Pathology of Sarcocystis campestris infection in Richardson''s ground squirrels (Spermophilus richardsoni).

Richardson's ground squirrels were infected with 1500 or 9000 sporocysts of Sarcocystis campestris from badgers. No lesions were found in animals killed one to three days postinfection (pi). Hepatitis and phlebitis of hepatic veins were present in animals killed between four and eight days pi. No meronts were detected in these animals, but the lesions suggested that a generation of merogony occurred in the hepatic veins. Meronts were found in endothelial cells in many tissues beginning on day 9 pi. They were most numerous on day 10 pi, and less so on day 11, and subsequently. Meronts were most numerous in the lung; none was found in liver or spleen. Four of ten squirrels infected with 1500 sporocysts in one trial died between days 11 and 13 pi. There were petechial hemorrhages in skeletal muscle, lung, serosal membranes, and brain in these animals, with microscopic evidence of pulmonary, myocardial, and brain injury. One animal infected with 9000 sporocysts had petechiae in the liver at six days pi. Foci of inflammation were visible in the myocardium and brain of animals killed to 64 days pi. This species may serve as an experimental model for sarcocystosis in domestic animals.     

The raccoon dog (Nyctereutes procyonoides) as definitive host for Sarcocystis spp. of reindeer (Rangifer tarandus)

A raccoon dog (Nyctereutes procyonoides; Family: Canidae), was given cardiac muscle of reindeer infected with S. grueneri, and started shedding Sarcocystis sporocysts 10 days post feeding. The sporocysts measured 13.9 (12.4–15.7) × 10.1 (9.2–11.2) µm, and were excreted for at least 16 days. The raccoon dog is thus an additional definitive host for S. grueneri (Yakimoff & Sokoloff, 1934) Gjerde, 1984.Another raccoon dog was given skeletal muscle infected with 4 species of Sarcocystis, none of which was S. grueneri. The raccoon dog started shedding Sarcocystis sporocysts on day 10 post feeding, and excreted sporocysts for at least 16 days. The sporocysts measured 14.0 (12.3–15.6) × 10.1 (9.2–11.2) µm, and are considered to be sporocysts of S. tarandivulpes Gjerde, 1984.This is the first record of the raccoon dog as an experimental definitive host for Sarcocystis.     

Heavy sarcocystosis in the ocular musculature of cattle and buffaloes

A high prevalence of 71.5 per cent and 69.7 per cent of sarcocystosis was observed in the ocular musculature of cattle and buffaloes respectively, in Bihar, India. The concentration of cysts in the eye muscle was also usually heavy. Ocular musculature appears to be a preferred site for the development ofSarcocystis in these intermediate hosts, second only to the heart muscle. The species ofSarcocystis involved in the present study were morphologically indistinguishable fromS. cruzi in cattle andS. levinei in buffaloes. This appears to be the first report on the occurrence ofS. cruzi andS. levinei in ocular musculature.     

Effects of heating and freezing on the viability of sarcocysts of Sarcocystis levinei from cardiac tissues of buffaloes

Dogs fed buffalo heart muscle containing sarcocysts of Sarcosystis levinei and heated at 65-75 degrees C did not shed sporocysts, whereas other dogs fed infected heart muscle heated between 40 and 60 degrees C shed sporocysts. Dogs fed infected heart muscle stored at -4 degrees C for 48 h did not shed sporocysts, but those fed similar infected tissues stored at -2 degrees C for 24 h shed sporocysts. The results indicate that sarcocysts of S. levinei are rendered noninfective by heating to 65 degrees C or by freezing at -4 degrees C.     

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。     

Transmission of Cowdria ruminantium by Amblyomma gemma from Infected African Buffalo (Syncerus caffer) and Eland (Taurotragus oryx) to sheep

Two African buffalo (Syncerus caffer), an eland (Taurotragus oryx) and a waterbuck (Kobus defassa) were intravenously inoculated with Cowdria ruminantium (Kiswani). Amblyomma gemma nymphs were fed on the animals at 3 weekly intervals. Jugular blood was also collected at 3 weekly intervals and inoculated into sheep. Nymphal ticks that fed on one buffalo on days 16 and 37 and on the other buffalo on day 58 after infection transmitted the disease as adults to sheep. Nymphs that were applied to the eland 16 days after infection also transmitted the disease to sheep. No nymphs that had fed on the waterbuck transmitted the disease. This is the first report of transmission of heartwater by Amblyomma gemma from infected wild ruminant species to a susceptible domestic ruminant species.     

Haematological and biochemical changes in buffalo calves inoculated with Sarcocystis fusiformis from cats

Two groups of buffalo calves were infected with Sarcocystis fusiformis sporocysts. Animals of the first group received each 5 X 10(5) sporocysts, those of the second group 5 million sporocysts. All calves were clinically normal during 6 weeks after infection. Minor changes were observed in the blood cytology, serum alkaline phosphatase, glutamic oxalacetic transaminase, glutamic pyruvic transaminase, total proteins, urea and glucose of infected buffalo calves.     

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.     

Infectious Pancreatic Necrosis Virus: Transmission from Infectious to Susceptible Rainbow Trout Fry

Abstract

Fry of rainbow trout Oncorhynchus mykiss were exposed to serotype VR-299 of infectious pancreatic necrosis virus (IPNV) by using a standardized immersion challenge. In concurrent experiments, fish were monitored for 11 d for excretion of IPNV or monitored for 9 d for excretion and transmission of IPNV to susceptible rainbow trout fry. Immersion-challenged fish began excreting virus within 2 d after challenge. The rate of IPNV excretion per fish increased steadily from about day 4 to day 8 and then decreased. Virus concentrations in tissues of immersion-challenged fish increased exponentially. Susceptible fish became infected with IPNV within 4 d after being introduced to immersion-challenged fish (e.g., 2 d after the challenged fish began excreting virus). By 9 d, 84% of the susceptible fish were infected with IPNV.     

Effect of temperature on the infectivity of Sarcocystis miescheriana cysts in pork

Sarcocysts of Sarcocystis miescheriana in the thigh muscles of pigs became non-infective to pups after heating infected pork in minute pieces at 60 degrees C for 20 min, 70 degrees C for 15 min and 100 degrees C for 5 min. Similar pieces of infected muscle tissues, when exposed to -4 degrees C for 2 days or -20 degrees C for 1 day, became non-infective to pups. The experiment suggests that pork containing sarcocysts of S. miescheriana, and possibly of S. suihominis, requires cooking at a minimum of 70 degrees C for 15 min or freezing at -4 degrees C for 2 days or -20 degrees C for 1 day for making it safe for consumption.     

Effect of intermittent oral administration of ponazuril on experimental Sarcocystis neurona infection of horses

OBJECTIVE: To evaluate the effect of intermittent oral administration of ponazuril on immunoconversion against Sarcocystis neurona in horses inoculated intragastrically with S neurona sporocysts. ANIMALS: 20 healthy horses that were seronegative for S neurona-specific IgG. PROCEDURES: 5 control horses were neither inoculated with sporocysts nor treated. Other horses (5 horses/group) each received 612,500 S neurona sporocysts via nasogastric tube (day 0) and were not treated or were administered ponazuril (20 mg/kg, PO) every 7 days (beginning on day 5) or every 14 days (beginning on day 12) for 12 weeks. Blood and CSF samples were collected on day - 1 and then every 14 days after challenge for western blot assessment of immunoconversion. Clinical signs of equine protozoal myeloencephalitis (EPM) were monitored, and tissues were examined histologically after euthanasia. Results: Sera from all challenged horses yielded positive western blot results within 56 days. Immunoconversion in CSF was detected in only 2 of 5 horses that were treated weekly; all other challenged horses immunoconverted within 84 days. Weekly administration of ponazuril significantly reduced the antibody response against the S neurona 17-kd antigen in CSF. Neurologic signs consistent with EPM did not develop in any group; likewise, histologic examination of CNS tissue did not reveal protozoa or consistent degenerative or inflammatory changes. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of ponazuril every 7 days, but not every 14 days, significantly decreased intrathecal anti-S neurona antibody responses in horses inoculated with S neurona sporocysts. Protocols involving intermittent administration of ponazuril may have application in prevention of EPM.     

Diclazuril preventive therapy of gamma interferon knockout mice fed Sarcocystis neurona sporocysts

Gamma interferon knockout (KO) mice (n=74) were fed a lethal dose of approximately 1000 sporocysts of the SN15-OP isolate of Sarcocystis neurona. Groups of mice were given pelleted rodent feed containing 50ppm of diclazuril at different times before and after feeding sporocysts. All mice were examined at necropsy and their tissues were examined immunohistochemically for S. neurona infection. Twenty mice were fed sporocysts and given diclazuril starting 5 days before feeding sporocysts and continuing 30-39 days post-infection (p.i.). One mouse died of causes unrelated to S. neurona with no demonstrable parasites; the remaining 19 mice remained clinically normal and S. neurona organisms were not found in their tissues. Sarcocystis neurona organisms were not demonstrable by bioassay of the brains of these 19 mice in uninfected KO mice. Sarcocystis neurona organisms were not found in tissues of five mice treated with diclazuril, starting 7 days after feeding sporocysts and continuing up to 39 days p.i. Therapy was less efficient when diclazuril was given 10 days p.i. Sarcocystis neurona organisms were found in two of 19 mice treated with diclazuril starting 10 days after feeding sporocysts, in two of five mice starting therapy 12 days p.i., and in 10 of 10 mice when treatment was delayed until 15 days p.i. All 15 mice fed S. neurona, but not given diclazuril, developed neural sarcocystosis and were euthanized 22-30 days after feeding sporocysts. Six mice not fed S. neurona, but given diclazuril for 44 days, remained clinically normal. Results indicate that diclazuril can kill the early stages of S. neurona.     

A comparative study of the pathogenic properties and transmissibility of a Greek and a Belgian encephalomyocarditis virus (EMCV) for piglets

Thirteen susceptible piglets, aged 40 days, were divided into two groups and were experimentally infected either with a Greek (myocardial) or a Belgian (reproductive) encephalomyocarditis virus (EMCV) strain (total dose 5 × 10⁶ TCID₅₀, intramuscularly and intranasally). Six piglets were placed in the same rooms, 24 h later, as contact controls. The following criteria were studied: ante mortem: clinical signs, serum cardiac isoenzyme activities (CK-MB and LD-1), viraemia, nasal and faecal virus excretion and serological response. Post mortem (after death or euthanasia): gross lesions, virus isolation from tissues, RT-PCR, as well as histopathological and immunohistochemical findings. The Greek strain was more pathogenic, producing mortality, with high cardiac isoenzyme activities and pronounced macroscopic myocardium lesions. The Belgian strain was able to induce mild heart lesions, as detected only by cardiac isoenzyme activity and histopathologically. All contact pigs were infected, within the first 1–2 days of their introduction, that coincided with the period of viral excretion by the experimentally infected pigs (up to the 3rd day post infection). Disease was mild, with no mortality.     

Development of sarcocysts of Sarcocystis levinei in water buffalo infected with sporocysts from dogs

Half a million sporocysts of Sarcocystis levinei obtained from experimentally infected dogs were fed to a buffalo calf, and sarcocysts of this species were recovered from its oesophageal muscles when the animal was killed on the 62nd day of inoculation, thus establishing a buffalo-dog-buffalo cycle.     

Infectivity of Cryptosporidium parvum isolated from asymptomatic adult goats to mice and goat kids.

An experimental study was carried out in neonatal goat kids to examine the infectivity of Cryptosporidium oocysts, pattern of oocyst shedding and morphological changes in the intestine during the infection. Cryptosporidium oocysts isolated from adult asymptomatic goats, and identified as C. parvum by polymerase chain reaction (PCR) were used in this study. Of three 4-day-old goat kids, two were orally infected with C. parvum oocysts (10(5) oocysts in 10 ml PBS/kid). One goat kid given 10 ml PBS only by the oral route served as a control. Cryptosporidium oocysts were detected in the faeces of one infected kid on day 3 post-inoculation (pi) whereas in the other 6 days pi. The faecal oocyst counts gradually increased and the peak counts in the two kids were 2 x 10(6)g(-1) (on day 12 pi) and 3.2 x 10(6)g(-1) (on day 14 pi). The increase in faecal oocyst output coincided with diarrhoea in an infected kid from days 10-17 pi. Although the oocyst excretion declined gradually after the peak, both infected kids excreted oocysts until euthanized on days 20 and 22 pi. Light and scanning electron microscopic investigations of the ileum revealed the endogenous stages on the brush border of the enterocytes, infiltration of neutrophils and mononuclear cells into the lamina propria, atrophy, stunting and fusion of villi. For purposes of comparison, goat Cryptosporidium oocysts were inoculated orally (10(3) oocysts/mouse) to eight, 1-week-old mice. All experimental mice excreted oocysts from day 3 pi, and four infected mice continued to excrete oocysts up to day 42 pi. The experimental infection described in goat kids resembled the natural disease in terms of oocyst excretion, clinical signs and intestinal pathology. The ability of oocysts excreted by asymptomatic goats, to infect goat kids and mice is likely to have a major impact on the epidemiology of cryptosporidiosis in livestock and man.     

Hosts of two canid genera, the red fox and the dog, as alternate vectors in the transmission of Sarcocystis tenella from sheep

Microscopic sarcocysts recovered from naturally infected sheep were infective to both the domestic dog (Canis familiaris) and the red fox (Vulpes vulpes). The parasite was passaged through experimental specific-parasite-free (SPF) sheep three times: infection was transmitted twice with sporocysts from foxes and subsequently with sporocysts from dogs. The sarcocysts from sheep muscle were infective to both dogs and foxes on each occasion. A cat was not infected. The prepatent period in individual canids ranged from 7 to 15 days. Sporocyst excretion was still detectable 60 days post infection. This study establishes that canids of two genera may act as vectors for a single isolate of the same Sarcocystis species from sheep.     

Comparative efficacy of santonin and piperazine against Neoascaris vitulorum in buffalo calves

An investigation was undertaken to evaluate the comparative efficacy of single dose treatment with santonin and piperazine against naturally acquired Neoascaris vitulorum in sixty-two buffalo calves of 20–60 days of age. Santonin was administered orally in doses of 5 mg, 10 mg and 15 mg/kg body weight to thirteen, eighteen, and sixteen buffalo calves, respectively. As a control, piperazine (88 mg/kg) was given by drench to a group of fifteen infected buffalo calves. Pretreatment and post treatment faecal eggs per gram (EPG) counts were determined by the Stoll's technique. The percentage reductions in EPG counts on the third and seventh days after administration of the two drugs were calculated. The percentage reduction in EPG counts in the piperazine treated group on the third day was 82 ± 15, 90.2 ± 3 and 91.3 ± 2.3% while on the seventh day these values were 88 ± 16, 97 ± 3, and 98 ± 2% in high, moderate and heavy infection calves, respectively. Treatment with santonin at 5, 10 and 15 mg/kg body weight also reduced the EPG counts. The percentage reduction in EPG counts in the calves treated with 15 mg/kg of santonin on the third day was 92.3 ± 18, 95.8 ± 7 and 93.5 ± 4% while on the seventh day these values were 100 ± 0, 100 ± 0 and 99.7 ± 2% in high, moderate and heavily infected calves, respectively. Both piperazine and santonin were associated with some side effects like diarrhoea, restlessness, etc. but their percentage incidence was not significantly different from each other. These findings suggest that santonin in a 15 mg/kg dose has an efficacy similar to piperazine given at the 88 mg/kg dose level for the treatment of ascariasis in buffalo calves.     

Resistance to Eimeria zuernii produced after chemotherapy of experimental infection in calves

Nineteen Holstein-Friesian bull calves were inoculated with 2.4 × 10⁶ sporocysts of Eimeria zuernii by stomach tube. The calves were divided into three groups 10 days after infection. The first group (seven calves) was treated with monensin (1 mg/kg body weight daily) from the 10th–20th day after infection; the second group (six calves) with amprolium (10 mg/kg body weight daily) for the same period of time and the third group (six calves) acted as infected controls. Both drugs were effective in preventing clinical signs, in reducing rates of weight gain and in suppressing oocyst production. The calves were reinfected with E. zuernii 35 days after the initial infection. The calves of all three groups were resistant to the second infection with E. zuernii as measured by rates of weight gain, fecal oocyst output and lack of clinical signs.     

Excretion of Eimeria alabamensis oocysts in grazing calves and young stock

The aims of the present study were to investigate the excretion of Eimeria alabamensis oocysts by young cattle during their first grazing season and during the first 16 days of their second grazing period. In trial 1, nine first-season grazing heifers were studied and found to have become infected with E. alabamensis shortly after turnout. The next grazing period they were turned out on to a permanent pasture together with two first-season grazing calves. Faecal samples were collected before turnout and then daily from day 3 to day 16. The second-season grazing heifers excreted insignificant numbers of E. alabamensis oocysts, whereas one of the two first-season grazing calves excreted up to 703,000 oocysts/g of faeces (OPG), indicating that the pasture was contaminated. In trial 2, faecal samples were collected from 12 calves before their first turnout in May, daily from day 2 to day 20 after turnout and then once a week until the end of September. The calves grazed pastures used in previous years by first-season grazing calves. Nine of the calves developed clinical E. alabamensis coccidiosis 4-7 days after turnout and excreted more than 950,000 OPG on days 9-10. By day 17 the oocyst excretion had decreased below 900 OPG and remained low throughout the rest of the grazing season. The results of the two studies indicate that reinfections with E. alabamensis are of little clinical importance in calves grazing contaminated pastures, and that young stock infected with E. alabamensis during their first grazing season may be used to cleanse contaminated pastures without risk of developing clinical coccidiosis.     

Development and evaluation of a Sarcocystis neurona-specific IgM capture enzyme-linked immunosorbent assay

Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses caused primarily by the protozoal parasite Sarcocystis neurona. Currently available antemortem diagnostic testing has low specificity. The hypothesis of this study was that serum and cerebrospinal fluid (CSF) of horses experimentally challenged with S neurona would have an increased S neurona-specific IgM (Sn-IgM) concentration after infection, as determined by an IgM capture enzyme linked immunoassay (ELISA). The ELISA was based on the S neurona low molecular weight protein SNUCD-1 antigen and the monoclonal antibody 2G5 labeled with horseradish peroxidase. The test was evaluated using serum and CSF from 12 horses experimentally infected with 1.5 million S neurona sporocysts and 16 horses experimentally infected with varying doses (100 to 100,000) of S neurona sporocysts, for which results of histopathologic examination of the central nervous system were available. For horses challenged with 1.5 million sporocysts, there was a significant increase in serum Sn-IgM concentrations compared with values before infection at weeks 2-6 after inoculation (P < .0001). For horses inoculated with lower doses of S neurona, there were significant increases in serum Sn-IgM concentration at various points in time after inoculation, depending on the challenge dose (P < .01). In addition, there was a significant increase between the CSF Sn-IgM concentrations before and after inoculation (P < .0001). These results support further evaluation of the assay as a diagnostic test during the acute phase of EPM.