed.
- Full Text:
- Date Issued: 2020
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
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
Production of Cydia pomonella granulovirus (CpGV) in a heteralogous host, Thaumatotibia Leucotreta (Meyrick) (False codling moth)
- Authors: Chambers, Craig Brian
- Date: 2015
- Subjects: Cryptophlebia leucotreta -- South Africa , Codling moth -- South Africa , Apples -- Diseases and pests -- South Africa , Codling moth -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Biological pest control agents -- South Africa , Baculoviruses -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5935 , http://hdl.handle.net/10962/d1017906
- Description: Cydia pomonella (Linnaeus) (Family: Tortricidae), the codling moth, is considered one of the most significant pests of apples and pears worldwide, causing up to 80% crop loss in orchards if no control measures are applied. Cydia pomonella is oligophagous feeding on a number of alternate hosts including quince, walnuts, apricots, peaches, plums and nectarines. Historically the control of this pest has been achieved with the use of various chemical control strategies which have maintained pest levels below the economic threshold at a relatively low cost to the grower. However, there are serious concerns surrounding the use of chemical insecticides including the development of resistance in insect populations, the banning of various insecticides, regulations for lowering of the maximum residue level and employee and consumer safety. For this reason, alternate measures of control are slowly being adopted by growers such as mating disruption, cultural methods and the use of baculovirus biopesticides as part of integrated pest management programmes. The reluctance of growers to accept baculovirus or other biological control products in the past has been due to questionable product quality and inconsistencies in their field performance. Moreover, the development and application of biological control products is more costly than the use of chemical alternatives. Baculoviruses are arthropod specific viruses that are highly virulent to a number of lepidopteran species. Due to the virulence and host specificity of baculoviruses, Cydia pomonella granulovirus has been extensively and successfully used as part of integrated pest management systems for the control of C. pomonella in Europe and around the world, including South Africa. Commercial formulations have been typically based on the Mexican strain of CpGV. However due to long-term multiple applications of CpGV and the reliance on CpGV in organic farming practices in Europe, resistance to the CpGV-M strain has developed in a number of field populations of C. pomonella. This study aimed to identify and characterize novel isolates of CpGV in South Africa and compare their virulence with the commercial standard CpGV-M. Secondly, since C. pomonella is difficult to culture on a large scale, an alternate method of CpGV production was investigated in order to determine if CpGV could be produced more efficiently and at a reduced cost without negatively impacting the quality of the product. Several isolates of CpGV were recovered either from field collected larvae or from a laboratory-reared C. pomonella colony. Characterisation of DNA profiles using a variety of restriction enzymes revealed that only a single isolate, CpGV-SA, was genetically different from the Mexican strain of the virus used in the commercially available CpGV based products in South Africa. In dose-response bioassays using CpGV-SA, LC₅₀ and LC₉₀ values for neonate C. pomonella larvae were 3.18 x 10³ OBs/ml and 7.33 x 10⁴ respectively. A comparison of these values with those of CpGV-M indicated no significant difference in the virulence of the two isolates under laboratory conditions. This is a first report of a genetically distinct CpGV isolate in South Africa. The biological activity and novelty of CpGV-SA makes this isolate a potentially important tool for CpGV resistance management in South Africa. In order to justify production of CpGV in an alternative host, studies on the comparative biological performance of C. pomonella and T. leucotreta based on oviposition, time to hatch, larval developmental times and rearing efficiency as well as production costs were performed. Thaumatotibia leucotreta was found to be more fecund and to have significantly shorter egg and larval developmental times. In addition, larval production per unit of artificial diet was significantly higher than for C. pomonella. This resulted in T. leucotreta being more cost effective to produce with implications for reduced insectary space, sanitation practices as well as the labour component of production. Virus yield data generated by inoculation both C. pomonella and T. leucotreta with nine concentrations of CpGV resulted in comparable virus yields, justifying the continuation of the research into production of CpGV in T. leucotreta. It was important to determine the LC and LT values required for mass production of CpGV in late instar T. leucotreta larvae. Dose- and time-response bioassays with CpGV-M were conducted on artificial diet to determine these values. Fourth instar LC₅₀ and LC₉₀ values were 5.96 x 10³ OBs/ml and 1.64 x 10⁵ OBs/ml respectively. LT50 and LT90 values were 81.10 hours and 88.58 hours respectively. Fifth instar LC₅₀ and LC₉₀ values were 6.88 x 10⁴ OBs/ml and 9.78 x 10⁶ OBs/ml respectively. LT₅₀ and LT₉₀ values were 111.56 hours and 137.57 hours respectively. Virus produced in fourth instar T. leucotreta larvae was bioassayed against C. pomonella neonate larvae and compared to CpGV-M to establish if production in the heterologous host negatively affected the virulence of the isolate. No significant difference in virulence was observed between virus produced in T. leucotreta and that produced in C. pomonella. The data generated in the bioassays was used in CpGV mass production trials to evaluate production. All production methods tested produced acceptable virus yields. To examine the quality of the virus product, genomic DNA was extracted from larval cadavers and subjected to REN analysis with HindIII. The resulting DNA profiles indicated that the virus product was contaminated with the homologous virus, CrleGV. Based on the above results, the use of T. leucotreta as an alternate host for the in vivo production of CpGV on a commercial basis is not at this stage viable and requires further investigation before this production methodology can be reliable used to produce CpGV. However, this study has shown that CpGV can be produced in a homologous host, T. leucotreta and significant strides have been made towards developing a set of quality control standards that are essential for further development of successful production methodology. Finally a novel isolate of CpGV has been identified with comparable virulence to the CpGV-M. This is an important finding as it has broad reaching implications for resistance management of CpGV products in South Africa.
- Full Text:
- Date Issued: 2015
- Authors: Chambers, Craig Brian
- Date: 2015
- Subjects: Cryptophlebia leucotreta -- South Africa , Codling moth -- South Africa , Apples -- Diseases and pests -- South Africa , Codling moth -- Biological control -- South Africa , Insect pests -- Biological control -- South Africa , Biological pest control agents -- South Africa , Baculoviruses -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5935 , http://hdl.handle.net/10962/d1017906
- Description: Cydia pomonella (Linnaeus) (Family: Tortricidae), the codling moth, is considered one of the most significant pests of apples and pears worldwide, causing up to 80% crop loss in orchards if no control measures are applied. Cydia pomonella is oligophagous feeding on a number of alternate hosts including quince, walnuts, apricots, peaches, plums and nectarines. Historically the control of this pest has been achieved with the use of various chemical control strategies which have maintained pest levels below the economic threshold at a relatively low cost to the grower. However, there are serious concerns surrounding the use of chemical insecticides including the development of resistance in insect populations, the banning of various insecticides, regulations for lowering of the maximum residue level and employee and consumer safety. For this reason, alternate measures of control are slowly being adopted by growers such as mating disruption, cultural methods and the use of baculovirus biopesticides as part of integrated pest management programmes. The reluctance of growers to accept baculovirus or other biological control products in the past has been due to questionable product quality and inconsistencies in their field performance. Moreover, the development and application of biological control products is more costly than the use of chemical alternatives. Baculoviruses are arthropod specific viruses that are highly virulent to a number of lepidopteran species. Due to the virulence and host specificity of baculoviruses, Cydia pomonella granulovirus has been extensively and successfully used as part of integrated pest management systems for the control of C. pomonella in Europe and around the world, including South Africa. Commercial formulations have been typically based on the Mexican strain of CpGV. However due to long-term multiple applications of CpGV and the reliance on CpGV in organic farming practices in Europe, resistance to the CpGV-M strain has developed in a number of field populations of C. pomonella. This study aimed to identify and characterize novel isolates of CpGV in South Africa and compare their virulence with the commercial standard CpGV-M. Secondly, since C. pomonella is difficult to culture on a large scale, an alternate method of CpGV production was investigated in order to determine if CpGV could be produced more efficiently and at a reduced cost without negatively impacting the quality of the product. Several isolates of CpGV were recovered either from field collected larvae or from a laboratory-reared C. pomonella colony. Characterisation of DNA profiles using a variety of restriction enzymes revealed that only a single isolate, CpGV-SA, was genetically different from the Mexican strain of the virus used in the commercially available CpGV based products in South Africa. In dose-response bioassays using CpGV-SA, LC₅₀ and LC₉₀ values for neonate C. pomonella larvae were 3.18 x 10³ OBs/ml and 7.33 x 10⁴ respectively. A comparison of these values with those of CpGV-M indicated no significant difference in the virulence of the two isolates under laboratory conditions. This is a first report of a genetically distinct CpGV isolate in South Africa. The biological activity and novelty of CpGV-SA makes this isolate a potentially important tool for CpGV resistance management in South Africa. In order to justify production of CpGV in an alternative host, studies on the comparative biological performance of C. pomonella and T. leucotreta based on oviposition, time to hatch, larval developmental times and rearing efficiency as well as production costs were performed. Thaumatotibia leucotreta was found to be more fecund and to have significantly shorter egg and larval developmental times. In addition, larval production per unit of artificial diet was significantly higher than for C. pomonella. This resulted in T. leucotreta being more cost effective to produce with implications for reduced insectary space, sanitation practices as well as the labour component of production. Virus yield data generated by inoculation both C. pomonella and T. leucotreta with nine concentrations of CpGV resulted in comparable virus yields, justifying the continuation of the research into production of CpGV in T. leucotreta. It was important to determine the LC and LT values required for mass production of CpGV in late instar T. leucotreta larvae. Dose- and time-response bioassays with CpGV-M were conducted on artificial diet to determine these values. Fourth instar LC₅₀ and LC₉₀ values were 5.96 x 10³ OBs/ml and 1.64 x 10⁵ OBs/ml respectively. LT50 and LT90 values were 81.10 hours and 88.58 hours respectively. Fifth instar LC₅₀ and LC₉₀ values were 6.88 x 10⁴ OBs/ml and 9.78 x 10⁶ OBs/ml respectively. LT₅₀ and LT₉₀ values were 111.56 hours and 137.57 hours respectively. Virus produced in fourth instar T. leucotreta larvae was bioassayed against C. pomonella neonate larvae and compared to CpGV-M to establish if production in the heterologous host negatively affected the virulence of the isolate. No significant difference in virulence was observed between virus produced in T. leucotreta and that produced in C. pomonella. The data generated in the bioassays was used in CpGV mass production trials to evaluate production. All production methods tested produced acceptable virus yields. To examine the quality of the virus product, genomic DNA was extracted from larval cadavers and subjected to REN analysis with HindIII. The resulting DNA profiles indicated that the virus product was contaminated with the homologous virus, CrleGV. Based on the above results, the use of T. leucotreta as an alternate host for the in vivo production of CpGV on a commercial basis is not at this stage viable and requires further investigation before this production methodology can be reliable used to produce CpGV. However, this study has shown that CpGV can be produced in a homologous host, T. leucotreta and significant strides have been made towards developing a set of quality control standards that are essential for further development of successful production methodology. Finally a novel isolate of CpGV has been identified with comparable virulence to the CpGV-M. This is an important finding as it has broad reaching implications for resistance management of CpGV products in South Africa.
- Full Text:
- Date Issued: 2015
The biology, behaviour and survival of pupating false codling moth, Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), a citrus pest in South Africa
- Authors: Love, Claire Natalie
- Date: 2015
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Larvae -- Behavior , Citrus -- Diseases and pests , Citrus -- Diseases and pests -- Biological control , Biological pest control agents , Entomopathogenic fungi , Insect nematodes , Pupae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5941 , http://hdl.handle.net/10962/d1018907
- Description: Control of the citrus pest, false codling moth (FCM), Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) is crucial for the South African citrus industry. The economic losses and phytosanitary status of this pest, coupled with increased consumer awareness and demands, has created a need for effective, IPM-compatible control measures for use against the soil-dwelling life stages of FCM. Promising developments in the field of microbial control through the use of entomopathogenic fungi (EPF) and entomopathogenic nematodes (EPNs) have highlighted the need for research regarding pupation biology, behaviour and survival of FCM, as a good understanding of biology of the target organism is an important component of any biological control programme. The aim of this study was to improve the current understanding of FCM pupation habits through the manipulation of soil texture class, ground cover, shading, soil compaction, air temperature, and soil moisture in the laboratory. These findings would then be used to aid the biological control programmes using EPF and EPNs against FCM in the soil. Three soil texture classes (sandy loam, silt loam and silty clay loam) were obtained from orchards for use in the study. FCM larvae were allowed to drop into the soil of their own accord and the pupation behaviour that followed was then captured on film with pupae formed in the soil being kept in order to measure adult eclosion. In general, very few abiotic factors had a clear influence on FCM pupation. Larval wandering time and distance was short, but also variable between individuals. Distance did increase when soils were moist. Pupation depth was shallow, with pupal cocoons generally being formed on the soil surface. Depth of pupation was less than one centimetre for all abiotic conditions, with little burrowing into soil. Eclosion success was higher for sandier soils when these were dry and uncompacted, but the addition of both moisture and soil compaction increased FCM eclosion success. FCM was sensitive to desiccation when the soils were dry and temperature limits of 15 °C and 32 °C had a strongly negative impact on eclosion success. Preferences for particular abiotic conditions were limited to only certain moisture conditions when interacting with soil texture class and a preference for pupating in soil when it is available. Limited preference was found for particular soil textures despite this having a strong influence on eclosion success, but individuals did appear to pupate in close proximity to one another. Viable direct habitat manipulation for FCM control could not be identified. These results and all of the abiotic variables measured have important implications for EPF and EPN application, survival and persistence in the soil in order to improve the ability of these biological control agents to control FCM. These are discussed in each chapter.
- Full Text:
- Date Issued: 2015
- Authors: Love, Claire Natalie
- Date: 2015
- Subjects: Cryptophlebia leucotreta -- South Africa , Cryptophlebia leucotreta -- Larvae -- Behavior , Citrus -- Diseases and pests , Citrus -- Diseases and pests -- Biological control , Biological pest control agents , Entomopathogenic fungi , Insect nematodes , Pupae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5941 , http://hdl.handle.net/10962/d1018907
- Description: Control of the citrus pest, false codling moth (FCM), Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) is crucial for the South African citrus industry. The economic losses and phytosanitary status of this pest, coupled with increased consumer awareness and demands, has created a need for effective, IPM-compatible control measures for use against the soil-dwelling life stages of FCM. Promising developments in the field of microbial control through the use of entomopathogenic fungi (EPF) and entomopathogenic nematodes (EPNs) have highlighted the need for research regarding pupation biology, behaviour and survival of FCM, as a good understanding of biology of the target organism is an important component of any biological control programme. The aim of this study was to improve the current understanding of FCM pupation habits through the manipulation of soil texture class, ground cover, shading, soil compaction, air temperature, and soil moisture in the laboratory. These findings would then be used to aid the biological control programmes using EPF and EPNs against FCM in the soil. Three soil texture classes (sandy loam, silt loam and silty clay loam) were obtained from orchards for use in the study. FCM larvae were allowed to drop into the soil of their own accord and the pupation behaviour that followed was then captured on film with pupae formed in the soil being kept in order to measure adult eclosion. In general, very few abiotic factors had a clear influence on FCM pupation. Larval wandering time and distance was short, but also variable between individuals. Distance did increase when soils were moist. Pupation depth was shallow, with pupal cocoons generally being formed on the soil surface. Depth of pupation was less than one centimetre for all abiotic conditions, with little burrowing into soil. Eclosion success was higher for sandier soils when these were dry and uncompacted, but the addition of both moisture and soil compaction increased FCM eclosion success. FCM was sensitive to desiccation when the soils were dry and temperature limits of 15 °C and 32 °C had a strongly negative impact on eclosion success. Preferences for particular abiotic conditions were limited to only certain moisture conditions when interacting with soil texture class and a preference for pupating in soil when it is available. Limited preference was found for particular soil textures despite this having a strong influence on eclosion success, but individuals did appear to pupate in close proximity to one another. Viable direct habitat manipulation for FCM control could not be identified. These results and all of the abiotic variables measured have important implications for EPF and EPN application, survival and persistence in the soil in order to improve the ability of these biological control agents to control FCM. These are discussed in each chapter.
- Full Text:
- Date Issued: 2015
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