The Poultry Red Mite, Dermanyssus gallinae, travels far but not frequently, and takes up permanent residence on farms.

Management of Dermanyssus gallinae, a cosmopolitan hematophagous mite responsible for damage in layer poultry farming, is hampered by a lack of knowledge of its spatio-temporal population dynamics. Previous studies have shown that the circulation of this pest between farms is of strictly anthropogenic origin, that a mitochondrial haplogroup has been expanding on European farms since the beginning of the 21st century and that its local population growth may be particularly rapid. To refine our understanding of how D. gallinae spreads within and among farms, we characterized the genetic structure of mite populations at different spatial scales and sought to identify the main factors interrupting gene flow between poultry houses and between mitochondrial haplogroups. To this end, we selected and validated the first set of nuclear microsatellite markers for D. gallinae and sequenced a region of the CO1-encoding mitochondrial gene in a subsample of microsatellite-genotyped mites. We also tested certain conditions required for effective contamination of a poultry house through field experimentation, and conducted a survey of practices during poultry transfers. Our results confirm the role of poultry transport in the dissemination of mite populations, but the frequency of effective contamination after the introduction of contaminated material into poultry houses seems lower than expected. The high persistence of mites on farms, even during periods when poultry houses are empty and cleaned, and the very large number of nodes in the logistic network (large number of companies supplying pullets or transporting animals) undoubtedly explain the very high prevalence on farms. Substantial genetic diversity was measured in farm populations, probably as a result of the mite's known haplodiploid mode of sexual reproduction, coupled with the dense logistic network. The possibility of the occasional occurrence of asexual reproduction in this sexually reproducing mite was also revealed in our analyses, which could explain the extreme aggressiveness of its demographic dynamics under certain conditions.

Nanoemulsion-based plant essential oil formulations: in vitro evaluation of pesticidal activity against ectoparasites in poultry.

Ectoparasite infestations significantly impact the health and productivity of poultry. Chemical applications, although common for pest control, lead to pesticide residues and parasite resistance in poultry. Nanoemulsion-based plant essential oil formulations (NEOFs) provide a promising alternative for controlling poultry ectoparasites. This study aimed to assess the efficacy of NEOFs from clove, cinnamon, and turmeric essential oils (EOs) against ectoparasites, Menopon gallinae and Megninia ginglymura, under laboratory conditions. The toxicity and repellent properties of the NEOFs were examined, with the major chemical compounds of the EOs analyzed using chromatography mass spectrometer. Results identified eugenol as the dominant component in clove and cinnamon EOs (84.60 and 75.19%, respectively), while turmerone (68.46%) was the major compound in turmeric EO. NEOFs with clove:cinnamon:turmeric ratios of 4:0:0, 2:2:0, and 2:0:2 had particle size of 20.76 nm, 20.66 nm, and 89.56 nm, respectively, while those based on eugenol and turmerone standards had sizes <21.0 nm. In addition, NEOFs at 0.3% concentration with ratios of 4:0:0 and 2:2:0 achieved full control of both ectoparasites. These formulas demonstrated exceptional potency in exterminating ectoparasites, with LC50 and LC90 at <0.160 and <0.250%, respectively, 6 h after treatments. Furthermore, both NEOFs showed higher repellence responses in M. gallinae compared to M. ginglymura. The toxicities of these NEOFs were comparably effective against both parasites, showing no significant difference compared with chemical insecticide treatment. Therefore, further research will explore the practicality of using clove and cinnamon-derived NEOFs under farm conditions.

Attractant Activity of Host-Related Chemical Blends on the Poultry Red Mite at Different Spatial Scales.

Many blood-feeding arthropods use volatile organic compounds (VOCs) to detect their vertebrate hosts. The role of chemical interactions in mediating the behavior of hematophagous insects and ticks has been investigated before but remains poorly understood in hematophagous mesostigmatic mites. The poultry red mite Dermanyssus gallinae is an obligatory blood-sucking mesostigmatic mite that feeds on birds and causes damage in poultry farms. We characterized the attractive response of D. gallinae to candidate VOCs previously reported from the odor emitted by living hens. We performed in-vitro choice-test bioassays as well as semi-field and field trials using baited and unbaited traps, in the presence and absence of hens. Among different tested combinations of VOCs, a blend of 5 VOCs (mix1.0) was significantly attractive to our reference population of D. gallinae in vitro, whereas the same individual compounds tested alone were not attractive. Ammonia was attractive on its own and increased the mix1.0 attractiveness. The attractiveness of mix1.0 was confirmed at 'natural' spatial scales in the absence of hens both at the lab and on the farm that provided the reference population. The presence of hens inhibited the mix1.0 attractiveness. The attractive power of mix1.0 was not found in other farms. This research is an important step to advance our understanding of host-parasite interactions in hematophagous mesostigmatic mites and paves the way for developing alternative control tools against D. gallinae by interfering with chemical interactions. Moreover, it underlines the importance of assessing kairomonal activity on different pest populations when developing attract-and-kill systems.

Research Note: Prevalence and zoonotic concern of Blastocystis in farmed chickens in southern China.

Blastocystis is a gastrointestinal protozoan parasite commonly reported in humans and animals globally, including poultry, and it can cause zoonotic transmission of blastocystosis. However, comprehensive information is not available on the prevalence, subtype distribution and zoonotic potential of Blastocystis in chickens in China. In this study, a total of 1,000 individual fecal samples of free-range broiler chickens of 4 breeds were collected from 43 farms in 5 cities of Guangdong Province and investigated for the occurrence of Blastocystis infection. Blastocystis was determined by nested PCR analysis of the small subunit ribosomal RNA (SSU rRNA) gene. The overall prevalence was 20.1% (201/1,000) in chicken samples and 69.8% (30/43) in screened farms, and considerable variation in prevalence between farms was evident, with a range of 0 to 76.9%. Population differences of Blastocystis in broilers among sites, breeds, and ages were assessed. The highest infection rates were observed in Yangjiang city (35.8%, 38/106), Sanhuang chickens (29.7%, 104/350), and the >80-day-old chicken group (30.5%, 40/131). DNA sequencing and phylogeny analyses identified 2 zoonotic subtypes, ST6 and ST7. A large predominance was observed for ST7, and genetic polymorphisms were confirmed at the intra-ST7 level with the identification of 5 divergent ST7 types. The incidence of both STs varied largely based on the breed, site, farm, and age. This is the first large-scale study to explore the prevalence and genetic characteristics of Blastocystis in chickens in China. The widespread distribution and avian adaptation of both zoonotic subtypes were demonstrated. The findings of this study highlight a potential threat to humans and will provide a better understanding of the epidemiology and public health impact of poultry Blastocystis.

Dermanyssus gallinae: the long journey of the poultry red mite to become a vector.

The possibility that Dermanyssus gallinae, the poultry red mite, could act as a vector of infectious disease-causing pathogens has always intrigued researchers and worried commercial chicken farmers, as has its ubiquitous distribution. For decades, studies have been carried out which suggest that there is an association between a wide range of pathogens and D. gallinae, with the transmission of some of these pathogens mediated by D. gallinae as vector. The latter include the avian pathogenic Escherichia coli (APEC), Salmonella enterica serovars Enteritidis and Gallinarum and influenza virus. Several approaches have been adopted to investigate the relationship between D. gallinae and pathogens. In this comprehensive review, we critically describe available strategies and methods currently available for conducting trials, as well as outcomes, analyzing their possible strengths and weaknesses, with the aim to provide researchers with useful tools for correctly approach the study of the vectorial role of D. gallinae.

Special Issue: Poultry Genetics, Breeding and Biotechnology.

Poultry products, meat and eggs, are important sources of protein in the human diet worldwide [...].

In vitro acaricidal activity of essential oil and crude extracts of Laurus nobilis, (Lauraceae) grown in Tunisia, against arthropod ectoparasites of livestock and poultry: Hyalomma scupense and Dermanyssus gallinae.

The current study assayed the toxicity of Laurus nobilis essential oil and crude extracts obtained using solvents of increasing polarity (cyclohexane, acetone and ethanol), on two ectoparasites of veterinary importance, i.e., Hyalomma scupense and Dermanyssus gallinae. The major components detected in bay laurel essential oil were dominated by 1.8-cineole (46.56 %), α-terpinenyl acetate (13.99 %), sabinene (7.69), α-pinene (5.75), linalool (5.50), methyleugenol (5.36 %) and ß-pinene (3.97). The highest total phenolic and flavonoids contents were present in the ethalonic extract of L. nobilis leaves at an amount of 152.88 mg gallic acid equivalents per gram of dry weight (GAE/g DW) and 21.77 mg quercetin equivalent per gram of dry weight (QE/g DW), respectively. In vitro acaricidal effects of essensial oil and crude extract of L. nobilis against H. scupense were ascertained by adult immersion test of engorged females (AIT) and larval packet test (LPT) compared with a reference drug amitraz. The essential oil exhibited strong acaricidal activity against tick engorged female and inhibition of hatching eggs. After 24 h of exposure, at the highest tested concentration (100 mg/mL) essential oil induced 90.67 % mortality of H. scupense larvae (LC50 = 10.69 mg/mL). Otherwise, essential oil exhibited high acaricidal activity compared to extracts, and among the extract, the ethanolic extract revealed the highest acaricidal efficacy (81.27 % female mortality). Results from mite contact toxicity showed that essential oil and extracts from L. nobilis were toxic to D. gallinae. Bay essential oil was both more toxic to mites, and faster in exerting this toxicity than other tested crude extracts. L. nobilis essential oil concentration leaded to enhance mortality of D. gallinae reaching the highest (100 %) mortality at 12 h with a concentration of 320 mg/mL. While, ethanolic extract acheived this rate after 24 h of exposure at same concentration. Cyclohexanic extract showed weak acaricidal activity.

Assessment of fluralaner as a treatment in controlling Dermanyssus gallinae infestation on commercial layer farms and the potential for resulting benefits of improved bird welfare and productivity.

BACKGROUND: Poultry red mite (PRM) (Dermanyssus gallinae) infestations are a cause of anaemia, impaired productivity and stress-related behaviours linked to reduced hen welfare. A study investigated the potential health, welfare and productivity benefits following fluralaner treatment to eliminate PRM from infested hens. METHODS: A PRM-infested layer house was selected on a free-range farm (5400 hens) and an aviary farm (42,400 hens). Fluralaner (Exzolt®; 0.5 mg/kg body weight) was administered twice, 7 days apart (Weeks 0 and 1), via drinking water. Mite populations were monitored by traps. Cameras recorded nighttime hen behaviours weekly, pre- and post-treatment. On the free-range farm, daytime behaviours were also recorded weekly. For pre- and post-treatment corticosterone assessments, eggs were randomly collected on both farms, and blood samples were collected from 50 randomly selected aviary farm hens. Production parameters were assessed using farm records. RESULTS: Throughout the post-treatment period, fluralaner efficacy against PRM was > 99% on both farms. On the aviary and free-range farms, treatment was followed by significant nighttime increases in the proportion of resting hens (P < 0.0001; P = 0.0175, respectively). Significant post-treatment versus pre-treatment nighttime reductions were observed in head shaking (aviary, P < 0.0001; free-range P = 0.0233) and preening (P = 0.0032; P = 0.0018) and on the aviary farm in bouts of body shaking (P = 0.0108), vertical wing shaking (P = 0.0002), head scratching (P = 0.0335), and gentle feather pecking (P < 0.0001). On the free-range farm there were significant daytime reductions in head scratching (P < 0.0001), head shaking (P = 0.0492) and preening (P = 0.0012). Relative to standard production parameters, no differences were detected on the aviary farm, but on the free-range farm the laying rate decline with increasing age was less than expected and the increase in egg weight greater than expected. Post-treatment increases in egg and plasma corticosterone were suggestive of stress factors in addition to mite infestation. Red blood cell counts and haematocrit increased following treatment. CONCLUSION: Fluralaner treatment eliminated mite challenge, leading to improved hen welfare and health, based on reductions in stress-related behaviours and restoration of the anaemia-inducing effects of mite blood feeding.

Designing and developing of high-resolution melting technique for separating different types of Toxoplasma gondii by analysis of B1 and ROP8 gene regions.

BACKGROUND: Determination of Toxoplasma gondii genotypes plays an important role in the health management and epidemiology of toxoplasmosis. We developed HRM analysis to differentiate genotypes of T. gondii using the B1 and ROP8 genes, through comparing the sensitivity and specificity of both genes and methods used for the detection of T. gondii. METHODS: A total of 96 DNA samples of muscle tissue of livestock and poultry brain tissue with three standard strains RH (type I), PRU (type II) and VEG (type III) were prepared and analyzed. Three methods of nested PCR, PCR-PCR and nested-qPCR-HRM were used. Specific new primers were designed and synthesized for developing HRM. Thirty positive samples obtained from nested-qPCR-HRM were sequenced (18 B1 and 12 ROP8). RESULTS: Overall, 87 infected samples were identified using both genes. Through the B1 gene, we could separate type I (Tm = 84.8 °C) from II/III types (Tm = 84.6 °C). Also, the ROP8 gene could separate type II (Tm = 84.5 °C) from I/III types (Tm = 84.12 °C). Highest sensitivity (100%) and specificity (78.72%) were observed by nested-qPCR-HRM assays of the B1 and ROP8 genes than by other methods, respectively. Thus, the B1 gene can be used to most accurately detect T. gondii, while the ROP8 gene was more appropriate for T. gondii genotyping. PCR-sequencing results were consistent with HRM results in most selected samples. CONCLUSION: HRM analysis is a powerful diagnostic tool for rapid detection and determination of main clonal lineages, and even unusual T. gondii genotypes.

Detection of larvae of Toxocara cati and T. tanuki from the muscles of free-ranging layer farm chickens.

Although raw or undercooked livestock meat or viscera has been suggested to be a source of human toxocariasis, there have been few reports on the prevalence of Toxocara larvae in the tissue of livestock animals. To investigate the presence of Toxocara larvae in chickens, we examined 50 culled chickens from a commercial layer farm. The liver, breast meat, and thigh meat were separated individually and artificially digested to examine for the presence of larvae. Nematode larvae were detected in 2 out of 50 chickens. One larva was detected from the breast meat, and it was molecularly identified as Toxocara tanuki. The other from the thigh meat of another chicken was molecularly identified as Toxocara cati. The present study demonstrated for the first time that T. tanuki larvae do infect chickens in the natural environment. The fact that Toxocara spp. larvae were found in muscles of farm chickens suggests that consumption of raw or undercooked chicken meat may present a risk for human toxocariasis.

Anthelmintic efficacy of fenbendazole and levamisole in native fowl in northern Iran.

BACKGROUND: With the increasing number of free-range domestic chickens, helminth parasites have potentially become more of a threat to commercial flocks in recent years, and routine poultry deworming is needed to improve the efficiency of chicken production. The present study deals with a field trial to study the efficacy of two generally used anthelmintics, fenbendazole and levamisole, against gastrointestinal nematodes of domestic chickens in northern Iran. METHODS: Of 45 domestic chicken flocks involved in the study, 20 flocks were selected to participate in fecal egg count reduction testing based on flock size from April 2017 to September 2018. The infected chickens were randomly divided into three equal groups of 30 each. Ninety chickens in the infected groups received one of the following treatments (d 0 of treatment): Group 1: 5 mg kg-1 body weight (BW) fenbendazole for 3 consecutive days; Group 2: 16 mg kg-1 BW levamisole; Group 3 control: placebo, water + DMSO (dimethylsulfoxide). The efficacy of the treatments was evaluated by comparing fecal egg counts in the treated and control groups. RESULTS: Examination of three flocks of chickens from the control group showed that 95.0% of the animals were infected with gastrointestinal nematodes with an average geometric value of 361 eggs per gram of feces. Fenbendazole at a dose of 5 mg kg-1 BW for 3 days showed an efficacy of 83.7% (P ≥ 0.05), and levamisole at a dose of 16 mg kg-1 BW showed 71.8% efficacy (P ≥ 0.05) with geometric mean eggs in a gram of feces of 100 and 199.6, respectively. In general, fenbendazole and levamisole treatment led to significantly lower activity. The result of this study revealed that fenbendazole was a better and more effective dewormer than levamisole on the three Iranian domestic chicken flocks, but the difference was not significant. Capillaria spp. were the most generally resistant nematodes followed by Trichostrongylus spp. and Amidostomum anseris. CONCLUSION: Our results indicated that fenbendazole and levamisole effectively reduced the number of nemathodes in three Iranian domestic chicken flocks. Given the results of our study, resistance can be expected in the parasitic helminths of poultry. Additional larger scale studies are required to determine the prevalence of anthelmintic resistance in the poultry industry.

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

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.

Epidemiologia e avaliação de risco associado à presença de ácaros hematófagos em galpões de granjas avícolas de postura / [Epidemiological and risk assessment associated with the presence of hematophagous mites in poultry farms]

Este estudo teve como objetivo caracterizar a epidemiologia da presença de ácaros hematófagos em granjas de postura no estado de Minas Gerais. Foi utilizado um banco de dados secundário, com informações de 402 galpões de 42 propriedades comerciais. As variáveis utilizadas para compor o modelo de correspondência foram selecionadas por meio do teste qui-quadrado (P≤0,05). Foi construído um índice para a presença de ácaros hematófagos, considerando alguns fatores de risco. Além disso, um estudo da análise espacial foi realizado para avaliar a presença de ácaros hematófagos em Minas Gerais. Observou-se a presença de ácaros hematófagos em 48% dos galpões, sendo a de O. sylviarum de 45,5%, O. bursa de 17,4% e D. gallinae de 2,7%. Houve associação entre o índice de risco com os intervalos de remoção de fezes e com a presença ou a ausência de aves sinantrópicas. Verificou-se que granjas mais tecnificadas não utilizam acaricida em seus galpões. A presença de ácaros hematófagos foi observada na mesorregião Sul/Sudoeste de Minas Gerais e entre as mesorregiões Oeste de Minas e Metropolitana de Belo Horizonte. Esses resultados fornecem conhecimento sobre a epidemiologia desses ectoparasitos e podem contribuir na tomada de decisões, reduzindo os riscos de possíveis infestações em aves de postura.(AU)

This study aimed to characterize the epidemiology of the presence of hematophagous mites in posture farms in the state of Minas Gerais. Was used a secondary database with information from 402 chicken houses in 42 commercial properties. The variables used to compose the correspondence model were selected through chi-square tests (P≤0.05). A risk index was built to the presence of hematophagous mites considering some risk factors. Additionally, a study of the spatial analysis was done to evaluate the presence of hematophagous mites in Minas Gerais. Was observed the presence of hematophagous mites in 48% of chicken houses, the presence of O. sylviarum was 45.5%, O. bursa 17.4% and D. gallinae 2.7%. There was an association between the risk index with the intervals for the removal of manure and the presence or absence of synanthropic birds. It was verified that more technified farms do not use acaricide in their chicken houses. The presence of hematophagous mites was observed in the Messoregions South/Southwest of Minas Gerais and between the West and Metropolitan Mesoregion of Belo Horizonte. These results provide knowledge about the epidemiology of these ectoparasites and may contribute to decision making by reducing the risks of possible infestations in poultry.(AU)

Assessment of laying-bird welfare following acaricidal treatment of a commercial flock naturally infested with the poultry red mite (Dermanyssus gallinae).

The poultry red mite (PRM), Dermanyssus gallinae, a potential vector of pathogens to animals and humans, causes impaired bird welfare. A study investigated changes in behavioural variables, physiological biomarkers, and health parameters following acaricidal treatment of PRM infestation of laying hens on a commercial farm. Mite traps determined the challenge to 12,700 hens before and after drinking water administration of the acaricide, fluralaner (Exzolt®, 0.5 mg/kg; Weeks 0 and 1). Weekly daytime direct observations and night-time video recordings monitored bird behaviours from Weeks -6 through +6. Blood samples were collected from randomly-selected birds (Weeks -6, -1, and +6). Following treatment, mite count reductions (>99%) were statistically significant (P < 0.0001), as were night-time reductions in the percent of hens showing activity, preening, head scratching (all P < 0.0001), and head shaking (P = 0.0007). Significant daytime reductions were observed in preening and head scratching (both P < 0.0001), head shaking (P = 0.0389), severe feather pecking (P = 0.0002), and aggressive behaviour (P = 0.0165). Post-treatment, comb wounds were significantly reduced (P = 0.0127), and comb colour was significantly improved (P < 0.0001). Heterophil/lymphocyte ratio was significantly reduced at Weeks 1 and 6 (P = 0.0009 and P < 0.0001, respectively). At Week 6, blood corticosterone (P = 0.0041) and total oxidant status (P < 0.0001) were significantly reduced, and haemoglobin and mean corpuscular haemoglobin significantly increased (P < 0.0001). Farm production records indicated that those post-treatment improvements were accompanied by significant reductions in weekly mortality rate (P = 0.0169), and significant recovery in mean weekly egg weights (P < 0.0001) and laying rate (P < 0.0001). The improvements in behavioural variables, physiological biomarkers, and health parameters that were observed following the elimination of PRM on a commercial farm indicate that infestations can be a cause of reduced hen welfare.

Comparative morphological and transcriptomic analyses reveal chemosensory genes in the poultry red mite, Dermanyssus gallinae.

Detection of chemical cues via chemosensory receptor proteins are essential for most animals, and underlies critical behaviors, including location and discrimination of food resources, identification of sexual partners and avoidance of predators. The current knowledge of how chemical cues are detected is based primarily on data acquired from studies on insects, while our understanding of the molecular basis for chemoreception in acari, mites in particular, remains limited. The poultry red mite (PRM), Dermanyssus gallinae, is one of the most important blood-feeding ectoparasites of poultry. PRM are active at night which suck the birds' blood during periods of darkness and hide themselves in all kinds of gaps and cracks during the daytime. The diversity in habitat usage, as well as the demonstrated host finding and avoidance behaviors suggest that PRM relies on their sense of smell to orchestrate complex behavioral decisions. Comparative transcriptome analyses revealed the presence of candidate variant ionotropic receptors, odorant binding proteins, niemann-pick proteins type C2 and sensory neuron membrane proteins. Some of these proteins were highly and differentially expressed in the forelegs of PRM. Rhodopsin-like G protein-coupled receptors were also identified, while insect-specific odorant receptors and odorant co-receptors were not detected. Furthermore, using scanning electron microscopy, the tarsomeres of all leg pairs were shown to be equipped with sensilla chaetica with or without tip pores, while wall-pored olfactory sensilla chaetica were restricted to the distal-most tarsomeres of the forelegs. This study is the first to describe the presence of chemosensory genes in any Dermanyssidae family. Our findings make a significant step forward in understanding the chemosensory abilities of D. gallinae.

Management of the poultry red mite, Dermanyssus gallinae, using silica-based acaricides.

Four silica-based acaricides were examined in laboratory tests for their effectiveness against poultry red mite, Dermanyssus gallinae. All acaricides resulted in 100% mite mortality. Two groups of active ingredients could be differentiated. The products Silicosec® and Ewazid®, based on naturally occurring diatomaceous earth (DE), killed 100% of adult D. gallinae within 48 h exposure time. The time to kill 50% of the mites (LT50) was calculated to be 31.7 and 34.9 h, respectively. The other two products, containing aggregates and agglomerates of pyrogenic synthetic amorphous silicon dioxide as active ingredients, killed the mites in a significantly shorter time: LT50 was 6.3 h for the liquid product Fossil Shield® Instant White and 11.8 h for the powdery product Fossil Shield 90.0 White. This is more remarkable as the quantities of active ingredients used for the DE treatments were several folds higher. The effectiveness of all tested products was also shown in practical tests. A professional company treated five chicken houses on one farm in the Berlin-Brandenburg region with the test products, three houses with Fossil Shield Instant White and one each with Ewazid and Silicosec. Over a period of 46 weeks after stocking, the mite development in the houses was assessed. Only in one of the houses, treated with Fossil Shield Instant White, the mite population remained permanently low. In two houses treated with Fossil Shield Instant White, small mite colonies appeared in week 36, which were controlled by a follow-up spot treatment in week 41. In the houses treated with DE, the first mite colonies appeared 12 weeks after stocking. The number increased continuously over the experimental period and in week 31 after stocking there were clearly visible colonies (2-3 cm diameter) and the first mites could also be detected on the chicken eggs. At this time both houses were treated again with a follow-up spot-treatment, which only led to a slight improvement in one house and to a stabilization of the infestation in the other house. In week 41, large mite colonies were detected in both houses. A spot treatment at this point was ineffective in reducing the infestation. The tests showed faster acaricidal action of the products with the synthetic active ingredients compared to the natural DE-based products. This matches the shorter killing times under laboratory conditions. The experiments in a commercial chicken farm showed that it is possible to control the mite population for a period of 46 weeks by using physically effective SiO2-based products. These products are therefore an effective alternative to the use of chemical acaricides.

Mesostigmatid mites (Acari: Mesostigmata) at the domestic-wildlife interface: Poultry and passerine birds of central Argentina.

Wild birds may be considered a possible source of parasitic mesostigmatid mites for poultry, but only few studies explored this hypothesis. In addition, there is very little information about the parasitic mites present in commercial poultry systems from southern South America. To contribute with data on parasitic mesostigmatid mites at the domestic-wildlife interface, we conducted a study in which samples were systematically collected from laying hens and wild birds (adults and nestlings), for two years at three commercial laying hen farms. The occurrence of mesostigmatid mites were compared among hosts. A proportion of the collected mites were morphologically identified to the species level, finding that host preference varied greatly depending on mite species: laying hens were only parasitized by Ornithonyssus sylviarum, wild bird nestlings were mostly parasitized by Ornithonyssus bursa, and in small proportion, by O. sylviarum, while adult passerines were parasitized by both Ornithonyssus species, and sporadically by Pellonyssus cf. reedi and Dermanyssus cf. triscutatus. In laying hens, there was intra- and inter-annual variability in mite occurrence, but no consistent seasonal pattern, whereas in adult wild birds, mites showed the highest prevalence in spring and the lowest in summer. Not coinciding with this general pattern, the occurrence of O. bursa matched the reproductive activity of wild birds. A phylogenetic analysis based on a fragment of the 16S rRNA gene was carried out for a subsample of the mites collected, showing that the O. sylviarum mites present on adult wild birds and laying hens had the same haplotype (100% identity). Additionally, mites obtained from wild birds morphologically identified as O. bursa presented two distinctive haplotypes (89.8% identity), one phylogenetically related to O. sylviarum and the other to O. monteiroi. These findings show that in central Argentina commercial laying hens are parasitized mainly by O. sylviarum while wild birds are also hosts to other mite species. Adult wild passerines, especially house sparrows, may be a source of O. sylviarum for commercial poultry.

Development of MALDI-TOF mass spectrometry for the identification of lice isolated from farm animals.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is now routinely used for the rapid identification of microorganisms isolated from clinical samples and has been recently successfully applied to the identification of arthropods. In the present study, this proteomics tool was used to identify lice collected from livestock and poultry in Algeria. The MALDI-TOF MS spectra of 408 adult specimens were measured for 14 species, including Bovicola bovis, B. ovis, B. caprae, Haematopinus eurysternus, Linognathus africanus, L. vituli, Solenopotes capillatus, Menacanthus stramineus, Menopon gallinae, Chelopistes meleagridis, Goniocotes gallinae, Goniodes gigas, Lipeurus caponis and laboratory reared Pediculus humanus corporis. Good quality spectra were obtained for 305 samples. Spectral analysis revealed intra-species reproducibility and inter-species specificity that were consistent with the morphological classification. A blind test of 248 specimens was performed against the in-lab database upgraded with new spectra and validated using molecular tools. With identification percentages ranging from 76% to 100% alongside high identification scores (mean = 2.115), this study proposes MALDI-TOF MS as an effective tool for discriminating lice species.

TITLE: Développement de la spectrométrie de masse MALDI-TOF MS pour l'identification de poux isolés d'animaux de ferme. ABSTRACT: La Spectrométrie de Masse à Temps de Vol par Désorption/Ionisation Laser Assistée après Matrice est maintenant utilisée pour l'identification rapide des microorganismes isolés à partir d'échantillons cliniques et a récemment été appliquée avec succès pour l'identification des arthropodes. Dans cette étude, cet outil protéomique a été utilisé pour identifier les poux prélevés sur le bétail et la volaille en Algérie. Les spectres MALDI-TOF MS de 408 spécimens adultes ont été mesurés pour 14 espèces, dont Bovicola bovis, B. ovis, B. caprae, Haematopinus eurysternus, Linognathus africanus, L. vituli, Solenopotes capillatus, Menacanthus stramineus, Menopon gallinae, Chelopistes meleagridis, Goniocotes gallinae, Goniodes gigas, Lipeurus caponis et Pediculus humanus corporis élevé en laboratoire. Des spectres de bonne qualité ont été obtenus pour 305 échantillons. L'analyse spectrale a révélé une reproductibilité intra-espèce et une spécificité inter-espèces qui concordaient avec la classification morphologique. Un test à l'aveugle de 248 échantillons a été effectué par rapport à la base de données de notre laboratoire mise à niveau avec de nouveaux spectres et validée à l'aide d'outils moléculaires. Avec des pourcentages d'identification allant de 76 à 100 % et des scores d'identification élevés (moyenne : 2,115), cette étude propose MALDI-TOF MS comme un outil efficace pour distinguer les espèces de poux.

A novel luciferase-linked antibody capture assay (LACA) for the diagnosis of Toxoplasma gondii infection in chickens.

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes the most common parasitic zoonosis worldwide in multiples species of mammals and birds. Although free-range chickens may play a role as an important reservoir for T. gondii, there is no reliable and commercially available diagnostic test for this disease in chickens. In this study, we aimed to develop a novel Luciferase-linked Antibody Capture Assay (LACA) for the serodiagnosis of Toxoplasma infection in chickens. Recombinant nanoluciferase fused-T. gondii dense granule antigen 8 (rNluc-GRA8) was produced and applied to LACA assay as a diagnostic antigen. GRA8-LACA was tested with the sera from uninfected and experimentally infected chickens with T. gondii and other parasitic pathogens and showed unexpectedly high sensitivity (90.5%) and specificity (95.4%). Interestingly, E. coli lysate expressing rNluc-GRA8 could be applied in GRA8-LACA with 85.7% sensitivity and an increased specificity (96.9%) that gave better diagnostic performance compared to conventional ELISA. We applied our diagnostic system to examine 267 free-range chicken sera collected from 12 farms and 100 closed-house broiler chicken sera from local poultry abattoirs. The overall seroprevalence of toxoplasmosis in free-range chickens was 10.9% (95% CI: 10.6%-11.1%), while no positive case was found in broiler chickens. GRA8-LACA could be a useful diagnostic technique for T. gondii infection in chickens. The detection of T. gondii seropositive chickens in this study warns a potential risk of Toxoplasma transmission by the consumption of raw or undercooked chicken meat.

Occurrence of Atypical and New Genotypes of Toxoplasma Gondii in Free-Range Chickens Intended for Human Consumption in Brazil.

PURPOSE: The aim of the present study was to detect and genotype T. gondii in free-range chickens destined to human consumption in Alagoas state, Brazil. METHODS: Two hundred blood samples were collected from free-range chickens and submitted to indirect immunofluorescence antibody test (IFAT). Brain tissue from 14 animals randomly selected were subjected to mouse bioassay. Positive samples in mouse bioassay were submitted to PCR and genotyped by PCR-RFLP. RESULTS: Out of two hundred blood samples from chickens, 72 (36%) samples were considered positive by IFAT. Two T. gondii strains were isolated, both being characterized as atypical and classified as #146 and a new genotype, named #279 in ToxoDB database. CONCLUSIONS: Our results showed a sero-occurrence of T. gondii in free-range chickens intended for humans, and the genetic diversity of the parasite in Brazil, with a new genotype described.