scholarly journals Transcriptomic Analysis of The Poultry Red Mite, Dermanyssus Gallinae, Across All Stages of The Lifecycle.

Author(s):  
Kathryn Bartley ◽  
Wan Chen ◽  
Richard Lloyd Mills ◽  
Francesca Nunn ◽  
Daniel Price ◽  
...  

Abstract Background: The blood feeding poultry red mite (PRM), Dermanyssus gallinae, causes substantial economic damage to the egg laying industry worldwide, and is 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 was 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 (2,725), whilst the protonymph supercluster the fewest (165). Fifteen pairwise comparisons performed between the different stages resulting 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 metallacarboxypeptidases) 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 described for the house dust mites indicated a bias in expression towards the non-feeding larval stage.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 an invaluable tool for researchers investigating the biology and novel interventions of this parasite.

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Kathryn Bartley ◽  
Wan Chen ◽  
Richard I. Lloyd Mills ◽  
Francesca Nunn ◽  
Daniel R. G. Price ◽  
...  

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.


2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Francesca Nunn ◽  
Jessica Baganz ◽  
Kathryn Bartley ◽  
Sarah Hall ◽  
Stewart Burgess ◽  
...  

Abstract 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.


2021 ◽  
Author(s):  
Eleanor Karp-Tatham ◽  
Dong Xia ◽  
Alasdair J Nisbet ◽  
Teresa Letra Mateus ◽  
Fiona M. Tomley ◽  
...  

Abstract The poultry red mite (Dermanyssus gallinae), an obligatory blood feeding ectoparasite, is primarily associated with egg laying hens where it is estimated to cause losses of ~€230 million per annum from European farmers. Current control strategies, including the use of acaricidal chemicals and desiccant dusts, are often ineffective and there is widespread resistance to acaricides across Europe. Alternative methods to control D. gallinae are urgently required and strategies include development of recombinant subunit vaccines and discovery of new potential acaricides. These strategies will benefit hugely from knowledge of the extent and rates of occurrence of genetic diversity within D. gallinae populations. In this study, genetic diversity of mites harvested from the UK and from sites across mainland Europe was studied at inter- and intra-farm levels. To achieve this, the genome analysis toolkit (GATK) best practices pipeline for single nucleotide polymorphism (SNP) and insertion/deletion variant calling was modified to be self-validating and used to identify 32,599 D. gallinae SNPs by comparing transcriptomic sequences (derived from mites harvested in Germany, Schicht et al.) with a D. gallinae genome assembly (derived from mites harvested in Scotland, Burgess et al.). Dermanyssus gallinae populations were sampled from 22 UK farms and 57 farms from 15 countries in mainland Europe. Analysis of 144 high-quality SNP markers across 117 pooled D. gallinae samples showed high spatial genetic diversity with significant linkage disequilibrium. Revisiting a subset of farms revealed notable temporal changes in genetic diversity.


2020 ◽  
Author(s):  
Francesca G Nunn ◽  
Jessica Baganz ◽  
Kathryn Bartley ◽  
Sarah Hall ◽  
Stewart Burgess ◽  
...  

Abstract Background Dermanyssus gallinae , or poultry red mite (PRM), is an important ectoparasite in laying hens, having a significant welfare and economic impact. 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. geese), could serve as a refinement in the use of animals in research. Methods The existing in vitro feeding device, employing a Parafilm™ M membrane [1] and adult females was 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; Parafilm™ M, Baudruche membrane, and a combination of these with an overlaid polyester mesh were then tested using goose blood. Results PRM fed equally well on goose or hen blood through a Parafilm™ M membrane, no significant differences in mortality of PRM fed with either blood type was demonstrated. A significant increase (t-test: t =3.467, df =4, P = 0.03). in eggs laid per fed mite when using goose blood was demonstrated. A 70% increase in PRM feeding was observed when mites were fed on goose blood through a Baudruche membrane when compared to the Parafilm™ M membrane. Addition of an overlaid polyester mesh did not improve feeding rates. A significant increase (ANOVA: F (3, 20) =3.193, P = 0.04) in PRM egg laying was observed in mites fed on goose blood through Baudruche membrane, compared to those fed through Parafilm™ M. A mean of 1.22 (SEM ± 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 had a polyester mesh overlay. Conclusion The in vitro feeding of adult female poultry red mite can be readily facilitated through the use of goose blood in feeding devices with Baudruche membrane.


Parasitology ◽  
2020 ◽  
Vol 147 (7) ◽  
pp. 740-746
Author(s):  
Chuanwen Wang ◽  
Xiaolin Xu ◽  
He Yu ◽  
Yu Huang ◽  
Hao Li ◽  
...  

AbstractThe poultry red mite, Dermanyssus gallinae, is currently the most common ectoparasite affecting egg-laying hens. Since continuous culture of D. gallinae on birds is a biologically and economically costly endeavour, storage techniques for mites are urgently needed. Effects of temperature on adult and nymph survival were first studied to optimize storage conditions. Then, fecundity of D. gallinae was studied after mites were stored at optimal storage conditions. Results showed the survival rates of protonymphs (42.11%), deutonymphs (8.19%) and females (19.78%) at 5°C after 84 days were higher than those at 0, 25 and 30°C. Thereafter the fecundity and the capability of re-establishing colonies of D. gallinae were evaluated after they were stored for 40 and 80 days at 5°C. After storage, the mean number of eggs showed no statistical difference between treated (5°C for 40 or 80 days) and control groups (25°C for 7 days), while the hatching rates of eggs were in all cases above 97%. The dynamic changes of mite populations and egg numbers showed similar trends to the control group after the stored adult or nymph mites were fed on chicks. Dermanyssus gallinae can be successfully stored at 5°C for 80 days with no interference with the fecundity of mites, and the stored mites could re-establish colonies successfully. Adults and nymphs were two main stages with capability for low temperature storage. These results suggest that low temperature storage is a viable option for colony maintenance of D. gallinae under laboratory conditions.


2018 ◽  
Vol 9 (2) ◽  
pp. 249 ◽  
Author(s):  
Chengjie Zhang ◽  
Yanbing Zhu ◽  
Song Wang ◽  
Zheng Zachory Wei ◽  
Michael Qize Jiang ◽  
...  

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 1266
Author(s):  
Francesca Nunn ◽  
Kathryn Bartley ◽  
Javier Palarea-Albaladejo ◽  
Alasdair J. Nisbet

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.


2017 ◽  
Vol 10 (2) ◽  
pp. 53-62
Author(s):  
Slobodan Knežević ◽  
Marko Pajić ◽  
Aleksandra Petrović ◽  
Suzana Vidaković ◽  
Jelena Babić ◽  
...  

Dermanyssus gallinae or the poultry red mite is currently the most im-portant ectoparasite aff ecting egg-laying hens in several countries causing reduced poultry welfare, mortality and even allergic reactions in poultry farms workers. Its short life cycle, which in optimal conditions can be com-pleted within 7 days, and ability to survive in extreme circumstances with-out a blood meal up to 13 months, and the ability to infest new fl ock, makes it even more diffi cult to eradicate. Dermanyssus gallinae prevalence rates in diff erent European countries, including Serbia, can reach up to 80-90%. Also, the poultry red mite is responsible in vector transmission of several bacterial and viral avian diseases, including Salmonella spp, Chlamydia spp., Escherichia coli, Staphylococcus spp., Pasteurella multocida, Newcas-tle disease and Fowl poxvirus. Besides that, the poultry red mite can also transfer antimicrobial resistance genes by carrying pathogenic bacterial fl ora. Control of Dermanyssus gallinae can be divided into conventional and alternative methods. Conventional methods are mostly focused on pre-venting infestations and/or killing Dermanyssus gallinae, while alternative methods include the use of essential oils, vaccines, light, odors, predatory mites, fungi, nematodes and bacterial endosymbionts, and temperature in order to eliminate the poultry red mite. Nevertheless, this small ectopara-site still makes millions worth damage to global poultry industry.


2020 ◽  
Author(s):  
Ercha Hu ◽  
Yuan Meng ◽  
Ying Ma ◽  
Ruiqi Song ◽  
Zhengxiang Hu ◽  
...  

Abstract Background: The ixodid tick Dermacentor marginatus is a vector of many pathogens wide spread in Eurasia. Studies of gene sequence on many tick species have greatly increased the information on tick protective antigen which might have the potential to function as effective vaccine candidates or drug targets for eco-friendly acaricide development. In the current study, RNA-seq was applied to identify D. marginatus sequences and analyze differentially expressed unigenes.Methods: To obtain a broader picture of gene sequences and changes in expression level, RNA-seq was performed to obtain the whole-body transcriptome data of D. marginatus adult female ticks after engorgement and long-term starvation. Subsequently, the real-time quantitative PCR (RT-qPCR) was applied to validate the RNA-seq data.Results: RNA-seq produced 30,251 unigenes, of which 32% were annotated. Gene expression was compared among groups that differed by status as newly molted, starved and engorged female adult ticks. Nearly one third of the unigenes in each group were differentially expressed compared to the other two groups, and the most numerous were genes encoding proteins involved in catalytic and binding activities and apoptosis. Selected up-regulated differentially expressed genes in each group were associated to protein, lipids, carbohydrate and chitin metabolism. Blood-feeding and long-term starvation also caused genes differentially expressed in the defense response and antioxidant response. RT-qPCR results indicated 6 differentially expressed transcripts showed similar trends in expression changes with RNA-seq results confirming that the gene expression profiles in transcriptome data is in consistent with RT-qPCR validation.Conclusions: Obtaining the sequence information of D. marginatus and characterizing the expression pattern of the genes involved in blood-feeding and during starvation would be helpful in understanding molecular physiology of D. marginatus and provides data for anti-tick vaccine and drug development for controlling the tick.


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