Molecular studies of Wolbachia and sex-determination genes in Australian Bactrocera species : complementary approaches to improved fruit fly control

Bactrocera tryoni (Diptera, Tephritidae), the Queensland fruit fly (Qfly), is the most significant horticultural pest species in Australia. The Sterile Insect Technique (SIT), which relies on the release of vast numbers of irradiated sterile insects to suppress field populations, has been used in this species and, globally, is a successful pest management strategy for many insect species. For Mediterranean fruit fly, Ceratitis capitata, the incorporation of a genetic sexing system (GSS), that allows male-only releases, can increase effectiveness of SIT by 3-5 times. However, there is currently no GSS for B. tryoni. Furthermore, field performance of sterile flies, mass-reared and irradiated, can be generally lower than of wild individuals. Novel strategies such as Incompatible Insect Technique (IIT) exploit the reproductive manipulations of the maternally-inherited common endosymbiotic bacterium of insects, Wolbachia, to induce unidirectionally incompatible matings between uninfected field-females and released males carrying Wolbachia. While this method circumvents irradiation, a male-only release cohort is essential because females carrying Wolbachia are fully fertile; accidental release of infected females and thus inheritance of released Wolbachia in field populations would soon lead to a breakdown of the mating incompatibility essential for pest suppression. This thesis details the results of three fundamental and independent, yet complementary, experiments designed to investigate prospects for IIT and improvement of SIT, namely: an assessment of the incidence, prevalence and type of Wolbachia strains in Australian tephritid fruit fly species across their geographic range; a survey of the general microbiome of tephritid fruit fly species from different ecological niches and both natural and laboratory sources; an examination of genes expressed in early embryos of two Australian Bactrocera pest species, to generate molecular tools for GSS. Moreover, this work represents a valuable contribution to the ecological and epidemiological understanding of Wolbachia infection life cycles in insect communities, through discovery of a new model system for examining the distribution of Wolbachia along a climatic gradient, and the horizontal transmission of Wolbachia among a community of tephritid fruit flies. This work also presents important molecular studies of sex-specific gene expression over early embryonic development, with the aim of clarifying the function of M in tephritid fruit flies as models for the many insect species with similar sex-determination systems

Substantial rearrangements, single nucleotide frameshift deletion and low diversity in mitogenome of Wolbachia‑infected strepsipteran endoparasitoid in comparison to its tephritid hosts

Insect mitogenome organisation is highly conserved, yet, some insects, especially with parasitic life cycles, have rearranged mitogenomes. Furthermore, intraspecific mitochondrial diversity can be reduced by fitness-affecting bacterial endosymbionts like Wolbachia due to their maternal coinheritance with mitochondria. We have sequenced mitogenomes of the Wolbachia-infected endoparasitoid Dipterophagus daci (Strepsiptera: Halictophagidae) and four of its 22 known tephritid fruit fly host species using total genomic extracts of parasitised flies collected across > 700 km in Australia. This halictophagid mitogenome revealed extensive rearrangements relative to the four fly mitogenomes which exhibited the ancestral insect mitogenome pattern. Compared to the only four available other strepsipteran mitogenomes, the D. daci mitogenome had additional transpositions of one rRNA and two tRNA genes, and a single nucleotide frameshift deletion in nad5 requiring translational frameshifting or, alternatively, resulting in a large protein truncation. Dipterophagus daci displays an almost completely endoparasitic life cycle when compared to Strepsiptera that have maintained the ancestral state of free-living adults. Our results support the hypothesis that the transition to extreme endoparasitism evolved together with increased levels of mitogenome changes. Furthermore, intraspecific mitogenome diversity was substantially smaller in D. daci than the parasitised flies suggesting Wolbachia reduced mitochondrial diversity because of a role in D. daci fitness

Characterization of body weight and composition changes during the sophomore year of college

<p>Abstract</p> <p>Background</p> <p>Years spent in college represents a critical time for obesity development though little information is known regarding how body weight and composition changes beyond the first year of college. The aim of this study was to investigate changes in body weight and composition and the factors influencing those changes among sophomore females.</p> <p>Methods</p> <p>Body composition by dual energy X-ray absorptiometry was obtained in participants beginning during their freshman year and continued through their sophomore year.</p> <p>Results</p> <p>No difference was observed between sophomore year fall and spring visits for body weight (60.4 versus 60.6 kg) or fat mass (19.3 versus 18.7 kg). However, a significant (<it>P </it>≤ 0.05) decrease was observed for body fat (31.9 versus 30.9 %fat) and a significant increase was observed for fat-free mass (37.7 versus 38.4 kg). Participants living off campus significantly (<it>P </it>≤ 0.05) declined in body fat (33.0 versus 31.0 %fat) and fat mass (19.4 versus 18.2 kg) and increased in fat-free mass (36.1 versus 37.2 kg) with no differences in those living on campus.</p> <p>Conclusion</p> <p>No change in body weight was observed in females during their sophomore year. However, an increase in fat-free mass accompanied with a decrease in fat mass resulted in a decrease in body fat. Participants living off campus had favorable changes in their body composition by means of decreasing %fat and fat mass while increasing fat-free mass. Participants living on campus did not demonstrate these favorable changes.</p

Bacterial Communities Are Less Diverse in a Strepsipteran Endoparasitoid than in Its Fruit Fly Hosts and Dominated by Wolbachia

Microbiomes play vital roles in insect fitness and health and can be influenced by interactions between insects and their parasites. Many studies investigate the microbiome of free-living insects, whereas microbiomes of endoparasitoids and their interactions with parasitised insects are less explored. Due to their development in the constrained environment within a host, endoparasitoids are expected to have less diverse yet distinct microbiomes. We used high-throughput 16S rRNA gene amplicon sequencing to characterise the bacterial communities of Dipterophagus daci (Strepsiptera) and seven of its tephritid fruit fly host species. Bacterial communities of D. daci were less diverse and contained fewer taxa relative to the bacterial communities of the tephritid hosts. The strepsipteran’s microbiome was dominated by Pseudomonadota (formerly Proteobacteria) (> 96%), mainly attributed to the presence of Wolbachia, with few other bacterial community members, indicative of an overall less diverse microbiome in D. daci. In contrast, a dominance of Wolbachia was not found in flies parasitised by early stages of D. daci nor unparasitised flies. Yet, early stages of D. daci parasitisation resulted in structural changes in the bacterial communities of parasitised flies. Furthermore, parasitisation with early stages of D. daci with Wolbachia was associated with a change in the relative abundance of some bacterial taxa relative to parasitisation with early stages of D. daci lacking Wolbachia. Our study is a first comprehensive characterisation of bacterial communities in a Strepsiptera species together with the more diverse bacterial communities of its hosts and reveals effects of concealed stages of parasitisation on host bacterial communities

Protozoal Coinfection in Horses with Equine Protozoal Myeloencephalitis in the Eastern United States

Background: Infection by 2 or more protozoa is linked with increased severity of disease in marine mammals with protozoan encephalitis. Hypothesis/Objectives: To assess whether horses with equine protozoal myeloencephalitis (EPM) caused by Sarcocystis neurona also have evidence of infection with Neospora hughesi or Toxoplasma gondii. We hypothesized that horses with EPM would be more likely than horses with cervical vertebral stenotic myelopathy (CVSM) to be positive for antibodies to multiple protozoan parasites. Animals: One hundred one horses with neurologic disease: 49 with EPM and 52 with CVSM. Methods: Case review. Archived serum and cerebrospinal fluid (CSF) from 101 horses were examined. Inclusion criteria included neurologic disease, antemortem or postmortem diagnosis of EPM or CVSM, and availability of serological results or archived samples for testing. Additional testing for antibodies was performed on serum for T. gondii, as well as serum and CSF for N. hughesi. Results: Horses with EPM were more likely than horses with CVSM to have positive immunologic results for S. neurona on serum (95.9% versus 76.9%, P = .0058), CSF (98.0% versus 44.2%, P \u3c .00001), and serum : CSF titer ratio (91.8% versus 0%, P \u3c .00001). Positive results for Neospora and Toxoplasma were uncommon, with total seroprevalence rates of 12.9% and 14.9%, respectively. The proportions of EPM cases testing positive for Neospora and Toxoplasma (16% and 12%) were not different from the proportions of CVSM cases testing positive (10% and 17%, P = .31 and .47, respectively). Conclusion: Results do not indicate an important role for protozoal coinfection in EPM in the eastern United States

Endosymbionts moderate constrained sex allocation in a haplodiploid thrips species in a temperature-sensitive way

Maternally inherited bacterial endosymbionts that affect host fitness are common in nature. Some endosymbionts colonise host populations by reproductive manipulations (such as cytoplasmic incompatibility; CI) that increase the reproductive fitness of infected over uninfected females. Theory predicts that CI-inducing endosymbionts in haplodiploid hosts may also influence sex allocation, including in compatible crosses, however, empirical evidence for this is scarce. We examined the role of two common CI-inducing endosymbionts, Cardinium and Wolbachia, in the sex allocation of Pezothrips kellyanus, a haplodiploid thrips species with a split sex ratio. In this species, irrespective of infection status, some mated females are constrained to produce extremely male-biased broods, whereas other females produce extremely female-biased broods. We analysed brood sex ratio of females mated with males of the same infection status at two temperatures. We found that at 20 °C the frequency of constrained sex allocation in coinfected pairs was reduced by 27% when compared to uninfected pairs. However, at 25 °C the constrained sex allocation frequency increased and became similar between coinfected and uninfected pairs, resulting in more male-biased population sex ratios at the higher temperature. This temperature-dependent pattern occurred without changes in endosymbiont densities and compatibility. Our findings indicate that endosymbionts affect sex ratios of haplodiploid hosts beyond the commonly recognised reproductive manipulations by causing female-biased sex allocation in a temperature-dependent fashion. This may contribute to a higher transmission efficiency of CI-inducing endosymbionts and is consistent with previous models that predict that CI by itself is less efficient in driving endosymbiont invasions in haplodiploid hosts

Phosphorylated neurofilament H (pNF-H) as a potential diagnostic marker for neurological disorders in horses

The current study aimed at the investigating the potential use of phosphorylated neurofilament H (pNF-H) as a diagnostic biomarker for neurologic disorders in the horse. Paired serum and cerebrospinal fluid (CSF) samples (n = 88) and serum only (n = 30) were obtained from horses diagnosed with neurologic disorders and clinically healthy horses as control. The neurologic horses consisted of equine protozoal myeloencephalitis (EPM) (38 cases) and cervical vertebral malformation (CVM) (23 cases). Levels of pNF-H were determined using an ELISA. The correlation between CSF and serum concentrations of pNF-H was evaluated using Spearman's Rank test and the significance of the difference among the groups was assessed using a nonparametric test. Horses had higher pNF-H levels in the CSF than serum. Horses afflicted with EPM had significantly higher serum pNF-H levels in comparison to controls or CVM cases. The correlation between CSF and serum pNF-H levels was poor in both the whole study population and among subgroups of horses included in the study. There was significant association between the likelihood of EPM and the concentrations of pNF-H in either the serum or CSF. These data suggest that pNF-H could be detected in serum and CSF samples from neurologic and control horses. This study demonstrated that pNF-H levels in serum and CSF have the potential to provide objective information to help in the early diagnosis of horses afflicted with neurologic disorders

Characterization of the bacterial communities of psyllids associated with Rutaceae in Bhutan by high throughput sequencing

Background: Several plant-pathogenic bacteria are transmitted by insect vector species that often also act as hosts. In this interface, these bacteria encounter plant endophytic, insect endosymbiotic and other microbes. Here, we used high throughput sequencing to examine the bacterial communities of five different psyllids associated with citrus and related plants of Rutaceae in Bhutan: Diaphorina citri, Diaphorina communis, Cornopsylla rotundiconis, Cacopsylla heterogena and an unidentified Cacopsylla sp. Results: The microbiomes of the psyllids largely comprised their obligate P-endosymbiont ‘Candidatus Carsonella ruddii’, and one or two S-endosymbionts that are fixed and specific to each lineage. In addition, all contained Wolbachia strains; the Bhutanese accessions of D. citri were dominated by a Wolbachia strain first found in American isolates of D. citri, while D. communis accessions were dominated by the Wolbachia strain, wDi, first detected in D. citri from China. The S-endosymbionts from the five psyllids grouped with those from other psyllid taxa; all D. citri and D. communis individuals contained sequences matching ‘Candidatus Profftella armatura’ that has previously only been reported from other Diaphorina species, and the remaining psyllid species contained OTUs related to unclassified Enterobacteriaceae. The plant pathogenic ‘Candidatus Liberibacter asiaticus’ was found in D. citri but not in D. communis. Furthermore, an unidentified ‘Candidatus Liberibacter sp.’ occurred at low abundance in both Co. rotundiconis and the unidentified Cacopsylla sp. sampled from Zanthoxylum sp.; the status of this new liberibacter as a plant pathogen and its potential plant hosts are currently unknown. The bacterial communities of Co. rotundiconis also contained a range of OTUs with similarities to bacteria previously found in samples taken from various environmental sources. Conclusions: The bacterial microbiota detected in these Bhutanese psyllids support the trends that have been seen in previous studies: psyllids have microbiomes largely comprising their obligate P-endosymbiont and one or two Sendosymbionts. In addition, the association with plant pathogens has been demonstrated, with the detection of liberibacters in a known host, D. citri, and identification of a putative new species of liberibacter in Co. rotundiconis and Cacopsylla sp