A fatal case of a captive snowy owl (Bubo scandiacus) with Haemoproteus infection in Japan.

Parasites of the genus Haemoproteus are vector-borne avian haemosporidia commonly found in bird species of the world. Haemoproteus infections are typically considered relatively benign in birds. However, some Haemoproteus species cause severe disease and mortality, especially for captive birds removed from their original habitat. In September 2018, a captive 15-year-old snowy owl (Bubo scandiacus), kept in a zoological garden of Japan, died subacutely after presenting leg dysfunction. This case showed significantly low PCV and elevated AST, ALT, CK, and LDH values. Many megalomeronts with prominent morphological characteristics of Haemoproteus were observed in the left leg muscles. Those megalomeronts exhibited multilocular structures and were internally filled with merozoites. A new lineage of Haemoproteus was detected by subsequent PCR for the cytochrome b (cytb) gene of avian haemosporidia from DNA extracted from several organ tissues. The detected lineage was classified in the subgenus Parahaemoproteus and was similar to those from the wild birds inhabiting the region including the study area, suggesting that this snowy owl likely acquired its infection from wild birds. This is the first report of a fatal case of a captive bird with a locally transmitted Haemoproteus infection in Japan. We considered the pathogenicity of this infection in conjunction with the clinical course and hematology results. We surmise that snowy owls may be particularly susceptible to infection with Haemoproteus parasites, and warming northern temperatures may exacerbate the overall health of these and other high latitude birds. Further research into the prevalence of Haemoproteus in wild birds near zoological gardens and potential biting midge vectors is necessary for the ex situ conservation of introduced birds.

Haplosporidium pinnae associated with mass mortality in endangered Pinna nobilis (Linnaeus 1758) fan mussels.

The fan mussel, Pinna nobilis (Linnaeus 1758), is an endemic bivalve of the Mediterranean basin, protected by international legislation as an endangered species. In the early summer of 2018, a mass mortality event (MME) of P. nobilis was recorded in the Gulf of Taranto (Southern Italy, Ionian Sea). Moribund specimens of P. nobilis were collected by scuba divers and processed by bacteriological, parasitological, histopathological and molecular analyses to investigate the causes of this MME. Different developmental stages (i.e., plasmodia, spores and sporocysts) of a presumptive haplosporidian parasite were observed during the histological analysis in the epithelium and in the lumen of the digestive tubules, where mature spores occurred either free or in sporocysts. The spores presented an operculum and an ovoid shape measuring 4.4 µm (±0.232) in length and 3.6 µm (±0.233) in width. BLAST analysis of an 18SrRNA sequence revealed a high nucleotide similarity (99%) with the reference sequence of Haplosporidium pinnae available in GenBank database. Phylogenetic analysis clustered the sequence of the pathogen in a paraphyletic clade with the reference sequence of H. pinnae, excluding other haplosporidians (i.e., Bonamia and Minchinia genera). Based on data reported, H. pinnae was the causative agent of MME in the populations of P. nobilis sampled in the Ionian Sea, where the conservation of this endangered species is heavily threatened by such a protozoan infection. Further investigations should contribute to knowledge about the life cycle of H. pinnae in order to reduce spread of the pathogen and to mitigate the burden of the disease where P. nobilis is facing the risk of extinction.

Two Tales of Cytauxzoon felis Infections in Domestic Cats.

Cytauxzoonosis is an emerging infectious disease that affects wild felids as well as the domestic cat; it is caused by the apicomplexan protozoan parasites belonging to the genus Cytauxzoon. Cytauxzoonfelis is the species of major concern, whose transmission occurs via the bite of an infected tick. Cytauxzoonosis of the domestic cat has historically been considered uniformly fatal, with a short course of illness, and most domestic cats die within 9 to 15 days postinfection. However, increasing evidence of domestic cats surviving C. felis infection suggests the existence of different strains with various levels of pathogenicity. Although wild felids are considered natural reservoirs for this parasite, a number of studies suggest that domestic cats that have survived nonlethal infections may serve as an additional reservoir. The current article comprehensively reviews the parasite and its life cycle, geographic distribution, genetic variability, and pathogenesis, as well as host immunology and the diagnosis, treatment, and prevention of infection in the domestic cat. This information should provide a basis for better understanding the parasite as well as the pathogenesis of the disease.

Protozoal-related mortalities in endangered Hawaiian monk seals Neomonachus schauinslandi.

Protozoal infections have been widely documented in marine mammals and may cause morbidity and mortality at levels that result in population level effects. The presence and potential impact on the recovery of endangered Hawaiian monk seals Neomonachus schauinslandi by protozoal pathogens was first identified in the carcass of a stranded adult male with disseminated toxoplasmosis and a captive monk seal with hepatitis. We report 7 additional cases and 2 suspect cases of protozoal-related mortality in Hawaiian monk seals between 2001 and 2015, including the first record of vertical transmission in this species. This study establishes case definitions for classification of protozoal infections in Hawaiian monk seals. Histopathology and immunohistochemistry were the primary diagnostic modalities used to define cases, given that these analyses establish a direct link between disease and pathogen presence. Findings were supported by serology and molecular data when available. Toxoplasma gondii was the predominant apicomplexan parasite identified and was associated with 100% of mortalities (n = 8) and 50% of suspect cases (n = 2). Incidental identification of sarcocysts in the skeletal muscle without tissue inflammation occurred in 4 seals, including one co-infected with T. gondii. In 2015, 2 cases of toxoplasmosis were identified ante-mortem and shared similar clinical findings, including hematological abnormalities and histopathology. Protozoal-related mortalities, specifically due to toxoplasmosis, are emerging as a threat to the recovery of this endangered pinniped and other native Hawaiian taxa. By establishing case definitions, this study provides a foundation for measuring the impact of these diseases on Hawaiian monk seals.

Apoptosis and cell proliferation in the mouse model of embryonic death induced by Tritrichomonas foetus infection.

Bovine tritrichomonosis is a sexually transmitted disease caused by the protozoon Tritrichomonas foetus and characterised by embryonic-death and abortion. During pregnancy, the processes of cell proliferation and death play a crucial role for blastocyst implantation and the subsequent maintenance of early pregnancy, and their misbalance may lead to the abortion. In this study, we aimed to investigate whether cell proliferation and death may be altered during tritrichomonosis. For this purpose, we used pregnant BALB/c mice as an alternative experimental animal model that has successfully reproduced the infection. We analysed the immunohistochemical expression of active caspase-3 and proliferating cell nuclear (PCNA) antigens in the endometrium of infected mice. We found an increase in the number of caspase-3 positive cells in infected mice that were not pregnant at the necropsy. Besides, the number of positive proliferating cells increased in the uterine luminal epithelium of infected animals killed at 5-7 days post coitum (dpc). Pregnant infected mice killed at 8-11 dpc showed higher proliferation than control animals. We suggest that the cytopathic effect induced by T. foetus in the uteri of infected mice may induce the apoptosis of the epithelial cells and, as a result, promote a compensatory proliferative response. The information described here will be helpful to further study the pathogenesis of the bovine tritrichomonosis.

A single strain of Tetratrichomonas gallinarum causes fatal typhlohepatitis in red-legged partridges (Alectoris rufa) to be distinguished from histomonosis.

Typhlohepatitis was observed in a flock of 2500 red-legged partridges in Great Britain, characterized by the sudden deaths of 15 birds within 2 days. Necropsy of five dead birds revealed severe lesions in the caeca with thickened caecal walls, a reddened lining and bloody contents. The livers contained multiple miliary lesions and similar pathological changes were found in the spleens of some birds. Microscopic examination of intestinal contents showed the occurrence of coccidial oocysts in two partridges. Different methods for the detection of bacteria from liver and intestine samples were conducted without positive results. Histopathological examination revealed the presence of protozoan parasites in the caecum, liver and spleen of the affected birds. In situ hybridization (ISH) for the detection of trichomonads resulted in positive findings and polymerase chain reaction (PCR) confirmed the presence of Tetratrichomonas gallinarum in the lesions. Additionally, archived tissues of red-legged partridges from different flocks suffering from severe typhlohepatitis in Great Britain in 2008 and 2009 were re-investigated by ISH and PCR. Beside the sporadic occurrence of histomonosis, in most of the cases trichomonads were detected by ISH in the caecum and liver of affected birds. Furthermore, dissemination of the flagellate into the lung and bursa of Fabricius could be demonstrated. Analyses of T. gallinarum DNA obtained from the different cases resulted in homologous nucleotide sequences. Altogether, the results demonstrate the circulation of a virulent strain of T. gallinarum in reared red-legged partridges.

Apparent mortality as a result of an elevated parasite infection in Presbytis rubicunda.

We provide the first parasite assessment of wild red langurs (Presbytis rubicunda). Seven helminth taxa and 4 protozoa were recovered from faecal samples. We report the details of the death of an adult female red langur with potential association of an elevated parasite infection. There is a paucity of published literature on red langurs and nothing is known of the factors affecting interspecific parasite transmission and disease spread between other primate species. Parasites may be substantial determinants of host health and may present a significant influence on the survival and reproduction of individuals and the subsequent fecundity of populations. Further research into the connection of parasite burden with individual deaths and potential population declines is recommended for monitoring population health.

Co-infection by alveolate parasites and frog virus 3-like ranavirus during an amphibian larval mortality event in Florida, USA.

A multispecies amphibian larval mortality event, primarily affecting American bullfrogs Lithobates catesbeianus, was investigated during April 2011 at the Mike Roess Gold Head Branch State Park, Clay County, Florida, USA. Freshly dead and moribund tadpoles had hemorrhagic lesions around the vent and on the ventral body surface, with some exhibiting a swollen abdomen. Bullfrogs (100%), southern leopard frogs L. sphenocephalus (33.3%), and gopher frogs L. capito (100%) were infected by alveolate parasites. The intensity of infection in bullfrog livers was high. Tadpoles were evaluated for frog virus 3 (FV3) by histology and PCR. For those southern leopard frog tadpoles (n = 2) whose livers had not been obscured by alveolate spore infection, neither a pathologic response nor intracytoplasmic inclusions typically associated with clinical infections of FV3-like ranavirus were noted. Sequencing of a portion (496 bp) of the viral major capsid protein gene confirmed FV3-like virus in bullfrogs (n = 1, plus n = 6 pooled) and southern leopard frogs (n = 1, plus n = 4 pooled). In July 2011, young-of-the-year bullfrog tadpoles (n = 7) were negative for alveolate parasites, but 1 gopher frog tadpole was positive. To our knowledge, this is the first confirmed mortality event for amphibians in Florida associated with FV3-like virus, but the extent to which the virus played a primary role is uncertain. Larval mortality was most likely caused by a combination of alveolate parasite infections, FV3-like ranavirus, and undetermined etiological factors.

An outbreak of blackhead disease (Histomonas meleagridis) in farm-reared bobwhite quail (Colinus virginianus).

An outbreak of blackhead disease (Histomonas meleagridis) in farm-reared flock of 13,500 bobwhite quail (Colinus virginianus) resulted in mortality totaling approximately 1500 in 4 wk. Necropsy of 56 dead birds at midoutbreak (from a total that day of 131) revealed that 55 had severe cecal lesions typical of blackhead, and only 3 had visible lesions in the liver. Necropsy of apparently healthy birds failed to detect any signs of infection. Presence of H. meleagridis in affected ceca was proved by culture in vitro and PCR tests.

Causes of death in beef cattle in southern Brazil.

We determined the prevalence of diseases and pathogens associated with mortality in beef cattle in the State of Rio Grande do Sul, Brazil, based on pathology laboratory submissions. Postmortem examinations were conducted on 1,277 beef cattle that died between 2008 and 2018. Information regarding age, time of the year, breed, and regional location were analyzed statistically. Most cattle were from the surrounding region of Porto Alegre, and 78.7% of the analyzed cases had diagnostic value. The diagnostic category with most cases was infectious and/or parasitic diseases (60%), followed by toxic and toxicoinfectious (25%). Most cases occurred in the fall. Major disease conditions identified included hemoprotozoal infection (18.2%), rabies (8.2%), and plant intoxications by Senecio spp. (8.5%) and Pteridium arachnoideum (4.6%). Hemoprotozoal infection occurred at a higher frequency in young cattle, mainly in animals up to 1 y old. Intoxication by Senecio spp. was more frequent in cattle 2-3 y old.

Prevalence of Leucocytozoon spp, in the endangered yellow-eyed penguin Megadyptes antipodes.

Yellow-eyed penguins on Stewart Island were identified with a Leucocytozoon spp. of a novel lineage in association with a high regional incidence of chick mortality (n=32, 100% mortality) during the November 2006 to January 2007 breeding season. Fourteen chicks from Stewart Island were examined post-mortem and histologically for Leucocytozoon infection. In addition, a survey of blood to detect Leucocytozoon spp. infections using PCR was performed on 107 yellow-eyed penguins from 4 distinct nesting areas on the South Island (Oamaru, Otago Peninsula, and Catlins) (n=95), and Stewart Island (n=12). The results of the study revealed that 2 of the 14 (14%) chicks necropsied showed severe, disseminated megaloschizont formation in the liver, spleen, lung, kidney and other tissues characteristic of leucocytozoonosis. Eighty-three percent (83%) of blood samples collected from Stewart Island penguins contained Leucocytozoon DNA, whereas samples from the 3 other nesting areas were negative for the blood parasite. Leucocytozoon spp. DNA sequences isolated from blood and tissues of adults (n=10) and chicks (n=7) were similar and grouped with other published Leucocytozoon spp. sequences but in a distinct cluster together with closely related isolates from a Western march harrier (Circus aerginosus) and common loon (Gavia immer). These findings suggest that yellow-eyed penguins on Stewart Island are infected with a regionally isolated, host-specific Leucocytozoon spp. which may contribute to the high chick mortality observed during this period.

Responding to Pfiesteria piscicida (the fish killer): phantomatic ontologies, indeterminacy, and responsibility in toxic microbiology.

Based on an analysis of an ongoing scientific-political controversy over the toxicity of a fish-killing microorganism, this paper explores the relationship between responsibility and nonhuman contributions to agency in experimental practices. Research into the insidious effects of the dinoflagellates Pfiesteria piscicida (the fish killer) that thrive in waters over-enriched with nutrients, has received considerable attention by both the media and government agencies concerned with public and environmental health. After nearly two decades of research, the question of whether Pfiesteria can be regarded the 'causative agent' of massive fish kills in the estuaries of the US mid-Atlantic could not be scientifically settled. In contrast to policymakers, who attribute the absence of a scientific consensus to gaps in scientific knowledge and uncertainties regarding the identity and behavior of the potentially toxic dinoflagellates, I propose that an inseparable entanglement of Pfiesteria's identities and their toxic activities challenges conventional notions of causality that seek to establish a connection between independent events in linear time. Building on Karen Barad's framework of agential realism, I argue for a move from epistemological uncertainties to ontological indeterminacies that follow from Pfiesteria's contributions to agency, as the condition for responsible and objective science. In tracking discrepant experimental enactments of Pfiesteria that have been mobilized as evidence for and against their toxicity, I investigate how criteria for what counts as evidence get built into the experimental apparatuses and suggest that the joint possibilities of causality and responsibility vary with the temporalities of the objects enacted. This discussion seeks to highlight a thorough entanglement of epistemic/ontological concerns with the ecological/political relevance of particular experiments. Finally, I introduce a new kind of scientific object that--borrowing from Derrida--I call phantomatic. Phantoms don't emerge as such, but appear as traces and are associated with specific matters of concern.

Oral infection of turkeys with in vitro-cultured Histomonas meleagridis results in high mortality.

In the present study a well-defined clonal culture of Histomonas meleagridis was used to investigate whether turkeys can be infected orally with in vitro-propagated parasites in the absence of any vector. Therefore, two subsequent experiments were arranged to study the morbidity and mortality of a virulent strain of histomonads in turkeys following crop instillation of protozoa and a feed restriction for 5 h. Soon after infection, the parasites triggered total morbidity and mortality in a group of 14 turkeys, of which 10 birds were orally infected and the others were kept as in-contact birds. Administration of the same number of histomonads into either the cloaca or the crop of individually housed birds was investigated subsequently, to avoid any bird-to-bird transmission. All eight cloacally infected turkeys and six out of eight orally infected turkeys contracted severe histomonosis, with only two birds surviving the infection expressing no clinical signs. During pathological investigations, all birds showed pathognomonic lesions in the livers and caeca indicative for histomonosis, except the two turkeys that survived the infection. The results of the experiments verify for the first time the effective oral infection of 1-day-old turkeys with clonal in vitro-cultured H. meleagridis.

Studies of trichomonad protozoa in free ranging songbirds: prevalence of Trichomonas gallinae in house finches (Carpodacus mexicanus) and corvids and a novel trichomonad in mockingbirds (Mimus polyglottos).

This study refutes the accepted dogma that significant pathogenic effects of Trichomonas gallinae are limited to columbiformes and raptors in free ranging bird populations in North America. Trichomonads were associated with morbidity and mortality amongst free ranging house finches (Carpodacus mexicanus), mockingbirds (Mimus polyglottos) and corvids (scrub jay: Aphelocoma californica; crow: Corvus brachyrhynchos; raven: Corvus corax) in northern California. Prevalence of trichomonad infection was 1.7% in house finches, 0-6.3% in corvids, and 0.9% in mockingbirds. Bird case fatality ratio was 95.5% in house finches, 0-100.0% in corvids, and 37.5% in mockingbirds. DNA sequences of parasites in house finches and corvids were identical to T. gallinae strain g7 (GeneBank AY349182.1) for the 5.8s ribosome. DNA sequences of parasites cultured from two mockingbirds were genetically distinct from that of available sequenced trichomonads. These isolates were clearly phylogenetically more closely related to the Trichomonadinae than the Tritrichomonadinae. While molecular techniques were required to differentiate between trichomonad species, wet mount preparations from the oral cavity/crop were a reliable and inexpensive method of screening for trichomonad infections in these species. Positive wet mount tests in house finches and corvids living in northern California were highly likely to indicate infection with T. gallinae, while in mockingbirds positive wet mounts most likely indicated a trichomonad other than T. gallinae.

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.

Mass Mortality of Green Frog ( Rana clamitans) Tadpoles in Wisconsin, USA, Associated with Severe Infection with the Pathogenic Perkinsea Clade.

We documented mortality of green frog ( Rana clamitans) tadpoles in Wisconsin, US, attributed to severe Perkinsea infection. Final diagnosis was determined by histopathology. followed by molecular detection of pathogenic Perkinsea clade (PPC) of frogs in the liver. To our knowledge, this represents the first detection of PPC in the midwestern US.

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.

A virulent mono-eukaryotic culture of Histomonas meleagridis is capable of inducing fatal histomonosis in different aged turkeys of both sexes, regardless of the infective dose.

Histomonosis (syn. histomoniasis) is a parasitic disease which affects predominately turkeys but also other avian species. Concurrent with the ban of therapeutic and prophylactic substances, the disease, caused by the flagellated protozoon Histomonas meleagridis, is more frequently reported. Due to somewhat diverse results reported in the past, a well-characterized culture was used in the present study to investigate the possible influence of certain parameters on the outcome of the disease. For this study, turkeys were infected with different doses of the mono-eukaryotic culture Histomonas meleagridis/Turkey/Austria/2922-C6/04 using birds of both sexes at various ages. All study groups consisted of 14 birds, of which 10 birds were directly infected via the cloacal route and four birds were kept as in-contact birds. This scheme was used to investigate the pathogenicity of the cloned isolate in 1-day-old and 14-day-old turkeys. In 8-week-old turkeys, only eight birds out of 12 were infected. When 1-day-old and 8-week-old turkeys were infected with 10(4) histomonads per bird, all turkeys died between 11 and 21 days postinfection or had to be euthanatized due to their poor condition. In a group of 14 poults, infective doses of either 10 histomonads (100 histomonads among 10 birds) or 10(3) histomonads per bird had hardly any influence on the first notification of clinical signs. However, even though the onset of clinical signs and mortality was delayed with the lower dose, none of the birds survived the infection. As a consequence, no differences were noticed between male and female turkeys using the mono-eukaryotic culture of Histomonas meleagrigis/Turkey/Austria/2922-C6/04 in the current experimental setting.

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.

Case Series: Virulent hemosporidiosis infections in juvenile great horned owls (Bubo virginianus) from Louisiana and California, USA.

A total of eight juvenile great horned owls (Bubo virginianus) were found lethargic and on the ground in spring 2015, 2016, and 2017, including one fledgling from Louisiana, USA and seven nestlings from California, USA. One bird survived to release after rehabilitation; seven birds died or were euthanized due to poor prognosis and were necropsied. Necropsy findings were similar and included general pallor of all tissues, particularly the subcutis and lungs, and enlarged liver and spleen. Histopathology revealed multi-organ necrosis, abundant meronts containing merozoites, and intracytoplasmic pigmented haemosporidian parasites in blood cells in one bird. Leucocytozoon lineages lSTOCC16 and BUVIR06 were identified by polymerase chain reaction and genetic sequencing. The systemic Leucocytozoon infections were likely associated with morbidity and mortality in these owls. A second parasite, Haemoproteus lineage hSTVAR01, was also identified in an owl from Louisiana. This is the first identification of Leucocytozoon lineages that have been associated with mortality in young great horned owls.