Flock management and histomoniasis in free-range turkeys in France: description and search for potential risk factors.

The relationship between flock management and histomoniasis, a re-emergent infection in poultry, was investigated by statistical techniques used in veterinary epidemiology to deal with various problems including: multicollinearity, confounding, interaction or sample size. Associations between the variables describing flock management were examined by multivariate descriptive analysis to reduce the number of independent variables, prior to investigating associations with the disease. No homogenous groups of farms were found in the 44 free-range turkey flocks sampled in France. Histomonas meleagridis was identified in 26/38 flocks and histomoniasis was confirmed in 19 flocks. Cleanliness of the building, wet litter and diarrhoea were linked with H. meleagridis and severity of histomoniasis. Sharing outdoor fields simultaneously with chickens was related to serious macroscopic lesions determined by post-mortem examinations. Contrary to general belief, acidification of drinking water with organic acid had consistent association with the presence of H. meleagridis in turkey caeca. These results confirm previous findings and provide several new hypotheses on the effects of hygiene and water management on H. meleagridis and histomoniasis.

Monitoring of Dermanyssus gallinae in free-range poultry farms.

Various methods for monitoring Dermanyssus gallinae infestations within free-range egg production units were compared. The study was carried out in five egg-producing free-range poultry buildings infested with D. gallinae. Each farm was divided into six zones (each zone including nest boxes, perches and duckboard) for placing two types of traps (corrugated cardboard and thick card traps) or examining dried droppings for presence of mites. Traps were removed 24 h later, placed into bags and mites were counted at the laboratory using binocular magnification. Droppings were also inspected by eye and mite numbers were estimated. All the methods used allowed us to detect mites although their efficacy differed. The number of mites collected was independent of the type of trap used. Examination of the droppings did not differentiate between buildings with differing mite populations. Placing traps in the nest boxes is a less reliable indicator than placing them on the perches. It appears that the most coherent method for evaluating the D. gallinae population within a free-range flock is to place thick card traps throughout the building, on perches favoured by birds.

Molecular phylogenetic assessment of host range in five Dermanyssus species.

Given that 14 out of the 25 currently described species of Dermanyssus Dugès, 1834, are morphologically very close to each another, misidentifications may occur and are suspected in at least some records. One of these 14 species is the red fowl mite, D. gallinae (De Geer, 1778), a blood parasite of wild birds, but also a pest in the poultry industry. Using molecular phylogenetic tools we aimed to answer two questions concerning host specificity and synanthropicity: (1) is D. gallinae the only species infesting European layer farms?, and (2) can populations of D. gallinae move from wild to domestic birds and vice versa? Mitochondrial cytochrome oxidase I gene sequences were obtained from 73 Dermanyssus populations collected from nests of wild European birds and from poultry farms and these were analyzed using maximum parsimony and Bayesian inference. Mapping of the observed host range on the obtained topology and correlation with behavioural observations revealed that (1) host range is strongly dependent on some ecological parameters (e.g. nest hygiene, exposure to pesticides and predators), that (2) out of five species under test, synanthropic populations were found only in lineages of D. gallinae, and that (3) at least some haplotypes found in wild birds were very close to those found in association with domestic birds.

Delimiting species boundaries within Dermanyssus Dugès, 1834 (Acari:Dermanyssidae) using a total evidence approach.

The genus Dermanyssus is currently composed of 24 hematophagous mite species and includes the Poultry Red Mite, Dermanyssus gallinae, a serious pest in poultry houses. Morphologically, Dermanyssus species fall into two groups corresponding to Moss'gallinae-group and to hirsutus-group+Microdermanyssus. Species of the gallinae-group exhibit high levels of morphological variability, and are nearly impossible to distinguish. Species of the second group display consistent characters and host associations and are easily distinguishable. Species of the gallinae-group tend to be the major problems in poultry houses and it is unknown whether D. gallinae is the only pest, or if there are numerous cryptic species present in the system. Twenty species of Dermanyssus were tested phylogenetically based on 46 morphological characters. A subset of species, mainly of the gallinae-group, represented each by several populations, was sequenced for two mitochondrial and one nuclear gene regions. This allowed testing their specific status and their interrelationships based and on morphological and molecular characters. The molecular data was analysed separately and in combination with morphological characters. As expected, morphology did a poor job resolving relationships. Molecular data proved more informative. The resulting phylogenetic hypotheses brought some information about interrelationships among species of the gallinae-group showing a split into two main clades. The invasion of human managed environments seems to occur only in taxa within one of the two clades. The host spectrum seems to get enlarged in more derived taxa in the same clade. A delineation of six species within the gallinae-group is provided. Additionally, a key for morphological identification of these species is provided. D. gallinae appears to be the only pest in poultry houses, but is composed of several different and more or less strongly isolated lineages. A new species found from the black swift is described.

Incidence of histomonosis in turkeys in France since the bans of dimetridazole and nifursol.

Between April 2003 and March 2005, 113 outbreaks of histomonosis were recorded in standard turkey farms in France, and 15 cases were recorded in turkey breeding centres. Most of the cases were in north-west France, the principal farming area for turkeys. The majority of the cases occurred during the hottest months, from April to September. Large numbers of cases occurred among birds from four to eight weeks of age, but there were some cases in three-week-old birds and some in birds up to 17 weeks of age. In most of the standard turkey flocks the mortality was less than 10 per cent, but it was above 30 per cent in nearly 20 per cent of the outbreaks. In the breeding flocks, the average mortality was 60.2 per cent. The size of the flocks, the sex of the birds and the age at which the first clinical signs appeared did not seem to influence the mortality.

Historical review of the genus Dermanyssus Dugès, 1834 (Acari: Mesostigmata: Dermanyssidae).

A synthetic review of the historical systematics of Dermanyssus Dugès, 1834 (Acari: Mesostigmata: Dermanyssidae) is provided. The classification at the specific level in this early genus has not really been clarified during more than a century despite its economic impact, and the history of the genus is complex and includes various stages. Moreover, Dermanyssus currently includes 23 species, whereas the last review took only 18 species into account. Changes in the species status and position in the genus Dermanyssus from 1834 until today are presented. The evolution of the generic definition is explored and compared with other genera of the group. How the discrimination between the different species evolved in the genus is also examined. Some difficulties in the specific definitions are discussed. A current diagnosis of the genus Dermanyssus is given. A table of the species included in this genus since its first description along with their respective current positions, a list of the currently included species in Dermanyssus with their hosts, and a world map presenting their geographic distribution are provided.

Detection of Salmonella sp. in Dermanyssus gallinae using an FTA filter-based polymerase chain reaction.

Salmonella spp. bacteria are responsible for some of the most important zoonoses worldwide. Because Dermanyssus gallinae (DeGeer) (Acari: Dermanyssidae) has been recently reported to be an experimental vector of Salmonella Enteritidis, it would be of benefit to evaluate the presence of this bacterium in mites. A molecular detection tool associating a simple filter-based DNA preparation with a specific 16S rDNA Salmonella sp. polymerase chain reaction (PCR) amplification was described. The limit of detection with this method was 2 x 10(4) bacteria per mite. To adapt this technique for large-scale studies, two sizes of mite pools were tested and a preliminary investigation was carried out on mites from 16 currently or previously contaminated farms. Mites sampled from one farm of each type were positive for Salmonella, suggesting that Dermanyssus could act as a reservoir between flocks. In further investigations, it will be necessary to carry out a large-scale study to assess the role of D. gallinae in the epidemiology of avian salmonellosis.

Comparison of the VIDAS system, FTA filter-based PCR and culture on SM ID for detecting Salmonella in Dermanyssus gallinae.

AIMS: To compare different analytical methods for detecting Salmonella in Dermanyssus gallinae. METHODS AND RESULTS: The detection limit of three Salmonella detection methods [Vitek immunodiagnostic assay (VIDAS) Salmonella immuno-concentration/immunoassay, FTA filter-based PCR, and Salmonella detection and identification medium (SM ID) preceded by a pre-enrichment step] was evaluated by crushing mites in serial dilutions of pure cultures of Salmonella enterica ssp. Enterica serotype Enteritidis. Each method was then compared for its ability to detect Salmonella in artificially contaminated mites. In 105 mites artificially engorged with Salm. Enteritidis-contaminated blood, Salmonella was isolated from 68 samples of the samples cultured on SM ID and tests were positive for Salmonella using FTA filter-based PCR and VIDAS in 77 and 65 samples, respectively. Using SM ID as our reference method, specificities and sensitivities were 97% and 94% and 73% and 98.5% for VIDAS and PCR, respectively. CONCLUSIONS: Each method allowed the detection of Salmonella in contaminated mites and is usable for screening mites. PCR is more sensitive but less specific than VIDAS for detecting Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time that the VIDAS has been used to detect pathogens in vectors. The development of analytical methods for Salmonella detection in mites is a necessary step in the study of the role of D. gallinae as a vector of salmonellae and to check the contamination of D. gallinae in poultry facilities.

Experimental infection of Salmonella Enteritidis by the poultry red mite, Dermanyssus gallinae.

Dermanyssus gallinae is an important ectoparasite of laying hens in Europe and it is suspected of being a vector of pathogens. We carried out an in vitro study to evaluate the role of D. gallinae as a vector of Salmonella enterica subsp. enterica serotype Enteritidis. Two means of infecting the mite were tested: through the blood meal and after cuticular contact. Mites became carriers of Salmonella immediately after the infection with 29% and 53%, respectively, for oral route and cuticular contact. This percentage increased over time until it reached 95% (D7) and 80% (D14). The numerical identification of bacteria on the selective medium SM ID demonstrated the multiplication of Salmonella inside previously infected mites. In addition, transovarial passage as well as transstadial passage (from N1 to N2 stages) were demonstrated. Moreover, the observation of a negative effect of Salmonella on Dermanyssus oviposition was also observed. Finally, previously infected mites were able to contaminate the blood during the blood meal. Therefore, it appears that D. gallinae may act as a biological vector of S. Enteritidis under experimental conditions. It may represent a suitable environment for the development of Salmonella and could be an additional factor for the persistence of salmonellosis infection between successive flocks.

Infectivity of Histomonas meleagridis in ducks.

The susceptibility of mule and muscovy ducks to "blackhead" disease caused by Histomonas meleagridis was studied, using an experimental intracloacal inoculation. Turkeys were used as controls. Morbidity, mortality and body weight gain were recorded regularly during the experiments. A direct examination of the caecal content was made to determine the absence or presence of the parasite. Gross and microscopic lesions were observed on days 7, 14, 21, 28 and 35 post infection to evaluate any clinical histomoniosis in ducks and to appraise the histomonad's carriage. A scoring system was developed both for gross and histological lesions of the caecum and liver. Infected mule and muscovy ducks (n = 83) never developed any clinical signs of histomoniasis. Weight gains of infected mule and muscovy ducks were similar to those of uninfected ducks. In 67% of the ducks (56/83), it was possible to demonstrate the parasite in the caecal content throughout the experiment. Typical macroscopic caecal lesions were observed in five of the ducks between days 7 and 21 post infection, with a caecal necropsy main lesion score (MLS = 1.6) less severe than that in turkeys (MLS = 2.9). Only caecal histological lesions occurred in six of the cases. Therefore, ducks do not seem to be a susceptible host for "blackhead" but may act as carrier animals for H. meleagridis. The virulence was apparently not changed, since 67% of turkeys (10/15) infected with the caecal content of positive ducks displayed classical signs of blackhead disease. Even if H. meleagridis alone does not represent a substantial danger in the duck production, its infectivity should to be taken into account in the transmission to more susceptible species.

Vectorial role of some dermanyssoid mites (Acari, Mesostigmata, Dermanyssoidea).

Among transmissible diseases, vectorial diseases represent a major problem for public health. In the group of acarina, while ticks are the most commonly implicated vectors, other arthropods and notably Dermanyssoidea are also involved in the transmission of pathogenic agents. Since the role of this superfamily is at present largely unknown, we have reviewed the vectorial role of these mites in the appearance, survival and propagation of pathogens. Various authors have shown that Dermanyssoidea are implicated in the transmission of both bacteria (Salmonella, Spirocheta, Rickettsia or Pasteurella) and viruses (equine encephalitis viruses, West Nile virus, Fowl pox virus, the virus causing Newcastle disease and tick borne encephalitis viruses or hantaviruses). Finally, some authors have also shown their role in the transmission of some protozoa and filaria. As the vectorial character of such mites has been more clearly demonstrated (Dermanyssus gallinae, Omithonyssus bacoti and Allodermanyssus sanguineus), it would be interesting to continue studies to better understand the role of this superfamily in the epidemiology of certain zoonoses.

Detection of Histomonas meleagridis in turkeys cecal droppings by PCR amplification of the small subunit ribosomal DNA sequence.

Histomonas meleagridis is a protozoan parasite that may cause histomoniasis, a disease of gallinaceous fowl characterized by necrotic typhlitis, hepatitis and high mortality. Diagnosis of this disease is based on direct identification or on cultivation of the parasite. With the aim of developing more sensitive, rapid and useful tools for parasite detection, PCR that amplified a DNA target of 209 pb of the 18S rRNA gene was designed to detect the genome of H. meleagridis and to differentiate it from the genome of Tetratrichomonas gallinarum, another common protozoan parasite of fowl. The sensitivity of the test was evaluated using serial diluted samples of cultured H. meleagridis and showed positive amplification for concentrations comprised between 10 and 10(-1)parasites/ml of culture. The sensitivity for cecal droppings samples was assessed using spiked material and was comprised between 3 x 10(3) and 3 x 10(5)parasites/ml of stool. The reliability of the PCR for the detection of Histomonas infection was also evaluated by experimental infection of turkeys. Results of the PCR appeared to be in agreement with the development of the clinical signs and of the cecal lesions. The PCR developed in this study may be a useful tool in the detection and identification of H. meleagridis for rapid, routine screening as a supplement to direct identification or cultivation of the parasite.

A mathematical model of the population dynamics of Heterakis gallinarum in turkeys (Meleagridis gallopavo).

Heterakis gallinarum is a relatively nonpathogenic organism, but it is important as the transport host for the pathogenic protozoan Histomonas meleagridis. A mathematical model was developed to describe the population dynamics of Heterakis gallinarum in a turkey flock to study its kinetics in a number of hosts. The model includes quantitative (parasite burden) and qualitative (number of hosts without mature parasite) descriptions of these dynamics. To understand the role of Heterakis as a transport host, the various elements that delay the beginning of development of the parasite population (e.g., necessary delay of larval stage, the probability of having a male and female in the same host) were taken into account. From published data, the negative binomial distribution parameter k = 0.24, which described the aggregated distribution of the Heterakis among the hosts, was calculated. The sensibility study showed that when the k parameter decreased (i.e., when the population was more aggregated), infestation increased quantitatively (mean parasite burden increased) but not qualitatively (the number of host without mature parasite increased). The model demonstrated that the population dynamics of Heterakis takes time; for instance, with an aggregated population of Heterakis at d 90, the host is mainly free of adult parasite. These results may be used in the future to test the role of Heterakis in the spread of Histomonas.

In vitro effect of essential oils from Cinnamomum aromaticum, Citrus limon and Allium sativum on two intestinal flagellates of poultry, Tetratrichomonas gallinarum and Histomonas meleagridis.

Essential oils may be effective preventive or curative treatments against several flagelated poultry parasites and may become primordial either to organic farms, or as more drugs are bannished. The anti-flagellate activity of essential oils obtained from fresh leaves of Cinnamomum aromaticum, Citrus limon pericarps and Allium sativum bulbs was investigated in vitro on Tetratrichomonas gallinarum and Histomonas meleagridis. On T. gallinarum, the minimal lethal concentration (MLC) at 24 hours was 0.25 microliter/ml for C. aromaticum oil, and 0.125 microliter/ml for C. limon and A. sativum oils. On H. meleagridis, MLC was 0.5 microliter/ml for C. aromaticum oil and 1 microliter/ml for C. limon and A. sativum oils at 24 and 48 hours. Moreover, no synergistic effects were evidenced in vitro. The essential oil constituents, based on their GC retention times have been also identified. The major component is trans-cinnamaldehyde (79%) for C. aromaticum; limonene for C. limon (71%) and diallyl tri- and disulfide (79%) for A. sativum. Even if concentration and protocol adaptations are required for successful in vivo treatments, it appears that these oils may be useful as chemotherapeutic agents against several poultry parasites.

In vitro activity of therapeutic drugs against Histomonas meleagridis (Smith, 1895).

Histomoniasis or blackhead is a life-threatening disease of turkeys that is caused by a flagellated protozoan, Histomonas meleagridis. The development of an assay to measure the sensitivity of drugs traditionally used against this parasite, as reputed to be effective against other protozoan parasites, is described. The in vitro minimum lethal concentrations (MLC), time for drug efficacy, and parasite viability after removal of residual drugs were determined. Three of the 10 tested drugs, fenbendazole, albendazole, and sulfadiazine, were found to be ineffective against H. meleagridis. Nifursol, the only compound still authorized as a feed additive in Europe, is an inhibiting agent but is not lethal in vitro. Roxarsone, an arsenical derivate similar to nitarsone (the only authorized drug in United States), is effective at high concentration (200 microg/mL) after a long exposure (48 h). The lethal activity of dimetridazole, metronidazole, ronidazole, tinidazole, and furazolidone in vitro was demonstrated. Dimetridazole (MLC = 25 microg/mL after 6 h of exposure), metronidazole (MLC = 50 microg/mL after 24 h), and furazolidone (MLC = 50 microg/mL after 24 h) are rapidly effective at low concentrations. These results confirm the effectiveness of dimetridazole, a drug that has been used in the treatment and prevention of blackhead. In May 2002 this compound was removed as feed additive in Europe.

Electron microscopic study on the endogenous development of Eimeria mulardi, Chauve, Reynaud and Gounel, 1994: a coccidium from the mule duck.

An electron microscopic study of the endogenous development of Eimeria mulardi Chauve, Reynaud and Gounel, 1994 was carried out in mule ducks which are hybrids of the domestic duck (Anas platyrhynchos) and the muscovy duck (Cairina moschata). All of the endogenous stages were seen within the nucleus of the host cell. Merozoites arose from ectomerogony and three mutually similar merogonies were noted. The asexual stages were found in leukocyte-like cells in the lamina propria of the jejunum, ileum and caecum, while the gamonts developed in glandular epithelial cells in the same part of the intestine.

First in vitro cycle of the chicken mite, Dermanyssus gallinae (DeGeer 1778), utilizing an artificial feeding device.

The red poultry mite, Dermanyssus gallinae, is one of the most economically deleterious ecto-parasites of layer hens worldwide. D. gallinae is difficult to eliminate from infested poultry farms, and even to study, because it resides on the host only during the bloodmeal at night, and hides in the crevices of poultry houses during the day. Here, the life-cycle of D. gallinae was reproduced entirely in vitro. Mites were incubated in a glass pipette at 30 degrees C, 60-95 degrees relative humidity and total darkness. A feeding apparatus, composed of a membrane, reservoir and blood was fitted on the pipette during bloodmeals. We tested feeding rates on blood mixed with 1 of 3 anti-coagulants (EDTA, heparin and trisodium citrate) at different concentrations, and biological and artificial membranes. The best engorgement and survival rates for all 3 haematophagous life-stages of the parasite were observed in 1-day-old chick membranes and heparinized (0.02 mmo/ml) blood. We then describe the steps in developing a complete self-sustaining in vitro life-cycle. A colony of mites was maintained in vitro for 7 generations. Losses in the first generation were heavy, but survival had multiplied 5-fold by the fifth generation. We hypothesize that heavy mortality rates during the first life-cycle correspond to selective pressure: only the mites which fed and survived in vitro were able to reproduce.