Concurrent Histomonas meleagridis and Hemorrhagic Enteritis Virus Infection in a Turkey Flock with Recurrent History of Blackhead Disease.

Intestinal health is one of the key factors required for the growth and production of turkeys. Histomoniasis (blackhead disease), caused by a protozoan parasite, Histomonas meleagridis, is a reemerging threat to the turkey industry. Increased incidences of histomoniasis have been reported in recent years due to withdrawal of antihistomonas treatments. H. meleagridis affects ceca and causes cecal inflammation and necrosis. H. meleagridis migrates from ceca to the liver and causes liver necrosis, resulting in high mortalities. Ironically, field outbreaks of histomoniasis are not always associated with high mortalities, while low mortalities have also been documented. There are several exacerbating factors associated with high mortality rates in histomoniasis outbreaks, with concurrent infection being one of them. Recurrent histomoniasis outbreaks in a newly constructed barn were documented, and concurrent infection of H. meleagridis and hemorrhagic enteritis virus was confirmed. Currently, neither commercial vaccines nor prophylactic or therapeutic solutions are available to combat histomoniasis. However, there are treatments, vaccines, and solutions to minimize or prevent concurrent infections in turkeys. In addition to implementing biosecurity measures, measures to prevent concurrent infections are critical steps that the turkey industry can follow to reduce mortality rates and minimize the production and economic losses associated with histomoniasis outbreaks.

Infección simultánea por Histomonas meleagridis y el virus de la enteritis hemorrágica en una parvada de pavos con antecedentes recurrentes de enfermedad de la cabeza negra. La salud intestinal es uno de los factores clave necesarios para el crecimiento y producción de los pavos. La histomoniasis (enfermedad de la cabeza negra), causada por un parásito protozoario, Histomonas meleagridis, es una amenaza reemergente para la industria del pavo. En los últimos años se ha informado de un aumento de la incidencia de histomoniasis debido al retiro de los tratamientos con antihistomonas. Histomonas meleagridis afecta los ciegos y causa inflamación y necrosis cecal. Histomonas meleagridis migra desde los ciegos al hígado y causa necrosis hepática, lo que resulta en una alta mortalidad. Irónicamente, los brotes de histomoniasis en el campo no siempre se asocian con una mortalidad elevada, aunque también se han documentado mortalidades bajas. Hay varios factores exacerbantes asociados con altas tasas de mortalidad en los brotes de histomoniasis, siendo la infección concurrente uno de ellos. Se documentaron brotes recurrentes de histomoniasis en un alojamiento avícola recién construido y se confirmó la infección concurrente de H. meleagridis y el virus de la enteritis hemorrágica. Actualmente no se dis-pone de vacunas comerciales ni soluciones profilácticas o terapéuticas para combatir la histomoniasis. Sin embargo, existen tratamientos, vacunas y soluciones para minimizar o prevenir infecciones concurrentes en los pavos. Además de implementar medidas de bioseguridad, las medidas para prevenir infecciones concurrentes son pasos críticos que la industria del pavo puede seguir para reducir las tasas de mortalidad y minimizar las pérdidas económicas y de producción asociadas con los brotes de histomoniasis.

Pathologic Characterization of Coinfection with Histomonas meleagridis, Marek's Disease Virus, and Subtype J Avian Leukosis Virus in Chickens.

Histomonas meleagridis is a trichomonad protozoan parasite that can cause an important poultry disease known as histomoniasis; Marek's disease virus (MDV) and subtype J avian leukosis virus (ALV-J) usually cause avian oncogenic diseases. Although these diseases have been reported in a single pathogen infection, information about their coinfection is scarce. This study reports a naturally occurring case of coinfection with H. meleagridis, MDV, and ALV-J in a local chicken flock at the age of 150 days. Necropsy revealed necrosis and swelling in the liver and spleen. Histologic analysis showed large areas of mild to severe necrosis of hepatocytes, with numerous intralesional trophozoites of H. meleagridis by H&E and periodic acid-Schiff staining; H&E staining showed pleomorphic and neoplastic lymphoid tumor cells in the liver and myeloid cells with eosinophilic cytoplasmic granules in the spleen. Coexpression of MDV and ALV-J antigens was detected in the liver by fluorescence multiplex immunohistochemistry staining. The 18S rRNA gene of H. meleagridis, meq gene of MDV, and gp85 gene of ALV-J were identified in mixed liver and spleen tissues by PCR and sequencing, respectively.

Reporte de caso­Caracterización patológica de la coinfección con Histomonas meleagridis, el virus de la enfermedad de Marek y el virus de la leucosis aviar subtipo J en pollos Histomonas meleagridis es un parásito protozoario tricomonial que puede causar una enfermedad avícola importante conocida como histomoniasis; El virus de la enfermedad de Marek (MDV) y el virus de la leucosis aviar subtipo J (ALV-J) suelen causar enfermedades oncogénicas aviares. Aunque estas enfermedades se han reportado como infecciones patógenas separadas, la información sobre coinfección es escasa. Este estudio reporta un caso natural de coinfección con H. meleagridis, el virus de la enfermedad de Marek y el virus de la leucosis aviar subtipo J en una parvada de pollos local a la edad de 150 días. La necropsia reveló necrosis e inflamación del hígado y el bazo. El análisis histológico mostró grandes áreas de necrosis de hepatocitos de leve a severa, con numerosos trofozoítos intralesionales de H. meleagridis por tinción de hematoxilina y eosina y por tinción de ácido periódico-Schiff. La tinción de hematoxilina y eosina mostró células linfoides neoplásicas y pleomórficas en el hígado y en el bazo presencia de células mieloides con gránulos citoplásmicos eosinofílicos. La coexpresión de antígenos del virus de Marek y de la leucosis aviar subtipo J se detectó en el hígado mediante tinción inmunohistoquímica de fluorescencia múltiple. El gene de ARNr 18S de H. meleagridis, el gene meq del virus de Marek y el gene gp85 del virus de la leucosis aviar subtipo J se identificaron en tejidos mixtos de hígado y bazo mediante PCR y secuenciación, respectivamente.

Sporadic isolation of Tetratrichomonas species from the cattle urogenital tract.

Tritrichomonas foetus is a venereal trichomonad parasite which causes reproductive issues in cattle. No other trichomonads are known to be urogenital pathogens in cattle, but there are several reports of Tetratrichomonas and Pentatrichomonas isolates of unclear origin from the cattle urogenital tract (UGT) in the Americas. This study reports the first case of a non-T. foetus cattle urogenital trichomonad isolate in Europe. Molecular analysis of the internal transcribed spacer (ITS) 1-5.8S ribosomal RNA-ITS 2 and 18S ribosomal RNA loci suggest that the isolate is a Tetratrichomonas species from a lineage containing other previously described bull preputial isolates. We identified close sequence similarity between published urogenital and gastrointestinal Tetratrichomonas spp., and this is reviewed alongside further evidence regarding the gastrointestinal origin of non-T. foetus isolates. Routine screening for T. foetus is based on culture and identification by microscopy, and so considering other trichomonad parasites of the bovine UGT is important to avoid misdiagnosis.

Excretion of Histomonas meleagridis following experimental co-infection of distinct chicken lines with Heterakis gallinarum and Ascaridia galli.

BACKGROUND: Histomonosis is a severe re-emerging disease of poultry caused by Histomonas meleagridis, a protozoan parasite which survives in the environment via the cecal worm Heterakis gallinarum. Following infection, the parasites reside in the ceca and are excreted via host feces. In the present work, male birds of conventional broiler (Ross 308, R), layer (Lohmann Brown Plus, LB) and a dual-purpose (Lohmann Dual, LD) chicken line were infected with 250 embryonated eggs of Ascaridia galli and Heterakis gallinarum, respectively, with the latter nematode harboring Histomonas meleagridis, to investigate a co-infection of nematodes with the protozoan parasite in different host lines. METHODS: In weekly intervals, from 2 to 9 weeks post infection (wpi), individual fecal samples (n = 234) from the chickens were collected to quantify the excretion of H. meleagridis by real-time PCR and to determine the number of nematode eggs per gram (EPG) in order to elucidate excretion dynamics of the flagellate and the nematodes. This was further investigated by indirect detection using plasma samples of the birds to detect antibodies specific for H. meleagridis and worms by ELISA. The infection with H. meleagridis was confirmed by histopathology and immunohistochemistry to detect the flagellate in the cecum of representing birds. RESULTS: The excretion of H. meleagridis could already be observed from the 2nd wpi in some birds and increased to 100% in the last week of the experiment in all groups independent of the genetic line. This increase could be confirmed by ELISA, even though the number of excreted H. meleagridis per bird was generally low. Overall, histomonads were detected in 60% to 78% of birds with temporary differences between the different genetic lines, which also showed variations in the EPG and worm burden of both nematodes. CONCLUSIONS: The infection with H. gallinarum eggs contaminated with H. meleagridis led to a permanent excretion of the flagellate in host feces. Differences in the excretion of H. meleagridis in the feces of genetically different host lines occurred intermittently. The excretion of the protozoan or its vector H. gallinarum was mostly exclusive, showing a negative interaction between the two parasites in the same host.

Lethal infection caused by Tetratrichomonas gallinarum in black swans (Cygnus atratus).

BACKGROUND: Tetratrichomonas gallinarum is parasitic protozoa with a wide host range. However, its lethal infection is rare reported. CASE PRESENTATION: Here, we described the first lethal cases of T. gallinarum infection in black swans in China. Five black swans died within a week in succession without obvious symptoms except mild diarrhea. At necropsy, severe lesions were observed in caeca with thickened caecal walls and hemorrhages in the mucosa. A large number of moving trophozoites were found in the contents of the cecum by microscopic examination. The livers were enlarged with multiple bleeding spots on the surface. Histopathology of the livers showed mononuclear cell infiltration and moderate hyperplasia of fibrous tissue. The histopathology of the cecum showed that the villi of the cecum were edematous. Finally, the presence of T. gallinarum was determined by specific PCR andin-situ hybridization assay. Additionally, common pathogens that can cause similar symptoms were excluded. CONCLUSIONS: The death of the black swan was caused by T. gallinarum, suggesting that the parasite might be a new threat to the Cygnus birds.

The Prevalence of Tetratrichomonas spp. in Reproductive Geese Flocks.

Tetratrichomonas is a genus of parasites that usually inhabits the lower digestive tract, especially the cecum, of various bird species. The infection might lead to birds' death, but in many cases it could be asymptomatic or with mild clinical signs which might be not observed. Subclinical infections can be undiagnosed, leading to production losses. To investigate the prevalence of Tetratrichomonas spp. in geese, 23 cloacal swabs were taken from each of 43 flocks of reproductive geese from five major geese production provinces in Poland after first, second, or third laying season. The obtained swabs were placed in culture medium for propagation of the parasite. All cultures were screened microscopically before PCR was applied to detect the parasites' DNA. After cultivation, the presence of genetic material of Tetratrichomonas was found in 430 out of 989 samples, which correspond to 38 (88.4%) of 43 flocks. The study shows how the number of laying seasons and the size of the flock in which the birds were kept affects the distribution of protozoa of the genus Tetratrichomonas in geese reproductive flocks in Poland.

Prevalence of Tetratrichomonas buttreyi and Pentatrichomonas hominis in yellow cattle, dairy cattle, and water buffalo in China.

The trichomonad species Tetratrichomonas buttreyi and Pentatrichomonas hominis have been reported in the bovine digestive tract in only a few studies, and the prevalence and pathogenicity of these two protists in cattle herds remain unknown. In this study, the prevalence of T. buttreyi and P. hominis in yellow cattle, dairy cattle, and water buffalo in Anhui Province, China, was determined with a PCR analysis of the small subunit ribosomal RNA genes. The overall infection rates for T. buttreyi and P. hominis were 8.1% and 5.4%, respectively. Double infections were found in 15 (1.6%) samples from four farms. The prevalence of P. hominis in cattle with abnormal feces was significantly higher than that in cattle with normal feces (χ2 = 13.0, p < 0.01), and the prevalence of T. buttreyi in the northern region of Anhui Province was also significantly higher than that in the mid region (χ2 = 16.6, p < 0.01). Minor allelic variations were detected in the T. buttreyi isolates from cattle in this study, as in other hosts in previous studies. Morphological observations, together with the PCR analysis, demonstrated that the trichomonads isolated in this study were P. hominis. The presence of T. buttreyi and P. hominis indicated that cattle are natural hosts of these two trichomonads and could be a potential source of P. hominis infections in humans and other animal hosts.

Morphological and molecular characterization of a species of Tetratrichomonas present in feces of Brazilian sheep (Ovis aries) and goats (Capra hircus).

The trichomonads form part of the phylum Parabasalia, a complex assemblage of diverse species of flagellated protists, with some members recognized as pathogens of men and/or animals. Associations, probably as commensals, between the species Tetratrichomonas ovis and sheep were reported in North America during the 1960s based on morphological and cultural characteristics. Intriguingly, no subsequent studies of this topic have been published. Feces, collected from sheep (n = 55) and goats (n = 14), reared on small-scale, production facilities in Southeastern Brazil, were examined for parabasalids. Protozoa, demonstrating morphologies and motility characteristic of trichomonads, were detected by direct microscopy in 64% of sheep and 43% of goat samples. In contrast to T. ovis, none of the samples could be cultured in Diamond's medium; however, cultures were obtained for three goat and seventeen sheep samples in peptonized broth. Based on morphological analyses, all isolates were classified as members of the genus Tetratrichomonas. Sequencing of the ITS1-5.8S rRNA gene-ITS2 region revealed three highly similar genotypes that were essentially identical to sequences reported for Tetratrichomonas spp. isolated from the preputial cavity of cattle in the USA and Southern Brazil. The findings of this study extend and enhance our knowledge of parasitism in small ruminants by parabasalids.

High prevalence of Pentatrichomonas hominis infection in gastrointestinal cancer patients.

BACKGROUND: Pentatrichomonas hominis is a flagellated protozoan that inhabits the large intestine of humans. Although several protozoans have been proposed to have a role in cancer progression, little is known about the epidemiology of P. hominis infection in cancer patients. METHODS: To determine the prevalence of P. hominis in patients with digestive system malignancies, we collected 195 and 142 fecal samples from gastrointestinal cancer patients and residents without any complaints related to the digestive system, respectively. Each sample was detected for the presence of P. hominis by nested PCR amplifying the internal transcribed spacer (ITS) region and partial 18S rRNA gene. RESULTS: A significantly higher prevalence of P. hominis was found in cancer patients than that in the control population (41.54 vs 9.15%, χ2 = 42.84, df = 1, P < 0.001), resulting in a 6.75-fold risk of gastrointestinal cancers (OR: 6.75, 95% CI: 3.55-12.83, P < 0.001). The highest prevalence of P. hominis infection was detected in small intestine cancer patients (60%, OR: 14.88, 95% CI: 0.82-4.58, P = 0.009) followed by liver (57.14%, χ2 = 10.82, df = 1, P = 0.001) and stomach cancer patients (45.1%, χ2 = 31.95, df = 1, P < 0.001). In addition, phylogenetic analysis provided some evidence supporting that human P. hominis infection might derive from animal sources. CONCLUSIONS: To our knowledge, this study is the first report presenting the high association between P. hominis and gastrointestinal cancers. Nevertheless, whether there is any possible pathological role of P. hominis infection in cancer patients needs to be further elucidated.

Metagenomics for broad and improved parasite detection: a proof-of-concept study using swine faecal samples.

Efficient and reliable identification of emerging pathogens is crucial for the design and implementation of timely and proportionate control strategies. This is difficult if the pathogen is so far unknown or only distantly related with known pathogens. Diagnostic metagenomics - an undirected, broad and sensitive method for the efficient identification of pathogens - was frequently used for virus and bacteria detection, but seldom applied to parasite identification. Here, metagenomics datasets prepared from swine faeces using an unbiased sample processing approach with RNA serving as starting material were re-analysed with respect to parasite detection. The taxonomic identification tool RIEMS, used for initial detection, provided basic hints on potential pathogens contained in the datasets. The suspected parasites/intestinal protists (Blastocystis, Entamoeba, Iodamoeba, Neobalantidium, Tetratrichomonas) were verified using subsequently applied reference mapping analyses on the base of rRNA sequences. Nearly full-length gene sequences could be extracted from the RNA-derived datasets. In the case of Blastocystis, subtyping was possible with subtype (ST)15 discovered for the first known time in swine faeces. Using RIEMS, some of the suspected candidates turned out to be false-positives caused by the poor status of sequences in publicly available databases. Altogether, 11 different species/STs of parasites/intestinal protists were detected in 34 out of 41 datasets extracted from metagenomics data. The approach operates without any primer bias that typically hampers the analysis of amplicon-based approaches, and allows the detection and taxonomic classification including subtyping of protist and metazoan endobionts (parasites, commensals or mutualists) based on an abundant biomarker, the 18S rRNA. The generic nature of the approach also allows evaluation of interdependencies that induce mutualistic or pathogenic effects that are often not clear for many intestinal protists and perhaps other parasites. Thus, metagenomics has the potential for generic pathogen identification beyond the characterisation of viruses and bacteria when starting from RNA instead of DNA.

PCR and serology confirm the infection of turkey hens and their resilience to histomonosis in mixed flocks following high mortalities in toms.

BACKGROUND: Histomonosis, caused by the protozoan parasite Histomonas meleagridis, is a severe disease especially in turkeys where it can cause high mortalities. Recently, outbreaks were described in which turkey hens showed no clinical signs despite high mortalities in toms, from which they were separated only by a wire fence. The present study investigated three similar outbreaks of histomonosis whereby in two of them only a few hens were being affected and none in the third. Hens from all flocks were kept until end of production and slaughtered as scheduled. However, in all three cases, the disease progressed in toms reaching nearly 100% within two weeks. METHODS: Following diagnosis of the disease, tissue samples were obtained from toms and hens at necropsy. Environmental dust, cloacal swabs and blood were taken on three successive farm visits within compartments of hens and toms and tested by real-time PCR or ELISA. The DNA from a total of 18 samples positive for H. meleagridis was further subjected to conventional PCR utilizing the 18S rRNA primers and sequenced for phylogenetic analysis. RESULTS: All tissue samples and some cloacal swabs were tested positive. Dust samples confirmed the presence of H. meleagridis DNA that spread within entire houses up to 6 weeks after the first clinical signs of histomonosis. Sequence analysis of the 18S rRNA locus demonstrated the presence of the same strain in birds of both sexes within each of the turkey houses. Investigation of serum samples two weeks post-initial diagnosis and prior to euthanasia resulted in antibody detection in 73% of toms and 70% of hens. Until the end of the investigation the number of positive hens per farm increased up to 100% with mean OD-values approaching those noticed in toms prior to euthanasia. CONCLUSIONS: For the first time it could be demonstrated that turkey hens kept in the same house as toms became infected during fatal outbreaks in toms. This highlights the value of different diagnostics methods in order to trace the parasite in connection with the host response. The strange phenomenon that only single hens succumb to the diseases despite being infected requires further investigations.

Tetratrichomonas in pyopneumothorax.

Pleural trichomonosis is clinically rare, and very few cases of trichomonal empyema have been reported so far. A rare case of an 81-year-old woman with pyopeumothorax presenting with recurrent fever and macroscopic pyuria was present. Microscopic examination of the pleural effusion showed mobile flagellated protozoa which molecular methods identified as Tetratrichomonas. In addition, Streptococcus anginosus was discovered in pleural fluid cultures. Treatment with imipenem/cilastatin and metronidazole successfully eliminated the pathogens and led to relief of clinical symptoms. In the context of a review of the relevant literature, the clinical application of molecular methods in the diagnosis of pleural trichomonosis is underlined.

Typhlohepatitis and Amyloidosis Associated with High Mortality in Chukar Partridges (Alectoris chukar).

Twelve chukar partridges (Alectoris chukar) from a farm experiencing poor uniformity and increased mortality of up to 65% were submitted for diagnosis. Several birds had mild to moderate multifocal white foci or multifocal petechial hemorrhages throughout the liver. Livers and spleens of older birds were moderate to severely diffusely enlarged. In addition, some birds had caseous cores mixed with blood within the ceca as well as segmentally thickened cecal walls. Histopathology showed acute, multifocal, severe, often coalescing foci of necrosis with accumulation of fibrin and/or fibrinosuppurative inflammation in livers and spleens. Scattered within exudate were protozoa that were spherical or round and measured 12-20 µm in diameter. In the ceca, acute necrosis of the mucosa was observed, often with ulceration and fibrinosuppurative inflammation. Immunohistochemistry using an antiserum against Tritrichomonas foetus revealed round protozoa in ceca, small intestines, liver, spleen, and lung. Quantitative PCR to detect DNA of Histomonas meleagridis was negative. Non-species-specific PCRs amplifying the partial rDNA, the internal transcribed spacer (ITS) region, and the partial beta-tubulin gene yielded products of the expected size. Sequences of the PCR products had the highest homology to sequences of Tetratrichomonas gallinarum and less homology to sequences of H. meleagridis. In addition there was accumulation of amyloid in the space of Disse in the liver, splenic sinuses, and walls of the blood vessels. The typhlohepatitis and other inflammatory processes that were diagnosed might be the underlying cause of the amyloidosis. Other findings were clusters of Clostridium perfringens associated with the lesions in the ceca; multifocal granulomas in the lungs, occasionally associated with fungal hyphae; hyperkeratosis associated with bacteria and Candida sp. cells in the crop; mild infection of the bursal mucosa with Cryptosporidium.

Tiflohepatitis y amiloidosis asociadas con alta mortalidad en perdices chukar (Alectoris chukar). Doce perdices chukar (Alectoris chukar) de una granja con baja uniformidad y alta mortalidad de hasta el 65% se presentaron para diagnóstico. Varias aves presentaron áreas blancas multifocales de leves a moderadas o hemorragias petequiales multifocales en todo el hígado. Los hígados y los bazos de las aves con mayor edad estuvieron agrandados de tamaño de manera difusa y de moderado a severo. Además, algunas aves tenían contenidos caseosos mezclados con sangre dentro de los ciegos, así como paredes cecales engrosadas de manera segmentaria. La histopatología mostró focos de necrosis agudos, multifocales, graves, a menudo coalescentes con acumulación de fibrina y/o inflamación fibrinosupurativa en hígados y bazos. Dispersos dentro del exudado se encontraban protozoarios que eran esféricos o redondos y que medían de 12 a 20 µm de diámetro. En el ciego, se observó necrosis aguda de la mucosa, a menudo con ulceración e inflamación fibrinosupurativa. La inmunohistoquímica con un antisuero contra Tritrichomonas foetus reveló protozoarios redondos en el ciego, intestino delgado, hígado, bazo y pulmón. El método de PCR cuantitativo para detectar el ADN de Histomonas meleagridis fue negativo. Los métodos de PCR no específicos de especie que amplifican parcialmente al rDNA de la región espaciadora transcrita interna (ITS) y el gene parcial de la beta-tubulina dieron productos del tamaño esperado. Las secuencias de los productos de PCR tuvieron la mayor similitud con las secuencias de Tetratrichomonas gallinarum y menos similitud con las secuencias de H. meleagridis. Además, se observó acumulación de amiloide en el espacio de Disse en el hígado, en senos esplénicos y paredes de los vasos sanguíneos. La tiflohepatitis y otros procesos inflamatorios que se diagnosticaron pueden ser la causa subyacente de la amiloidosis. Otros hallazgos incluyeron grupos de Clostridium perfringens asociados con las lesiones en el ciego; granulomas multifocales en los pulmones, ocasionalmente asociados con hifas fúngicas; hiperqueratosis asociada a bacterias y Candida spp. en el buche, e infección leve de la mucosa bursal con Cryptosporidium.

Epidemic Characteristics of Clinical Histomoniasis in Chicken Flocks in Eastern China.

Preliminary diagnosis of clinical symptoms and gross lesions with subsequent histopathologic and PCR analyses revealed histomoniasis in 276 chicken flocks in Jiangsu Province, China, and surrounding areas from January 2012 to December 2015. Detailed statistical analysis was performed to explore the occurrence and epidemic characteristics of histomoniasis in chicken flocks. The results indicated that histomoniasis usually occurred in free-range flocks of local broilers and laying hens. Also, 2- to 3-mo-old chickens were most susceptible to infection, and adult chickens rarely developed infection. The morbidity rate of chickens was generally 10%-30%, with mortality rates of less than 10%. Moreover, histomoniasis is a seasonal disease, occurring most often from April to June, and the rate of coinfection with heterakids in the ceca of infected chicken was 50.8%. The symptoms of diseased chickens included mental fatigue, bowing of the head and wings, and yellowish green droppings, with bloody stool in very limited cases. Most of the pathologic changes were characteristic of the disease, but there were also some atypical lesions confirmed by laboratory techniques. In the current study, the histomoniasis epidemic was first investigated in Chinese chicken flocks, and the results provided a useful reference for further study of this disease.

Retrospective Study of Histomoniasis (Blackhead) in California Turkey Flocks, 2000-2014.

The flagellate parasite Histomonas meleagridis causes a severe disease in turkeys. Since nitarsone, the last antiflagellate feed additive, was withdrawn from the market, H. meleagridis has gained increasing clinical and economic importance, and frequently entire turkey flocks are lost to the disease. Even before the antiflagellate market withdrawal, H. meleagridis has been recognized as a noteworthy disease in commercial turkeys. The aim of this study was to compile and analyze data on histomoniasis outbreaks diagnosed by the California Animal Health and Food Safety System in the years 2000 through 2014 while nitarsone was still available. Sixty-two cases were included in the study. In all cases, the parasite was detected by histopathology. Five cases were from breeder flocks, 44 from commercial meat turkey flocks, and 13 from other flocks such as backyard flocks or pet turkeys. There were between one and eight cases each year with no clear trend over time. Most cases occurred in the warmer months between April and October with a clear peak in August. Turkeys in the flocks with the disease were aged between 2 wk and 15 mo, with a median age of 9 wk. In cases for which mortality was reported, daily mortality ranged between 0.04% and 2% with an average of 0.34% and a median of 0.15%. Five-week mortality in infected houses on three infected ranches was between 1.3% and 68%. In 12 cases, the parasite was observed in organs other than ceca and liver. These included spleen, kidney, bursa of Fabricius, proventriculus, lung, pancreas, and crop. In 58 cases, histomoniasis was considered the most significant and primary diagnosis; in four cases histomoniasis was regarded as an incidental finding. In 14 other outbreaks, detailed information was available. In 11 cases, more than one house on the ranch was affected; in five cases the disease spread to all houses. Total mortality in turkeys from affected flocks ranged between 12% and 65%. In eight flocks, the birds were marketed with a weight that was considered too light. In conclusion, the present study shows that even with nitarsone available as antiflagellate feed, additive histomoniasis was a serious disease of turkeys in the United States with an epidemiology similar to that observed in Europe.

Tetratrichomonas and Trichomonas spp.-Associated Disease in Free-Ranging Common Eiders ( Somateria mollissima) from Wellfleet Bay, MA and Description of ITS1 Region Genotypes.

During an outbreak of Wellfleet Bay virus (WFBV) in common eiders ( Somateria mollissima) from the Cape Cod region of Massachusetts, several birds were diagnosed with trichomonosis consisting of multiple trichomonad species. Six birds were examined, with trichomonads found in ceca in four birds and associated typhlitis in three of these four birds. PCR and DNA sequencing utilizing trichomonad-specific primers targeting the ITS1 region of the ribosomal DNA (rDNA) revealed the presence of Tetratrichomonas gallinarum in the gastrointestinal tracts of five birds and Trichomonas spp. in the livers of two birds, one of which had a dual Te. gallinarum-Trichomonas gallinae infection. Sequence analysis revealed no variation between Te. gallinarum sequences whereas the ITS1 sequences obtained from the other Trichomonas spp. demonstrated the presence of multiple genotypes. One sequence had 100% identity to a Trichomonas sp. previously isolated from a Cooper's hawk ( Accipiter cooperii) and the other sequence was 100% identical to a previously described Tr. gallinae isolate obtained from a Pacific Coast band-tailed pigeon ( Patagioenas fasciata monilis). These findings suggest Te. gallinarum and other Trichomonas spp. possibly contributed to morbidity and mortality in this species. Furthermore, to the authors' knowledge, this is the first report of trichomonad-associated disease in a free-ranging sea duck.

Occurrence of Blastocystis sp. and Pentatrichomonas hominis in sheep and goats in China.

BACKGROUND: Global data regarding the molecular epidemiology of Blastocystis sp. and Pentatrichomonas hominis in sheep and goats are sparse. China has one of the largest sheep and goat populations in the world. In this study we investigated the occurrence of Blastocystis sp. and P. hominis in domestic sheep and goats in China, and analyzed the genetic characterization of these two parasite species. METHODS: In total, we collected fresh fecal samples from 832 sheep and 781 goats located on seven and ten farms, respectively, in the central eastern region of China. The corresponding sequences obtained in this study were subject to molecular analysis for subtype and allele identification of Blastocystis sp., and species and genotype confirmation of P. hominis. RESULTS: The occurrence of Blastocystis sp. was 6.0% (50/832) in sheep and 0.3% (2/781) in goats. The most predominant subtype (ST) of Blastocystis sp. in sheep was ST10 (50.0%), followed by ST14 (20%), ST5 (16%), novel sequence 1 (6%), novel sequence 4 (4%), novel sequence 2 (2%) and novel sequence 3 (2%). However, only ST1 was observed in goats. No mixed infections with different subtypes were found in this study. The 18S alleles showed allele 2 (100%) for ST1; allele 115 (75%) for ST5; and no match allele for ST5 (25%), ST10 (100%), ST14 (100%), novel sequence 1 (100%), novel sequence 2 (100%), novel sequence 3 (100%), and novel sequence 4 (100%) on the Blastocystis subtype (18S) and Sequence Typing (MLST) database. For P. hominis, two goats (0.3%) and zero sheep (0%) were identified as positive in this study. The 18S rRNA gene sequences of two P. hominis isolates from goats displayed 100% identity to type CC1, found previously in dogs, monkeys and humans. CONCLUSIONS: These results provide the detailed data on the occurrence and molecular epidemiology of Blastocystis sp. and P. hominis in sheep and goats in China. They also contribute to and expand our knowledge of the Blastocystis sp. and P. hominis epidemiology around the world.

An Outbreak of Histomoniasis in Backyard Sanhuang Chickens.

Histomonas meleagridis is a facultative anaerobic parasite, which can cause a common poultry disease known as histomoniasis. The species and age of the birds impacts on the susceptibility, with turkey being the most susceptible species. Chickens are less susceptible to H. meleagridis than turkeys and usually serve as reservoir hosts. Here, the diagnosis of an outbreak of histomoniasis in backyard Sanhuang chickens is described. The primary diagnosis was made based on clinical symptoms, general changes at necropsy, histopathology, and the isolation and cultivation of parasites. The pathogen was further confirmed by cloning, PCR identification, and animal inoculation tests. A strain of H. meleagri- dis, named HM-JSYZ-C, with a higher pathogenicity level in chickens was obtained. The study lays a foundation for further investigations into H. meleagridis and histomoniasis in chickens.

Systemic Histomoniasis in a Leucistic Indian Peafowl (Pavo cristatus) from Southern Brazil.

The pathological and molecular findings associated with Histomonas meleagridis are described in a leucistic Indian peafowl (Pavo cristatus) from Southern Brazil. The most significant gross findings were multifocal necrotizing hepatitis and diphtheric typhlitis. Histopathologic evaluation of the liver, ceca, kidney, spleen, and small intestine revealed systemic histomoniasis (SH) associated with intralesional and intravascular accumulations of histomonad organisms consistent with H. meleagridis. PCR was used to amplify the DNA of H. meleagridis from the liver, ceca, small intestine, spleen, lungs, and kidneys. Direct sequencing and phylogenetic analyses confirmed that the isolate of the flagellated trichomonad identified from this investigation is more phylogenetically related to H. meleagridis than Tetratrichomonas gallinarum, Tritrichomonas foetus, and Dientamoeba fragilis. These results confirmed the occurrence of SH in this peafowl and add to the diagnosis of this disease in birds from Brazil. This report might represent the first complete identification of spontaneous histomoniasis in a peafowl due to pathological and molecular characteristics and one of the few documented cases of SH in non-commercial birds.

Prevalence of Pentatrichomonas hominis infections in six farmed wildlife species in Jilin, China.

Pentatrichomonas hominis is an anaerobic flagellated protozoan that primarily parasitizes the gastrointestinal tract and is a conditional pathogen. It has an extensive host range and is well known as a potential causative agent of zoonotic disease. The objective of this study was to provide the first findings of the prevalence of P. hominis in six farmed wildlife species, sika deer (S.D.), Rex rabbits (R.R.), blue foxes (B.F.), silver foxes (S.F.), raccoon dogs (R.D.) and minks (M.), that are commercially important in Jilin Province, China. In this study, 450 faecal samples were tested for P. hominis infection by culturing and nested PCR assays. The average prevalence of P. hominis infections were as follows: S.D. 20% (26/130), R.R. 16.25% (13/80), B.F. 45% (27/60), S.F. 43.33% (26/60), R.D. 53.33% (32/60) and M. 48.33% (29/60). The prevalence in herbivores (18.57% for S.D. and R.R.) was significantly lower than that in non-herbivores (47.5%). PCR product sequencing indicated that infections were mainly caused by the P. hominis strain Changchun Canine 1, and we found a P. hominis strain with a mutated sequence, Changchun-RR, which had three mutations compared with the referenced homologous P. hominis sequences. Morphological observations of the Changchun-RR strain showed that it was similar to P. hominis. Our study suggests that P. hominis is widespread in six farmed wildlife species in Jilin Province and provides baseline information for the presence of this parasite in these animals.