ABSTRACT
BACKGROUND: The blood feeding poultry red mite (PRM), Dermanyssus gallinae, causes substantial economic damage to the egg laying industry worldwide, and is a serious welfare concern for laying hens and poultry house workers. In this study we have investigated the temporal gene expression across the 6 stages/sexes (egg, larvae, protonymph and deutonymph, adult male and adult female) of this neglected parasite in order to understand the temporal expression associated with development, parasitic lifestyle, reproduction and allergen expression. RESULTS: RNA-seq transcript data for the 6 stages were mapped to the PRM genome creating a publicly available gene expression atlas (on the OrcAE platform in conjunction with the PRM genome). Network analysis and clustering of stage-enriched gene expression in PRM resulted in 17 superclusters with stage-specific or multi-stage expression profiles. The 6 stage specific superclusters were clearly demarked from each other and the adult female supercluster contained the most stage specific transcripts (2725), whilst the protonymph supercluster the fewest (165). Fifteen pairwise comparisons performed between the different stages resulted in a total of 6025 Differentially Expressed Genes (DEGs) (P > 0.99). These data were evaluated alongside a Venn/Euler analysis of the top 100 most abundant genes in each stage. An expanded set of cuticle proteins and enzymes (chitinase and metallocarboxypeptidases) were identified in larvae and underpin cuticle formation and ecdysis to the protonymph stage. Two mucin/peritrophic-A salivary proteins (DEGAL6771g00070, DEGAL6824g00220) were highly expressed in the blood-feeding stages, indicating peritrophic membrane formation during feeding. Reproduction-associated vitellogenins were the most abundant transcripts in adult females whilst, in adult males, an expanded set of serine and cysteine proteinases and an epididymal protein (DEGAL6668g00010) were highly abundant. Assessment of the expression patterns of putative homologues of 32 allergen groups from house dust mites indicated a bias in their expression towards the non-feeding larval stage of PRM. CONCLUSIONS: This study is the first evaluation of temporal gene expression across all stages of PRM and has provided insight into developmental, feeding, reproduction and survival strategies employed by this mite. The publicly available PRM resource on OrcAE offers a valuable tool for researchers investigating the biology and novel interventions of this parasite.
Subject(s)
Mite Infestations , Mites , Poultry Diseases , Animals , Chickens , Female , Humans , Male , Mites/genetics , Poultry , TranscriptomeABSTRACT
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.
Subject(s)
Allergens/genetics , Gene Expression Regulation, Developmental , Psoroptidae/genetics , Animals , Cluster Analysis , Feeding Behavior , Female , Gene Expression Profiling , Genomics , Life Cycle Stages/genetics , Male , Multigene Family , Phylogeny , Psoroptidae/classification , Psoroptidae/growth & development , Psoroptidae/physiology , Sex Factors , Sheep/parasitologyABSTRACT
Dermanyssus gallinae, also known as the poultry red mite (PRM), is a blood-feeding ectoparasite of poultry and sylvatic birds. This mite is endemic in many parts of the globe and poses a threat to the egg industry, while compromising the health and welfare of hens, both directly and as a vector of diseases. In addition, people attacked by D. gallinae may develop gamasoidosis. Despite the high prevalence in several European countries, epidemiological information on D. gallinae in Portugal is scarce. This study aimed to assess the prevalence and infestation levels in laying farms in Portugal and evaluate the perception and attitudes of producers regarding D. gallinae. A survey was performed between August 2016 - November 2017, which included 24 farms in the NUTS2 regions Centro and Norte. Mites were sampled with corrugated cardboard traps and the perception and attitudes of farmers regarding the PRM were evaluated with the European COREMI questionnaire prepared by WG 1 of the COST action FA1404. D. gallinae was detected in 95.8% of farms (95% CI: 79.8-99.3%). The average number of trapped mites among farms was 5200 ± 16,522, with a median of 359 mites (interquartile range = 46-3135). Results from the questionnaire show that insufficient monitoring, under-detection and late and suboptimal treatment may contribute to the maintenance of significant infestation levels. The present data highlight the need for adequate monitoring of D. gallinae, timely action and effective treatment in order to improve poultry productivity and ensure human and animal health and welfare. RESEARCH HIGHLIGHTS A survey on the prevalence of D. gallinae in Portuguese layer farms is presented The perceived importance of D. gallinae was assessed with a questionnaire D. gallinae was detected in 95.8% of farms The results emphasize the need for adequate monitoring and treatment optimization.
Subject(s)
Chickens/parasitology , Mite Infestations/veterinary , Mites/physiology , Poultry Diseases/epidemiology , Animals , Farms , Female , Mite Infestations/epidemiology , Mite Infestations/parasitology , Portugal/epidemiology , Poultry Diseases/parasitology , PrevalenceABSTRACT
Dermanyssus gallinae, the poultry red mite, is a global threat to the commercial egg-laying industry. Control of D. gallinae is difficult, with only a limited number of effective pesticides and non-chemical treatments available. Here, we characterize the candidate vaccine antigen D. gallinae cathepsin D-1 (Dg-CatD-1) and demonstrate that purified refolded recombinant Dg-Cat-D1 (rDg-CatD-1) is an active aspartyl proteinase which digests haemoglobin with a pH optimum of pH 4. Soluble protein extracts from D. gallinae also have haemoglobinase activity, with a pH optimum comparable to the recombinant protein, and both proteinase activities were inhibited by the aspartyl proteinase inhibitor Pepstatin A. Enzyme activity and the ubiquitous localization of Dg-CatD-1 protein in sections of adult female mites is consistent with Dg-CatD-1 being a lysosomal proteinase. Using Dg-CatD-1 as a model vaccine antigen, we compared vaccine delivery methods in laying hens via vaccination with: (i) purified rDg-CatD-1 with Montanide™ ISA 71 VG adjuvant; (ii) recombinant DNA vaccines for expression of rDg-CatD-1 and (iii) transgenic coccidial parasite Eimeria tenella expressing rDg-CatD-1. In two independent trials, only birds vaccinated with rDg-CatD-1 with Montanide™ ISA 71 VG produced a strong and long-lasting serum anti-rDg-Cat-D1 IgY response, which was significantly higher than that in control birds vaccinated with adjuvant only. Furthermore, we showed that egg-laying rates of D. gallinae mites fed on birds vaccinated with rDg-CatD-1 in Montanide™ ISA 71 VG was reduced significantly compared with mites fed on unvaccinated birds. RESEARCH HIGHLIGHTS Dermanyssus gallinae cathepsin D-1 (Dg-CatD-1) digests haemoglobin Vaccination of hens with rDg-CatD-1 in Montanide™ ISA 71 VG results in long-lasting IgY levels Serum anti-rDg-CatD-1 antibodies reduce egg laying in D. gallinae after a single blood meal.
Subject(s)
Chickens/immunology , Mite Infestations/veterinary , Mites/immunology , Poultry Diseases/prevention & control , Vaccination/veterinary , Vaccines/administration & dosage , Adjuvants, Immunologic , Animals , Antibody Formation , Chickens/parasitology , Female , Mite Infestations/parasitology , Mite Infestations/prevention & control , Recombinant ProteinsABSTRACT
The gammaherpesvirus alcelaphine herpesvirus 1 (AlHV-1) causes fatal malignant catarrhal fever (MCF) in susceptible species including cattle, but infects its reservoir host, wildebeest, without causing disease. Pathology in cattle may be influenced by virus-host cell interactions mediated by the virus glycoproteins. Cloning and expression of a haemagglutinin-tagged version of the AlHV-1 glycoprotein B (gB) was used to demonstrate that the AlHV-1-specific monoclonal antibody 12B5 recognised gB and that gB was the main component of the gp115 complex of AlHV-1, a glycoprotein complex of five components identified on the surface of AlHV-1 by immunoprecipitation and radiolabelling. Analysis of AlHV-1 virus particles showed that the native form of gB was detected by mAb 12B5 as a band of about 70 kDa, whilst recombinant gB expressed by transfected HEK293T cells appeared to be subject to additional cleavage and incomplete post-translational processing. Antibody 12B5 recognised an epitope on the N-terminal furin-cleaved fragment of gB on AlHV-1 virus particles. It could be used to detect recombinant and virus-expressed gB on western blots and on the surface of infected cells by flow cytometry, whilst recombinant gB was detected on the surface of transfected cells by immunofluorescence. Recombinant gB has potential as an antigen for ELISA detection of MCF virus infection and as a candidate vaccine antigen.
Subject(s)
Antibodies, Viral/immunology , Cattle Diseases/diagnosis , Gammaherpesvirinae/immunology , Glycoproteins/immunology , Malignant Catarrh/diagnosis , Viral Structural Proteins/immunology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/isolation & purification , Antibodies, Viral/isolation & purification , Cattle , Gammaherpesvirinae/chemistry , Glycoproteins/analysis , Immunoprecipitation , Membrane Proteins/analysis , Membrane Proteins/immunology , Radioimmunoassay , Viral Structural Proteins/analysis , Virion/chemistryABSTRACT
We wished to determine the effect of of CpG ODN adjuvant on the magnitude and duration of protective immunity against alcelaphine herpesvirus-1 (AlHV-1) malignant catarrhal fever (MCF), a fatal lymphoproliferative disease of cattle. Immunity was associated with a mucosal barrier of virus-neutralising antibody. The results showed that CpG ODN included either with emulsigen adjuvant and attenuated AlHV-1 (atAlHV-1) or alone with atAlHV-1 did not affect the overall protection from clinical disease or duration of immunity achieved using emulsigen and atAlHV-1. This is in contrast to other similar studies in cattle with BoHV-1 or cattle and pigs with various other immunogens. In addition to this, several other novel observations were made, not reported previously. Firstly, we were able to statistically verify that vaccine protection against MCF was associated with virus-neutralising antibodies (nAbs) in nasal secretions but was not associated with antibodies in blood plasma, nor with total virus-specific antibody (tAb) titres in either nasal secretions or blood plasma. Furthermore, CpG ODN alone as adjuvant did not support the generation of virus-neutralising antibodies. Secondly, there was a significant boost in tAb in animals with MCF comparing titres before and after challenge. This was not seen with protected animals. Finally, there was a strong IFN-γ response in animals with emulsigen and atAlHV-1 immunisation, as measured by IFN-γ secreting PBMC in culture (and a lack of IL-4) that was not affected by the inclusion of CpG ODN. This suggests that nAbs at the oro-nasal-pharyngeal region are important in protection against AlHV-1 MCF.
Subject(s)
Adjuvants, Immunologic/administration & dosage , Cattle Diseases/immunology , Gammaherpesvirinae/immunology , Herpesviridae Infections/veterinary , Malignant Catarrh/immunology , Oligodeoxyribonucleotides/administration & dosage , Viral Vaccines/immunology , Animals , Antibodies, Viral/analysis , Antibodies, Viral/blood , Cattle , Cattle Diseases/virology , Enzyme-Linked Immunosorbent Assay/veterinary , Gammaherpesvirinae/physiology , Herpesviridae Infections/immunology , Herpesviridae Infections/virology , Immunity, Active/drug effects , Male , Malignant Catarrh/virology , Methylation , Nose/virology , Oligodeoxyribonucleotides/chemistry , Toll-Like Receptor 9/agonists , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/immunology , Viral Vaccines/administration & dosageABSTRACT
Vaccination is a feasible strategy for controlling the haematophagous poultry red mite Dermanyssus gallinae. A cDNA library enriched for genes upregulated after feeding was created to identify potential vaccine antigens. From this library, a gene (Dg-CatD-1) encoding a 383 amino acid protein (Dg-CatD-1) with homology to cathepsin D lysosomal aspartyl proteinases was identified as a potential vaccine candidate. A second gene (Dg-CatL-1) encoding a 341 amino acid protein (Dg-CatL-1) with homology to cathepsin L cysteine proteinases was also selected for further study. IgY obtained from naturally infested hens failed to detect Dg-CatD-1 suggesting that it is a concealed antigen. Conversely, Dg-CatL-1 was detected by IgY derived from natural-infestation, indicating that infested hens are exposed to Dg-CatL-1. Mortality rates 120 h after mites had been fed anti-Dg-CatD-1 were significantly higher than those fed control IgY (PF<0·01). In a survival analysis, fitting a proportional hazards model to the time of death of mites, anti-Dg-CatD-1 and anti-Dg-CatL-1 IgY had 4·42 and 2·13 times higher risks of dying compared with controls (PF<0·05). Dg-CatD-1 and L-1 both have potential as vaccine antigens as part of a multi-component vaccine and have the potential to be improved as vaccine antigens using alternative expression systems.
Subject(s)
Cathepsin D/immunology , Cathepsin L/immunology , Mite Infestations/veterinary , Mites/enzymology , Poultry Diseases/prevention & control , Vaccines/immunology , Amino Acid Sequence , Animals , Antibodies/blood , Antigens/genetics , Antigens/immunology , Cathepsin D/genetics , Cathepsin L/genetics , Chickens/parasitology , Female , Mite Infestations/immunology , Mite Infestations/parasitology , Mite Infestations/prevention & control , Mites/immunology , Molecular Sequence Data , Poultry Diseases/immunology , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Sequence Analysis, DNAABSTRACT
Introduction: Sheep scab caused by Psoroptes ovis, is a disease of concern to many stakeholders in Wales due to its welfare implications. There are good diagnostic tests and treatments available to deal with the disease. Even so, it remains a problem in Welsh flocks. As such a coordinated approach is required to deal with this issue in a more sustainable manner. Pilot design: Sheep scab positive 'index' farms were initially diagnosed using a skin scrape to identify P. ovis mites. Contiguous farms were identified and antibody responses used to confirm onward infestation. All infested farms were treated by either dipping with an organophosphate (OP) dip or injecting with a licensed macrocyclic lactone (ML) product depending on farmer choice. Results: Three positive 'index' farms were identified along with 12 contiguous properties. Positive serological responses were observed in seven of the 12 contiguous farms; four of which were treated by OP dip and three by an injectable ML product. Discussion: To avoid reinfestation of treated farms, dealing with disease on contiguous properties is crucial. Through the project coordinating team, three local outbreaks of scab were dealt with in a short space of time with appropriate diagnosis and treatment being carried out. Some farmers were uncooperative and strategies such as providing additional external support and veterinary involvement might alleviate these issues in the future. This coordinated approach is recommended to veterinary surgeons in the field when dealing with scab on farm.
ABSTRACT
The minor capsid protein of ovine herpesvirus 2, identified as a potential antigen for serological testing, was over-expressed and purified to allow its assessment in ELISA. The corresponding gene sequence (OvHV-2 orf65, Ov65) was modified to incorporate epitope tags and internal restriction enzyme sites in an E. coli codon-optimised version of the gene. This codon-optimised gene was then subject to internal deletions to identify regions of the protein that could be removed while maintaining protein solubility and antigenicity. It was found that a derivative with deletion of the conserved 5'-end of the gene (Ov65delB) expressed a polypeptide that was soluble when over-expressed in bacteria and was detected by OvHV-2 specific sera. Proteomic analysis of the affinity purified Ov65delB showed that it contained multiple predicted Ov65 tryptic peptides but also showed contamination by co-purifying E. coli proteins. An indirect ELISA, based on this affinity-purified OV65delB, was optimised for use with sheep and cattle samples and cut-off values were established based on known negative serum samples. Analysis of groups of samples that were either presumed infected (UK sheep) or tested OvHV-2 positive or negative by PCR (cattle MCF diagnostic samples) showed that the assay had 95 % sensitivity and 96 % specificity for sheep serum; and 80 % sensitivity and 95 % specificity for cattle serum. The lower sensitivity with cattle samples appeared to be due to a lack of serological response in some MCF-affected cattle. This recombinant antigen therefore shows promise as the basis of an inexpensive, simple and reliable test that can be used to detect OvHV-2-specific antibody responses in both MCF-affected animals and in OvHV-2 reservoir hosts.
Subject(s)
Malignant Catarrh , Sheep Diseases , Animals , Cattle , Enzyme-Linked Immunosorbent Assay/veterinary , Escherichia coli/genetics , Gammaherpesvirinae , Malignant Catarrh/diagnosis , Proteomics , Sheep , Sheep Diseases/diagnosisABSTRACT
Many obligate blood-sucking arthropods rely on symbiotic bacteria to provision essential B vitamins that are either missing or at sub-optimal levels in their nutritionally challenging blood diet. The poultry red mite Dermanyssus gallinae, an obligate blood-feeding ectoparasite, is a serious threat to the hen egg industry. Poultry red mite infestation has a major impact on hen health and welfare and causes a significant reduction in both egg quality and production. Thus far, the identity and biological role of nutrient provisioning bacterial mutualists from D. gallinae are little understood. Here, we demonstrate that an obligate intracellular bacterium of the Rickettsiella genus is detected in D. gallinae mites collected from 63 sites (from 15 countries) across Europe. In addition, we report the genome sequence of Rickettsiella from D. gallinae (Rickettsiella - D. gallinae endosymbiont; Rickettsiella DGE). Rickettsiella DGE has a circular 1.89Mbp genome that encodes 1,973 proteins. Phylogenetic analysis confirms the placement of Rickettsiella DGE within the Rickettsiella genus, related to a facultative endosymbiont from the pea aphid and Coxiella-like endosymbionts (CLEs) from blood feeding ticks. Analysis of the Rickettsiella DGE genome reveals that many protein-coding sequences are either pseudogenized or lost, but Rickettsiella DGE has retained several B vitamin biosynthesis pathways, suggesting the importance of these pathways in evolution of a nutritional symbiosis with D. gallinae. In silico metabolic pathway reconstruction revealed that Rickettsiella DGE is unable to synthesize protein amino acids and, therefore, amino acids are potentially provisioned by the host. In contrast, Rickettsiella DGE retains biosynthetic pathways for B vitamins: thiamine (vitamin B1) via the salvage pathway; riboflavin (vitamin B2) and pyridoxine (vitamin B6) and the cofactors: flavin adenine dinucleotide (FAD) and coenzyme A (CoA) that likely provision these nutrients to the host.
ABSTRACT
BACKGROUND: The avian haematophagous ectoparasite Dermanyssus gallinae, commonly known as the poultry red mite, causes significant economic losses to the egg-laying industry worldwide and also represents a significant welfare threat. Current acaricide-based controls are unsustainable due to the mite's ability to rapidly develop resistance, thus developing a novel sustainable means of control for D. gallinae is a priority. RNA interference (RNAi)-mediated gene silencing is a valuable tool for studying gene function in non-model organisms, but is also emerging as a novel tool for parasite control. METHODS: Here we use an in silico approach to identify core RNAi pathway genes in the recently sequenced D. gallinae genome. In addition we utilise an in vitro feeding device to deliver double-stranded (ds) RNA to D. gallinae targeting the D. gallinae vATPase subunit A (Dg vATPase A) gene and monitor gene knockdown using quantitative PCR (qPCR). RESULTS: Core components of the small interfering RNA (siRNA) and microRNA (miRNA) pathways were identified in D. gallinae, which indicates that these gene silencing pathways are likely functional. Strikingly, the P-element-induced wimpy testis (PIWI)-interacting RNA (piRNA) pathway was absent in D. gallinae. In addition, feeding Dg vATPase A dsRNA to adult female D. gallinae resulted in silencing of the targeted gene compared to control mites fed non-specific lacZ dsRNA. In D. gallinae, dsRNA-mediated gene knockdown was rapid, being detectable 24 h after oral delivery of the dsRNA, and persisted for at least 120 h. CONCLUSIONS: This study shows the presence of core RNAi machinery components in the D. gallinae genome. In addition, we have developed a robust RNAi methodology for targeting genes in D. gallinae that will be of value for studying genes of unknown function and validating potential control targets in D. gallinae.
Subject(s)
Arthropod Proteins/genetics , Gene Knockdown Techniques/methods , Genomics/methods , Mite Infestations/veterinary , Mites/genetics , Poultry/parasitology , RNA Interference , Adenosine Triphosphatases/genetics , Animals , Chickens/parasitology , Computer Simulation , Female , MicroRNAs/genetics , MicroRNAs/metabolism , Mite Infestations/parasitology , Mite Infestations/prevention & control , Poultry Diseases/parasitologyABSTRACT
A study was performed to examine any effect of hen age on the feeding ability and mortality of different life-stages of Dermanyssus gallinae [Poultry Red Mite (PRM)] when fed using a high welfare, on-hen mite feeding device. Mite feeding assays were carried out every two weeks on a cohort of five Lohman Brown hens with devices containing adult and deutonymph PRM or adult and protonymph PRM. Feeding rates and mortality of each PRM life stage and oviposition of adult female PRM were evaluated over an 18-week period. There was a significant reduction in oviposition rates of female PRM as they fed on hens of increasing age. However, no clear trend was detected between the feeding rates of all three haematophagous life stages and hen age. The same conclusion was reached regarding mite mortality post-feeding in both deutonymph and adult female PRMs, although a weak positive association was apparent between hen age and protonymph PRM mortality. This study shows that the on-hen feeding device can be used both for short term studies to assess novel anti-PRM products (new acaricides, vaccines etc.) and longer, longitudinal studies to determine longevity of the effects of such novel anti-PRM products. It also demonstrates that blood feeding by mites on older hens is less able to sustain PRM populations than feeding on younger hens. This on-hen mite feeding device directly impacts upon reduction and refinement by greatly reducing the numbers of birds required per experimental group compared to traditional PRM challenge infestation models and by eliminating the need for birds to be exposed to large numbers of mites for extended periods of time that can cause welfare concerns. This paper describes the methodology for these studies and how to assemble pouches and handle mites both before and after feeding assays.
Subject(s)
Mites , Poultry Diseases , Aging , Animals , Chickens , Female , Oviposition , PoultryABSTRACT
BACKGROUND: Dermanyssus gallinae, or poultry red mite (PRM), is an important ectoparasite in laying hen, having a significant effect on animal welfare and potentially causing economic loss. Testing novel control compounds typically involves in vitro methodologies before in vivo assessments. Historically, in vitro methods have involved PRM feeding on hen blood through a membrane. The use of hen blood requires multiple procedures (bleeds) to provide sufficient material, and the use of a larger species (e.g. goose) could serve as a refinement in the use of animals in research. METHODS: The in vitro feeding device used was that which currently employs a Parafilm™ M membrane (Bartley et al.: Int J Parasitol. 45:819-830, 2015). Adult female PMR were used to investigate any differences in mite feeding, egg laying and mortality when fed goose or hen blood. Effects on these parameters when PRM were fed through either the Parafilm™ M membrane or the Baudruche membrane alone or through a combination of the membrane with an overlaid polyester mesh were tested using goose blood. RESULTS: Poultry red mites fed equally well on goose or hen blood through the Parafilm™ M membrane, and there were no significant differences in mortality of PRM fed with either blood type. A significant increase (t test: t = 3.467, df = 4, P = 0.03) in the number of eggs laid per fed mite was observed when goose blood was used. A 70% increase in PRM feeding was observed when the mites were fed on goose blood through a Baudruche membrane compared to when they were fed goose blood through the Parafilm™ M membrane. The addition of an overlaid polyester mesh did not improve feeding rates. A significant increase (analysis of variance: F(3, 20) = 3.193, P = 0.04) in PRM egg laying was observed in mites fed on goose blood through the Baudruche membrane compared to those fed goose blood through the Parafilm™ M membrane. A mean of 1.22 (standard error of the mean ± 0.04) eggs per fed mite was obtained using the Baudruche feeding device compared to only 0.87 (SEM ± 0.3) eggs per fed mite using the Parafilm™ M device when neither was combined with a polyester mesh overlay. CONCLUSION: The in vitro feeding of adult female PRM can be readily facilitated through the use of goose blood in feeding devices with the Baudruche membrane.
Subject(s)
Membranes, Artificial , Mites/physiology , Skin/parasitology , Animals , Blood , Chickens/parasitology , Feeding Behavior , Female , Geese , Mite Infestations/parasitology , Paraffin , Poultry Diseases/parasitologyABSTRACT
Dermanyssus gallinae (De Geer), the poultry red mite, is a blood-feeding ectoparasite that infests many bird species. We have used an in vitro feeding assay to allow the identification of protective D. gallinae antigens that may have potential as vaccine candidates. Homogenised mites were extracted sequentially with PBS, Tween 20, Triton X100 and urea giving four protein fractions. Five experimental groups of Lohmann Brown hens were used to generate antibodies; four groups were injected with one of each of the protein fractions in QuilA adjuvant and a control group was injected with adjuvant only. Booster injections were administered 2 and 4 weeks after initial immunisation. Eggs were collected throughout the experiment and soluble IgY antibodies were extracted from a pool of egg yolks collected at week six post-injection. Western blots, performed using post vaccination antibodies from test and control groups, revealed a strong antibody response against a range of injected proteins. Fresh chicken blood, supplemented with antibodies raised against these protein fractions, was fed to mites in an in vitro feeding assay in order to determine whether the antibodies had an anti-mite effect. Although there was variability in the numbers of feeding mites, it was found that the strongest anti-mite effect was seen with the PBS protein fraction, which had a cumulative average mortality of 34.8% 14 days after feeding compared with 27.3% for the control group (P = 0.043).
Subject(s)
Antibodies/pharmacology , Bird Diseases/prevention & control , Chickens/parasitology , Mite Infestations/veterinary , Mites/immunology , Vaccines/immunology , Animals , Antibody Formation , Antigens , Chemical Fractionation , Chickens/blood , Chickens/immunology , Mite Infestations/prevention & control , Mites/drug effects , Mites/physiologyABSTRACT
The poultry red mite (PRM) is one of the most economically important ectoparasites of laying hens globally. This mite can have significant deleterious effects on its fowl host including distress, anemia, reduced egg production, and reduced egg quality. This study was conducted to evaluate the influence of PRM on the serum protein profile in laying hens and its effect on the acute phase proteins (APPs) to assess their potential as biomarkers for mite infestation. Three APPs: alpha-1 acid glycoprotein (AGP), serum amyloid-A (SAA), and ceruloplasmin (CP) were measured in serum samples collected from laying hens at 12 and 17 wk of age, and then for up to 4 mo after a challenge with PRM (starting at 18.5 wk of age). The serum protein profile (SDS-PAGE/nanoflow HPLC electrospray tandem mass spectrometry) and concentration of individual serum proteins (SDS-PAGE-band densitometry) were also compared. Post challenge there was a positive correlation (r = 0.489; P < 0.004) between the levels of SAA and the PRM numbers. The levels of SAA steadily increased after the PRM challenge and were significantly different than the pre-challenge levels at 28, 32, and 36 wk of age (P < 0.01). The PRM numbers also peaked around 31-33 wk of age. The results for AGP and CP in comparison were inconsistent. Proteomics revealed the presence of 2 high molecular weight proteins in the serum between 12 and 17 wk of age. These were identified as Apolipoprotein-B and Vitellogenin-2, and their increase was commensurate with the onset of lay. No other major differences were detected in the protein profiles of blood sera collected pre and post challenge. We conclude that SAA could be used as a useful biomarker to monitor PRM infestation in commercial poultry flocks and that PRM infestation does not disrupt the production of the major proteins in the serum that are associated with egg formation.
Subject(s)
Blood Proteins/metabolism , Chickens , Mite Infestations/veterinary , Poultry Diseases/parasitology , Proteome/metabolism , Acute-Phase Proteins/metabolism , Animals , Avian Proteins , Female , Mite Infestations/metabolism , Mite Infestations/parasitology , Mites/physiology , Poultry Diseases/metabolism , ReproductionABSTRACT
Optimisation and use of a device for the on-hen in vivo feeding of all hematophagous stages of Dermanyssus gallinae is described. The sealed mesh device contains the mites and is applied to the skin of the hen's thigh where mites can feed on the bird through a mesh which has apertures large enough to allow the mites' mouth-parts to access to the bird but small enough to contain the mites. By optimising the depth and width of the mesh aperture size we have produced a device which will lead to both reduction and refinement in the use of animals in research, allowing the pre-screening of new vaccines and systemic acaricides/insecticides which have been developed for the control of these blood-feeding parasites before progressing to large field trials. For optimal use, the device should be constructed from 105 µm aperture width, 63 µm depth, polyester mesh and the mites (irrespective of life stage) should be conditioned with no access to food for 3 weeks at 4 °C for optimal feeding and post-feeding survival.
Subject(s)
Animal Welfare , Mite Infestations/prevention & control , Mite Infestations/veterinary , Poultry Diseases/parasitology , Poultry/parasitology , Animal Experimentation , Animal Feed , Animals , Feeding Methods/instrumentation , Mites/physiologyABSTRACT
The poultry red mite (PRM), Dermanyssus gallinae, is a hematophagous ectoparasite of birds with worldwide distribution that causes economic losses in the egg-production sector of the poultry industry. Traditional control methods, mainly based on acaricides, have been only partially successful, and new vaccine-based interventions are required for the control of PRM. Vaccination with insect Akirin (AKR) and its homolog in ticks, Subolesin (SUB), have shown protective efficacy for the control of ectoparasite infestations and pathogen infection/transmission. The aim of this study was the identification of the akr gene from D. gallinae (Deg-akr), the production of the recombinant Deg-AKR protein, and evaluation of its efficacy as a vaccine candidate for the control of PRM. The anti-Deg-AKR serum IgY antibodies in hen sera and egg yolk were higher in vaccinated than control animals throughout the experiment. The results demonstrated the efficacy of the vaccination with Deg-AKR for the control of PRM by reducing mite oviposition by 42% following feeding on vaccinated hens. A negative correlation between the levels of serum anti-Deg-AKR IgY and mite oviposition was obtained. These results support Deg-AKR as a candidate protective antigen for the control of PRM population growth.
ABSTRACT
The poultry red mite (PRM), Dermanyssus gallinae, is a hematophagous ectoparasite considered as the major pest in the egg-laying industry. Its pesticide-based control is only partially successful and requires the development of new control interventions such as vaccines. In this study, we follow a vaccinology approach to identify PRM candidate protective antigens. Based on proteomic data from fed and unfed nymph and adult mites, we selected a novel PRM protein, calumenin (Deg-CALU), which is tested as a vaccine candidate on an on-hen trial. Rhipicephalus microplus Subolesin (Rhm-SUB) was chosen as a positive control. Deg-CALU and Rhm-SUB reduced the mite oviposition by 35 and 44%, respectively. These results support Deg-CALU and Rhm-SUB as candidate protective antigens for the PRM control.
ABSTRACT
Sheep scab, caused by infestation with Psoroptes ovis, is highly contagious, results in intense pruritus, and represents a major welfare and economic concern. Here, we report the first draft genome assembly and gene prediction of P. ovis based on PacBio de novo sequencing. The â¼63.2-Mb genome encodes 12,041 protein-coding genes.
ABSTRACT
The poultry red mite, Dermanyssus gallinae, is a major worldwide concern in the egg-laying industry. Here, we report the first draft genome assembly and gene prediction of Dermanyssus gallinae, based on combined PacBio and MinION long-read de novo sequencing. The â¼959-Mb genome is predicted to encode 14,608 protein-coding genes.