The role of Clostridium botulinum in the aetiology of equine grass sickness and other dysautonomias

Equine grass sickness is a primary dysautonomy of unproven aetiology that is mainly fatal, untreatable and unpreventable. Much circumstantial evidence has been demonstrated that supports the hypothesis that the disease is caused by a toxicoinfection with Clostridium botulinum type C/D. Further evidence supporting this hypothesis and an understanding of the immune response of the horse to this organism and its toxins will facilitate vaccine research and design.Serum samples from 6-month old ponies (n = 26) were assayed for antibodies against botulinum neurotoxins type C and D (BoNT/C & D) and against surface antigens (SA). Statistically significant rises in specific lgG levels against all three antigens were shown across the sampling period, demonstrating specific antibody acquisition with age. Adult horses (n = 40) were sampled fortnightly for twelve months and assayed for specific IgG against the same antigens. In this group fluctuations in specific IgG levels were observed that corresponded with changes in management.Seven acute cases of grass sickness that went to postmortem within 36h of onset of clinical signs and four control horses that went to post-mortem had both serum and gut samples taken. Specific IgA was assayed and cases demonstrated significantly more specific IgA against BoNT/C in the jejunum, ileum and caecum (P = 0.04, 0.02 and 0.006 respectively). Against BoNT/D, cases demonstrated significantly more IgA in the pharynx, duodenum and jejunum (P = 0.01, 0.01 and 0.02 respectively) and against surface antigens cases demonstrated significantly more specific IgA in the duodenum (P = 0.01). Serum samples from these acute cases were assayed for specific IgG and IgG subclasses against the same antigens and compared to co-grazing controls. Cases demonstrated lower specific IgG against surface antigens than co-grazing controls (P = 0.06).Sera from twelve chronic cases of grass sickness were assayed for specific IgG. Six horses that were subsequently euthanased demonstrated significantly lower initial IgG levels against SA (P = 0.05) than those that survived and almost significantly lower IgG levels to BoNT/C (P = 0.06) indicating that immune status at disease onset may be important in survival / prognosis. Longitudinal analysis showed that rising or falling levels of specific IgG had no influence on disease outcome.An outbreak of feline dysautonomia was investigated by bacteriological and serological techniques. BoNT/C was detected in faeces in seven of eight affected cats after enrichment and also detected in their food. No toxin was detected in the faeces of controls. Specific IgA was also present in significantly higher amounts in cases than in controls (P = <0.001) against BoNT/C and SA. In serological studies, associations were found between low antibody levels and disease in cats but not in dogs, which lacked an age-matched control group.EGS and dysautonomias of other species appear to be associated with toxicoinfections with C. botulinum types C and/or type D. As such, a vaccine, consisting of both surface antigen and toxin components, would play a vital role in protecting animals against this distressing disease

RNAi gene knockdown in the poultry red mite, Dermanyssus gallinae (De Geer 1778), a tool for functional genomics

The authors gratefully acknowledge funding for this project from the Scottish Government Rural Affairs, Food and the Environment (RAFE) Strategic Research Portfolio 2016-2021. DRGP is supported by a research fellowship provided by the Moredun Foundation. WC is supported by a studentship provided by the University of Aberdeen and the Moredun Foundation.Peer reviewedPublisher PD

A genomic analysis and transcriptomic atlas of gene expression in Psoroptes ovis reveals feeding- and stage-specific patterns of allergen expression

Background: Psoroptic mange, caused by infestation with the ectoparasitic mite, Psoroptes ovis, is highly contagious, resulting in intense pruritus and represents a major welfare and economic concern for the livestock industry Worldwide. Control relies on injectable endectocides and organophosphate dips, but concerns over residues, environmental contamination, and the development of resistance threaten the sustainability of this approach, highlighting interest in alternative control methods. However, development of vaccines and identification of chemotherapeutic targets is hampered by the lack of P. ovis transcriptomic and genomic resources. Results: Building on the recent publication of the P. ovis draft genome, here we present a genomic analysis and transcriptomic atlas of gene expression in P. ovis revealing feeding- and stage-specific patterns of gene expression, including novel multigene families and allergens. Network-based clustering revealed 14 gene clusters demonstrating either single- or multi-stage specific gene expression patterns, with 3075 female-specific, 890 male-specific and 112, 217 and 526 transcripts showing larval, protonymph and tritonymph specific-expression, respectively. Detailed analysis of P. ovis allergens revealed stage-specific patterns of allergen gene expression, many of which were also enriched in "fed" mites and tritonymphs, highlighting an important feeding-related allergenicity in this developmental stage. Pair-wise analysis of differential expression between life-cycle stages identified patterns of sex-biased gene expression and also identified novel P. ovis multigene families including known allergens and novel genes with high levels of stage-specific expression. Conclusions: The genomic and transcriptomic atlas described here represents a unique resource for the acarid-research community, whilst the OrcAE platform makes this freely available, facilitating further community-led curation of the draft P. ovis genome

Transcriptomic analysis of the poultry red mite, Dermanyssus gallinae, across all stages of the lifecycle

Acknowledgements Thanks go to the Centre for Genomic Research (CGR) at the University of Liverpool performing the TruSeq RNA-seq analysis and to our local layer farmers for their continued support and provision of mite material. Funding The authors gratefully acknowledge funding for this project from BBRSC (grant reference BB/J01513X/1), Zoetis and Akita Co. Ltd. and The British Egg Marketing Board Trust.Peer reviewedPublisher PD

A genomic analysis and transcriptomic atlas of gene expression in Psoroptes ovis reveals feeding- and stage-specific patterns of allergen expression

Background: Psoroptic mange, caused by infestation with the ectoparasitic mite, Psoroptes ovis, is highly contagious, resulting in intense pruritus and represents a major welfare and economic concern for the livestock industry Worldwide. Control relies on injectable endectocides and organophosphate dips, but concerns over residues, environmental contamination, and the development of resistance threaten the sustainability of this approach, highlighting interest in alternative control methods. However, development of vaccines and identification of chemotherapeutic targets is hampered by the lack of P. ovis transcriptomic and genomic resources. Results: Building on the recent publication of the P. ovis draft genome, here we present a genomic analysis and transcriptomic atlas of gene expression in P. ovis revealing feeding- and stage-specific patterns of gene expression, including novel multigene families and allergens. Network-based clustering revealed 14 gene clusters demonstrating either single- or multi-stage specific gene expression patterns, with 3075 female-specific, 890 male-specific and 112, 217 and 526 transcripts showing larval, protonymph and tritonymph specific-expression, respectively. Detailed analysis of P. ovis allergens revealed stage-specific patterns of allergen gene expression, many of which were also enriched in "fed" mites and tritonymphs, highlighting an important feeding-related allergenicity in this developmental stage. Pair-wise analysis of differential expression between life-cycle stages identified patterns of sex-biased gene expression and also identified novel P. ovis multigene families including known allergens and novel genes with high levels of stage-specific expression. Conclusions: The genomic and transcriptomic atlas described here represents a unique resource for the acarid-research community, whilst the OrcAE platform makes this freely available, facilitating further community-led curation of the draft P. ovis genome

Transcriptomic analysis of the temporal host response to skin infestation with the ectoparasitic mite Psoroptes ovis

<p>Abstract</p> <p>Background</p> <p>Infestation of ovine skin with the ectoparasitic mite <it>Psoroptes ovis </it>results in a rapid cutaneous immune response, leading to the crusted skin lesions characteristic of sheep scab. Little is known regarding the mechanisms by which such a profound inflammatory response is instigated and to identify novel vaccine and drug targets a better understanding of the host-parasite relationship is essential. The main objective of this study was to perform a combined network and pathway analysis of the <it>in vivo </it>skin response to infestation with <it>P. ovis </it>to gain a clearer understanding of the mechanisms and signalling pathways involved.</p> <p>Results</p> <p>Infestation with <it>P. </it>ovis resulted in differential expression of 1,552 genes over a 24 hour time course. Clustering by peak gene expression enabled classification of genes into temporally related groupings. Network and pathway analysis of clusters identified key signalling pathways involved in the host response to infestation. The analysis implicated a number of genes with roles in allergy and inflammation, including pro-inflammatory cytokines (<it>IL1A, IL1B, IL6, IL8 </it>and <it>TNF</it>) and factors involved in immune cell activation and recruitment (<it>SELE, SELL, SELP, ICAM1, CSF2, CSF3, CCL2 </it>and <it>CXCL2</it>). The analysis also highlighted the influence of the transcription factors NF-kB and AP-1 in the early pro-inflammatory response, and demonstrated a bias towards a Th2 type immune response.</p> <p>Conclusions</p> <p>This study has provided novel insights into the signalling mechanisms leading to the development of a pro-inflammatory response in sheep scab, whilst providing crucial information regarding the nature of mite factors that may trigger this response. It has enabled the elucidation of the temporal patterns by which the immune system is regulated following exposure to <it>P. ovis</it>, providing novel insights into the mechanisms underlying lesion development. This study has improved our existing knowledge of the host response to <it>P. ovis</it>, including the identification of key parallels between sheep scab and other inflammatory skin disorders and the identification of potential targets for disease control.</p

Development of a serodiagnostic test for sheep scab using recombinant protein Pso o 2

a b s t r a c t Early stages of sheep scab, the disease caused by the non-burrowing mite Psoroptes ovis, are often subclinical, or can be mis-diagnosed. A diagnostic test capable of detecting early disease and latent infestations is therefore highly desirable in disease control. This paper describes the design and validation of an ELISA, which incorporates a recombinant P. ovis antigen (Pso o 2), for the early detection of anti-P. ovis serum antibodies in sheep. This ELISA was evaluated using sera from sheep infested with P. ovis (n ¼ 58) and sheep (n ¼ 433) with no P. ovis infestation as well as sheep infected with other parasites including gastrointestinal nematodes (GIN), or chewing lice. A receiver operating characteristic (ROC) curve analysis was generated using the ELISA results for 491 sheep sera with the area under the curve (AUC) being 0.97. An optimal OD 450 cut-off of &gt;0.06 absorbance units gave a test sensitivity of 0.93 and specificity of 0.90. The Pso o 2-based ELISA was able to detect specific antibodies to P. ovis during early experimental infestation prior to disease patency, indicating its utility for detecting sub-clinical infestation. Crow

End-tidal versus manually-controlled low-flow anaesthesia

During low-flow manually-controlled anaesthesia (MCA) the anaesthetist needs constantly adjust end-tidal oxygen (EtO2) and anaesthetic concentrations (EtAA) to assure an adequate and safe anaesthesia. Recently introduced anaesthetic machines can automatically maintain those variables at target values, avoiding the burden on the anaesthetist. End-tidal-controlled anaesthesia (EtCA) and MCA provided by the same anaesthetic machine under the same fresh gas flow were compared. Eighty patients were prospectively observed: in MCA group (n = 40) target end-tidal sevoflurane (1 %) and EtO2 concentrations ( 6535 %) were manually controlled by the anaesthetist. In EtCA group (n = 40) the same anaesthetic machine with an additional end-tidal control feature was used to reach the same targets, rendering automatic the achievement and maintenance of those targets. Anaesthetic machine characteristics, amount of consumed gases, oxygen and sevoflurane efficiencies, and the amount of interventions by the anaesthetist were recorded. In EtCA group EtAA was achieved later (145 s) than in MCA (71 s) and remained controlled thereafter. Even though the target expired gas fractions were achieved faster in MCA, manual adjustments were required throughout anaesthesia for both oxygen and sevoflurane. In MCA patients the number of manual adjustments to stabilize EtAA and EtO2 were 137 and 107, respectively; no adjustment was required in EtCA. Low-flow anaesthesia delivered with an anaesthetic machine able to automatically control EtAA and EtO2 provided the same clinical stability and avoided the continuous manual adjustment of delivered sevoflurane and oxygen concentrations. Hence, the anaesthetist could dedicate more time to the patient and operating room activitie