Life cycle of Frenkelia. IV. Pathomorphological findings in the organs of experimentally infected bank voles]

In 1975 the buzzard (Buteo buteo) was found to be the final host of Frenkelia clethrionomyobuteonis. After this discovery it became possible to investigate systematically the pathomorphology of the infection in the intermediate host, the bank vole (Clethrionomys glareolus). Fifty bank voles were infected orally with a suspension of sporocysts recovered from the faeces of experimentally infected buzzards. Each rodent receive 7000 sporocysts. Six controls each were given a faecal suspension from a non-infected buzzard. The voles were killed between 1 and 140 days after infection and examined histologically. Between the 5th and 8th day of the infection during the schizogonic multiplication of the parasite a focal necrosis of liver cells and of the liver parenchyma is observed followed by a reversible resorptive inflammation associated with siderophagia and the occurrence of giant cells. The spleen was spodogenously enlarged up to twice its normal size. There also was haemosiderosis of the bone marrow, the liver and the spleen up to 25 days after infection. At the same time the erythropoiesis in the bone morrow, the spleen and in the lymph nodes increased; there also was a lymphoid hyperplasia in spleen and lymph nodes. About 10 days after infection a reversible infiltration with lymphocytes and plasma cells developed in the liver, heart and brain. This infiltration was again detectable as perivascular and meningeal reactions in the brain after the 49th day after infection. The second asexual multiplication of the parasite was seen histologically in the grey and white matter of the central nervous system after the 18th day of infection. The developing cysts increased in size continuously thereby compressing the surrounding nervous tissue. Disseminated focal necrosis with resorptive inflammatory components was prominent in the parenchyma of the brain after the 49th day of infection. It was possible to differentiate between damage in single organs and systemic pathological lesions. The lesions in single organs were directly connected with the development of parasitic stages in the liver (schizonts) and in the brain (cysts). The generalized lesions occurred in the haemopoietic system after an impairment of the blood during the first asexual multiplication. They also occurred in the immunocytic systems after the first and during the second asexual multiplication and during the relatively late cystic phase of the parasite in the brain. The pathogenesis of the disintegration of blood cells is not clear. The immunocytic reaction can be considered an immunological response of the host against the parasite. The effect of the development of the cysts on the function and structure of the central nervous system is expected to lead to an increasing impairment of the motility of the intermediate host.     

Host resistance and faecal sporocyst excretion in dogs exposed to repeated infection with Sarcocystis levinei

A marked reduction in the faecal excretion of sporocysts was observed in experimental pups, following the repeated oral administration to them of buffalo cardiac muscle infected with Sarcocystis levinei. Sporocysts excreted from days 9 to 25 post-infection (pi) exhibited a gradual reduction in the quantum. Maximum intensity of excretion of sporocysts was recorded between days 9 and 16 pi, becoming moderate after day 16, light after day 21 and completely absent after day 36. After the subsequent feeding to pups of S. levinei infected buffalo cardiac tissues at 40 day intervals the quantity of sporocysts shed was less, the prepatent period was prolonged and the patent period was considerably shortened. The peak period of excretion varied depending upon the number of exposures of the pups to the infected S. levinei tissues from buffaloes (Bubalus bubalis).     

Abortion and death in goats inoculated with Sarcocystis sporocysts from coyote feces

Ten 75- to 105-day-pregnant does each were inoculated orally within 1 million (2 does), 10,000 (4 does), or 1,000 (4 does) sporocysts of Sarcocystis from coyote feces. Two does not inoculated with sporocysts served as controls. The 2 does inoculated with 1 million sporocysts died from acute sarcocystosis 21 and 22 days after inoculation (DAI), and each had 2 dead fetuses. The 4 does inoculated with 10,000 sporocysts were ill 19 to 33 DAI but survived; 1 aborted at 33 DAI, 1 had a live kid that died within 2 hours of birth 31 DAI, 1 aborted 2 dead fetuses 23 DAI, and 1 had a normal kid 56 DAI. The 4 does inoculated with 1,000 sporocysts and the 2 control does remained clinically normal and had normal kids. Does and their offspring were killed within 24 hours of parturition, and their tissues were examined histologically and microbiologically. Meronts of Sarcocystis were found in the maternal placenta of does inoculated with 1 million sporocysts. Sarcocystis was not found in the placenta, fetuses, or tissues of kids from does inoculated with 10,000 or 1,000 sporocysts, or from control does. Other abortifacient agents were not found in the placenta, fetuses, or kids from any does.     

Effect of a single dose of ponazuril on neural infection and clinical disease in Sarcocystis neurona-challenged interferon-gamma knockout mice

Interferon gamma-knockout mice were challenged with 5000 Sarcocystis neurona sporocysts acquired from a naturally infected opossum. Ponazuril was administered once, by gavage, at day 1, 3, 7, 10, or 14 post-infection (pi). Ponazuril was given at either 20 or 200mg/kg. Mice that survived to day 30 pi were euthanized. Severity of CNS infection was quantified as schizont density in the cerebellum. Unchallenged mice in treatment and non-treatment groups remained free of disease and gained weight throughout the experiment. All challenged mice, regardless of treatment, developed histologic evidence of CNS infection even though clinical signs were prevented in some groups. The greatest treatment benefits were seen in mice given 200mg/kg ponazuril between days 4 and 14 pi. Weight gain over the course of the experiment occurred only in mice that were given 200mg/kg ponazuril on day 7 or 10 pi. With the exception of groups given 200mg/kg ponazuril on day 7 or 14 pi, mice in groups that got sporocysts developed abnormal neurologic signs. No deaths before day 30 pi occurred in mice given ponazuril at 20mg/kg on day 7 pi or 200mg/kg on day 1, 7, 10, or 14 pi. This effect was not significant. Mice given 200mg/kg on day 7 pi had significantly fewer cerebellar schizonts than did those of the control group that was not given ponazuril. These results indicate that single-dose administration of ponazuril for prevention of CNS infection is partially protective when given between days 4 and 14 pi.     

Experimental Sarcocystis levinei infection in buffalo calves

Six buffalo calves were orally inoculated with 3 graded doses of sporocysts of Sarcocystis levinei (0.5, 1.0 and 2.0 million sporocysts; 2 calves for each dose) while two more calves were kept as uninoculated controls. One calf from each group was killed at 30 days post infection (DPI) and the other at 80 DPI. Inoculated calves showed a dose dependent response. The calves inoculated with 0.5 and 1.0 million sporocysts did not manifest any clinical signs of disease up to 80 DPI. One of the two calves inoculated with 2.0 million sporocysts showed clinical signs of weakness, emaciation and anaemia during the 5th week post infection. The other calf remained healthy until it was killed at 30 DPI. Pale liver tissue, gelatinization of fat and haemorrhages in the heart were observed in one calf inoculated with 2.0 million sporocysts; only microscopic lesions were seen in other calves. Schizonts and merozoites were not observed in any calf. Mature sarcocysts were observed in cardiac and skeletal muscle of calves killed at 80 DPI whereas no sarcocysts were seen in calves killed at 30 DPI.     

Early hepatic immune response in rats infected with Fasciola hepatica

We investigated the phenotype of the T cells (CD4+ and CD8+) that produced Th1 (IFN-gamma) and Th2 cytokines (IL-4 and IL-10) during the firsttwo weeks of experimental fasciolosis in rats. We also followed the kinetics of the cytokine and proliferative responses of hepatic mononuclear cells (HMNC) over the same period. We found that HMNC were more numerous in the infected animals than in the controls. The percentage of CD4+ cells increased significantly after infection, whereas the percentage of CD8+ cells did not change. Moreover, the frequency of the cells producing (CP) cytokine changed after infection. The frequency of CP IFN-gamma on 7 days postinfection (pi) was similar to that in control animals. However, the frequency of CP IFN-gamma was clearly lower on day 14 pi, whereas the frequency of CP IL-4 and CP IL-10 had increased. The CP IL-10-were mostly CD4+. Mitogenic stimulation (phorbol myristate acetate/ionomycin) of HMNC led to an increase in the amounts of the Th2 cytokines in the supernatant on days 7 and 14 pi, with the increase more pronounced on day 14. In contrast, IFN-gamma levels also increased by day 7 pi but then decreased to below control levels by day 14. In addition, HMNC proliferation in response to mitogen followed a similar pattern to IFN-gamma production. These findings suggested that, during the first 2 weeks of infection, F hepatica induced a transient ThO cytokine profile followed by downregulation of the cellular response and the induction of a Th2 cytokine profile.     

Evaluation of the shedding of Sarcocystis falcatula sporocysts in experimentally infected Virginia opossums (Didelphis virginiana)

Five Virginia opossums (Didelphis virginiana) were fed muscles of brown-headed cowbirds (Molothrus ater) containing sarcocysts of Sarcocystis falcatula. Shedding of sporocysts was confirmed in all five opossums by fecal flotation. Counts were conducted daily for 2 weeks and then biweekly until the animals were euthanized and necropsied. The average prepatent period was 9.8 (7-16) days. The number of sporocysts shed varied greatly between the opossums with maximum mean shedding occurring at 71.6 (26-112) days post-infection (DPI). Average sporocyst production was 1480 sporocysts/gram of feces (SPG). Maximum output was 37,000 SPG. Average fecal yield in captivity was 17.5g of feces/day. Opossums shed 25,900 sporocysts/day (average) and a maximum of 647,500 sporocysts/day. All opossums shed sporocysts until time of euthanasia (46-200 DPI). Histologically, numerous sporocysts were present in the lamina propria at necropsy, primarily in the proximal half of the small intestine. Sporocysts were generally in clusters within the lamina propria of the luminal two-thirds of the villi. Sporocysts were found less frequently in the epithelium. No evidence of ongoing gametogony or other development was evident.     

Pathologic features of polybrominated biphenyl toxicosis in the rat and guinea pig.

Young male rats were fed a diet containing 0, 1, 10, 100, or 500 ppm of a commercial mixture of polybrominated biphenyls (PBB) that had been accidentally incorporated into a mineral mixture and fed to Michigan livestock and poultry. After 30 days, 9 of the 12 rats in each group were killed and tissues were examined. Liver weight to body weight ratios were significantly increased at all feeding levels; at 500 ppm, liver weight had more than doubled. Kidney weight was not affected. Microscopic lesions were mostly confined to the liver and consisted of extensive swelling and vacuolation of hepatocytes in rats fed diets containing 100 and 500 ppm of PBB. Slight swelling and vacuolation were seen in rats fed the diet containing 10 ppm, and lesions were not found at 0 or 1 ppm. There was a significant increase in hepatic mitochondrial size at 1 ppm, and smooth endoplasmic reticulum was markedly increased at 100 and 500 ppm. Myelin bodies were present at 100 and 500 ppm, and vacuoles were numerous. Rats killed at 60 days had similar lesions. The activity of hepatic microsomal enzymes increased at all levels of feeding of PBB. Rat pups nursing dams fed a diet containing 10 ppm of PBB had microscopic and ultrastructural hepatic lesions. When guinea pigs were fed PBB at the same amounts as were rats, the results were strikingly different. Guinea pigs fed a diet containing 500 ppm of PBB died within 15 days; at 100 ppm, only 2 of 6 survived for 30 days. Effects on liver weight were inconsistent, but 2 of 6 fed a diet containing 10 ppm had enlarged livers.     

Pathologic findings in rabies-suspect, random-source, and accidentally killed skunks

To evaluate sampling biases, pathologic findings in accidentally killed skunks (ie, killed by motor vehicles) were compared with those in random-source skunks (live-trapped and euthanatized, or trap-killed during research) and skunks submitted to a public health laboratory as rabies-suspect. Presence or absence of microscopic lesions in the brain, kidneys, liver, and lungs were used to test the null hypothesis that prevalence of disease did not differ by source of collection. Brain lesions differed with the source; rabid and nonrabid skunks submitted to a public health laboratory had higher prevalences of lesions than did other skunks. Kidney, liver, and lung lesions did not differ among skunks by source of collection. Liver and lung lesions were attributed mainly to parasitism, were not severe, and did not cause debilitated condition. Lesions were seen more often in the kidneys than in other tissues. Usually, lesions were mild to severe, focal, chronic, nonsuppurative, interstitial nephritis (possibly a consequence of leptospirosis). Six of 177 skunks necropsied appeared cachectic. Aleutian disease was diagnosed in one skunk and histoplasmosis was diagnosed in another, but rabies and canine distemper virus infection were the only diseases found with the potential to cause the high population mortality. Public health surveillance cases were biased toward diseased animals (rabies and canine distemper virus infection), but random-source or accidentally killed animals provided unbiased data. Although other factors must be considered, accidentally killed skunks provided cost-effective and useful data for the evaluation of enzootic rabies.     

Demonstration of schizogonous stages of Sarcocystis gigantea in experimentally infected sheep

Sarcocystis-free lambs were orally dosed with 1 X 10(6) sporocysts of Sarcocystis gigantea. Schizonts were found in endothelial cells of capillaries and arterioles of the brain, lung and kidney of lambs 7 and 14 days post-inoculation (d.p.i.). Between 21 and 35 d.p.i. there was extensive multi-focal encephalitis; however no organisms were detected in association with these lesions.     

Development of Sarcocystis alceslatrans Dubey, 1980, in the small intestine of dogs

Laboratory-reared dogs were fed moose musculature infected with Sarcocystis alceslatrans. These dogs shed sporocysts [15.6 X 11.4 microns (14.4 to 15.8 X 10.8 to 11.5)] 11 to 15 days after inoculation. The prepatent period was 10 to 14 days. Two cats and 1 coyote that also ate infected moose musculature did not pass sporocysts. Histologic examination of intestinal tissue from experimentally infected dogs revealed microgamonts, macrogametes, and oocysts. All stages were present in the lamina propria of the small intestine, usually in the luminal third of the villi. Infections were concentrated in the proximal half of the small intestine. Oocysts were first noticed in dogs killed 7 days after inoculation and a sequence of sporogonic development occurred in dogs killed on subsequent days. Ultrastructural observations were made on the oocyst and sporocyst walls during sporogony.     

交叉感染后黄牛与水牛枯氏住肉孢子虫包囊超微结构的比较研究

选用4头7日龄奶牛和4头4～5月龄水牛,用水牛源孢子囊感染黄牛及黄牛源孢子囊感染水牛,同时设感染对照和不感染对照,对交叉感染后黄牛与水牛体内包囊的超微结构进行了比较研究,结果发现两者无结构区别,所有包囊的超微结构均与前人对黄牛和水牛枯氏住肉孢子虫包囊的描述一致,证实水牛与黄牛同是枯氏住肉孢子虫的中间宿主。作者还首次在枯民住肉孢子虫包囊的母细胞和缓殖子发现晶状体。     

Unusual biphasic disease in domestic pigeons (Columba livia f. domestica) following experimental infection with Sarcocystis calchasi

A novel Sarcocystis species has recently been reported in the domestic pigeon (Columba livia f. domestica) as intermediate host, causing severe central nervous signs similar to Paramyxovirus-1 or Salmonella Typhimurium var. cop. infection. Transmission of the parasite via the northern goshawk (Accipiter gentilis) as definitive host has been established. Experimental infection of domestic pigeons with sporocysts excreted by experimentally infected northern goshawks reproduced the natural infection in the pigeon, proving the causative role of the parasite in the disease. Here, we describe in greater detail the course of the fulminant biphasic disease depending on the infectious dose. Pigeons infected with 10(3) or 10(4) sporocysts showed clinical signs of polyuria and apathy around 10-11 days postinfection (dpi) and sudden neurological signs 51-57 dpi as a second phase of disease. Pigeons infected with higher doses died within 7-12 dpi, also showing polyuria and apathy but without nervous signs. At necropsy, livers and spleens had multifocal necroses and infestations with parasitic stages, namely, schizonts. Moreover, lesions and schizonts were also found in the lung, bone marrow, and next to blood vessels in the connective tissue of various organs. Pigeons infected with 102 sporocysts remained symptomless until 58-65 dpi, when sudden central nervous signs occurred. Major histopathologic findings of pigeons with neurological signs were encephalitis and myositis of virtually every skeletal muscle with high infestations of sarcocysts. Only mild myocarditis and very few cysts were found in the heart muscles. Importantly, a sentinel pigeon developed identical lesions when compared to those of low-dose infected pigeons, suggesting a risk of mechanical transmission of sporocysts from freshly infected to uninfected pigeons in a flock. By contrast, chickens failed to develop any clinical signs or pathologic lesions in the same experiment. The findings further characterize the new highly pathogenic disease in domestic pigeons, which clinically mimics paramyxovirosis and salmonellosis in both phases of the disease and exclude chickens as further intermediate host species.     

Experimental microcyst sarcocystis infection in lambs: pathology

Six 34- to 42-day-old lambs raised in coccidia-free conditions were inoculated with 70,000 sporocysts derived from sheep heart with microscopic sarcocysts. Fever and mild anorexia occurred between 25 and 33 days after inoculation. A transient anaemia was most marked 32 days after inoculation. Lambs were killed and examined 14, 25, 33, 42, 60 and 81 days after inoculation. Gross lesions were absent. First and second generation meronts were present in endothelial cells at 25 and 33 days after inoculation. Meronts were most numerous in kidney glomeruli. Developing sarcocysts were rare at 42 days after inoculation. Sarcocysts with a primary cyst wall 2 to 3 micron thick composed of palisade projections were common at 60 and 81 days after inoculation in striated muscle and brain. Mild to severe striated muscle myositis and non-suppurative encephalitis or encephalomyelitis with glial nodules were observed 25 to 81 days after inoculation. Sarcocyst frequency varied considerably; it was highest in myocardium, M vastus intermedius, M vastus medialis, M extensor carpi radialis and tongue muscle and was lowest in M masseter.     

Mycoplasma hyopneumoniae increases the susceptibility of pigs to experimental Pasteurella multocida pneumonia.

The interaction between Mycoplasma hyopneumoniae and Pasteurella multocida in experimental pneumonia was investigated in conventional pigs. The experimental animals were 49 days old when inoculated with M. hyopneumoniae; they were inoculated with P. multocida after 23 days, and killed 13 days later. In pigs inoculated only with P. multocida, clinical signs and lung lesions were not observed, and the agent was not recovered. Pigs inoculated with M. hyopneumoniae developed fever, moderate cough and dyspnea which tended to disappear, and small proliferative lung lesions from which M. hyopneumoniae was isolated. Pigs inoculated with both agents had higher fever, severe cough and dyspnea which tended to aggravate, and extensive exudative lung lesions from which organisms were isolated. All animals had similar growth rates, but the group infected with both agents consumed 60% more food. Therefore, M. hyopneumoniae causes mild pneumonia, whereas P. multocida is not pathogenic alone but aggravates the pneumonia initiated by M. hyopneumoniae.     

Larval toxocarosis in sheep: the immunohistochemical characterization of lesions in some affected organs

Morphological and immunohistochemical responses of lambs following oral infection with 10,000 infective Toxocara canis (T. canis) eggs were studied up to 28 days post-infection (pi). The small intestine, liver, lungs and brain of both infected and control lambs were examined using the routine histological methods for paraffin sections and immunohistochemical techniques for frozen tissue sections. Eosinophil-rich hepatic granulomas and diffuse T. canis-induced pulmonary inflammation were the most prominent pathological features. CD2+, CD4+, CD8+, IgM bearing cells, and macrophages were demonstrated in the liver granulomas. The observed histomorphologic changes were similar to other paratenic hosts. It was concluded that larval toxocarosis followed the classical migratory path in the infected lambs.     

Lesions of the Gastrointestinal Tract of Pigs Infected with Transmissible Gastroenteritis

Gross, subgross and histological lesions were studied in 103 pigs infected with transmissible gastroenteritis virus and killed at daily intervals for 14 days. Twenty-three pigs served as controls. Thirty-six pigs were given colchicine four hours prior to being killed in order to determine the mitotic activity in the gastrointestinal tract. The gross lesions consisted of dehydration, excessive milk curd in the stomach, focal hemorrhage in the submucosa of the diverticulum ventriculi of the stomach, fundic and pyloric congestion in severly dehydrated animals and thinning of the small intestinal wall. The major subgross lesion was a marked shortening of the villi in the lower duodenum, jejunum and ileum within 24 hours after exposure to the virus. Regrowth of the villi became evident on about the sixth day after infection. Histological examination of the small intestine revealed that the villus-height/crypt-depth ratio in the jejunum was reduced from 7:1 in normal pigs to less than 1:1 in infected pigs. Villous atrophy was less severe in the proximal duodenum and ileum. Cells covering the atrophic villi were flatened or cuboidal and did not have well defined brush borders. Inflammatory changes in the gastrointestinal tract were minimal at all stages of infection. Goblet cell numbers increased slightly in the recovery stage of the disease and small numbers of mononuclear cells accumulated in the lamina propria during regrowth of the villi. The number of metaphase nuclei in the small intestinal crypts of infected pigs was greater than in normal pigs.     

Residues of endosulfan in the tissues of lactating goats

SUMMARY: The sites of tissue accumulation in lactating goats of the organochlorine insecticide endosulfan were studied. Twelve lactating goats were dosed orally with endosulfan (1 mg/kg body weight per day) for 28 days. Groups of 3 animals were killed on days 1, 8, 15, and 21 after endosulfan treatment ended and their tissues examined for the presence of endosulfan. Total residues of α and β endosulfan and endosulfan sulphate (mg/kg) were detected in kidney (0.29), gastro-intestinal tract (0.20), liver (0.12), brain (0.06), muscle and spleen (0.04), lung and heart (0.01) and milk (0.02) on the flrst sampllng day but within 15 days, concentrations had fallen to < 0.01 mg/kg in all tissues except kidney (0.20). Endosulfan could not be detected in animals 21 days after dosing had ceased. The residue in milk could only be detected on day 1 of sampling. This study indicates that kidney rather than fatty tissue should be used to monitor the presence of endosulfan in animals intended for human consumption.     

The pathology of experimental Lasiospermum bipinnatum (Thunb.) Druce (Asteraceae) poisoning in sheep. I. Hepatic lesions

Poisoning with the plant Lasiospermum bipinnatum was studied in 9 lambs. Intraruminal doses, varying from 1-12 g/kg/day of dried plant, were administered to 8 animals and 1 was fed 2.5 g/kg/day of the material mixed with maize meal for 13 days. Periodic serum analyses were done to monitor liver function. Lambs given 6-12 g/kg/day died or were killed in extremis. Clinical signs included progressive anorexia and depression in all these lambs and icterus in 2 animals. Lambs given 1-4 g/kg/day were sacrificed after about 2 weeks. Clinical signs in these animals were minimal or absent. Hepatosis was found in all the lambs, the severity of which correlated with levels of plant administered. Centrilobular necrosis and haemorrhage occurred in 2 of the 4 lambs given high doses; single cell necrosis of hepatocytes was observed with intermediate doses, and diffuse degeneration, which was more severe peripherally, was seen at various doses. In 1 lamb, degeneration was most severe midzonally. Bile ductule epithelial proliferation was observed in 7 of the 9 poisoned animals. Marked hypertrophy of hepatocellular smooth endoplasmic reticulum was seen in 3 lambs given low doses. The hepatic lesions were compared with those reported for poisoning by other hepatotoxic plants belonging to the family Asteraceae and found to be indistinguishable.     

ENCEPHALOMYOCARDITIS VIRUS INFECTION OF PIGS 2. Experimental Disease

Encephalomycarditis virus recovered from a pig mortality in New South Wales was used to produce experimental infections. Of 34 pigs exposed, 17 died and a further 7 were found to have severe heart lesions when killed. Deaths occurred from 2 to 11 days after exposure with a mode of approximately 3 days. Ten of 11 pigs exposed by intramuscular injection died and the remaining pig was killed after 28 days and found to have severe resolving heart lesions. Of 15 pigs exposed per os to various doses of virus, 6 died, 4 were killed and found to have severe heart lesions and 5 were apparently not infected. Intranasal exposure of 8 pigs resulted in 1 death and 4 pigs with mild to severe heart lesions. Different doses of virus and routes of exposure did not substantially influence the character of the lesions. Lesions were similar to those found previously in field cases, and virus was recovered from all of 19 animals examined with severe acute lesions of 10 days standing or less.