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ABSTRACT: Dermanyssus gallinae is a major hematophagous ectoparasite in layer hens. Although the acaricide β-cypermethrin has been used to control mites worldwide, D. gallinae has developed resistance to this compound. Carboxylesterases (CarEs) are important detoxification enzymes that confer resistance to β-cypermethrin in arthropods. However, CarEs associated with β-cypermethrin resistance in D. gallinae have not yet been functionally characterized. Here, we isolated a CarE gene (Deg-CarE) from D. gallinae and assayed its activity. The results revealed significantly higher expression of Deg-CarE in the β-cypermethrin-resistant strain (RS) than in the susceptible strain (SS) toward α-naphthyl acetate (α-NA) and β-naphthyl acetate (β-NA). These findings suggest that enhanced esterase activities might have contributed to β-cypermethrin resistance in D. gallinae. Quantitative real-time PCR analysis revealed that Deg-CarE expression levels were significantly higher in adults than in other life stages. Although Deg-CarE was upregulated in the RS, significant differences in gene copy numbers were not observed. Additionally, Deg-CarE expression was significantly induced by β-cypermethrin in both the SS and RS. Moreover, silencing Deg-CarE via RNA interference decreased the enzyme activity and increased the susceptibility of the RS to β-cypermethrin, confirming that Deg-CarE is crucial for β-cypermethrin detoxification. Finally, recombinant Deg-CarE (rDeg-CarE) expressed in Escherichia coli displayed high enzymatic activity toward α/β-NA. However, metabolic analysis indicated that rDeg-CarE did not directly metabolize β-cypermethrin. The collective findings indicate that D. gallinae resistance to β-cypermethrin is associated with elevated CarEs protein activity and increased Deg-CarE expression levels. These findings provide insights into the metabolic resistance of D. gallinae and offer scientific guidance for the management and control of D. gallinae.

ABSTRACT: Poultry red mite (PRM), the ectoparasitic mite Dermanyssus gallinae found in laying hen farms, is a significant threat to poultry production and human health worldwide. It is a suspected disease vector and attacks hosts’ other than chickens, including humans, and its economic importance has increased greatly. Different strategies to control PRM have been widely tested and investigated. In principle, several synthetic pesticides have been applied to control PRM. However, recent alternative control methods to avoid the side effects of pesticides have been introduced, although many remain in the early stage of commercialization. In particular, advances in material science have made various materials more affordable as alternatives for controlling PRM through physical interactions between PRM. This review provides a summary of PRM infestation, and then includes a discussion and comparison of different conventional approaches: 1) organic substances, 2) biological approaches, and 3) physical inorganic material treatment. The advantages of inorganic materials are discussed in detail, including the classification of materials, as well as the physical mechanism-induced effect on PRM. In this review, we also consider the perspective of using several synthetic inorganic materials to suggest novel strategies for improved monitoring and better information regarding treatment interventions.

ABSTRACT: The poultry red mite (Dermanyssus gallinae, PRM) is a blood-sucking ectoparasite in chickens and is one of the most serious threats to poultry farms. Mass infestation with PRMs causes various health problems in chickens, resulting in significant productivity reduction in the poultry industry. Infestation with hematophagous ectoparasites, such as ticks, induces host inflammatory and hemostatic reactions. On the other hand, several studies have reported that hematophagous ectoparasites secrete various immunosuppressants from their saliva to suppress host immune responses to maintain blood sucking. Here, we examined the expression of cytokines in peripheral blood cells to investigate whether PRM infestation affects immunological states in chickens. In PRM-infested chickens, anti-inflammatory cytokines, IL-10 and TGF-β1, and immune checkpoint molecules, CTLA-4 and PD-1, were highly expressed compared to noninfested chickens. PRM-derived soluble mite extracts (SME) upregulated the gene expression of IL-10 in peripheral blood cells and HD-11 chicken macrophages. In addition, SME suppressed the expression of interferons and inflammatory cytokines in HD-11 chicken macrophages. Moreover, SME induces the polarization of macrophages into anti-inflammatory phenotypes. Collectively, PRM infestation could affect host immune responses, especially suppress the inflammatory responses. Further studies are warranted to fully understand the influence of PRM infestation on host immunity.

ABSTRACT: Dermanyssus gallinae is an obligatory ectoparasite of birds which feeds on blood and significantly compromise the well-being of commercially raised laying hens. In this study, the mortality rates and responses of D. gallinae to 2 acaricides with a physical mode of action (Dergall and Mite Max) and 2 acaricides with a chemical mode of action (Milben Ex and Bio PK) were evaluated in tree dilutions (S1–3) and compared at 8-time intervals after application. The evaluation involved a novel method that simulates real-world conditions in a commercial poultry farm. Tested products have shown high efficacy (84.3–100%) against D. gallinae in the producer recommended solution (S1). Acaricides with a physical mechanism of action were as effective as chemical agents in eradicating poultry red mites. The compared preparations differed only in the onset of action which was longer in acaricides with a physical mode of action (1–6 h for chemical 24 h for physical in S1). An increase in the concentration of the active ingredient did not significantly speed up the onset of action of the evaluated preparations. However, the efficacy of Dergall and Bio PK decreased when the applied dose was halved, to 12% and 0% respectively. A decrease in the dose Mite Max led to a somewhat smaller, but not statistically significant decrease in mite mortality rates (74%). The proposed method for evaluating acaricide efficacy can be helpful in selecting the most effective preparations and the optimal concentration of the working solution to be applied in commercial layer farms, thus reducing the costs associated with the eradication of D. gallinae. The developed method enables a reliable evaluation of acaricides with both a physical and chemical mode of action, and it supports observations of the parasites’ responses to the applied treatment.

Although it has rapidly decreased since the early 2000s, fowl typhoid still occurs in commercial layer chickens, causing a significant economic loss in Korea. There is growing concern about the emergence of new pathogenic strains of the causative agent, Salmonella Gallinarum, which is able to overcome vaccine immunity. It has also been suspected that the poultry red mite, Dermanyssus gallinae, which is commonly found in layer chicken farms, may be an important cause of the recurrence of fowl typhoid in the farms. This study was conducted to examine changes in the virulence of recent isolates of S. Gallinarum obtained from layer farms and estimate the potential of the disease transmission of D. gallinae in the farms. Clinical and environmental samples and mites collected from layer farms affected by fowl typhoid between 2013 and 2018 were tested for S. Gallinarum. The isolates were characterized by genotypic analyses and in vitro virulence assays with chicken-derived cell lines. Vaccine protection against recent isolates was examined in the chickens. A total of 45 isolates of S. Gallinarum were collected and there was no evidence of changes in their virulence. It has also been demonstrated that the S. Gallinarum 9R vaccine strain widely used in Korea is still effective in controlling fowl typhoid if the susceptibility of birds to the disease is not increased by stress. Salmonella Gallinarum isolated from the outer and inner parts of D. gallinae, environmental dust, and dead birds of the same farm showed the same or closely related genotypes. Consequently, the present study indicated that the horizontal transmission and environmental persistence of S. Gallinarum and the increased disease susceptibility of chickens in layer farms could be mediated by D. gallinae, causing persistent outbreaks of fowl typhoid.

ABSTRACT: Poultry red mites (PRMs, Dermanyssus gallinae) are hematophagous ectoparasites that negatively affect egg production, which causes serious economic losses to the poultry industry worldwide. Currently, the emergence of acaricide-resistant PRMs has impeded PRM control in poultry farms. Several alternatives for acaricide use have been described for managing PRM-caused problems. Vaccination is among the methods for controlling PRMs in poultry houses. Currently, several candidates for vaccine antigens have been identified. This study identified a cysteine protease, Deg-CPR-2, which differs from 2 other previously reported cysteine proteases in PRMs, from previously obtained data from RNA-sequencing (RNA-seq) analysis. We investigated the characteristics of Deg-CPR-2 and assessed its efficacy as a vaccine antigen in vitro. Phylogenetic analysis revealed that Deg-CPR-2 belonged to a different cluster from those of other cysteine proteases in PRMs. This cluster also included cathepsin L-like proteases, enzymes thought to be involved in hemoglobin digestion in ticks. Expression analysis revealed Deg-CPR-2 expression in midguts and all the life-stages; however, there were differences in the expression levels across the life-stages. The enzyme activity of recombinant Deg-CPR-2 was inhibited in the presence of a cysteine protease inhibitor, which suggests that Deg-CPR-2 functions as a cysteine protease in PRMs. Finally, there was an in vitro increase in the mortality of PRMs, mainly protonymphs that were artificially fed with plasma from chickens immunized with Deg-CPR-2. These findings suggest that Deg-CPR-2 may contribute to protein digestion in the midgut of PRMs and is crucially involved in physiological processes in PRMs. Additionally, immunization with Deg-CPR-2 may reduce the number of protonymphs, and Deg-CPR-2 should be considered as a candidate antigen for anti-PRM vaccine development.

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. [ABSTRACT FROM AUTHOR]

Dermanyssus gallinae (D.gallinae) infestation causes economic losses due to impaired health and production of hens and costs of parasite control across the world. Moreover, infestations are associated with reduced welfare of hens and may cause itching in humans. To effectively implement control methods it is crucially important to have high quality information about the D.gallinae populations in poultry houses in space and time. At present no validated tool is available to quantitatively monitor the dynamics of all four stages of D.gallinae (i.e., eggs, larvae, nymphs, and adults) in poultry houses. This article describes the experimental validation of the AVIVET trap, a device to quantitatively monitor dynamics of D.gallinae infestations. We used the device to study D.gallinae in fully equipped cages with two white specific pathogen free Leghorn laying hens experimentally exposed to three different infestation levels of D.gallinae (low to high). The AVIVET trap was successfully able to detect D.gallinae at high (5,000 D.gallinae), medium (2,500 D.gallinae), and low (50 D.gallinae) level of D.gallinae infestation. The linear equation Y = 10∧10∧(0.47 + 1.21X) with Y = log10 (Total number of D.gallinae nymphs and adults) in the cage and X = log10 (Total number of D.gallinae nymphs and adults) in the AVIVET trap explained 93.8% of the variation. The weight of D.gallinae in the AVIVET trap also appears to be a reliable parameter for quantifying D.gallinae infestation in a poultry house. The weight of D.gallinae in the AVIVET trap correlates 99.6% (P < 0.000) to the counted number of all stages of D.gallinae in the trap (i.e., eggs, larvae, nymphs, and adults) indicating that the trap is highly specific. From this experiment it can be concluded that the AVIVET trap is promising as quantitative tool for monitoring D.gallinae dynamics in a poultry house. [ABSTRACT FROM AUTHOR]