Prevalence pattern and biology of Sarcocystis capracanis infection in the Egyptian goats: a light and ultrastructural study

Cysts of Sarcocystis capracanis obtained from infected goats were examined to clarify the effect of the parasite on the host. Muscle tissues from fresh oesophagus, tongue, diaphragm and skeletal muscles of 680 goats were slaughtered in the main abattoir of Cairo, Egypt and they were examined microscopically for Sarcocystis infection for the first time in Egypt. 540 out of 680 (79.4%) of examined goats were found to be infected with Sarcocystis sp. The infection was recorded firstly by light microscopy as spindle shaped cysts embedded in the muscle tissues. The validity of this species as S. capracanis was confirmed by means of ultrastructural characteristics of the primary cyst wall which revealed the presence of thick-radially striated wall with finger like projections, underlined by a thick layer of ground substance enclosing the developing metrocytes and merozoites that usually contain nearly all the structures of the apical complex and fill the interior cavity of the cyst. The cyst cavity is divided by many septa extending from the ground substance and producing large number of chambers. An experimental infection using the highly infected muscles was carried out to determine the final host, which is dog. Smears of intestinal epithelium were taken to examine the endogenous stages (gamogony and sporogony) by means of light microscopy. These stages were mainly observed as to infect the lamina propria of the posterior third of the small intestine. Gamogony and zygote formation (fertilization) occurred 2-8 days post infection, while sporulation took place within the final host 13-15 days and sporocysts were passed within faeces of the infected puppies at that time. The prepatent period of S. capracanis was 12-15 days, while the patent period was extended to 37 days. In goats, infection with S. capracanis led to the loss of weight, anaemia, abortion and even death in cases of heavy infection. While bleeding, watery faeces filled with mucous on 5th and 8th day p.i. as well as intestinal lesions are the pathogenic effects occurred in puppies after experimental infection.     

旱獭住肉孢子虫感染的调查

住肉孢子虫是球虫类的寄生原虫 ,其中某些种类为人兽共患的寄生虫 ,其宿主范围很广。目前已发现的住肉孢子虫约有 1 0 0多种。旱獭住肉孢子虫 ,尚无报道 ,本地区旱獭住肉孢子虫的感染情况报道如下。1　材料与方法1 .1　调查对象　来自天峻地区国家计划猎杀的 66只旱獭 ,在每只旱獭上采取膈肌、咬肌、心肌、骨骼肌作为被检肉样 ,编号待检。1 .2　调查方法　每份肉样 ,弃去包膜后沿肌纤维走向 ,分别剪取如燕麦粒大小肌肉薄块 2 4粒 ,排列于载皮片上 ,滴加 50 %甘油水溶液 ,盖上同一大小玻片一块 ,加压制成半透明压片 ,在生物显微镜下检查 ,发…     

Characterization of a Sarcocystis neurona isolate from a Missouri horse with equine protozoal myeloencephalitis

Little information is available about antigenic variation of Sarcocystis neurona isolated from horses with equine protozoal myeloencephalitis, nor is there much information available on the specific antibody pattern to S. neurona antigens of horses from different geographic regions where S. neurona isolates have been obtained. This communication reports on the characterization of a new S. neurona isolate, SN-MU1. The isolate was obtained from a 3-year old Thoroughbred that had asymmetrical neurological signs and localized skeletal muscle atrophy. This S. neurona isolate is similar to other S. neurona isolates by molecular analysis of the internal transcribed spacer (ITS-1) region and a random-amplified polymorphic DNA marker, but is phenotypically distinct from the other S. neurona isolates examined. Evaluation of the antibodies from the affected horse and immunohistochemical results suggested that antigenic variation of S. neurona can result in variable antibody-antigen reactivity observed in the S. neurona immunoblot test.     

Comparison of Sarcocystis neurona isolates derived from horse neural tissue

Sarcocystis neurona is a protozoan parasite that can cause neurological deficits in infected horses. The route of transmission is by fecal-oral transfer of sporocysts from opossums. However, the species identity and the lifecycle are not completely known. In this study, Sarcocystis merozoites from eight isolates obtained from Michigan horses were compared to S. neurona from a California horse (UCD1), Sarcocystis from a grackle (Cornell), and five Sarcocystis isolates from feral opossums from Michigan.Comparisons were made using several techniques. SDS-PAGE analysis with silver staining showed that Sarcocystis spp. from the eight horses appeared the same, but different from the grackle isolate. One Michigan horse isolate (MIH6) had two bands at 72 and 25kDa that were more prominent than the UCD1 isolate and other Michigan horse isolates. Western blot analysis showed that merozoites of eight of eight equine-derived isolates, and the UCD1 S. neurona isolate had similar bands when developed with serum or CSF of an infected horse. Major bands were seen at 60, 44, 30, and 16kDa. In the grackle (Cornell) isolate, bands were seen at 60, 44, 29, and 16kDa. DNA from merozoites of each of the eight equine-derived isolates and the grackle-derived isolate produced a 334bp PCR product (Tanhauser et al., 1999). Restriction fragment length polymorphism (RFLP) analysis of these horse isolates showed banding patterns characteristic for S. neurona. The grackle (Cornell) isolate had an RFLP banding pattern characteristic of other S. falcatula species. Finally, electron microscopy examining multiple merozoites of each of these eight horse isolates showed similar morphology, which differed from the grackle (Cornell) isolate. We conclude that the eight Michigan horse isolates are S. neurona species and the grackle isolate is an S. falcatula species.     

Naturally occurring Sarcocystis neurona-like infection in a dog with myositis

Tissue stages similar to those of Sarcocystis neurona, the causative agent of equine protozoal myeloencephalitis, were identified in skeletal muscles of a dog. The dog, a 6-year-old Labrador retriever, was seropositive for Toxoplasma gondii infection and euthanized due to a history of polymyositis and progressive muscular atrophy. Histologically, 30, variably sized, microscopic, intracellular sarcocysts were observed in 60 sections of skeletal muscles taken from the neck, fore limbs and hind limbs. The cysts were only observed in inflamed skeletal muscles, but were mostly in myocytes at the periphery of areas infiltrated with leukocytes. Ultrastructurally, the cyst wall had villar protrusions consistent with sarcocysts. Immunohistochemistry with monoclonal S. neurona antibodies demonstrated positive labeling of zoites in merozoites or schizonts in the skeletal muscle interstitium, but no labeling of the sarcocysts. Initial PCR analysis with primers amplifying a genetic sequence encoding Apicomplexan 18s rRNA, and subsequent PCR analysis with differentiating primers indicated that the genetic sequences had 100% identity with sequences reported for S. neurona.     

Parasitemia in an immunocompetent horse experimentally challenged with Sarcocystis neurona sporocysts

Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in Americans. Most cases are attributed to infection of the central nervous system with Sarcocystis neurona. Parasitemia has not been demonstrated in immunocompetent horses, but has been documented in one immunocompromised foal. The objective of this study was to isolate viable S. neurona from the blood of immunocompetent horses. Horses used in this study received orally administered S. neurona sporocysts (strain SN 37-R) daily for 112 days at the following doses: 100/day for 28 days, followed by 500/day for 28 days, followed by 1000/day for 56 days. On day 98 of the study, six yearling colts were selected for attempted culture of S. neurona from blood, two testing positive, two testing suspect and two testing negative for antibodies against S. neurona on day 84 of the study. Two 10 ml tubes with EDTA were filled from each horse by jugular venipuncture and the plasma fraction rich in mononuclear cells was pipetted onto confluent equine dermal cell cultures. The cultures were monitored weekly for parasite growth for 12 weeks. Merozoites grown from cultures were harvested and tested using S. neurona-specific PCR with RFLP to confirm species identity. PCR products were sequenced and compared to known strains of S. neurona. After 38 days of in vitro incubation, one cell culture from a horse testing positive for antibodies against S. neurona was positive for parasite growth while the five remaining cultures remained negative for parasite growth for all 12 weeks. The Sarcocystis isolate recovered from cell culture was confirmed to be S. neurona by PCR with RFLP. Gene sequence analysis revealed that the isolate was identical to the challenge strain SN-37R and differed from two known strains UCD1 and MIH1. To our knowledge this is the first report of parasitemia with S. neurona in an immunocompetent horse.     

Acremonium strictum pulmonary infection in a horse

A 10-year-old Thoroughbred gelding was admitted to the Veterinary Medical Teaching Hospital of the University of California-Davis with a 2-week history of intermittent fever and acute onset of lethargy, anorexia, and ataxia. Although the clinical signs were nonspecific, the results of initial hematologic and biochemical analysis were consistent with a chronic inflammatory process. Thoracic radiographs revealed an increased fine reticulonodular interstitial opacity throughout the dorsal caudal lung fields. Cytologic examination of bronchoalveolar lavage (BAL) fluid showed mixed inflammation with many mononuclear phagocytes containing single, spherical, intracytoplasmic fungal organisms. Four mold species were cultured in low numbers from the BAL fluid. One of the fungal elements observed on the culture plates was identified as Acremonium strictum by real-time polymerase chain reaction (PCR). A diagnosis of fungal pneumonia due to A strictum was made based on the results of thoracic imaging, cytologic evaluation, culture, and PCR testing. The horse made an uneventful recovery with supportive treatment and was disease-free based on normal physical, radiographic, and cytologic findings at 21 days after presentation. To our knowledge, this is the first report of isolation of A strictum from the BAL fluid of a horse with interstitial pneumonia.     

Experimental infection with Sarcocystis medusiformis in sheep

The development of the parasite was studied in 48 sheep killed between 188 and 1132 days after experimental inoculation with Sarcocystis medusiformis sporocysts from cats. Immature sarcocysts were present at 188 days post inoculation (d.p.i.). At 331 d.p.i. macroscopic sarcocysts with an elongate fusiform appearance were seen in the laryngeal, abdominal and diaphragm musculature. The largest cyst measured 2 mm in length by 0.5 mm in width at 331 d.p.i.; histologically they contained metrocytes at the periphery of the cyst with more densely staining merozoites in the central region. By 443 d.p.i. typical 'thin' cysts 2-3.5 mm in length were seen in the flank and external thoracic muscles. By 765 d.p.i. sarcocysts were 5 mm in length. The ultrastructure of the cyst wall of these cysts resembled that of S. medusiformis. At 1132 d.p.i. sarcocysts measured 4 mm X 0.5 mm.     

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.     

Blastomycotic osteomyelitis associated with severe lameness in a horse

A 12-year-old Quarter horse gelding was presented for evaluation of severe right forelimb lameness, 2 draining tracts over the lateral aspect of the right proximal antebrachium, and weight loss. A presumptive diagnosis of blastomycotic osteomyelitis was established based on radiographs and cytology of the exudate. This diagnosis was confirmed at necropsy.     

Suspected Sarcocystis infection in an aborted bovine foetus

Extract

Sir:- Samples from a case of sporadic abortion, two weeks before term in a two-year-old heifer, were submitted to the Palmerston North Animal Health Laboratory. They consisted of fresh foetal stomach contents, fixed foetal lung, and maternal urine and serum. No significant pathogens were isolated from the stomach contents or urine and serological tests for brucellosis and leptospirosis were negative.