Studies on existing and new isolates of Cryptophlebia leucotreta granulovirus (CrleGV) on Thaumatotibia leucotreta populations from a range of geographic regions in South Africa
- Authors: Opoku-Debrah, John Kwadwo
- Date: 2012
- Subjects: Cryptophlebia leucotreta Cryptophlebia leucotreta -- South Africa Cryptophlebia leucotreta -- Biological control Cryptophlebia leucotreta -- Life cycles Baculoviruses Lepidoptera -- Biological control Tortricidae -- Biological control Microbial insecticides Pests -- Integrated control
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5778 , http://hdl.handle.net/10962/d1005466
- Description: Baculoviruses are arthropod-specific DNA viruses that are highly virulent to most lepidopteran insects. Their host specificity and compatibility with IPM programmes has enabled their usage as safe microbial insecticides (biopesticides). Two baculovirus-based biopesticides, Cryptogran and Cryptex, which have been formulated with Cryptophlebia leucotreta granulovirus (CrleGV) have been registered for the control of false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) in South Africa and have been successfully incorporated into IPM programmes. However, several studies have indicated that insects can develop resistance to baculovirus-based biopesticide as was shown with field populations of codling moth (CM), Cydia pomonella (L.), which developed resistance to the biopesticide Cydia pomonella granulovirus (CpGV-M) in Europe. Other studies have shown that, under laboratory conditions, FCM populations differ in their susceptibility to Cryptogran and Cryptex. In order to investigate difference in susceptibility as well as protect against any future resistance by FCM to Cryptogran and Cryptex, a search for novel CrleGV-SA isolates from diseased insects from different geographic regions in South Africa was performed. Six geographic populations (Addo, Citrusdal, Marble Hall, Nelspruit, Baths and Mixed colonies) of FCM were established and maintained in the laboratory. Studies on the comparative biological performance based on pupal mass, female fecundity, egg hatch, pupal survival, adult eclosion and duration of life cycle of the Addo, Citrusdal, Marble Hall, Nelspruit and Mixed colonies revealed a low biological performance for the Citrusdal colony. This was attributed to the fact that FCM populations found in the Citrusdal area are not indigenous and may have been introduced from a very limited gene pool from another region. When insects from five colonies, excluding the Baths colony, were subjected to stress by overcrowding , a latent baculovirus resident in the Addo, Nelspruit, Citrusdal, Marble Hall and Mixed colonies was brought into an overt lethal state. Transmission electron micrographs revealed the presence of GV occlusion bodies (OBs) in diseased insects. DNA profiles obtained by single restriction endonuclease analysis of viral genomic DNA using BamH 1, Sa/1, Xba1 , Pst1, Xh01 , Kpn1, Hindlll and EcoR1 revealed five CrleGV-SA isolates latent within the insect populations. The new isolates were named CrleGV-SA Ado, CrleGV-SA Cit, CrleGV-SA Mbl, CrleGVSA Nels and CrleGV-SA Mix isolates. The novelty of the five CrleGV-SA isolates was confirmed by the presence of unique submolar bands, indicating that each isolate was genetically different. PCR amplification and sequencing of the granulin and egt genes from the five isolates revealed several single nucleotide polymorph isms (SNPs) which, in some cases, resulted in amino acid substitutions. DNA profiles from RFLPs, as well as phylogenetic analysis based on granulin and egt sequencing showed the presence of two CrleGV-SA genome types for the CrleGV-SA isolate. Cryptex and CrleGV-SA Ado, CrleGV-SA Cit, CrleGV-SA Mbl and CrleGV-SA Mix were placed as members of Group one CrleGV-SA, and Cryptogran and CrleGV-SA Nels isolate were placed into Group two CrleGV-SA. In droplet feeding bioassays, the median survival time (STso) for neonate larvae inoculated with Group one and two CrleGV-SA were determined to range from 80 - 88 hours (3.33 - 3.67 days), for all five colonies. LDso values for Group one and two CrleGV-SA against neonates from the Addo, Citrusdal, Marble Hall, Nelspruit and Mixed colonies varied between some populations and ranged from 0.80 - 3.12 OBs per larva, indicating some level of variation in host susceptibility. This is the first study reporting the existence of genetically distinct CrleGV baculovirus isolates infecting FCM in different geographical areas of South Africa. The results of this study have broad-ranging implications for our understanding of baculovirus-host interactions and for the application of baculovirus basedbiopesticides.
- Full Text:
- Date Issued: 2012
- Authors: Opoku-Debrah, John Kwadwo
- Date: 2012
- Subjects: Cryptophlebia leucotreta Cryptophlebia leucotreta -- South Africa Cryptophlebia leucotreta -- Biological control Cryptophlebia leucotreta -- Life cycles Baculoviruses Lepidoptera -- Biological control Tortricidae -- Biological control Microbial insecticides Pests -- Integrated control
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5778 , http://hdl.handle.net/10962/d1005466
- Description: Baculoviruses are arthropod-specific DNA viruses that are highly virulent to most lepidopteran insects. Their host specificity and compatibility with IPM programmes has enabled their usage as safe microbial insecticides (biopesticides). Two baculovirus-based biopesticides, Cryptogran and Cryptex, which have been formulated with Cryptophlebia leucotreta granulovirus (CrleGV) have been registered for the control of false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) in South Africa and have been successfully incorporated into IPM programmes. However, several studies have indicated that insects can develop resistance to baculovirus-based biopesticide as was shown with field populations of codling moth (CM), Cydia pomonella (L.), which developed resistance to the biopesticide Cydia pomonella granulovirus (CpGV-M) in Europe. Other studies have shown that, under laboratory conditions, FCM populations differ in their susceptibility to Cryptogran and Cryptex. In order to investigate difference in susceptibility as well as protect against any future resistance by FCM to Cryptogran and Cryptex, a search for novel CrleGV-SA isolates from diseased insects from different geographic regions in South Africa was performed. Six geographic populations (Addo, Citrusdal, Marble Hall, Nelspruit, Baths and Mixed colonies) of FCM were established and maintained in the laboratory. Studies on the comparative biological performance based on pupal mass, female fecundity, egg hatch, pupal survival, adult eclosion and duration of life cycle of the Addo, Citrusdal, Marble Hall, Nelspruit and Mixed colonies revealed a low biological performance for the Citrusdal colony. This was attributed to the fact that FCM populations found in the Citrusdal area are not indigenous and may have been introduced from a very limited gene pool from another region. When insects from five colonies, excluding the Baths colony, were subjected to stress by overcrowding , a latent baculovirus resident in the Addo, Nelspruit, Citrusdal, Marble Hall and Mixed colonies was brought into an overt lethal state. Transmission electron micrographs revealed the presence of GV occlusion bodies (OBs) in diseased insects. DNA profiles obtained by single restriction endonuclease analysis of viral genomic DNA using BamH 1, Sa/1, Xba1 , Pst1, Xh01 , Kpn1, Hindlll and EcoR1 revealed five CrleGV-SA isolates latent within the insect populations. The new isolates were named CrleGV-SA Ado, CrleGV-SA Cit, CrleGV-SA Mbl, CrleGVSA Nels and CrleGV-SA Mix isolates. The novelty of the five CrleGV-SA isolates was confirmed by the presence of unique submolar bands, indicating that each isolate was genetically different. PCR amplification and sequencing of the granulin and egt genes from the five isolates revealed several single nucleotide polymorph isms (SNPs) which, in some cases, resulted in amino acid substitutions. DNA profiles from RFLPs, as well as phylogenetic analysis based on granulin and egt sequencing showed the presence of two CrleGV-SA genome types for the CrleGV-SA isolate. Cryptex and CrleGV-SA Ado, CrleGV-SA Cit, CrleGV-SA Mbl and CrleGV-SA Mix were placed as members of Group one CrleGV-SA, and Cryptogran and CrleGV-SA Nels isolate were placed into Group two CrleGV-SA. In droplet feeding bioassays, the median survival time (STso) for neonate larvae inoculated with Group one and two CrleGV-SA were determined to range from 80 - 88 hours (3.33 - 3.67 days), for all five colonies. LDso values for Group one and two CrleGV-SA against neonates from the Addo, Citrusdal, Marble Hall, Nelspruit and Mixed colonies varied between some populations and ranged from 0.80 - 3.12 OBs per larva, indicating some level of variation in host susceptibility. This is the first study reporting the existence of genetically distinct CrleGV baculovirus isolates infecting FCM in different geographical areas of South Africa. The results of this study have broad-ranging implications for our understanding of baculovirus-host interactions and for the application of baculovirus basedbiopesticides.
- Full Text:
- Date Issued: 2012
The effects of sustained attention, workload and task-related fatigue on physiological measures and performance during a tracking task
- Authors: De Gray Birch, Casey
- Date: 2012
- Subjects: Fatigue -- Physiological aspects -- Research , Attention -- Physiological aspects -- Research , Human engineering -- Research
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5120 , http://hdl.handle.net/10962/d1005198 , Fatigue -- Physiological aspects -- Research , Attention -- Physiological aspects -- Research , Human engineering -- Research
- Description: Despite extensive research into the concept of mental fatigue there is as yet no “gold standard” definition or measurement technique available. Because of this a large amount of fatigue-related errors are still seen in the workplace. The complexity of the problem lies with the inability to directly measure mental processes as well as the various endogenous and exogenous factors that interact to produce the experienced fatigue. Fatigue has been divided into sleep-related and task-related fatigue; however the task-related aspect is evident both during normal waking hours as well as during periods of sleep deprivation, therefore this aspect is considered important in the understanding of fatigue in general. The concept of task-related fatigue has further been divided into active and passive fatigue states; however differentiation between the two requires careful consideration. Various physiological measures have been employed in an attempt to gain a better understanding of the mechanisms involved in the generation of fatigue, however often studies have produced dissociating results. The current study considered the task-related fatigue elicited by a tracking task requiring sustained attention, in order to evaluate the usefulness of various cardiovascular and oculomotor measures as indicators of fatigue. A secondary aim was to determine whether the behavioural and physiological parameter responses could be used to infer the type of fatigue incurred (i.e. an active versus passive fatigue state) as well as the energetical mechanisms involved during task performance. A simple driving simulator task was used as the main tracking task, requiring constant attention and concentration. This task was performed for approximately two hours. Three experimental groups (consisting of 14 subjects each) were used: a control group that performed the tracking task only, a group that performed a five minute auditory memory span task concurrently with the driving task after every 20 minutes of pure driving, and a group that performed a visual choice reaction task for five minutes following every 20 minute driving period. The secondary tasks were employed in order to evaluate the extent of resource allocation as well as arousal level. Performance measures included various driving performance parameters, as well as secondary task performance. Physiological measures included heart rate frequency (HR) and various time- and frequency-domain heart rate variability (HRV)parameters, pupil dilation, blink frequency and duration, fixations, and saccadic parameters as well as critical flicker fusion frequency (CFFF). The Borg CR-10 scale was used to evaluate subjective fatigue during the task, and the NASA-TLX was completed following the task. A decline in driving performance over time was supplemented by measures such as HR, HRV and pupil dilation indicating an increase in parasympathetic activity (or a reduction in arousal). An increase in blink frequency was considered as a sign of withdrawal of attentional resources over time. Longer and faster saccades were also evident over time, coupled with shorter fixations. With regards to the secondary task influence, the choice RT task did not affect any behavioural or physiological parameters, thereby contesting the active fatigue theory of resource depletion, as well as implying that the increase in demand for the same resources used by the primary task was insufficient to affect the state of the subjects. The increased load elicited by the memory span task improved driving performance and increased measures of HR, HRV, pupil dilation and blink frequency. Some of these measures produced opposite effects to what was expected; an attempt to explain the dissociation of the various physiological parameters was expressed in terms of arousal, effort and resource theories. Overall, the results indicate that the fatigue and/or reduced arousal accompanying a monotonous sustained attention task can, to some degree, be alleviated through intermittent performance of a secondary task engaging mental resources other than the ones used for the primary task. The degree to which such a task is beneficial, however, requires careful consideration as while an immediate increase in arousal and primary task performance is noted, the impact of the task on general attentional resources may be detrimental in the case of reacting should an emergency situation occur.
- Full Text:
- Date Issued: 2012
- Authors: De Gray Birch, Casey
- Date: 2012
- Subjects: Fatigue -- Physiological aspects -- Research , Attention -- Physiological aspects -- Research , Human engineering -- Research
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5120 , http://hdl.handle.net/10962/d1005198 , Fatigue -- Physiological aspects -- Research , Attention -- Physiological aspects -- Research , Human engineering -- Research
- Description: Despite extensive research into the concept of mental fatigue there is as yet no “gold standard” definition or measurement technique available. Because of this a large amount of fatigue-related errors are still seen in the workplace. The complexity of the problem lies with the inability to directly measure mental processes as well as the various endogenous and exogenous factors that interact to produce the experienced fatigue. Fatigue has been divided into sleep-related and task-related fatigue; however the task-related aspect is evident both during normal waking hours as well as during periods of sleep deprivation, therefore this aspect is considered important in the understanding of fatigue in general. The concept of task-related fatigue has further been divided into active and passive fatigue states; however differentiation between the two requires careful consideration. Various physiological measures have been employed in an attempt to gain a better understanding of the mechanisms involved in the generation of fatigue, however often studies have produced dissociating results. The current study considered the task-related fatigue elicited by a tracking task requiring sustained attention, in order to evaluate the usefulness of various cardiovascular and oculomotor measures as indicators of fatigue. A secondary aim was to determine whether the behavioural and physiological parameter responses could be used to infer the type of fatigue incurred (i.e. an active versus passive fatigue state) as well as the energetical mechanisms involved during task performance. A simple driving simulator task was used as the main tracking task, requiring constant attention and concentration. This task was performed for approximately two hours. Three experimental groups (consisting of 14 subjects each) were used: a control group that performed the tracking task only, a group that performed a five minute auditory memory span task concurrently with the driving task after every 20 minutes of pure driving, and a group that performed a visual choice reaction task for five minutes following every 20 minute driving period. The secondary tasks were employed in order to evaluate the extent of resource allocation as well as arousal level. Performance measures included various driving performance parameters, as well as secondary task performance. Physiological measures included heart rate frequency (HR) and various time- and frequency-domain heart rate variability (HRV)parameters, pupil dilation, blink frequency and duration, fixations, and saccadic parameters as well as critical flicker fusion frequency (CFFF). The Borg CR-10 scale was used to evaluate subjective fatigue during the task, and the NASA-TLX was completed following the task. A decline in driving performance over time was supplemented by measures such as HR, HRV and pupil dilation indicating an increase in parasympathetic activity (or a reduction in arousal). An increase in blink frequency was considered as a sign of withdrawal of attentional resources over time. Longer and faster saccades were also evident over time, coupled with shorter fixations. With regards to the secondary task influence, the choice RT task did not affect any behavioural or physiological parameters, thereby contesting the active fatigue theory of resource depletion, as well as implying that the increase in demand for the same resources used by the primary task was insufficient to affect the state of the subjects. The increased load elicited by the memory span task improved driving performance and increased measures of HR, HRV, pupil dilation and blink frequency. Some of these measures produced opposite effects to what was expected; an attempt to explain the dissociation of the various physiological parameters was expressed in terms of arousal, effort and resource theories. Overall, the results indicate that the fatigue and/or reduced arousal accompanying a monotonous sustained attention task can, to some degree, be alleviated through intermittent performance of a secondary task engaging mental resources other than the ones used for the primary task. The degree to which such a task is beneficial, however, requires careful consideration as while an immediate increase in arousal and primary task performance is noted, the impact of the task on general attentional resources may be detrimental in the case of reacting should an emergency situation occur.
- Full Text:
- Date Issued: 2012
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