Ultrastructure of sarcocysts from water buffalo in India

The ultrastructure of sarcocysts of macro- and microscopic species of Sarcocystis was compared from naturally infected water buffalo from India. Grossly visible sarcocysts had walls consisting of cauliflower-like villar protrusions, typical of S. fusiformis. The sarcocyst wall of the microscopic species of Sarcocystis was 6.4 microns thick and consisted of tightly packed conical villar protrusions that were 9.6 microns long and 3.7 microns wide at the base. At approximately 3 microns above the base, the distal two-thirds of the villar protrusion became conical shaped and was bent laterally at an angle of 45 degrees to the sarcocyst surface. The granular layer beneath the villar protrusions was 0.9 microns thick. In S. levinei the granular layer was 1.9 microns thick, the villar protrusions were narrow and it had a highly undulating primary cyst wall. Whether the microscopic S. levinei-like sarcocysts of Indian and Malaysian water buffalo are distinct species of Sarcocystis will require further investigation.     

Seroprevalence of and Risk Factors for Toxoplasma gondii in the US Swine Herd Using Sera Collected During the National Animal Health Monitoring Survey (Swine 2006)

The United States Department of Agriculture (USDA) initiated the National Animal Health Monitoring System (NAHMS) in 1983 to collect, analyse and disseminate data on animal health, management and productivity in US domestic livestock populations, including swine. The programme includes an on‐farm serological sampling component which can be used to monitor seroprevalence of various pathogens, including Toxoplasma gondii. The purpose of this study was to determine the seroprevalence of T. gondii in grower/finisher pigs using sera collected during NAHMS Swine 2006 and to determine farm level factors associated with differences in seroprevalence on farms where sera was collected during the Swine 2006 survey. Sera and data on management practices for this study were collected from 185 grower/finisher swine production sites located in 16 states accounting for >90% of US swine production (Arkansas, Colorado, Iowa, Illinois, Indiana, Kansas, Michigan, Minnesota, Missouri, Nebraska, North Carolina, Ohio, Pennsylvania, South Dakota, Texas and Wisconsin). A total of 6238 sera were tested for T. gondii antibodies using a commercial ELISA assay (Vet. Parasitol. 128 , 2005, 177). Seroprevalence in this study, as determined by ELISA, was 2.6%, with a herd prevalence of 21.6% and a mean within‐herd prevalence of 2.7%. Analysis of swine management practices indicated that rodent control methods and carcass disposal methods were associated with differences in the number of T. gondii positive samples on farm. These results are consistent with current epidemiological knowledge of the transmission of Toxoplasma on the farm (ingestion of organic matter containing oocysts, or ingestion of infected animal tissues). Production practices which eliminate these sources of exposure can reduce the risk of Toxoplasma infection in pigs, and reduce the likelihood of human infection from consumption of infected pork.     

Identification of bovine viral diarrhea virus type 1 in yaks (Bos poephagus grunniens) in the Himalayan region

Since cattle are widely infected by bovine viral diarrhea virus (BVDV) in India, we searched for pestivirus infection in yaks. Of 71 pure and crossbred yaks from Himalayan region, pestivirus antigen was detected by Ag-ELISA in three animals. Pestivirus in leukocyte and cell culture isolated virus samples originating from positive yaks was also confirmed by RT-PCR using panpestivirus specific primers selected from 5'-untranslated region (5' UTR). The 5' UTR, N(pro) and E2 regions were sequenced and used for genetic typing. Phylogenetic analysis revealed that pestiviruses detected in three Himalayan yaks were similar genetically, belonging to BVDV-1. Antigenic characterisation of yak pestivirus also confirmed the typing as BVDV-1. This is the first report on the identification of BVDV type 1 in yaks.     

Relationships among Sarcocystis species transmitted by New World opossums (Didelphis spp.)

At least three species of Sarcocystis (S. neurona, S. falcatula, S. speeri) have recently been shown to use opossums of the genus Didelphis as their definitive host. In order to evaluate the evolutionary relationships among Sarcocystis spp. isolates from the Americas, and to determine whether organisms representing the same parasite lineages are transmitted north and south of the Panamanian isthmus, we inferred the phylogenetic relationships from nucleotide sequence variation in parasites isolated from three opossum species (D. virginiana, D. albiventris, D. marsupialis). In particular, we used variation in the 25/396 marker to compare several isolates from Brazil, Argentina, and the United States to each other and to cloned S. neurona and S. falcatula whose morphology and host affinities have been defined in the laboratory. S. neurona was identified from a Brazilian D. albiventris, as well as from North American D. virginiana. Parasites resembling the Cornell isolate of S. falcatula are transmitted both south and north of the Panamanian isthmus by D. albiventris and D. virginiana, respectively. Distinct attributes at two genetic loci differentiated a Brazilian isolate of S. falcatula from all other known parasite lineages. We confirm S. neurona as the causative agent of recently reported neurologic disease in Southern sea otters, Enhydra lutris nereis. And we found that S. speeri could not be compared to the other opossum-derived Sarcocystis isolates on the basis of nucleotide variation at the 25/396 locus. The widespread distribution of certain species of Sarcocystis may derive from their ability to parasitize migratory bird hosts in their intermediate stage.     

Dual Sarcocystis neurona and Toxoplasma gondii infection in a Northern sea otter from Washington state, USA

Dual Sarcocystis neurona and Toxoplasma gondii infection was observed in a Northern sea otter from Washington, USA. The animal was found stranded, convulsed, and died shortly thereafter. Encephalitis caused by both S. neurona and T. gondii was demonstrated in histological sections of brain. Immunohistochemical examination of sections with S. neurona specific antisera demonstrated developmental stages that divided by endopolygeny and produced numerous merozoites. PCR of brain tissue from the sea otter using primer pairs JNB33/JNB54 resulted in amplification of a 1100 bp product. This PCR product was cut in to 884 and 216 bp products by Dra I but was not cut by Hinf I indicating that it was S. neurona [J. Parasitol. 85 (1999) 221]. No PCR product was detected in the brain of a sea otter which had no lesions of encephalitis. Examination of brain sections using T. gondii specific antisera demonstrated tachyzoites and tissue cysts of T. gondii. The lesions induced by T. gondii suggested that the sea otter was suffering from reactivated toxoplasmosis. T. gondii was isolated in mice inoculated with brain tissue. A cat that was fed infected mouse brain tissue excreted T. gondii oocysts which were infective for mice. This is apparently the first report of dual S. neurona and T. gondii in a marine mammal.     

Toxoplasma-like sporozoa in an aborted equine fetus

Multifocal areas of necrosis and infiltrations of mononuclear cells were seen in lung specimens of an equine fetus aborted 2 months before term. Extracellular and intracellular protozoa were seen in the alveolar tissue. Individual organisms were 4 microns by 2.5 microns, and cyst-like structures were 25 microns by 18 microns. Organisms did not stain with periodic acid-Schiff or by use of the immunoperoxidase and peroxidase-antiperoxidase method for Toxoplasma gondii. Twelve days after abortion, the mare had serum antibody titer of less than 1:10 against T gondii.     

Attempeted transmission of feline coccidia from chronically infected queens to their kittens

Eight female, 12- to 34-month-old, specific-pathogen free cats were inoculated orally with Toxoplasma gondii cysts on day 0, then with Isospora felis and Isospora rivolta oocysts on day 39, and cysts of Hammondia hammondi on day 86 after inoculation with Toxoplasma. All cats shed oocysts of all 4 of these coccidia within 11 postinoculation days. The female cats were caged with 4 male Toxoplasma-free cats, starting 66 days after inoculation with Toxoplasma, until they were 5 to 6 weeks pregnant. Kittens that were born were housed with their mothers until necropsied or weaned. One 42-day-old kitten shed T gondii oocysts in feces. It was necropsied 2 days later and asexual stages of Toxoplasma (types D and E), gametocytes, and oocysts were demonstrated in sections of superficial epithelial cells of its small intestine. Lesions or forms of Toxoplasma were not demonstrated histologically in tis extraintestinal organs. Toxoplasma was not isolated from feces or tissues of the remaining 47 kittens born to these 8 queens. Toxoplasma was not isolated from the 4 male cats that were caged with infected females for 53, 59, 217, and 217 days. The source of toxoplasma infection in the kitten remained unknown but was considered unlikely to be congenital or through fecal contamination. Oocysts of I felis, I rivolta, and H hammondi were not found in the feces of any kittens, indicating that these coccidia are unlikely to be transmitted congenitally.     

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.