The repellent and persistent toxic effects of essential oils against the poultry red mite, Dermanyssus gallinae.

The economic impact of the poultry red mite, Dermanyssus gallinae, the lack of new acaricides, the occurrence of resistance and tighter legislation have all led to the need to find new ways to control this pest. One promising alternative method of control focuses on employing repellent and/or toxic effects of selected plant essential oils against D. gallinae. Ten essential oils (basil, thyme, coriander, eucalyptus, lavender, lemon, fir tree, oregano, mint, and juniper) were tested for the persistence of toxic and repellent effects. In filter-paper toxicity bioassays against D. gallinae, the best results were observed for lavender (more than 97% mortality after 48 and 72 h) and thyme (84% at 72 h) at a dose of 0.12 mg/cm(2). In addition, two oils showed significant persistent toxic effects 15 and 30 days post application to filter papers. Thyme was the most effective (100% mortality at 72 h), followed by lavender (nearly 80% mortality after 72 h). Out of the ten oils tested for their repellent effect, thyme was the strongest, with nearly 80% of the tested area avoided by mites; oregano caused a 60% avoidance and lavender exhibited an effect close to 40%. All other oils exhibited a repellent effect of less than 30%. None of the experiments showed a repellent effect for HM (commercial alimentary oil) or negative controls. We found that the thyme and lavender essential oils exhibited promising results when tested in vitro for toxic and repellent effects against D. gallinae; thus, we suggest that future experiments focus on in vivo tests using these oils in farm units.

Modelling population dynamics and response to management options in the poultry red mite Dermanyssus gallinae (Acari: Dermanyssidae).

The poultry red mite Dermanyssus gallinae is a major pest and widespread ectoparasite of laying hens and other domestic and wild birds. Under optimal conditions, D. gallinae can complete its lifecycle in less than 10 days, leading to rapid proliferation of populations in poultry systems. This paper focuses on developing a theoretical model framework to describe the population dynamics of D. gallinae. This model is then used to test the efficacy and residual effect of different control options for managing D. gallinae. As well as allowing comparison between treatment options, the model also allows comparison of treatment efficacies to different D. gallinae life stages. Three different means for controlling D. gallinae populations were subjected to the model using computer simulations: mechanical cleaning (killing once at a given time all accessible population stages), sanitary clearance (starving the mite population for a given duration, e.g. between flocks) and acaricide treatment (killing a proportion of nymphs and adults during the persistence of the treatment). Simulations showed that mechanical cleaning and sanitary clearance alone could not eradicate the model D. gallinae population, although these methods did delay population establishment. In contrast, the complete eradication of the model D. gallinae population was achieved by several successive acaricide treatments in close succession, even when a relatively low treatment level was used.

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.

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.

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.

Histomonas meleagridis in turkeys: dissemination kinetics in host tissues after cloacal infection.

Histomonas meleagridis is a flagellated protozoa causing histomoniasis, a disease of gallinaceous fowl. This disease is characterized by necrotic typhlitis, hepatitis, and high mortality, especially in turkeys. In an attempt to detect the progression of H. meleagridis in the turkey, birds were infected via the cloaca. Between d 0 and 19, a group of 4 turkeys was killed and autopsied every 3 d. Cecal and hepatic lesion scores were used to measure severity of infection. For each turkey, 15 tissue samples were taken. Another group of 3 infected turkeys were placed separately, and samples of cecal and intestinal stool were collected every 3 d. Samples were analyzed by PCR. For samples of cecal droppings, cecum, cecal content, rectum, proventriculus, and bursa of Fabricius, the number of birds detected as positive by PCR followed the evolution of the lesion scores. Within the liver, the parasite DNA was detected only in some severe lesions. The parasite DNA was also detected in duodenum, jejuno-ileum, spleen, heart, lungs, and brain samples. The parasite DNA was not detected in the blood, kidneys, pancreas, or muscle of the thigh. Results of the PCR were in agreement with the evolution of the clinical signs and of the cecal and liver lesions.

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.

Phylogenetic position of the trichomonad parasite of turkeys, Histomonas meleagridis (Smith) Tyzzer, inferred from small subunit rRNA sequence.

The phylogenetic position of the trichomonad, Histomonas meleagridis was determined by analysis of small subunit rRNAs. Molecular trees including all identified parabasalid sequences available in data bases were inferred by distance, parsimony, and likelihood methods. All reveal a close relationship between H. meleagridis, and Dientamoeba fragilis. Moreover, small subunit rRNAs of both amoeboid species have a reduced G + C content and increased chain length relative to other parabasalids. Finally, the rRNA genes from H. meleagridis and D. fragilis share a recent common ancestor with Tritrichomonasfoetus, which exhibits a more developed cytoskeleton. This indicates that Histomonas and Dientamoeba secondarily lost most of the typical trichomonad cytoskeletal structures and hence, do not represent primitive morphologies. A global phylogeny of parabasalids revealed significant discrepancies with morphology-based classifications, such as the polyphyly of most of the parabasalid families and classes included in our study.