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Light microscopic and three-dimensional morphology of the human muscular sarcocyst

Wong KT, Clarke G, Pathmanathan R, Hamilton PW

Parasitol Res, 1994;80(2):138-40.
PMID: 8202453

Established criteria for morphological typing of sarcocysts was applied to a large series of cases of human skeletal muscle sarcocystosis in Malaysia to determine the type of sarcocyst present. We also wanted to test the general usefulness of this classification and to determine if there are any new cyst types. Three-dimensional (3-D) reconstruction was done to see if the sarcocyst has a distinct 3-D morphology. A total of 66 sarcocysts from 21 cases of human muscle sarcocystosis obtained from a previous prevalence study were examined. Tissue sections (5 microns thick) were stained with haematoxylin and eosin and studied under the light microscope. For 3-D reconstruction, an image analyser was used to align and reconstruct the sarcocyst after microscopic images had been captured with a charge-coupled device (CCD) camera. All the cysts best fit into the type 4 category. This classification is generally useful, although cyst wall characteristics and zoite size appear to be the most reliable criteria for classification. The cyst width averaged 77 microns (range, 30-137.5 microns). Cyst walls were smooth, had no cytophaneres and were less than 1 micron thick. No secondary cyst wall or surrounding inflammation was evident. Numerous cyst merozoites with diameters averaging 1 micron filled the cyst lumen. Although septa were not apparent, in many cysts, zoites were arranged in a unique, curvilinear fashion that suggested their presence. 3-D reconstruction showed the sarcocyst to be a long, tortuous "cylinder" with no branching or other distinguishing feature.

Matched MeSH terms: Sarcocystis/isolation & purification
Scanning electron microscopy of the human muscular sarcocyst

Wong KT, Yusoff M

Parasitol Res, 1995;81(4):359-60.
PMID: 7624297
Matched MeSH terms: Sarcocystis/isolation & purification
Letter: Sarcocystis in the Malaysian long-tailed monkey, Macaca irus

Prathap K

Trans R Soc Trop Med Hyg, 1973;67(4):615.
PMID: 4206526
Matched MeSH terms: Sarcocystis/isolation & purification*
Further stages of the "sporozoan" from the moon rat Echinosorex gymnurus, and its possible connection with Sarcocystis

Dissanaike AS, Lim BL, Poopalachelvam M

Southeast Asian J Trop Med Public Health, 1974 Sep;5(3):456-7.
PMID: 4215149
Matched MeSH terms: Sarcocystis/isolation & purification
Parasites of some rodents in Malaysia

Sinniah B

Southeast Asian J Trop Med Public Health, 1979 Mar;10(1):115-21.
PMID: 113889

A total of 2,337 rodents trapped from various parts of Peninsular Malaysia were dissected and studied for the distribution and prevalence of parasitic infections. Four new rodent hosts for Sarcocystis in Malaysia are reported (Bandicota indica, Rattus sabanus Rattus argentiventer and Rattus norvegicus). Sarcocystis was found in 17.2 percent of the rodents examined. Rattus annandalei, Rattus tiomanicus and Rattus norvegicus are new hosts of Syphacia muris in Peninsular Malsysia. Rattus sabanus was found to be infected with Zonorchis borneonenis. Brachylaima ratti Baugh, 1962 was recovered from the small intestine of Rattus rattus diardii for the first time in Malaysia. The prevalence and distribution of other parasites are also discussed.

Matched MeSH terms: Sarcocystis/isolation & purification
Fulltext Genetic assemblage of Sarcocystis spp. in Malaysian snakes

Lau YL, Chang PY, Subramaniam V, Ng YH, Mahmud R, Ahmad AF, et al.

Parasit Vectors, 2013 Sep 09;6(1):257.
PMID: 24010903 DOI: 10.1186/1756-3305-6-257

BACKGROUND: Sarcocystis species are protozoan parasites with a wide host range including snakes. Although there were several reports of Sarcocytis species in snakes, their distribution and prevalence are still not fully explored.
METHODS: In this study, fecal specimens of several snake species in Malaysia were examined for the presence of Sarcocystis by PCR of 18S rDNA sequence. Microscopy examination of the fecal specimens for sporocysts was not carried as it was difficult to determine the species of the infecting Sarcocystis.
RESULTS: Of the 28 snake fecal specimens, 7 were positive by PCR. BLASTn and phylogenetic analyses of the amplified 18S rDNA sequences revealed the snakes were infected with either S. nesbitti, S. singaporensis, S. zuoi or undefined Sarcocystis species.
CONCLUSION: This study is the first to report Sarcocystis infection in a cobra, and S. nesbitti in a reticulated python.

Matched MeSH terms: Sarcocystis/isolation & purification
Fulltext Sarcocystosis among wild captive and zoo animals in Malaysia

Latif B, Vellayan S, Omar E, Abdullah S, Mat Desa N

Korean J Parasitol, 2010 Sep;48(3):213-7.
PMID: 20877499 DOI: 10.3347/kjp.2010.48.3.213

Sarcocystis sp. infection was investigated in 20 necropsied captive wild mammals and 20 birds in 2 petting zoos in Malaysia. The gross post-mortem lesions in mammals showed marbling of the liver with uniform congestion of the intestine, and for birds, there was atrophy of the sternal muscles with hemorrhage and edema of the lungs in 2 birds. Naked eye examination was used for detection of macroscopic sarcocysts, and muscle squash for microscopic type. Only microscopically visible cysts were detected in 8 animals and species identification was not possible. Histological examination of the sections of infected skeletal muscles showed more than 5 sarcocysts in each specimen. No leukocytic infiltration was seen in affected organs. The shape of the cysts was elongated or circular, and the mean size reached 254 x 24.5 µm and the thickness of the wall up to 2.5 µm. Two stages were recognized in the cysts, the peripheral metrocytes and large numbers of crescent shaped merozoites. Out of 40 animals examined, 3 mammals and 5 birds were positive (20%). The infection rate was 15% and 25% in mammals and birds, respectively. Regarding the organs, the infection rate was 50% in the skeletal muscles followed by tongue and heart (37.5%), diaphragm (25%), and esophagus (12.5%). Further ultrastructural studies are required to identify the species of Sarcocystis that infect captive wild animals and their possible role in zoonosis.

Matched MeSH terms: Sarcocystis/isolation & purification*
High prevalence of human skeletal muscle sarcocystosis in south-east Asia

Wong KT, Pathmanathan R

Trans R Soc Trop Med Hyg, 1992 11 1;86(6):631-2.
PMID: 1287922

The prevalence of human skeletal muscle sarcocystosis in Malaysia was determined by serial examination of formalin-fixed, paraffin-embedded sections of tongue tissues obtained from consecutive, routine autopsies of subjects aged 12 years or more. Of 100 tongues examined, 21% were found to contain Sarcocystis; 66 cysts were found. The number of cysts per case varied from 1 to 13. In one case, 5 cysts were found in a single tissue section. The age range of positive cases was from 16 to 57 years (mean 37.7 years). Prevalence did not differ with regard to race, sex or occupation. The prevalence of human muscular sarcocystosis in our study was higher than that reported elsewhere. Preferential localization of Sarcocystis in tongue or head and neck and/or genuinely high prevalence in south-east Asia are possible explanations for this observation.

Matched MeSH terms: Sarcocystis/isolation & purification
Fulltext Human and animal invasive muscular sarcocystosis in Malaysia--recent cases, review and hypotheses

Tappe D, Abdullah S, Heo CC, Kannan Kutty M, Latif B

Trop Biomed, 2013 Sep;30(3):355-66.
PMID: 24189667

Sarcocystosis, an unusual parasitic zoonotic disease, is caused by coccidian/ apicomplexan protozoa in humans and animals. The parasites usually develop in a heteroxenous predator-prey life-cycle involving final (carnivore) and intermediate (omnivore/herbivore) hosts. Besides the intestinal, non-invasive form of the disease in which humans and animals are the definitive hosts for certain Sarcocystis spp., the invasive form has come to recent attention. In the latter, humans and animals serve as intermediate host harbouring sarcocysts in their muscle tissue. Already in 1991 sarcocystosis was seen as a potential emerging food borne zoonosis in Malaysia, and in 2011 and 2012 the largest cluster of symptomatic human muscular sarcocystosis world-wide was reported from Tioman Island, Pahang state. In this review, we focus on invasive sarcocystosis in humans and animals in Malaysia, review the recorded cases and epidemiology, and present hypotheses.

Matched MeSH terms: Sarcocystis/isolation & purification*
Fulltext Examination of Sarcocystis spp. of giant snakes from Australia and Southeast Asia confirms presence of a known pathogen - Sarcocystis nesbitti

Wassermann M, Raisch L, Lyons JA, Natusch DJD, Richter S, Wirth M, et al.

PLoS One, 2017;12(11):e0187984.
PMID: 29131856 DOI: 10.1371/journal.pone.0187984

We examined Sarcocystis spp. in giant snakes from the Indo-Australian Archipelago and Australia using a combination of morphological (size of sporocyst) and molecular analyses. We amplified by PCR nuclear 18S rDNA from single sporocysts in order to detect mixed infections and unequivocally assign the retrieved sequences to the corresponding parasite stage. Sarcocystis infection was generally high across the study area, with 78 (68%) of 115 examined pythons being infected by one or more Sarcocystis spp. Among 18 randomly chosen, sporocyst-positive samples (11 from Southeast Asia, 7 from Northern Australia) the only Sarcocystis species detected in Southeast Asian snakes was S. singaporensis (in reticulated pythons), which was absent from all Australian samples. We distinguished three different Sarcocystis spp. in the Australian sample set; two were excreted by scrub pythons and one by the spotted python. The sequence of the latter is an undescribed species phylogenetically related to S. lacertae. Of the two Sarcocystis species found in scrub pythons, one showed an 18S rRNA gene sequence similar to S. zamani, which is described from Australia for the first time. The second sequence was identical/similar to that of S. nesbitti, a known human pathogen that was held responsible for outbreaks of disease among tourists in Malaysia. The potential presence of S. nesbitti in Australia challenges the current hypothesis of a snake-primate life cycle, and would have implications for human health in the region. Further molecular and biological characterizations are required to confirm species identity and determine whether or not the Australian isolate has the same zoonotic potential as its Malaysian counterpart. Finally, the absence of S. nesbitti in samples from reticulated pythons (which were reported to be definitive hosts), coupled with our phylogenetic analyses, suggest that alternative snake hosts may be responsible for transmitting this parasite in Malaysia.

Matched MeSH terms: Sarcocystis/isolation & purification*
Fulltext Outbreak of human infection with Sarcocystis nesbitti, Malaysia, 2012

Abubakar S, Teoh BT, Sam SS, Chang LY, Johari J, Hooi PS, et al.

Emerg Infect Dis, 2013 Dec;19(12):1989-91.
PMID: 24274071 DOI: 10.3201/eid1912.120530

An outbreak of fever associated with myalgia and myositis occurred in 2012 among 89 of 92 college students and teachers who visited Pangkor Island, Malaysia. The Sarcocystis nesbitti 18S rRNA gene and sarcocysts were obtained from muscle tissues of 2 students. Our findings indicate emergence of S. nesbitti infections in humans in Malaysia.

Matched MeSH terms: Sarcocystis/isolation & purification
Fulltext Detection of sarcocystosis in goats in Malaysia by light microscopy, histology, and PCR

Kutty MK, Latif B, Muslim A, Hussaini J, Daher AM, Heo CC, et al.

Trop Anim Health Prod, 2015 Apr;47(4):751-6.
PMID: 25740651 DOI: 10.1007/s11250-015-0789-4

A number of methods have been used for the detection of the presence of microsarcocysts in animals, but little information exists on the value between the various methods. This study therefore examined for Sarcocystis spp. using three different methods in 105 samples of skeletal muscle collected from goats slaughtered in an abattoir in Selangor, Malaysia from January to February 2014. Three methods were used, direct light microscopy of squashed fresh muscle tissues; histological examination of fixed, sectioned, and hematoxylin and eosin (H&E)-stained samples of muscle; and molecular identification by polymerase chain reaction (PCR). Of the 105 tissue samples, 55 (52.38 %) were positive by light microscopy (LM), 46 (43.8 %) by histology, and 95 (90.48 %) by PCR. Only 29 (27.6 %) and 5 (4.76 %) samples were positive and negative, respectively, by all three methods. The cysts were elongated to a spindle shape with a mean size of 393.30 × 81.6 μm and containing banana-shaped bradyzoites of size 12.32 × 2.08 μm. The wall of the cyst was radially striated with a thickness of 2.83 μm. Samples were tested for the presence of Sarcocystis-specific 18S rRNA and were identified as Sarcocystis capracanis. Of the three methods used, the PCR test appears to be the most useful method for the diagnosis of sarcocystosis especially for species identification.

Matched MeSH terms: Sarcocystis/isolation & purification*
Suspected new wave of muscular sarcocystosis in travellers returning from Tioman Island, Malaysia, May 2014

Tappe D, Stich A, Langeheinecke A, von Sonnenburg F, Muntau B, Schäfer J, et al.

Euro Surveill, 2014 May 29;19(21).
PMID: 24906376

In May 2014, six patients presented in Germany with a Sarcocystis-associated febrile myositis syndrome after returning from Tioman Island, Malaysia. During two earlier waves of infections, in 2011 and 2012, about 100 travellers returning to various European countries from the island were affected. While the first two waves were associated with travel to Tioman Island mostly during the summer months, this current series of infections is associated with travel in early spring, possibly indicating an upcoming new epidemic.

Matched MeSH terms: Sarcocystis/isolation & purification*
Molecular evidence of Sarcocystis nesbitti in water samples of Tioman Island, Malaysia

Shahari S, Tengku-Idris TI, Fong MY, Lau YL

Parasit Vectors, 2016 11 23;9(1):598.
PMID: 27881179

BACKGROUND: Sarcocystis are intracellular protozoan parasites that are characterised by their ability to invade muscle tissue and form intramuscular sarcocysts. A muscular sarcocystosis outbreak was reported by travellers returning from Tioman Island in 2011 and 2012 where Sarcocystis nesbitti was identified as the main cause. The source of the S. nesbitti that was involved has remained elusive, although water is hypothesised to be the main cause of transmission. A surveillance study was therefore undertaken in the northern regions of Tioman Island to identify the source of S. nesbitti by screening rivers, water tanks, wells and seawater.
METHODS: Water samples were collected from rivers, water tanks, wells and seawater on Tioman Island over the course of April to October 2015. Water samples were indirectly screened for Sarcocystis species by obtaining sediment from respective water sources. PCR amplification of the 18S rRNA gene region was conducted to identify positive samples. Microscopy was used in an attempt to reappraise PCR results, but no sporocysts were detected in any of the samples.
RESULTS: A total of 157 water samples were obtained and 19 were positive for various Sarcocystis species. Through BLASTn and phylogenetic analysis, these species were found to be S. singaporensis, S. nesbitti, Sarcocystis sp. YLL-2013 and one unidentified Sarcocystis species.
CONCLUSIONS: This is the first positive finding of S. nesbitti in water samples on Tioman Island, which was found in a water tank and in river water samples. This finding supports the hypothesis that water was a potential medium for the transmission of S. nesbitti during the outbreak. This will potentially identify areas in which preventive measures can be taken to prevent future outbreaks.

Matched MeSH terms: Sarcocystis/isolation & purification*
Notes from the field: acute muscular sarcocystosis among returning travelers - Tioman Island, Malaysia, 2011

Centers for Disease Control and Prevention (CDC)

MMWR Morb Mortal Wkly Rep, 2012 Jan 20;61(2):37-8.
PMID: 22258418

GeoSentinel (the surveillance program of the International Society of Travel Medicine and CDC) has identified 32 cases of suspected acute muscular sarcocystosis in travelers returning from Tioman Island off the east coast of peninsular Malaysia. All the patients traveled to Tioman Island during the summer of 2011. Within days or weeks of returning home, all experienced fever and muscle pain, often severe and prolonged. All had peripheral eosinophilia, and most had elevated serum creatinine phosphokinase levels. Most were tested for acute trichinosis and toxoplasmosis by serology, and all of these tests were negative. Approximately half of the patients were identified in Germany; others were reported elsewhere in Europe, and in North America and Asia. Muscle biopsy from two patients demonstrated organisms consistent with sarcocystosis, one from a group of five ill travelers and one from a group of three.

Matched MeSH terms: Sarcocystis/isolation & purification*
Fulltext Acute muscular sarcocystosis: an international investigation among ill travelers returning from Tioman Island, Malaysia, 2011-2012

Esposito DH, Stich A, Epelboin L, Malvy D, Han PV, Bottieau E, et al.

Clin Infect Dis, 2014 Nov 15;59(10):1401-10.
PMID: 25091309 DOI: 10.1093/cid/ciu622

BACKGROUND: Through 2 international traveler-focused surveillance networks (GeoSentinel and TropNet), we identified and investigated a large outbreak of acute muscular sarcocystosis (AMS), a rarely reported zoonosis caused by a protozoan parasite of the genus Sarcocystis, associated with travel to Tioman Island, Malaysia, during 2011-2012.
METHODS: Clinicians reporting patients with suspected AMS to GeoSentinel submitted demographic, clinical, itinerary, and exposure data. We defined a probable case as travel to Tioman Island after 1 March 2011, eosinophilia (>5%), clinical or laboratory-supported myositis, and negative trichinellosis serology. Case confirmation required histologic observation of sarcocysts or isolation of Sarcocystis species DNA from muscle biopsy.
RESULTS: Sixty-eight patients met the case definition (62 probable and 6 confirmed). All but 2 resided in Europe; all were tourists and traveled mostly during the summer months. The most frequent symptoms reported were myalgia (100%), fatigue (91%), fever (82%), headache (59%), and arthralgia (29%); onset clustered during 2 distinct periods: "early" during the second and "late" during the sixth week after departure from the island. Blood eosinophilia and elevated serum creatinine phosphokinase (CPK) levels were observed beginning during the fifth week after departure. Sarcocystis nesbitti DNA was recovered from 1 muscle biopsy.
CONCLUSIONS: Clinicians evaluating travelers returning ill from Malaysia with myalgia, with or without fever, should consider AMS, noting the apparent biphasic aspect of the disease, the later onset of elevated CPK and eosinophilia, and the possibility for relapses. The exact source of infection among travelers to Tioman Island remains unclear but needs to be determined to prevent future illnesses.

Matched MeSH terms: Sarcocystis/isolation & purification
Fulltext High prevalence of muscular sarcocystosis in cattle and water buffaloes from Selangor, Malaysia

Latif B, Vellayan S, Heo CC, Kannan Kutty M, Omar E, Abdullah S, et al.

Trop Biomed, 2013 Dec;30(4):699-705.
PMID: 24522140 MyJurnal

The prevalence of sarcocystosis in cattle and water buffaloes from peninsular Malaysia was investigated in abattoirs in Selangor state, February, 2011, to March, 2012. Fresh muscle samples were collected from the tongue, heart, oesophagus, diaphragm and skeletal muscles of 102 cattle and 18 water buffaloes. Each sample was initially screened by light microscopy and then fixed for further histopathological analysis. Out of 120 animals examined, 49 (40.8%) harboured the microscopic type of Sarcocystis spp. The positivity rate for cattle was 36.2% and for water buffaloes 66.7%. In cattle, the organs highly infected were the skeletal muscles and diaphragm (27% each), followed by tongue and esophagus (24.3% each), and the heart (8%). In water buffaloes, the heart was most often infected (66.7%), followed by the oesophagus (50%) and skeletal muscle (33.3%); no sarcocysts were detected in the tongue and diaphragm. The shape of the sarcocyst was fusiform to oval with a mean cyst size of 151.66 x 75.83 μm and wall thickness of 2.47 μm in cattle, and 114 x 50.81 μm cyst size and the wall thickness of 1.11 μm in water buffaloes, consistent with Sarcocystis cruzi and Sarcocystis levinei, respectively. Remaining tissue from cattle was subjected to parasite specific 18S rRNA gene PCR and Sarcocystis cruzi was confirmed, at least exemplarily. The peripheral metrocytes and the banana-shaped bradyzoites (15.23 x 2.2 μm in cattle and 11.49 x 2.45 μm in water buffalo hosts) were easily recognized. In conclusion, a high positivity rate was found in Malaysian meat-producing animals with possible implications for meat consumption and human health.

Matched MeSH terms: Sarcocystis/isolation & purification*
Ongoing outbreak of an acute muscular Sarcocystis-like illness among travellers returning from Tioman Island, Malaysia, 2011-2012

Esposito DH, Freedman DO, Neumayr A, Parola P

Euro Surveill, 2012 Nov 08;17(45).
PMID: 23153473

As of 4 November, 2012, 100 patients with an acute muscular Sarcocystis-like illness associated with travel to Tioman Island, Malaysia, have been identified. Thirty-five travelled there mostly during July and August 2011 and 65 mostly during July and August 2012, suggesting an ongoing outbreak. Epidemiological investigations are ongoing. Public health agencies and practicing clinicians should be aware of this rarely-reported disease in humans and consider it as differential diagnosis in travellers returning from Tioman Island.

Matched MeSH terms: Sarcocystis/isolation & purification
Sarcocystis booliati n.sp. and a parasite of undetermined taxonomic position, Octoplasma garnhami n. gen. n. sp., from the moonrat, Echinosorex gymnurus

Dissanaike AS, Poopalachelvam M

Southeast Asian J Trop Med Public Health, 1975 Jun;6(2):175-85.
PMID: 809845

Sarcocystis booliati n.sp. is described from the moonrat Echinosorex gymnurus (Mammalia, Insectivora) from West Malaysia. The cysts are very thin-walled, not visible to the naked eye, and have no trabeculae or cytophaneres. They are found in skeletal but not heart muscle. The zoites are small, 5-8 by 2-3 mum with a mean of 6.5 by 2.2 mum, in dry fixed smears. Octoplasma garnhami n.gen. n.sp., a parasite of undetermined taxonomic status but belonging to the Coccidiasina, Apicomplexa, is also described from the same host. Only schizononts and pseudocysts with typically 8 zoites, have so far been seen in monocytes of the spleen and liver. The zoites are large, 15 by 3 mum and have a distinct nucleolus even in dry-fixed smears.

Matched MeSH terms: Sarcocystis/isolation & purification*
Fulltext Neglected tropical diseases among two indigenous subtribes in peninsular Malaysia: highlighting differences and co-infection of helminthiasis and sarcocystosis

Lee SC, Ngui R, Tan TK, Muhammad Aidil R, Lim YA

PLoS One, 2014;9(9):e107980.
PMID: 25248116 DOI: 10.1371/journal.pone.0107980

Soil-transmitted helminth (STH) infections have been documented among these minority groups since 1938. However the prevalence of STH is still high among these communities. Most studies tend to consider the Orang Asli (indigenous) as a homogenous group. In contrary, different subtribes have their own cultural practices. To understand this variation better, we studied the prevalence and associated factors of STH and other gut parasitic infections among two common subtribes (i.e. Temuan and Temiar). Results showed that the prevalence of the overall STH infections was higher in the Temuan subtribe (53.2% of 171) compared to the Temiar subtribe (52.7% of 98). Trichuris trichiura (46.2%) was the most prevalent parasite in the Temuan subtribe, followed by Ascaris spp. (25.7%) and hookworm (4.1%). In contrast, Ascaris spp. (39.8%) was more prevalent among the Temiar subtribe, preceded by T. trichiura (35.7%) and finally hookworm (8.3%). There were also co-infections of helminthiasis and intestinal protozoa among both Temuan and Temiar subtribes with rates being three times higher among the Temiar compared to Temuan. The most common co-infection was with Entamoeba histolytica/dispar/moshkovskii (n = 24; 24.5%, 16.0-33.0), followed by Giardia spp. (n = 3; 3.1%, -0.3-6.5). In Temuan, STH infection individuals were also infected with Entamoeba histolytica/dispar/moshkovskii (n = 11; 6.4%, 5.0-13.8), Cryptosporidium spp. (n = 3, 1.8%, -0.2-3.8) and Giardia spp. (n = 2, 1.2%, -0.4-2.8). In comparison, there was no Cryptosporidium spp. detected among the Temiar. However, it was interesting to note that there was an occurrence of co-infection of intestinal helminthiasis and sarcocystosis (intestinal) in a Temiar individual. The last report of sarcocystosis (muscular) among the Orang Asli was in 1978. The present study highlighted the importance of understanding the variation of infections amongst the different Orang Asli subtribes. It is vital to note these differences and use this knowledge to customise effective control measures for the various subtribes.

Matched MeSH terms: Sarcocystis/isolation & purification

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