Histopathologic study of experimental Sarcocystis hemionilatrantis infection in fawns.

Mule deer fawns (Odocoileus hemionus hemionus) inoculated with sporocysts of Sarcocystis hemionilatrantis became infected, developed clinical signs of disease, and died, due to the infection itself or from intercurrent pneumonia. Clinical signs were first noticed 18 days after infection and fawns died from postinfection days 27 to 63. Histopathologic examination revealed early lesions in skeletal muscle which consisted of perivascular necrosis with mononuclear and neutrophilic cell infiltration, accompanied by edema, degeneration, and focal necrosis of muscle. Subsequently, this reaction subsided and the cellular infiltrate dissipated. An infected macrophage usually remained in the vacuolated muscle space; each macrophage was surrounded by a clear halo. Developing Sarcocystis schizonts were identified in the cytoplasm of the macrophages, and the cytoplasmic membrane eventually ruptured releasing merozoites. The merozoites then developed into typical muscle cysts. Results of the present study indicated that S hemionilatrantis is a pathogen of mule deer under experimental conditions. Pathogenicity should be investigated to determine if S hemionilatrantis causes death or debilitation in wild mule deer under natural conditions.     

Experimental transmission of chronic wasting disease (CWD) of elk (Cervus elaphus nelsoni), white-tailed deer (Odocoileus virginianus), and mule deer (Odocoileus hemionus hemionus) to white-tailed deer by intracerebral route

To compare clinical and pathologic findings of chronic wasting disease (CWD) in a natural host, 3 groups (n = 5) of white-tailed deer (WTD) fawns were intracerebrally inoculated with a CWD prion of WTD, mule deer, or elk origin. Three other uninoculated fawns served as controls. Approximately 10 months postinoculation (MPI), 1 deer from each of the 3 inoculated groups was necropsied and their tissues were examined for lesions of spongiform encephalopathy (SE) and for the presence of abnormal prion protein (PrP(d)) by immunohistochemistry (IHC) and Western blot (WB). The remaining deer were allowed to live until they developed clinical signs of the disease which began approximately 18 MPI. By 26 MPI, all deer were euthanatized on humane grounds. Obvious differences in clinical signs or the incubation periods were not observed between the 3 groups of deer given CWD. In 1 of 3 nonclinical deer euthanatized at 10 MPI, minimal microscopic lesions of SE were seen in the central nervous system (CNS) tissues, and PrP(d) was observed by IHC in tissues of all 3 deer. In the clinical deer, CNS lesions of SE and PrP(d) accumulations were more severe and extensive. It is concluded that the 3 sources of CWD prion did not induce significant differences in time to clinical disease or qualitative differences in signs or lesions in WTD. However, this observation does not imply that these CWD agents would necessarily behave similarly in other recipient species.     

Sarcocystis infections in mule deer (Odocoileus hemionus) in Montana and the descriptions of three new species

Four structural types of sarcocysts of Sarcocystis were found in skeletal muscles of mule deer in Montana. Type I sarcocysts were thin walled (1 to 3 micron) and belonged to S hemionilatrantis. Types II to IV sarcocysts were thick walled (2 to 10 microns) and new names were proposed for them. Type II sarcocysts with long villar projections were named S hemioni. Type III sarcocysts with club-shaped villi of uneven thickness were named S youngi. Type IV sarcocysts with walls of uneven thickness and containing hair-like protrusions were named S americana.     

Prevalence, transmission, and pathogenicity of Sarcocystis gigantea of sheep

Between March and May 1983, tongues and esophagi of 355 adult ewes from Colorado and Idaho were examined for grossly visible sarcocysts. Sarcocysts of Sarcocystis gigantea were found in 35 sheep. Cats fed sarcocysts from these naturally infected sheep shed sporocysts in their feces. Two adult ewes and 12 lambs inoculated with 1,000 to 1,000,000 sporocysts were euthanatized at postinoculation days (PID) 146, 230, 265, 391, 721, and 882, and their tissues were fed to Sarcocystis-free cats. All inoculated sheep remained clinically normal except for mild pyrexia between PID 12 and 18. Sarcocysts first became grossly visible at PID 391 and sarcocysts from sheep first became infectious for cats at PID 230.     

Sarcocystosis in a Barasingha deer (Cervus duvauceli branderi).

An adult female hard-ground swamp deer (Barasingha), Cervus duvauceli branderi, was found dead of unknown cause in Kanha National Park. A necropsy of this animal failed to reveal any significant gross lesions. However, histopathologic examination revealed mature sarcocysts of Sarcocystis in the cardiac muscles. The morphology of the sarcocysts was similar to that of different species of Sarcocystis from wild and domestic animals. Final identification of Sarcocystis in this case could not be made. This is the first recorded case of Sarcocystis infection in hard-ground Barasingha deer.     

Lesions and transmission of experimental adenovirus hemorrhagic disease in black-tailed deer fawns

Adenovirus infection was the cause of an epizootic of hemorrhagic disease that is believed to have killed thousands of mule deer (Odocoileus hemionus) in California during the latter half of 1993. A systemic vasculitis with pulmonary edema and hemorrhagic enteropathy or a localized vasculitis associated with necrotizing stomatitis/pharyngitis/glossitis or osteomyelitis of the jaw were common necropsy findings in animals that died during this epizootic. To study transmission of adenovirus infection in deer and susceptibility of black-tailed deer (Odocoileus hemionus columbianus) fawns to adenovirus infection, six 3-6-month-old black-tailed fawns were divided into two treatment groups. One group was inoculated intravenously and the other group was inoculated through the mucous membranes of the eyes, nose and mouth with purified adenovirus. Each treatment group also included two additional fawns (four total) that were not inoculated but were exposed to inoculated animals (contact animals). One fawn served as a negative control. Between 4 and 16 days postinoculation, 8/10 fawns developed systemic or localized infection with lesions identical to lesions seen in animals with natural disease that died during the epizootic. Transmission was by direct contact, and the route of inoculation did not affect the incubation period or the distribution of the virus (systemic or the localized infection). Immunohistochemical analysis using polyclonal antiserum against bovine adenovirus type 5 demonstrated staining in endothelial cells of vessels in numerous tissues in animals with systemic infection and endothelial staining only in vessels subtending necrotic foci in the upper alimentary tract in animals with the localized form of the disease. All inoculated or exposed animals had staining in the tonsillar epithelium. Transmission electron microscopic examination of lung and ileum from two fawns with pulmonary edema and hemorrhagic enteropathy demonstrated endothelial necrosis and adenovirus virions in endothelial cell nuclei. Adenovirus was reisolated in black-tailed deer pulmonary artery endothelial cells using lung homogenate of the first fawn that developed systemic adenovirus infection. Serum virus neutralization test results suggest that this deer adenovirus is a new serotype.     

Prevalence of Sarcocystis sarcocysts in nine-banded armadillos (Dasypus novemcinctus) from Florida.

The prevalence and identity of Sarcocystis spp. sarcocysts in the skeletal muscles of nine-banded armadillos (Dasypus novemcinctus) collected from Alachua County, FL, were determined. H & E stained sections of skeletal muscle from tongue and thigh were examined. Thirty nine of 63 (61.9%) armadillos examined contained Sarcocystis sarcocysts. Two species were identified, Sarcocystis dasypi and Sarcocystis diminuta. Sarcocystis dasypi sarcocysts were found in 38 of 63 (60.3%) and S. diminuta sarcocysts were found in 6 of 63 (9.5%). Sarcocysts of S. dasypi were larger, more densely packed with bradyzoites, and bradyzoites contained within the sarcocyst were smaller than those of S. diminuta. Mixed infections occurred in 5 of 63 (7.9%) armadillos examined.     

Merogony and gametogony of Eimeria mccordocki (Protozoa-Eimeriidae) in the mule deer, Odocoileus h. hemionus

Eimeria mccordocki and E. madisonensis oocysts were isolated from feces of 21 of 40 captive mule deer in Fort Collins, Colorado. The two species were separated from each other by infecting one mule deer fawn, and the life cycle of E. mccordocki was studied for the first time. Four to six-weeks-old mule deer fawns were inoculated orally with E. mccordocki and killed 9, 13 and 15 days after infection. Asexual and sexual stages of life cycle developed in the ileum of mule deer, only in the surface epithelial cells of the villi. The asexual stages consisted of two generations of meronts.     

Sarcocystis infection in the white-tailed deer (Odocoileus virginianus) in Montana: intensity and description of Sarcocystis odoi n sp

Sarcocystis infection was diagnosed in 27 of 36 (75%) samples of meat from white-tailed deer (Odocoileus virginianus) in Montana. Two structurally distinct thin- and thick-walled sarcocysts were found in the white-tailed deer; the thin-walled sarcocysts were those of S odocoileocanis. A new name, S odoi, was proposed for the thick-walled sarcocysts. Sarcocysts of S odoi were up to 1,050 microns long and 260 microns wide and contained a 6.5- to 12-microns thick wall. The ultrastructure of sarcocysts of S odocoileocanis was compared with that of S odoi. A cat fed meat containing thin- and thick-walled sarcocysts shed oocysts and sporocysts 24 days later. The sporocysts in the cat's feces were 13.3 X 9.6 microns and probably belonged to S odoi.     

Prevalence of Sarcocystis species in sheep and goats in northern Nigeria.

Tissue samples comprising the oesophagus and diaphragm were collected from 400 sheep and 400 goats slaughtered at the abattoirs in the study area. Out of this number, 36 were positive for Sarcocystis cysts (sarcocysts) in sheep and 56 in goats. The sarcocysts in sheep measured 35.7 to 500 microns lengthwise and the cyst-wall 2.4 microns. They were identified to be Sarcocystis tenella. The cysts in goats measured 98 to 700 microns and the cyst-wall 2.7 microns. They were identified to be Sarcocystis capracanis. In both animals species, the sarcocysts were more frequent in the oesophagus than in the diaphragm. All sarcocysts seen were microscopic.     

Morphological and molecular identification of three species of Sarcocystis in reindeer (Rangifer tarandus tarandus) in Iceland

Six Sarcocystis species have previously been described from reindeer in Norway based on sarcocyst morphology and DNA sequencing. The aim of this study was to determine whether reindeer in Iceland, which descend from reindeer imported from Norway in 1787, also were infected with Sarcocystis, and to identify and genetically characterise any species present. Muscle tissue from the heart, diaphragm and/or oesophagus was collected from 36 reindeer in Iceland. Pieces of all tissue samples were examined histologically. Frozen/thawed samples of cardiac muscle, oesophagus and/or diaphragm from 11 of the 36 reindeer were also examined under a stereoscopic microscope and sarcocysts present were identified to species either in situ or under a light microscope. Two cysts of each species, originating from two different reindeer were randomly selected for DNA analyses. The complete ssu rRNA gene was amplified by the polymerase chain reaction (PCR) and sequenced. In addition, two sarcocysts that could not be classified by microscopic examination were selected for partial ssu rRNA gene sequence analysis. By histology, sarcocysts were found in the diaphragm and/or oesophagus of 8 of 36 (22.2%) animals. By examination of fresh tissue, sarcocysts of Sarcocystis rangi, S. tarandivulpes and S. hardangeri were found in the oesophagus of seven of nine (77.8%) animals, suggesting a high prevalence of Sarcocystis in the Icelandic reindeer population. Cyst morphology and the ssu rRNA gene sequence of each of the three species were identical to isolates of the same species from Norwegian reindeer. DNA sequencing was useful in order to identify cysts with an ambiguous morphology. This is the first record of these Sarcocystis species in reindeer outside Norway.     

Studies on the pathogenesis and interspecies transmission of respiratory syncytial virus isolated from sheep

Inoculation of lambs with an ovine isolate of respiratory syncytial virus (RSV) by a combined intranasal and intratracheal route resulted in mild respiratory tract illness, with respiratory tract lesions. Lung lesions were characterized by bronchitis and bronchiolitis, hyperplasia of bronchial and bronchiolar epithelium, peribronchiolar and perivascular accumulations of lymphocytes, alveolar septal thickening, and collapse. Respiratory syncytial virus was recovered from the respiratory tract of inoculated lambs, and RSV antigen was demonstrated by immunoperoxidase staining of bronchiolar and alveolar epithelial cell in pneumonic lesions of lambs euthanatized on post-inoculation days 5 and 6. Other primary respiratory tract pathogens were not isolated. Clinical signs of respiratory tract illness or respiratory tract lesions did not develop in the in-contact control lamb. Inoculation of the ovine RSV isolate into calves and deer fawns resulted in infection in both species, and at necropsy, pneumonic lesions were present. A mild to moderate respiratory tract illness developed in the calves, but clinical disease was not seen in the fawns. Lung lesions in fawns were similar to those seen in lambs; lesions in calves were characterized by collapse, scattered areas of parenchymal necrosis, and bronchiolitis. Respiratory syncytial virus was reisolated from the lower respiratory tract of inoculated calves and fawns, and immunoperoxidase-positive epithelial cells were seen in pneumonic lesions. Other primary respiratory pathogens were not detected. Respiratory syncytial virus infection was not demonstrable in control animals that were in contact with inoculated animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Seasonal changes in digestibility, passage rate and rumen fermentation of alfalfa hay in sika deer (Cervus nippon) under restricted feeding

To investigate seasonal variations in the digestive functions of sika deer, five female sika deer were provided with an amount of alfalfa hay cubes equivalent to voluntary food intake during winter. We measured the rate at which the food passed through the digestive tract, digestibility and rumen fermentation during the summer (August), autumn (November), winter (February) and spring (May). Total mean retention time in the digestive tract during summer and autumn was numerically longer than that in winter and spring, but the difference did not reach significance. Organic matter and fiber were less digestible in summer and autumn than in winter and spring (P < 0.05), whereas the digestibility of the dry matter tended to vary with the seasons (P < 0.1). Ruminal pH values seasonally changed (P < 0.01), and were the lowest in autumn. The concentration of ruminal ammonia‐nitrogen differed significantly among the seasons (P < 0.05), increasing in winter and decreasing during spring and summer. The numbers of protozoa changed significantly among the seasons (P < 0.05), being higher in autumn than in winter and spring, and intermediate in summer. The concentration of total volatile fatty acids was not seasonally affected, but the molar percentages of propionic acid and butyric acid significantly changed according to season (P < 0.05 and P < 0.01, respectively), and the ratio of acetic to propionic acid tended to change with the seasons (P < 0.1). The results of this study suggested that the digestive functions in sika deer, fed a commercial diet at a restricted level, differed notably among the seasons and these variations might partially be due to environmental effects.     

The fox as definitive host for Sarcocystis sp. Gjerde, 1984 from skeletal muscle of reindeer (Rangifer tarandus). With a proposal for Sarcocystis tarandivulpes n. sp. as replacement name

Skeletal muscle of 5 wild reindeer was examined for sarcocysts and used for experimental infection of 6 foxes. Skeletal and cardiac muscle of another reindeer were only examined for sarcocysts. The skeletal muscle of all animals was infected with Sarcocystis sp.. In 2 of the animals cysts of S. hardangeri were also present. The single heart examined contained only cysts of S. grueneri.Four foxes given skeletal muscle containing apparently only cysts of Sarcocystis sp., started shedding Sarcocystis sporocysts, measuring on average 13.6×9.8 µm, after a prepatent period of 10–12 days. Two foxes given skeletal muscle containing cysts of both Sarcocystis sp. and S. hardangeri shed similar sporocysts, measuring on average 13.5×9.7 µm, after a prepatent period of 10–12 days.Based on the results from the present and previous investigations, Sarcocystis sp. is considered to have foxes (Vulpes vulpes and Alopex lagopus) and dogs (Ganis familiaris) as definitive hosts, becoming the second species of Sarcocystis with a known reindeer/Canidae life cycle. The name Sarcocystis tarandivulpes n. sp. is proposed as a replacement name for Sarcocystis sp. Gjerde, 1984 from skeletal muscle of reindeer.     

Tolazoline-induced apnea in mule deer (Odocoileus hemionus)

Eighteen mule deer (Odocoileus hemionus) and six Columbia black-tailed deer (Odocoileus hemionus columbianus) were held in pens and repeatedly anesthetized from April 2004 through June 2005 as part of an external parasite study. Deer were anesthetized using a combination of Telazol and xylazine hydrochloride (HCL) administered intramuscularly. Tolazoline HCL was slowly administered at 4 mg/kg intravenously to reverse the effects of xylazine with good results. For 17 of the 19 mule deer anesthesias in the fall of 2004, a mean dose of 7.3 mg/kg of intravenous tolazoline (range 6.1-8.4 mg/kg) was given by mistake. This paper describes clinical signs of apnea, muscle tensing, and fasciculations immediately following intravenous administration of tolazoline HCL in mule deer (O. hemionus) at 1.5-3 times the recommended dose. Mean dose for black-tailed deer during this time was 8.1 mg/kg (range 5.5-12.4 mg/kg) with no clinical signs as seen in the mule deer. Based on these findings, intravenous tolazoline use in mule deer is recommended at < or = 4 mg/kg.     

Clinical Sarcocystis neurona encephalomyelitis in a domestic cat following routine surgery

Sarcocystis neurona is an important cause of equine protozoal myeloencephalitis (EPM) in horses in the Americas. An EPM-like neurological disease also has been reported from other mammals but it is difficult to induce this disease in the laboratory. A 4-month-old male domestic cat developed neurological signs 3 days following castration. The cat was euthanized 12 days later because of paralysis. Encephalomyelitis was the only lesion and was associated with numerous Sarcocystis schizonts and merozoites in the brain and spinal cord. The protozoa reacted positively with S. neurona-specific polyclonal rabbit antibody. Two unidentified sarcocysts were present in the cerebellum. It may be possible that stress of surgery triggered relapse of S. neurona infection in this cat.     

Investigation of the transmission of Mycobacterium bovis from deer to cattle through indirect contact

OBJECTIVE: To investigate the infection of calves with Mycobacterium bovis through oral exposure and transmission of M. bovis from experimentally infected white-tailed deer to uninfected cattle through indirect contact. ANIMALS: 24 11-month-old, white-tailed deer and 28 6-month-old, crossbred calves. PROCEDURE: In the oral exposure experiment, doses of 4.3 x 10(6) CFUs (high dose) or 5 x 10(3) CFUs (low dose) of M. bovis were each administered orally to 4 calves; as positive controls, 2 calves received M. bovis (1.7 x 10(5) CFUs) via tonsillar instillation. Calves were euthanatized and examined 133 days after exposure. Deer-to-cattle transmission was assessed in 2 phases (involving 9 uninfected calves and 12 deer each); deer were inoculated with 4 x 10(5) CFUs (phase I) or 7 x 10(5) CFUs (phase II) of M. Bovis. Calves and deer exchanged pens (phase I; 90 days' duration) or calves received uneaten feed from deer pens (phase II; 140 days' duration) daily. At completion, animals were euthanatized and tissues were collected for bacteriologic culture and histologic examination. RESULTS: In the low- and high-dose groups, 3 of 4 calves and 1 of 4 calves developed tuberculosis, respectively. In phases I and II, 9 of 9 calves and 4 of 9 calves developed tuberculosis, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that experimentally infected deer can transmit M. bovis to cattle through sharing of feed. In areas where tuberculosis is endemic in free-ranging white-tailed deer, management practices to prevent access of wildlife to feed intended for livestock should be implemented.     

Experimental deer-to-deer transmission of Mycobacterium bovis

OBJECTIVE: To determine whether Mycobacterium bovis can be transmitted from experimentally infected deer to uninfected in-contact deer. ANIMALS: Twenty-three 6-month-old white-tailed deer. PROCEDURE: On day 0, M bovis (2 X 10(8) colony-forming units) was administered by intratonsillar instillation to 8 deer; 3 control deer received saline (0.9% NaCl) solution. Eight in-contact deer were comingled with inoculated deer from day 21. On day 120, inoculated deer were euthanatized and necropsied. On day 180, 4 in-contact deer were euthanatized, and 4 new in-contact deer were introduced. On day 360, all in-contact deer were euthanatized. Rectal, oral, and nasal swab specimens and samples of hay, pelleted feed, water, and feces were collected for bacteriologic culture. Tissue specimens were also collected at necropsy for bacteriologic culture and histologic analysis. RESULTS: On day 90, inoculated and in-contact deer developed delayed-type hypersensitivity (DTH) reactions to purified protein derivative of M bovis. Similarly, new in-contact deer developed DTH reactions by 100 days of contact with original in-contact deer. Tuberculous lesions in in-contact deer were most commonly detected in lungs and tracheobronchial and medial retropharyngeal lymph nodes. Mycobacterium bovis was isolated from nasal secretions and saliva from inoculated and in-contact deer, urine and feces from in-contact deer, and hay and pelleted feed. CONCLUSIONS AND CLINICAL RELEVANCE: Mycobacterium bovis is efficiently transmitted from experimentally infected deer to uninfected in-contact deer through nasal secretions, saliva, or contaminated feed. Wildlife management practices that result in unnatural gatherings of deer may enhance both direct and indirect transmission of M bovis.     

Prevalence of dog-derived Sarcocystis spp. in some New Zealand lambs

Pepsin: hydrochloric acid digestion and histological examination of the myocardium of 51 hearts from five to nine month-old lambs from the southern half of the North Island, showed that 47 (92%) were infected with microscopic sarcocysts that are presumed to be dog-derived. Of 157 sarcocysts detected histologically, 32 corresponded morphologically with Sarcocystis ovicanis and 120 with Sarcocystis tenella (Syn. S. arieticanis). The remainder could not be identified as to type.     

Prevalence of Sarcocystis in domestic pigs in India.

Muscle samples from 890 slaughtered pigs were examined for the presence of sarcocysts. A high prevalence rate of 67.98% was observed. Two types of microsarcocysts were recorded. The sarcocyst wall of one type had redial striations and the other possessed hair-like villar protrusions. The species were identified as Sarcocystis miescheriana and Sarcocystis suihominis; there was a slightly higher incidence of the latter species (47.11%) than of the former (43.14%). S. suihominis has been identified for the first time from pigs in India.