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.
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- Date Issued: 2015
Evaluation of a plant-herbivore system in determining potential efficacy of a candidate biological control agent, cornops aquaticum for water hyacinth, eichhornia crassipes
- Authors: Bownes, Angela
- Date: 2009
- Subjects: Water hyacinth -- Control -- South Africa , Eichhornia crassipedes , Pontederiaceae , Grasshoppers , Biological pest control agents -- South Africa , Weeds -- Biological control -- South Africa , Invasive plants -- Biological control -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5687 , http://hdl.handle.net/10962/d1005373
- Description: Water hyacinth, Eichhornia crassipes Mart. Solms-Laubach (Pontederiaceae), a freefloating aquatic macrophyte of Neotropical origin, was introduced into South Africa as an ornamental aquarium plant in the early 1900’s. By the 1970’s it had reached pest proportions in dams and rivers around the country. Due to the sustainability, cost efficiency and low environmental risk associated with biological control, this has been a widely used method in an attempt to reduce infestations to below the threshold where they cause economic and ecological damage. To date, five arthropod and one pathogen biocontrol agents have been introduced for the control of water hyacinth but their impact has been variable. It is believed that their efficacy is hampered by the presence of highly eutrophic systems in South Africa in which plant growth is prolific and the negative effects of herbivory are therefore mitigated. It is for these reasons that new, potentially more damaging biocontrol agents are being considered for release. The water hyacinth grasshopper, Cornops aquaticum Brüner (Orthoptera: Acrididae), which is native to South America and Mexico, was brought into quarantine in Pretoria, South Africa in 1995. Although the grasshopper was identified as one of the most damaging insects associated with water hyacinth in its native range, it has not been considered as a biocontrol agent for water hyacinth anywhere else in the world. After extensive host-range testing which revealed it to be safe for release, a release permit for this candidate agent was issued in 2007. However, host specificity testing is no longer considered to be the only important component of pre-release screening of candidate biocontrol agents. Investigating biological and ecological aspects of the plant-herbivore system that will assist in determination of potential establishment, efficacy and the ability to build up good populations in the recipient environment are some of the important factors. This thesis is a pre-release evaluation of C. aquaticum to determine whether it is sufficiently damaging to water hyacinth to warrant its release. It investigated interactions between the grasshopper and water hyacinth under a range of nutrient conditions found in South African water bodies as well as the impact of the grasshopper on the competitive performance of water hyacinth. Both plant growth rates and the response of water hyacinth to herbivory by the grasshopper were influenced by nutrient availability to the plants. The ability of water hyacinth to compensate for loss of tissue through herbivory was greater under eutrophic nutrient conditions. However, a negative linear relationship was found between grasshopper biomass and water hyacinth performance parameters such as biomass accumulation and leaf production, even under eutrophic conditions. Water hyacinth’s compensatory ability in terms of its potential to mitigate to detrimental effects of insect feeding was dependent on the amount of damage caused by herbivory by the grasshopper. Plant biomass and the competitive ability of water hyacinth in relation to another freefloating aquatic weed species were reduced by C. aquaticum under eutrophic nutrient conditions, in a short space of time. It was also found that grasshopper feeding and characteristics related to their population dynamics such as fecundity and survival were significantly influenced by water nutrient availability and that environmental nutrient availability will influence the control potential of this species should it be released in South Africa. Cornops aquaticum shows promise as a biocontrol agent for water hyacinth but additional factors that were not investigated in this study such as compatibility with the South African climate and the current water hyacinth biocontrol agents need to be combined with these data to make a decision on its release. Possible management options for this species if it is to be introduced into South Africa are discussed.
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- Date Issued: 2009