Sarcocystis cruzi infection in wood bison (Bison bison athabascae).

Endangered wood bison (Bison bison athabascae) is the largest terrestrial mammal in the American continent. Animal health is an important issue in their conservation, and Sarcocystis cruzi may be a cause of clinical disease in Bovidae. Hearts of eight wood bison from Alaska, USA were examined for sarcocysts by histology, transmission electron microscopy, pepsin digestion, and molecularly. Sarcocystis bradyzoites were found in pepsin digests of all eight and sarcocysts were found in histologic sections of myocardium of four bison. Sarcocysts were thin-walled and ultrastructurally consistent with S. cruzi. Characterization of DNA obtained from lysis of pepsin liberated bradyzoites by PCR-RFLP and subsequent phylogenetic analyses matched with that previously reported for S. cruzi infecting cattle in the USA. Collectively, data indicate that wood bison is a natural intermediate host for S. cruzi.

Phylogenetic analysis of Sarcocystis nesbitti (Coccidia: Sarcocystidae) suggests a snake as its probable definitive host.

Sarcocystis nesbitti was first described by Mandour in 1969 from rhesus monkey muscle. Its definitive host remains unknown. 18S rRNA gene of S. nesbitti was amplified, sequenced, and subjected to phylogenetic analysis. Among those congeners available for comparison, it shares closest affinity with those species of Sarcocystis which use snakes as definitive hosts. We therefore hypothesize that a snake may serve as the definitive host for S. nesbitti.

Sarcocystis cruzi: comparative studies confirm natural infections of buffaloes.

Controversy exists concerning whether cattle and water buffalo sustain infections with cysts of distinct arrays of species in the genus Sarcocystis. In particular, morphologically similar parasites have been alternately ascribed to Sarcocystis cruzi or to Sarcocystis levinei, depending on their occurrence in cattle or water buffalo. We used light and transmission electron microscopy, genetic analysis, and experimental infections of definitive canine hosts to determine whether consistent differences could be identified from parasites derived from several natural infections of each host, examining several tissue types (esophagus, skeletal muscles, and heart). Cysts derived from cattle and water buffalo shared similar structure; variation among 18S rRNA sequences did not segregate consistently according to intermediate host type; parasites derived from cattle and water buffalo induced similar outcomes in the canine definitive host. One cattle specimen harbored unusually large (macroscopic) sarcocysts which nonetheless conformed to previously reported ultrastructural and genetic features of S. cruzi. Finding no consistent basis to differentiate between them, we conclude that the parasites infecting each host and tissue type correspond to S. cruzi. In our sample, no phylogenetically distinct taxon was sampled which might correspond to a distinct taxon previously described as S. levinei. Either that taxon was missed by our sampling effort, or it may represent a junior synonym to S. cruzi, which would then cycle between dogs and a broader range of intermediate bovine hosts than was previously considered.

Detection of sarcocystis parasites in retail beef: a regional survey combining histological and genetic detection methods.

Sarcocystis spp. are parasitic protists acquired when undercooked, cyst-laden meat is consumed. While both Sarcocystis hominis and S. cruzi encyst in beef, only S. hominis is pathogenic to humans. In this study, we used histological methods and novel molecular techniques to determine the regional prevalence and identity of Sarcocystis spp. in retail beef. Of 110 samples, 60 supported amplification of parasite rRNA by PCR. All 41 sequenced representatives were identified as S. cruzi. To compare detection methods, 48 samples were then examined in parallel by histology and PCR, and 16 and 26 samples, respectively, were positive. Five samples positive by initial histologic sections were not amplified by PCR. Fifteen PCR-positive samples did not contain sarcocysts on initial histologic section, but additional sections from these samples revealed sarcocysts in an additional 12 samples. When combined, histology with additional sections and PCR detected 31 positive specimens of the 48 total specimens. We found no evidence of human pathogen S. hominis and confirm that cattle pathogen S. cruzi is highly prevalent in this regional sample. PCR assays may increase the detection sensitivity of Sarcocystis spp. and contribute diagnostic precision.

Human dispersal of Trichinella spiralis in domesticated pigs.

To investigate the human impact on the evolutionary ecology of animal pathogens, we compared genetic diversity of severe foodborne parasites contracted by eating infected pork or wild game. In particular, we characterized Trichinella spp. from twenty-eight countries and four continents by genotyping nine microsatellite loci and sequencing one-fifth of the mitochondrial genome. All specimens of Trichinella spiralis, a swine parasite that can infect many species of wildlife, were remarkably uniform across Europe, North Africa, and the Americas. Far greater diversity characterized a comparable sample of Trichinella britovi, which parasitizes various sylvatic mammals endemic to Eurasia and North-Western Africa. A limited sample of T. spiralis in Asia, where swine were first domesticated, encompassed greater genetic variability than those in the West, as did small samples of Trichinella nativa and Trichinella murrelli, which parasitize wildlife hosts. We conclude that European lineages of T. spiralis originated several thousand years ago, approximately when pigs were first domesticated there. These data also imply that Europeans inadvertently introduced T. spiralis to the Americas via infected pigs and/or rats. Despite evidence that early hominid hunters ingested foodborne parasites by hunting wild game millions of years earlier, swine husbandry has governed the subsequent transmission, dissemination, and evolutionary diversification of T. spiralis. Where viable parasites have been eliminated from their diet, the residual risk posed to swine by exposure to wildlife or rats should be more precisely defined because breaking the cycle of transmission would confer enduring economic and health benefits.

Restricted genetic diversity in the ubiquitous cattle parasite, Sarcocystis cruzi.

Although parasites of the genus Sarcocystis have likely cycled between bovine herbivores and canine carnivores for tens of millions of years, humans may have profoundly influenced the ecology and evolution of those prevalent in domesticated dogs and cattle. To preliminarily assess the possibility of such anthropogenic effects, we surveyed genetic variation in conserved (18S small subunit) and variable (ITS-1) portions of ribosomal DNA from a large sample of Sarcocystis cruzi occurring in taurine beef cattle raised in the United States and Uruguay, and compared these data to available homologues, including those reported from zebu cattle, water buffalo, and bison. For additional context, we compared the apparent diversity of cattle parasites to that reported from congeneric parasites in other hosts. We find that the S. cruzi of taurine cattle, whether derived from the Americas or Asia, are devoid of variability in the sequenced portion (80%) of the small subunit rDNA. By contrast, geographically limited samples of related parasites in other hosts, including those of wildlife, are more variable. At the adjacent ITS-1 locus, allelic distribution patterns did not indicate any regional barriers to gene flow, suggesting that the parasite may have been introduced to the Americas via a common source such as domesticated dogs or cattle. Thus, human impact on this parasite's distribution and diversification would seem to have been great.

Parelaphostrongylus odocoilei in Columbian black-tailed deer from Oregon.

Documenting the occurrence of Parelaphostrongylus odocoilei has historically relied on the morphological examination of adult worms collected from the skeletal muscle of definitive hosts, including deer. Recent advances in the knowledge of protostrongylid genetic sequences now permit larvae to be identified. Dorsal-spined larvae (DSLs) collected in 2003-2004 from the lung and feces of six Columbian black-tailed deer (Odocoileus hemionus columbianus) from Oregon were characterized genetically. The sequences from unknown DSLs were compared to those from morphologically validated adults and larvae of P. odocoilei at both the second internal transcribed spacer (ITS-2) of ribosomal DNA and the mitochondrial cytochrome oxidase II gene. We provide the first unequivocal identification of P. odocoilei in Columbian black-tailed deer from Oregon. The broader geographic distribution, prevalence, and pathology of P. odocoilei are not known in populations of Oregon deer.

Population genetics of duplicated disease-defense genes, hm1 and hm2, in maize (Zea mays ssp. mays L.) and its wild ancestor (Zea mays ssp. parviglumis).

Plant defense genes are subject to nonneutral evolutionary dynamics. Here we investigate the evolutionary dynamics of the duplicated defense genes hm1 and hm2 in maize and its wild ancestor Zea mays ssp. parviglumis. Both genes have been shown to confer resistance to the fungal pathogen Cochliobolus carbonum race 1, but the effectiveness of resistance differs between loci. The genes also display different population histories. The hm1 locus has the highest nucleotide diversity of any gene yet sampled in the wild ancestor of maize, and it contains a large number of indel polymorphisms. There is no evidence, however, that high diversity in hm1 is a product of nonneutral evolution. In contrast, hm2 has very low nucleotide diversity in the wild ancestor of maize. The distribution of hm2 polymorphic sites is consistent with nonneutral evolution, as indicated by Tajima's D and other neutrality tests. In addition, one hm2 haplotype is more frequent than expected under the equilibrium neutral model, suggesting hitchhiking selection. Both defense genes retain >80% of the level of genetic variation in maize relative to the wild ancestor, and this level is similar to other maize genes that were not subject to artificial selection during domestication.