Heritabilities and quantitative trait loci for blood gases and blood pH in swine

Maintaining pH and blood gases in a narrow range is essential to sustain normal biochemical reactions. Decreased oxygenation, poor tissue perfusion, disturbance to CO₂ expiration, and shortage of HCO₃⁻ can lead to metabolic acidosis. This is a common situation in swine, and originates from a broad range of medical conditions. pH and blood gases appear to be under genetic control, and populations with physiological traits closer to the pathological thresholds may be more susceptible to developing pathological conditions. However, little is known about the genetic basis of such traits. We have therefore estimated phenotypic and genetic variability and identified quantitative trait loci (QTL) for pH and blood gases in blood samples from 139 F₂ pigs from the Meishan/Pietrain family. Samples were taken before and after challenge with Sarcocystis miescheriana , a protozoan parasite of muscle. Twenty-seven QTL influencing pH and blood gases were identified on nine chromosomes. Five of the QTL were significant on a genome-wide level; 22 QTL were significant on a chromosome-wide level. QTL for pH-associated traits have been mapped to SSC3, 18 and X. QTL associated with CO₂ have been detected on SSC6, 7, 8 and 9, and QTL associated with O₂ on SSC2 and SSC8. QTL showed specific health/disease patterns that were related to the physiological state of the pigs from day 0, to acute disease (day 14), convalescence (day 28) and chronic disease (day 42). The results demonstrate that pH and blood gases are influenced by multiple chromosomal areas, each with relatively small effects.     

Molecular phylogeny of Sarcocystis fayeri (Apicomplexa: Sarcocystidae) from the domestic horse Equus caballus based on 18S rRNA gene sequences and its prevalence

Sarcocystosis is a parasitic disease caused by an intracellular protozoan parasite Sarcocystis belonging to the phylum Apicomplexa. These parasites have a requisite two-host life cycle. Recently, there are many Sarcocystis species that identified morphologically. In the present study, diaphragmatic muscle samples from the domestic horse (Equus caballus) were examined for Sarcocystis infection. The natural infection with sarcocysts was recorded to be 62·5% for only microcysts in the infected muscles. Molecular analysis using the 18S rRNA gene was conducted to swiftly and accurately identify the recovered species. Studies on the expression of the 18S rRNA gene have confirmed that the present parasite isolates belong to the Sarcocystis genus. The sequence data showed significant identities (>80%) with archived gene sequences from species within the Sarcocystidae family, and a dendrogram showing the phylogenetic relationship was constructed. The most closely related species were the previously described Sarcocystis fayeri and Sarcocystis bertrami. The current data showed that the present species was identified as S. fayeri and deposited in GenBank (accession number MF614956.1). This study highlights the importance of the genetic data in the exact taxonomy within sarcocystid species.     

Identification of an Apically-Located Antigen That Is Conserved in Sporozoan Parasites

ABSTRACT. Sporozoan parasites of the phylum Apicomplexa all possess common apical structures. The current study used a monoclonal antibody (mAb-E12) to identify a conserved antigen in the apical region of merozoites of seven species of Plasmodium (including rodent, primate and human pathogens), tachyzoites of Toxoplasma gondii , bradyzoites of Sarcocystis bovis , and sporozoites and merozoites of Eimeria tenella and E. acervulina. The antigen was also present in sporozoites of haemosporinid parasites. Immunofluorescence studies showed that the antigen was restricted to the apical 3rd of these invasive stages. Using immunoelectron microscopy, labeling was demonstrated in the region of the polar ring, below the paired inner membranes of the parasite pellicle, and near the subpellicular microtubules radiating from the polar ring of merozoites and sporozoites of E. tenella . The majority of the antigen could be extracted with 1% Triton-X 100, but a portion remained associated with the cytoskeletal elements. The molecule has a relative rate of migration (M_r) of 47,000 in Plasmodium spp. and 43–46,000 in coccidian species. Since the epitope recognized by mAb-El 2 is highly conserved, restricted to motile stages, and appears to be associated with microtubules, this antigen could be involved in cellular motility and cellular invasion.     

Expression, purification, and biochemical characterization of a recombinant Iectin of Sarcocystis muris (Apicomplexa) cyst merozoites

The mature major microneme protein of Sarcocystis muris cyst merozoites, which is known as a dimeric lectin with high affinity to galactose and some of its derivatives, was expressed in Escherichia coli as a histidine-tagged fusion protein. The recombinant polypeptide, which was recognized by a monoclonal antibody directed against the native lectin, was purified from inclusion bodies after solubilization and refolding, using a combination of metal chelate and lactose affinity chromatography. The apparent molecular mass of the refolded polypeptide as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoreses was 16 kDa, whereas gel filtration chromatography clearly demonstrated that the recombinant protein, like its native counterpart, exists as a homodimer of two non-covalently associated subunits. Inhibition of haemagglutination suggests that the combining site of the recombinant lectin recognizes N-acetyl-galactosamine as the dominant sugar, thus confirming the correct folding of the monosaccharide combining site in the renatured lectin. To the best of our knowledge, this work represents the first reported detailed characterization of a recombinant lectin from apicomplexan parasites, and may contribute to a better understanding of the process of host cell recognition and invasion by these obligate intracellular protozoa.     

Investigations on the Host Specificity of Dihomoxenous Sarcosporidia in the Intermediate and Definitive Host

ABSTRACT. Cross-transmission experiments were performed in order to determine the host specificity in the intermediate and definitive hosts of the four described dihomoxenous Sarcocystis species, S. gallotiae, S. stehlinii, S. simonyi , and S. dugesii from lacertid lizards of the genera Gallotia and Podarcis from the Macaronesian Islands. Sarcocysts of either species from experimentally infected lizards were fed to a variety of laboratory-bred lizard species of the genera Gallotia, Lacerta , and Podarcis . These sarcocysts proved to be infectious to all examined animals, showing no definitive host specificity in the tested genera. Lizards of the genera Chalcides and Tarentola , however, were not susceptible definitive hosts for S. gallotiae . The inoculation of experimentally obtained sporocysts of each of the four Sarcocystis species to various lacertid lizard species revealed varying degrees of intermediate host specificity, generally demonstrating each native host to be the most susceptible.     

Fine Structure of Sarcocystis cruzi Schizonts

SYNOPSIS Schizogony of Sarcocystis cruzi Hasselmann (syn. S. fusiformis Railliet) takes place in vascular endothelial cells 26 to 33 days after cattle ingest sporocysts from dogs. Kidney cortex from a heavily infected, dexamethasone-treated bovine was fixed for electron microscopy to determine the method of schizogonie development. Schizogony takes place by endopolygeny characterized by marked enlargement of the parasite nucleus, formation of nuclear lobes, presence of numerous spindles with adjacent pairs of centrioles along the nucleus, and simultaneous formation of daughter merozoites in the cytoplasm adjacent to the spindle poles. Endopolygeny in S. cruzi differs from that in other Sporozoa in that merozoite anlagen are seen in the cytoplasm before any nuclei divide. The resultant merozoites continue development and, when mature, resemble other sporozoan zoites. Upon release from the host cell into capillaries, they travel to muscle tissue to continue the life cycle by forming sarcocysts.     

Sarcocystis neurona: molecular characterization of enolase domain I region and a comparison to other protozoa

Sarcocystis neurona causes protozoal myeloencephalitis and has the ability to infect a wide host range in contrast to other Sarcocystis species. In the current study, five S. neurona isolates from a variety of sources, three Sarcocystis falcatula, one Sarcocystis dasypi/S. neurona-like isolate, and one Besnoitia darlingi isolate were used to compare the enolase 2 gene segment containing the domain I region to previously sequenced enolase genes from Neospora caninum, Neospora hughesi, Toxoplasma gondii, Plasmodium falciparum, and Trypanosoma cruzi; enolase 2 segment containing domain I region is highly conserved amongst these parasites of veterinary and medical importance. Immunohistochemistry results indicates reactivity of T. gondii enolase 1 and 2 antibodies to S. neurona merozoites and metrocytes, but no reactivity of anti-enolase 1 to the S. neurona bradyzoite stage despite reactivity to T. gondii bradyzoites, suggesting expression differences between organisms.     

Immunity of sheep to homologous challenge with dog-borne Sarcocystis species following varying levels of prior exposure

Ford G. E. 1985. Immunity of sheep to homologous challenge with dog-borne Sarcocystis species following varying levels of prior exposure. International Journal for Parasitology15: 629–634. Grazing sheep in which sarcocysts develop (sarcosporidiosis) survive the pre-cyst stages of the parasite infection which cause pathophysiological changes (sarcocystosis). A peak temperature rise occurs at 3–4 weeks post-infection in experimental animals. A maximum fall in packed cell volume occurs at 5–6 weeks post-infection. Groups of sheep maintained specific pathogen free were experimentally infected with sporocysts of dog-origin to determine the dose level required for protection or resistance, and 13 weeks later challenged with a pathogenic dose of 50,000 sporocysts. Prior exposure levels (PEL) were used of nil, 5, 50, 500 and 5000 sporocysts. All prior infections elicited an antibody response to a complement fixation test in all sheep. A peak antibody level was detected within 2 weeks of challenge, with a higher titre for all previously exposed groups than for the nil PEL controls. Resistance of the sheep to the challenge dose, based on reduction of counts of parasites developing (sarcosporidiosis), was shown by significantly lower counts for the 50 PEL group (85%), as well as for the 500 (92%) and 5000 PEL (98%) groups. The 5 PEL group was not resistant. The protection of sheep, based on reduction of the above pathophysiological changes (sarcocystosis), was highly correlated with the level of prior exposure (r = ?0.80, P < 0.001). The changes in body temperature and packed cell volume were significantly less in groups with 500 and 5000 PEL than with nil and 5 PEL. The 50 PEL group was intermediate.     

寄生于褐家鼠的肉孢子虫一新种(孢子纲,真球虫目)

描记于云南省安宁市褐家鼠Rattus norvegicus肌肉内检获肉孢子虫1新种:左氏肉孢子虫Sarcocystis zuoisp.nov..新种肌肉内的包囊乳白色,呈梭形,大小为1 058μm×117μm.包囊壁具有长指状突起,突起大小为10.0μm×2.5μm.透射电镜下,包囊壁突起在基部高度分枝,突起通过分枝的基部相互连接在一起.     

Naturally occurring Sarcocystis infection in domestic cats (Felis catus)

Equine protozoal myeloencephalitis is an important neurological disease of horses in the United States. Consequently, there is an active research effort to identify hosts associated with the primary causative agent, Sarcocystis neurona. The purpose of this study was to determine whether the domestic cat (Felis catus) is a natural host for S. neurona. Muscle sections from 50 primarily free-roaming domestic cats were examined for the presence of sarcocysts. Serum from cats in this group and another group of 50 free-roaming cats were evaluated for the presence of S. neurona antibody. Sarcocysts were found in five of 50 (10%) cats, and S. neurona antibody in five of 100 (5%) cats. Morphological, molecular (including ribosomal RNA genes), and biological characterisation of these sarcocysts showed that they were not S. neurona or S. neurona-like. Sarcocysts found in the cats were identified morphologically as Sarcocystis felis, a common parasite of wild felids. The life cycle of S. felis is not known, and prior to this study, no molecular marker for S. felis existed. Although cats were found to be infected with S. felis sarcocysts, serological data provided evidence of possible infection with S. neurona as well. Further work is needed to determine the role of the domestic cat in the life cycle of S. neurona.