Prevalence of Sarcocystis spp. in meat-producing animals in Iraq.

The prevalence of Sarcocystis spp. infection was investigated in 605 sheep, 826 goats, 1080 cattle, 580 water buffaloes and 36 camels slaughtered from 1992 to 1996 in the Baghdad area (Iraq) using naked eye examination for macroscopic sarcocysts, and peptic digestion, muscle squash, squeezing methods and indirect fluorescent antibody test (IFAT) for microscopic types. The intestinal stages of the parasite were also studied in dogs experimentally fed with tissues containing microscopic cysts. The percentage prevalence of macroscopic cysts were 4.1, 33.6, 0.2, 15.6 and 0, and of the microscopic type, 97.0, 97.4, 97.8, 82.9 and 91.6 for the above-mentioned hosts, respectively. Among the different organs examined, macroscopic cysts were found to be highest in the oesophagus and the lowest in the heart. Peptic digestion method gave the highest rate (93.3%) followed by indirect fluorescent antibody test (IFAT) (88.6%), squeezing (81.3%), and muscle squash (81.2%). Each infected dog shed a total of about 150-200 million sporocysts. Histologically, developmental stages of the parasite were detected in the small intestinal mucosa of the dogs on Days 7 and 13 post-infection.     

Prevalence of Sarcocystis spp. in water buffaloes in Vietnam.

Muscles from heart, tongue, oesophagus, neck and abdomen from 502 adult water buffaloes (Bubalus bubalis) slaughtered in Ho Chi Minh City, Vietnam, between 1996 and 1997 were examined for Sarcocystis cysts by a combination of ocular and histological examination. Sarcocysts were present in 396 (79%) of the animals and the prevalence increased with age from a 57% infection rate among 2-3 year old animals to 93% among 6-7 year olds. The prevalence was higher in animals originating from the northern part (89%) than in those from the southern part (69%) of the country. Four species of Sarcocystis were identified. S. levinei (74%) was the most common species found, followed by S. fusiformis (41%), S. buffalonis (33%) and S. dubeyi (12%). All four species were present in 8% of the infected animals. The most common site for sarcocyst location was oesophagus, followed by cervical muscles, tongue and heart.     

Prevalence of dog-derived Sarcocystis spp. in some New Zealand lambs

Pepsin: hydrochloric acid digestion and histological examination of the myocardium of 51 hearts from five to nine month-old lambs from the southern half of the North Island, showed that 47 (92%) were infected with microscopic sarcocysts that are presumed to be dog-derived. Of 157 sarcocysts detected histologically, 32 corresponded morphologically with Sarcocystis ovicanis and 120 with Sarcocystis tenella (Syn. S. arieticanis). The remainder could not be identified as to type.     

Differential molecular identification of Taeniid spp. and Sarcocystis spp. cysts isolated from infected pigs and cattle

In the present work, the species-specific identification of Taeniid spp. cysticerci and sarcocystis cysts isolated from infected pigs and cattle was achieved by PCR. In particular: (i) multiplex-PCR derived from HDP2 DNA fragment, specific for Taenia saginata/Taenia solium; (ii) PCRs and PCR-RFLPs of the rDNA internal transcribed spacers 1 and 2 (ITS1 and ITS2) for the differential diagnosis of taeniids; (iii) PCR derived from the 18S rRNA gene and sequencing, specific for Sarcoystis spp. The combined application of these three PCR protocols provided an unequivocally specific diagnosis of T. saginata, T. solium, T. hydatigena, Sarcocystis hominis and Sarcocystis suihominis, and may have practical application in the identification of calcified degenerating or morphologically dubious cysts, for example in the slaughter house situation or in human biopsy samples.     

Sarcocystis sp. from cattle slaughtered in Japan

Sarcocystis sp. was detected from cattle slaughtered in Saitama Prefecture, Japan. The cysts were 3,400-4,400 x 198-238 microm in size and had the thick cyst wall which was 7 to 10 microm thick and provided with finger-like villar protrusions. The protrusions were 8-9.5 x 2-2.5 microm in size and had microtubules in the core.     

Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle

Cattle are intermediate host for several species of Sarcocystis, including S. cruzi, S. hirsuta, and S. hominis with high prevalence worldwide. The present study aimed to determine the prevalence of Sarcocystis infection, species identification, and phylogenetic analysis of the parasite in cattle in Northwest Iran. The samples of diaphragm and esophagus from 290 cattle were collected from slaughterhouses in Northwest Iran and subjected to macroscopic, microscopic, and histopathology examinations, PCR-RFLP, sequencing and phylogenetic analyses. Tissue cysts of Sarcocystis spp. were detected in 92% of cattle by digestion and microscopic tests. Based on the PCR–RFLP and specific PCR, 87.9% and 1.03% of isolates were identified as S. cruzi, and S. hominis, respectively. Macrocyst was seen in a single sample that was identified as S. gigantea. The haplotype network exhibited the extension of the various haplotypes of S. cruzi between neighboring provinces in Northwest Iran. Heterogeneity analysis of S. cruzi 18S-rRNA sequences indicated genetic diversity among S. cruzi isolates (Haplotype diversity: 0.733-0.854) consisting 16 haplotypes; however, the nucleotide differences showed low diversity (0.01481 to 0.03351). Pair wise sequence distance matrix amongst S. cruzi sequences indicated an intra-species divergence of 0%-7.8% and identity of 92.6%-100%. Sarcocystis infection is highly prevalent in cattle in Northwest Iran, with the predominance of S. cruzi, and genetic variants of this species are unequivocally distributing in Northwest provinces. First global detection of S. gigantea in cattle reflects new insights of transmission dynamic and biology of this parasite in Iran.     

Prevalence of Sarcocystis in domestic pigs in India.

Muscle samples from 890 slaughtered pigs were examined for the presence of sarcocysts. A high prevalence rate of 67.98% was observed. Two types of microsarcocysts were recorded. The sarcocyst wall of one type had redial striations and the other possessed hair-like villar protrusions. The species were identified as Sarcocystis miescheriana and Sarcocystis suihominis; there was a slightly higher incidence of the latter species (47.11%) than of the former (43.14%). S. suihominis has been identified for the first time from pigs in India.     

Evaluation of an enzyme immunoassay for the detection of antibodies against Sarcocystis spp. in naturally infected cattle in China

An enzyme immunoassay was used to detect anti-Sarcocystis antibodies in the sera of 159 cattle sent to slaughter in Jilin Province, north China. The musculature of each carcase was closely examined for the presence of parasitic cysts and aliquots of muscle were digested with pepsin and examined microscopically for cystozoites. Specific antibodies were detected in 126 (79.25%) cattle whereas cystozoites were detected in 123 (77.36%) and cysts in 103 (64.78%). The accuracy, sensitivity and specificity of the enzyme immunoassay were high (0.97, 0.99 and 0.91, respectively) and false reactions were only detected in four cases. The assay exhibited a high level of reproducibility when samples were re-tested and the soluble Sarcocystis spp. antigens did not cross-react with anti-Toxoplasma antibodies raised in rabbits. This report presents the first successful application of an enzyme immunoassay in the diagnosis of Sarcocystis spp. infections in naturally infected cattle in China. The assay, however, failed to detect specific antibodies in four dogs experimentally infected with S. cruzi from cattle.     

Prevalence and first genetic identification of Cryptosporidium spp. in cattle in central Viet Nam

We investigated the prevalence of Cryptosporidium infection in relation to age and clinical status in cattle in the central region of Viet Nam. A total of 266 fecal samples from diarrheic and non-diarrheic cattle were examined by the modified Ziehl-Neelsen staining method. Prevalence of Cryptosporidium parvum type infections, those of the Cryptosporidium andersoni type, and mixed infection of both types was 33.5% (89/266), 5.6% (15/266), and 3.4% (9/266), respectively. The infection rate of 44.3% (35/79) of C. parvum in calves less than 6 months old was significantly higher than that of 28.9% (54/187) in cattle greater than 6 months old (P < 0.01). Although no C. andersoni oocysts were detected in calves less than 3 months old, no significant difference was observed between the age groups in the prevalence of C. andersoni infection and mixed infection. The percentage of diarrheic and non-diarrheic cattle identified to be shedding C. parvum oocysts was 46.5% (74/159) and 14.0% (15/107), respectively (P < 0.0001). The risk of diarrhea was 1.7 times greater in C. parvum-infected calves than in their non-infected counterparts. DNA sequences of 18S rRNA genes of C. parvum type and C. andersoni type indicated that they were C. parvum bovine genotype and C. andersoni, respectively. This is the first genetic identification of C. parvum bovine genotype and C. andersoni from cattle in Viet Nam.     

Prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle from farms in China

Fecal samples of 2,056 dairy cattle from 14 farms were collected in three geographical regions of China and stained using a modified acid-fast staining technique to identify Cryptosporidium oocysts. A total of 387 (18.82%) positive samples were identified and further analyzed by polymerase chain reaction (PCR) using primers designed to amplify DNA fragments from the small subunit ribosomal RNA. The PCR products were sequenced and the sequences were deposited in the GenBank database under accession numbers EU369377-84 and GU070730-33. Phylogenetic analysis was performed and a distances matrix generated from these sequences confirmed the existence of Cryptosporidium (C.) parvum ''mouse'' genotype, C. bovis, C. andersoni, C. hominis, and C. serpentis in cattle. These results represent the first report on the prevalence and genetic identification of Cryptosporidium species, and may contribute to a better understanding of the epidemiology of Cryptosporidium in cattle in China.     

First isolation of Sarcocystis hominis from cattle in Japan.

Sarcocystis hominis was first isolated from slaughtered cattle raised in Japan. Cysts were 1,220-4,460 x 80-384 microns in size and their wall was 3 to 6 microns thick and appeared radially striated in the histopathological sections because of the presence of palisade-like villar protrusions on the surface. The protrusions were 3.1-4.3 x 0.7-1.1 microns in size and had many microtubules in the core. Two cynomolgus monkeys, Macaca fascicularis, fed with the Sarcocystis cysts began to pass sporocysts, which measured a size of 14.3-15 x 9.5-10 microns, in the feces 10 days after ingestion.     

The prevalence of Sarcocystis spp in dogs and red foxes.

Protozoan parasites of the genus Sarcocystis have been recognised for many years as intramuscular cysts of numerous vertebrates. It is only comparatively recently that the two-host nature of the life cycle has been recognised and that the intramuscular cysts are a stage in the developmental cycle of coccidian parasites of flesh eating mammals (Fayer 1974, Fayer and Johnson 1973, 1974, Rommel and others 1972, Dubey 1976). Carnivores ingest the intramuscular cysts from herbivores and presumably from other animals too and eventually shed sporulated tetrazoic sporocysts in their faeces. The cystic stages which occur in the flesh of herbivores are probably non-pathogenic but the earlier stages in which schizonts develop in vascular endothelium may be severely pathogenic. Sarcocystis cruzi, S ovicanis and S porcifelis are known to be severely pathogenic in cattle, sheep and pigs respectively (Dubey 1976). Observations on the prevalence of Sarcocystis spp in the faeces of working farm dogs, greyhounds and foxes (Vulpes vulpes) are recorded.     

Prevalence of Campylobacter spp. in slaughtered cattle and buffaloes in Vientiane, Lao People's Democratic Republic

This is the first report regarding isolation of Campylobacter in caecum and bile samples obtained from ruminants in Vientiane, Lao PDR. Campylobacter was isolated from 3 (1.6%) of the 184 caecum samples and 1 (1.0%) of the 100 bile samples obtained from buffaloes. Three of the 4 isolates were determined to be C. jejuni, which was detected in 2 caecum samples and 1 bile sample; the other caecum sample contained C. fetus. Campylobacter was not isolated from any of the 82 cattle caecum samples. Our results suggest that cattle and buffaloes may not be important sources of Campylobacter food poisoning in Lao PDR.     

Prevalence of Cryptosporidium spp. oocysts in cattle and wildlife in Morogoro region, Tanzania

Prevalences of Cryptosporidium spp. oocysts in cattle (n = 486) on five selected farms in Morogoro municipality and three species of herbivorous wildlife (n = 87) from Mikumi National Park, Morogoro, Tanzania, were determined using the modified Ziehl-Neelsen staining technique. Of 486 bovine faecal samples, 5.3% were positive for Cryptosporidium spp. The prevalence of Cryptosporidium was higher in calves less than 3 months of age compared to weaned calves and adults. Cryptosporidium spp. oocysts were detected in both diarrhoeic and non-diarrhoeic animals, but there was a significantly higher prevalence (P < 0.001) of oocyst shedding in diarrhoeic than in non-diarrhoeic animals. Of the 87 faecal specimens from the wildlife species, 36 were from the African buffaloes (Syncerus caffer), 25 from zebra (Equus zebra) and 26 from the wildebeest (Connochaetes gnou). Cryptosporidium spp. oocysts were detected in eight (22%) buffaloes, seven (28%) zebras and seven (27%) wildebeests. Confirmation of the diagnosis was performed using anti-Cryptosporidium monoclonal antibody specific for Cryptosporidium muris, Cryptosporidium parvum and Cryptosporidium baileyi (Pathasure Cryptosporidium test kit).     

Prevalence,quantitative load and genetic diversity of Campylobacter spp. in dairy cattle herds in Lithuania

Background

Campylobacteriosis is a zoonotic disease, and animals such as poultry, pigs and cattle may act as reservoirs for Campylobacter spp. Cattle shed Campylobacter spp. into the environment and they can act as a reservoir for human infection directly via contact with cattle or their faeces or indirectly by consumption of contaminated food. The aim of this study was to determine the prevalence, the quantitative load and the genetic strain diversity of Campylobacter spp. in dairy cattle of different age groups.

Results

Faecal samples of 200 dairy cattle from three farms in the central part of Lithuania were collected and examined for Campylobacter. Cattle herds of all three farms were Campylobacter spp. positive, with a prevalence ranging from 75% (farm I), 77.5% (farm II) to 83.3% (farm III). Overall, the highest prevalence was detected in calves (86.5%) and heifers (86.2%). In contrast, the lowest Campylobacter prevalence was detectable in dairy cows (60.6%). C. jejuni, C. coli, C. lari and C. fetus subsp. fetus were identified in faecal samples of dairy cattle. C. upsaliensis was not detectable in any sample. The high counts of Campylobacter spp. were observed in faecal material of dairy cattle (average 4.5 log₁₀ cfu/g). The highest numbers of Campylobacter spp. were found in faecal samples from calves (average 5.3 log₁₀ cfu/g), whereas, faecal samples from cows harboured the lowest number of Campylobacter spp. (average 3.7 log₁₀ cfu/g). Genotyping by flaA PCR-RFLP analysis of selected C. jejuni isolates showed that some genotypes were present in all farms and all age groups. However, farm or age specific genotypes were also identified.

Conclusions

Future studies are needed to investigate risk factors related to the degree of colonisation in cattle. Based on that, possible measures to reduce the colonisation and subsequent shedding of Campylobacter in cattle could be established. It is important to further investigate the epidemiology of Campylobacter in the cattle population in order to assess associated risks to public health.     

Characterization of monoclonal antibodies to Sarcocystis spp

Monoclonal antibodies have become powerful tools in immunology and biotechnology during the last decade. They are successfully used in a lot of fields in parasitology. This paper gives an overview of difficulties currently associated with immunodiagnosis of Sarcocystis infections and of the production and characterization of monoclonal antibodies for the investigation of problems that could not be solved by classical methods.