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.

The complexity of parasites and their life cycles makes vaccination against parasitic diseases challenging. This review highlights this by discussing vaccination against four relevant parasites of poultry. Coccidia, i.e., Eimeria spp., are the most important parasites in poultry production, causing multiple billions of dollars of damage worldwide. Due to the trend of antibiotic-free broiler production, use of anticoccidia vaccines in broilers is becoming much more important. As of now, only live vaccines are on the market, almost all of which must be produced in birds. In addition, these live vaccines require extra care in the management of flocks to provide adequate protection and prevent the vaccines from causing damage. Considerable efforts to develop recombinant vaccines and related work to understand the immune response against coccidia have not yet resulted in an alternative. Leucozytozoon caulleryi is a blood parasite that is prevalent in East and South Asia. It is the only poultry parasite for which a recombinant vaccine has been developed and brought to market. Histomonas meleagridis causes typhlohepatitis in chickens and turkeys. The systemic immune response after intramuscular vaccination with inactivated parasites is not protective. The parasite can be grown and attenuated in vitro, but only together with bacteria. This and the necessary intracloacal application make the use of live vaccines difficult. So far, there have been no attempts to develop a recombinant vaccine against H. meleagridis. Inactivated vaccines inducing antibodies against the poultry red mite Dermanyssus gallinae have the potential to control infestations with this parasite. Potential antigens for recombinant vaccines have been identified, but the use of whole-mite extracts yields superior results. In conclusion, while every parasite is unique, development of vaccines against them shares common problems, namely the difficulties of propagating them in vitro and the identification of protective antigens that might be used in recombinant vaccines.
Estudio recapitulativo- Vacunación contra los parásitos de las aves de corral. La complejidad de los parásitos y sus ciclos de vida hace que la vacunación contra las enfermedades parasitarias sea un desafío. Esta revisión destaca este concepto al discutir la vacunación contra cuatro parásitos relevantes en la avicultura. Las coccidias, como, Eimeria spp., son los parásitos más importantes en la producción avícola y causan daños por miles de millones de dólares en todo el mundo. Debido a la tendencia de la producción de pollos de engorde sin antibióticos, el uso de vacunas anticoccidianas en pollos de engorde se está volviendo mucho más importante. Por el momento, sólo hay en el mercado vacunas vivas y casi todas ellas deben producirse en aves. Además, estas vacunas vivas requieren un cuidado especial en el manejo de las parvadas para brindar una protección adecuada y evitar que las vacunas causen daños. Los esfuerzos considerables para desarrollar vacunas recombinantes y los trabajos relacionados para comprender la respuesta inmune contra coccidias aún no han dado como resultado una alternativa. Leucozytozoon caulleryi es un parásito sanguíneo que prevalece en el este y el sur de Asia. Es el único parásito de las aves de corral para el que se ha desarrollado y comercializado una vacuna recombinante. El parásito Histomonas meleagridis causa tiflohepatitis en pollos y pavos. La respuesta inmune sistémica después de la vacunación intramuscular con parásitos inactivados no es protectora. El parásito se puede cultivar y atenuar in vitro, pero sólo junto con bacterias. Esto y la necesaria aplicación intracloacal dificultan el uso de vacunas vivas. Hasta el momento no ha habido intentos de desarrollar una vacuna recombinante contra H. meleagridis. Las vacunas inactivadas que inducen anticuerpos contra el ácaro rojo de las aves Dermanyssus gallinae tienen el potencial de controlar las infestaciones por este parásito. Se han identificado antígenos potenciales para vacunas recombinantes, pero el uso de extractos completos de ácaros produce resultados superiores. En conclusión, si bien cada parásito es único, el desarrollo de vacunas contra ellos comparte problemas comunes, por ejemplo, las dificultades de propagarlos in vitro y la identificación de antígenos protectores que podrían usarse en vacunas recombinantes.

Histomoniasis is a deadly disease of turkeys causing devastating economic losses to the poultry industry. In field outbreaks, a presumptive diagnosis is made based on gross pathology lesions and confirmed by histopathology. An early detection tool with quick turnaround time is needed to prevent the spread of histomoniasis. With this objective, two studies were conducted in turkeys. In Study 1, 40 poults were housed in two pens (20 poults/pen) and challenged at 14 days of age with Histomonas meleagridis by intracloacal route. Blood samples were collected 4 days postchallenge. Fifty-five percent (22/40) of the blood samples tested positive for H. meleagridis based on PCR using primers targeted against the 18S rRNA gene and confirmed by sequencing. In Study 2, 40 poults were housed in two groups and raised in floor pens. Groups 1 and 2 served as negative and challenge controls, respectively. At 14 days of age, the birds in Group 2 were challenged with H. meleagridis by intracloacal route. Blood samples were collected 2 days postchallenge. Five percent (1/20) of the blood samples tested positive for H. meleagridis, based on PCR and confirmed by sequencing. The results from both studies indicate that H. meleagridis DNA can be detected in the blood samples by PCR and confirmed by sequencing as early as 4 days postchallenge. This early detection method could be applied in field outbreaks to detect and confirm histomoniasis as early as possible.
Detección temprana de histomoniasis en muestras de sangre mediante PCR y secuenciación La histomoniasis es una enfermedad mortal de los pavos que causa pérdidas económicas devastadoras a la industria avícola. En los brotes de campo, se realiza un diagnóstico presuntivo basado en lesiones patológicas macroscópicas y se confirma mediante histopatología. Se necesita una herramienta de detección temprana con un tiempo de respuesta rápido para prevenir la propagación de la histomoniasis. Con este objetivo, se realizaron dos estudios en pavos. En el Estudio 1, se alojaron 40 pavipollos en dos corrales (20 pavipollos/corral) y se desafiaron a los 14 días de edad con Histomonas meleagridis por vía intracloacal. Se recolectaron muestras de sangre a los cuatro días después del desafío. El cincuenta y cinco por ciento (22/40) de las muestras de sangre resultaron positivas para H. meleagridis según el método de PCR utilizando iniciadores dirigidos contra el gene 18S rRNA y confirmado mediante secuenciación. En el Estudio 2, se alojaron 40 pavipollos en dos grupos y se criaron en corrales en piso. Los grupos 1 y 2 sirvieron como controles negativos y de desafío, respectivamente. A los 14 días de edad, las aves del Grupo 2 fueron expuestas a H. meleagridis por vía intracloacal. Se recolectaron muestras de sangre dos días después del desafío. El cinco por ciento (1/20) de las muestras de sangre dieron positivo para H. meleagridis, según el método de PCR y confirmado mediante secuenciación. Los resultados de ambos estudios indican que el ADN de H. meleagridis puede detectarse en las muestras de sangre mediante PCR y confirmarse mediante secuenciación tan pronto como cuatro días después de la exposición. Este método de detección temprana podría aplicarse en brotes de campo para detectar y confirmar la histomoniasis lo antes posible.

Intestinal health plays a major role in profitable and efficient turkey production. Blackhead disease (histomoniasis) is caused by Histomonas meleagridis, an anaerobic protozoan parasite. Histomonas meleagridis disrupts intestinal integrity and may cause systemic infection. Some field outbreaks of blackhead disease are associated with low morbidity and mortality, while in some instances, it may cause severe morbidity and mortality. In the current study, a presumptive diagnosis of blackhead disease was made based on the characteristic gross lesions in the liver and ceca. The cecal culture, PCR, and sequencing confirmed the presence of H. meleagridis and Pentatrichomonas hominis. Pentatrichomonas hominis has been reported in enteritis cases of several other species, such as dogs, cats, and cattle. The impact of P. hominis on intestinal health of turkeys has not previously been studied, and to the best of our knowledge, this is the first case report of concurrent H. meleagridis and P. hominis infection in turkeys.
Reporte de caso- Infección simultánea de Histomonas meleagridis y Pentatrichomonas hominis en un brote de enfermedad de la cabeza negra en pavos. La salud intestinal juega un papel importante en la producción rentable y eficiente de pavos. La enfermedad de la cabeza negra (histomoniasis) es causada por Histomonas meleagridis, que es un parásito protozoario anaeróbico. Histomonas meleagridis altera la integridad intestinal y puede causar una infección sistémica. Algunos brotes de campo de la enfermedad de la cabeza negra están asociados con una baja morbilidad y mortalidad, mientras que en algunos casos puede causar una morbilidad y mortalidad severas. En el presente estudio, se realizó un diagnóstico presuntivo de la enfermedad de la cabeza negra con base a las lesiones macroscópicas características en el hígado y el ciego. El cultivo cecal, un método de PCR y secuenciación confirmaron la presencia de H. meleagridis y Pentatrichomonas hominis. Se ha reportado la presencia de Pentatrichomonas hominis en casos de enteritis de varias otras especies, como perros, gatos y ganado. El impacto de P. hominis en la salud intestinal de los pavos no se había estudiado previamente y según nuestro conocimiento, este es el primer reporte de un caso de infección simultánea por H. meleagridis y P. hominis en pavos.

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The objectives of this study were to evaluate whether a preinfection of Eimeria adenoeides (EAD) or Eimeria tenella (ET) could affect the severity of subsequent histomoniasis in turkeys (Experiment 1) and if previous exposure to EAD infection, when a single or multiple inoculations of EAD were administered with sufficient time for complete cecal recovery, would affect the severity of HM incidence and lesions (Experiment 2). In Experiment 1, 200 poults were assigned to 1 of 5 groups, as follows: unchallenged negative control, positive challenge control inoculated with 105 HM, EAD at 500 oocysts/bird and Histomonas meleagridis (HM), EAD at 2500 oocysts/bird and HM, or ET at 9 × 106 oocysts/bird and HM. ET and EAD were inoculated on day 15 and HM on day 20. In Experiment 2, the trial consisted of two different challenge ages to evaluate short- or long-term EAD effects before HM challenge. Poults (n = 260) were assigned to either early-HM-challenged groups (HM on day 19 challenge control or EAD at 2500 oocysts/bird on day 14 with HM on day 19) or late-HM-challenged groups (HM on day 35 challenge control, EAD at 2500 oocysts/bird on day 14 and HM on day 35, or EAD at 100 oocysts/bird every 2-3 days during the first 3 weeks and HM on day 35). An unchallenged negative-control group was used for both the early- and late-challenge phases in Experiment 2. Mortalities were recorded, and surviving poults were scored for histomoniasis-related hepatic and cecal lesions. In Experiment 1, preinfection with both doses of EAD reduced the mortality as well as the cecal and hepatic lesions caused by histomoniasis. In Experiment 2, neither short- nor long-term preinfection with EAD had an effect on histomoniasis-related mortality or lesions. Differences between Experiments 1 and 2 may be due to the level of infection caused by the prechallenge with EAD and the resulting destruction of cecal tissue.
Evaluación de la preinfección por Eimeria adenoeides sobre la severidad de la histomoniasis en pavos. Los objetivos de este estudio fueron evaluar si una preinfección por Eimeria adenoeides (EAD) o Eimeria tenella (ET) podría afectar la severidad de la histomoniasis subsequente en pavos (Experimento 1); y si la exposición previa a la infección por E. adenoeides, cuando se administraron una o varias inoculaciones de E. adenoeides con tiempo suficiente para la completa recuperación cecal, afectaría la gravedad de la incidencia y las lesiones de Histomonas meleagridis (Experimento 2). En el Experimento 1, se asignaron 200 pavipollos en cinco grupos, de la siguiente manera: control negativo no desafiado, control de desafío positivo inoculado con 105 de H. meleagridis, un grupo con E. adenoeides a 500 ooquistes/ave e H. meleagridis (HM), otro grupo con E. adenoeides a 2500 ooquistes/ave y H. meleagridis, o E. tenella a 9×106 ooquistes/ave y H. meleagridis. Se inocularon E. tenella y E. adenoeides el día 15 y H. meleagridis el día 20. En el Experimento 2, el ensayo consistió en dos edades de exposición diferentes para evaluar los efectos de E. adenoeides a corto o largo plazo antes del desafío con H. meleagridis. Los pavipollos (n = 260) se asignaron a los grupos de desafío temprano con H. meleagridis (H. meleagridis en el día 19 en el grupo control de desafío o E. adenoeides con 2500 ooquistes/ave el día 14 y con H. meleagridis en el día 19) o los grupos de desafío tardío con H. meleagridis (H. meleagridis en el día 35 del control de desafío, E. adenoeides a 2,500 ooquistes/ave el día 14 y H. meleagridis en el día 35, o E. adenoeides con 100 ooquistes/ave cada 2-3 días durante las primeras 3 semanas y H. meleagridis en el día 35). En el Experimento 2, se utilizó un grupo de control negativo no desafiado para ambas fases de exposición temprana y tardía. Se registraron la mortalidad y los pavipollos supervivientes se asignaron puntuaciones en cuanto a lesiones hepáticas y cecales relacionadas con histomoniasis. En el Experimento 1, la preinfección con ambas dosis de E. adenoeides redujo la mortalidad, así como las lesiones cecales y hepáticas causadas por histomoniasis. En el Experimento 2, ni la preinfección a corto ni a largo plazo con E. adenoeides tuvo un efecto sobre la mortalidad o las lesiones relacionadas con la histomoniasis. Las diferencias entre los Experimentos 1 y 2 pueden deberse al nivel de infección causado por el desafío previo con E. adenoeides y la destrucción resultante del tejido cecal.

Over the last couple of years, the number of histomoniasis cases in commercial turkeys has increased substantially in British Columbia, particularly in the Fraser Valley. Due to a lack of approved efficacious preventive or curative drugs in Canada, histomoniasis outbreaks have had significant economic and animal welfare impacts on the commercial turkey industry. In July 2020, Health Canada conditionally approved the treatment use of paromomycin sulfate on a case-by-case basis via an emergency drug release authorization. Three flocks infected with Histomonas meleagridis were treated with labeled-dose paromomycin sulfate in the feed shortly after presumptive diagnosis. Despite the treatment, two out of three flocks suffered significant losses. One flock suffered over 67% mortality by the eighth day of treatment. Due to significant production loss and animal welfare concerns, the flock was shipped early for mercy cull; thus sustained 100% production loss. Another flock experienced over 38% mortality by the end of the fourth week of treatment and was slaughtered early to minimize production loss. The treatment response in two out of three cases suggests that any curative effect of paromomycin is limited. Thus, future field evaluation should carefully consider the prophylactic use of paromomycin sulfate, especially on farms with recurrent outbreaks.

Repeated serial in vitro passage of Histomonas meleagridis, the etiological agent of histomoniasis (blackhead) of turkeys, was demonstrated to markedly achieve attenuation and reduction of virulence as compared to the original wild-type isolate. Four experiments were performed to evaluate the route (oral vs. intracloacal) and age (day-of-hatch vs. d 14) for administration of attenuated H. meleagridis isolates as vaccine candidates against homologous or heterologous wild-type challenge. Attenuated H. meleagridis were developed from 2 different strains (Buford strain originating in Georgia; PHL2017 strain originating in Northwest Arkansas). Buford P80a (passage 80, assigned as isolate lineage \"a\" following repeated passage) was selected as the primary vaccine candidate and was evaluated in Experiments 1-3. Experiment 4 evaluated selected candidates of attenuated PHL2017 (P67, P129) and Buford (P80a, P200a, P138b, P198c) strains against Buford wild-type challenge. As has been demonstrated previously, wild-type H. meleagridis cultures administered orally after 1 day of age were not infective in the current studies, but infection with wild-type cultures could be induced orally at day-of-hatch. Infection was effectively achieved via the intracloacal route at day-of-hatch and in older turkeys (d 21, d 28-29, d 35). Intracloacal inoculation of turkeys with the attenuated passaged isolates as vaccine candidates at d 14 was shown to produce significant (P < 0.05) protection from mortality, reduction in body weight gain, as well as reduction in hepatic and cecal lesions in these experiments following challenge with either the homologous wild-type isolate or from a wild-type strain obtained years later from a geographically disparate area of the United States. Inoculation with the attenuated H. meleagridis isolates at day-of-hatch, either orally or cloacally, did not produce significant protection against subsequent wild-type challenge. While offering significant protection with minimal vaccine-related negative effects, the protection from cloacal vaccine administration was neither significantly robust nor encouraging for industry application using the methods evaluated in the present manuscript since mortalities and lesions were not completely reduced which could thereby potentially allow transmission from residual infection and shedding within a flock.

Histomoniasis, also commonly referred to as blackhead disease, is caused by the protozoan parasite Histomonas meleagridis. Since the removal of nitarsone in 2015, no approved prophylactics are available for mitigating histomoniasis. Disease incidence and high mortalities are frequently associated with turkey flocks, although infection of broiler breeders also occurs. Quinine is a naturally occurring alkaloid with antimalarial properties. In vitro assays have shown strong antihistomonal properties of quinine, leading to our hypothesis that quinine inclusion within the feed could prevent histomoniasis in turkeys. Selected concentrations of quinine were included within a turkey starter diet to evaluate effects on body weight gain (BWG), liver lesions, cecal lesions, and mortality of H. meleagridis-challenged turkeys. On day-of-hatch, poults were randomly assigned to either the basal diet or a quinine diet. Groups consisted of a non-challenged control (NC; basal diet), 0.022% quinine + challenge, 0.067% quinine + challenge, 0.2% quinine + challenge, or a positive-challenged control (PC; basal diet). On d 10, challenged groups were intracloacally inoculated with 105H. meleagridis cells/turkey, and lesions were evaluated on d 21 post-infection. Individual body weights were recorded on d 0, d 10, and d 31 to calculate the pre-challenge and post-challenge BWG. No significant differences (P > 0.05) were observed between the d 0 to 10 pre-challenged BWG between quinine treatment diets and the basal diet. Similarly, no differences (P > 0.05) were observed in post-challenge d10-31 BWG of the quinine dietary treatments as compared to the PC. Cumulative mortalities, liver lesions, and cecal lesions related to histomoniasis were not reduced (P > 0.05) in any of the quinine treatment groups as compared to the PC. Although quinine successfully reduced H. meleagridis cells in vitro, results from the in vivo experiment indicated no reduction in histomoniasis severity as evidenced by similar lesions and mortality as the PC. Taken together, these data indicate that quinine inclusion within the feed at these concentrations and under these experimental conditions was not efficacious in the prevention or treatment of histomoniasis.

Field visits at two different farms suggest a correlation between commercial turkey (Meleagridis gallopavo) flocks having increased mortality from blackhead disease (histomoniasis) if they suffer from poor poult quality at placement and coccidiosis (Eimeria spp.) before age 6 wk. In both cases, the flocks were all-in/all-out with curtain-sided houses and received a coccidiosis vaccine on day of hatch. At Farm I 2018, poults from different hatcheries were placed in two houses on the same farm (Houses 1 and 2). House 2 had poults considered poor quality and suffered from mortality associated with coccidiosis at 2 and 4 wk of age. At 8 wk, blackhead disease was diagnosed in both houses by postmortem examination. House 2 had mortality of >2000 poults, and the subpopulation of necropsied poults had gross lesions characteristic of histomoniasis. Gross lesions associated with blackhead disease were only found in eight poults in House 1, which was populated with good-quality poults and did not have a second spike in mortality due to coccidiosis. The Farm II 2020 poults were delivered from the same hatchery onto a three-house farm (Houses A, B, and C). House C had poults that were considered poor quality and had mortality associated with coccidiosis at 3 wk of age. At 8-9 wk, House C had mortality approaching 1000 birds, with all poults examined postmortem having clinical signs of blackhead disease. Houses A and B were populated with good-quality poults and had no diagnosed mortality from coccidiosis or blackhead disease. The similarity of these two cases suggest that poult quality at placement coupled with coccidiosis before 6 wk of age can influence the severity of blackhead disease in commercial turkey flocks.