Transcriptome dynamics of blood-fed and starved poultry red mites, Dermanyssus gallinae

Fujisawa, Sotaro; Shiro, Motoo; Isezaki, Masayoshi; Oishi, Eiji; Taneno, Akira; Maekawa, Naoya; Okagawa, Tomohiro; Konnai, Satoru; Ohashi, Kazuhiko

doi:10.1016/j.parint.2020.102156

Cited by 15 publications

(30 citation statements)

References 25 publications

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“…The candidate reference genes were extracted based on data obtained from RNA-seq analysis (Accession No. : SAMD00228960, SAMD00229086) [ 8 ]. We searched for contigs with the following properties: having high expression intensity without a significant difference between fed and starved PRMs, having being annotated using the InterProScan, and showing high identities with genes in other mite-species.…”

Section: Resultsmentioning

confidence: 99%

“…Candidate reference genes were identified based on data obtained from RNA-seq analysis (Accession No. SAMD00228960, SAMD00229086) [ 8 ]. Initially, we searched the contigs with >500 fragments per kilobase of exon per million reads mapped (FPKM) value in both fed and starved PRMs to extract genes with sufficient expression intensity.…”

Section: Methodsmentioning

confidence: 99%

“…Elucidation of the biological status of PRMs is valuable for developing alternative strategies for PRM control. We previously reported that blood feeding greatly affects the PRM transcriptome through RNA sequencing (RNA-seq) analysis [ 8 ]. However, for data validation, additional multi-sample analyses are desirable; furthermore, there is a need to establish a quantitative polymerase chain reaction (qPCR) method for quantifying the expression levels of PRM transcripts.…”

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confidence: 99%

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Selection of reference genes for quantitative PCR analysis in poultry red mite (<i>Dermanyssus gallinae</i>)

Ariizumi

Shiro

Fujisawa

et al. 2021

The Journal of Veterinary Medical Science

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Poultry red mites (PRMs, Dermanyssus gallinae ) are harmful ectoparasites that affect farmed chickens and cause serious economic losses in the poultry industry worldwide. Acaricides are used for PRM control; however, some PRMs have developed acaricide-resistant properties, which have indicated the need for different approaches for PRM control. Therefore, it is necessary to elucidate the biological status of PRMs to develop alternative PRM control strategies. Quantitative polymerase chain reaction (qPCR) allows analysis of the biological status at the transcript level. However, reference genes are preferable for accurate comparison of expression level changes given the large variation in the quality of the PRM samples collected in each farm. This study aimed to identify candidate reference genes with stable expression levels in the different blood feeding states and life stages of PRMs. First, we selected candidates based on the following criteria: sufficient expression intensity and no significant expression difference between fed and starved states. We selected and characterized seven candidate reference genes. Among them, we evaluated the gene expression stability between the starved and fed states using RefFinder; moreover, we compared their expression levels in each life-stage and identified two reference genes, Elongation factor 1-alpha ( ELF1A )-like and apolipophorins -like. Finally, we evaluated the utility of the candidates as reference genes, and the use of ELF1A -like and apolipophorins -like successfully normalized ATP synthase subunit g -like gene expression. Thus, ELF1A -like and apolipophorins -like could be suitable reference genes in PRMs.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Selection of reference genes for quantitative PCR analysis in poultry red mite (<i>Dermanyssus gallinae</i>)

Ariizumi

Shiro

Fujisawa

et al. 2021

The Journal of Veterinary Medical Science

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“…gene silencing by RNAi) have recently been developed and optimised in PRM [16,43,99] and some limited transcriptomic and bioinformatics studies on differential gene expression in PRM in different physiological states are starting to emerge (e.g. [3,30,41]).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2021

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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.

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“…gene silencing by RNAi) have recently been developed and optimised in PRM (Kamau et al, 2013;Wang et al, 2020, Chen et al, 2020 and some limited transcriptomic and bioinformatics studies on differential gene expression in PRM in different physiological states are starting to emerge (e.g. Bartley et al, 2012;Fujisawa et al, 2020;Huang et al, 2020).…”

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confidence: 99%

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

Bartley

Chen

Mills

et al. 2021

Preprint

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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.

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Transcriptome dynamics of blood-fed and starved poultry red mites, Dermanyssus gallinae

Cited by 15 publications

References 25 publications

Selection of reference genes for quantitative PCR analysis in poultry red mite (<i>Dermanyssus gallinae</i>)

Selection of reference genes for quantitative PCR analysis in poultry red mite (<i>Dermanyssus gallinae</i>)

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

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

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