Detection of human Sarcocystosis using dried blood on filter papers: An Immunofluorescent Antibody Test.

Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats' muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis.

Detection of human Sarcocystosis using dried blood on filter papers: An Immunofluorescent Antibody Test

@#Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats’ muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis.

Light microscopy and molecular identification of Sarcocystis spp. in meat producing animals in Selangor, Malaysia.

One thousand and forty-five tissue samples of skeletal muscles, tongue, heart, diaphragm and esophagus were collected from 209 animals (43 sheep, 89 goats and 77 cattle) from an abattoir in Selangor between February and October, 2013. Each sample was divided into three pieces with each piece measuring 2-3 mm3. Each piece was then squeezed between two glass slides and examined microscopically at x 10 magnification for the presence of sarcocystosis. Three positive samples from each animal species were then fixed in 10% formalin for histological processing. Seven positive samples collected from each animal species were preserved at -80°C or 90% ethanol for gene expression studies. Microsarcocysts were detected in 114 (54.5%) animals by light microscopy (LM). The infection rates in sheep, goat and cattle were 86, 61.8 and 28.6% respectively. The highest rate of infection was in the skeletal muscles of sheep (64.9%) and goats (63.6%) and in the heart of cattle (63.6%). The cysts were spindle to oval in shape and two stages were recognized, the peripheral metrocytes and centrally located banana-shaped bradyzoites. 18S rRNA gene expression studies confirmed the isolates from the sheep as S. ovicanis, goats as S. capracanis and cattle as S. bovicanis. This, to the best of our knowledge, is the first molecular identification of an isolate of S. ovicanis and S. capracanis in Malaysia. Further studies with electron microscopy (EM) are required in the future to compare the features of different types of Sarcocysts spp.

Morphological descriptions on the larvae of Hypopygiopsis fumipennis (Walker, 1856) (Diptera: Calliphoridae).

The study on biodiversity of forensically important Diptera in the tropical rain forest in Malaysia is scarce. Thus, a preliminary survey was conducted at a jungle fringe near Kampung Bahagia Bukit Lagong, Sungai Buloh, Selangor. A rat carcass was offered to attract carrion flies and we collected an adult female calliphorid, Hypopygiopsis fumipennis (Walker, 1856) during the fresh stage of carcass decomposition. The female fly was allowed to oviposit on chicken liver in a container and the resulting larvae were reared to the adult stage. Along the developmental process, several individuals from each instar were collected and preserved in 70% ethanol and then processed on the slides. We recorded the duration of development for each instar and described its larval features for the first time. The third instar larvae of H. fumipennis showed accessory oral sclerite present, anterior spiracle with 13-15 papillae, intersegmental spines mostly unicuspid with pointed end, and posterior spiracles heavily sclerotized with inter-slit projections. Some larval differences between H. fumipennis and Hypopygiopsis violacea were noted.

Light microscopy and molecular identification of Sarcocystis spp. in meat producing animals in Selangor, Malaysia

One thousand and forty-five tissue samples of skeletal muscles, tongue, heart, diaphragm and esophagus were collected from 209 animals (43 sheep, 89 goats and 77 cattle) from an abattoir in Selangor between February and October, 2013. Each sample was divided into three pieces with each piece measuring 2-3 mm3. Each piece was then squeezed between two glass slides and examined microscopically at x 10 magnification for the presence of sarcocystosis. Three positive samples from each animal species were then fixed in 10% formalin for histological processing. Seven positive samples collected from each animal species were preserved at -80°C or 90% ethanol for gene expression studies. Microsarcocysts were detected in 114 (54.5%) animals by light microscopy (LM). The infection rates in sheep, goat and cattle were 86, 61.8 and 28.6% respectively. The highest rate of infection was in the skeletal muscles of sheep (64.9%) and goats (63.6%) and in the heart of cattle (63.6%). The cysts were spindle to oval in shape and two stages were recognized, the peripheral metrocytes and centrally located banana-shaped bradyzoites. 18S rRNA gene expression studies confirmed the isolates from the sheep as S. ovicanis, goats as S. capracanis and cattle as S. bovicanis. This, to the best of our knowledge, is the first molecular identification of an isolate of S. ovicanis and S. capracanis in Malaysia. Further studies with electron microscopy (EM) are required in the future to compare the features of different types of Sarcocysts spp.

Morphological descriptions on the larvae of Hypopygiopsis fumipennis (Walker, 1856) (Diptera: Calliphoridae)

The study on biodiversity of forensically important Diptera in the tropical rain forest in Malaysia is scarce. Thus, a preliminary survey was conducted at a jungle fringe near Kampung Bahagia Bukit Lagong, Sungai Buloh, Selangor. A rat carcass was offered to attract carrion flies and we collected an adult female calliphorid, Hypopygiopsis fumipennis (Walker, 1856) during the fresh stage of carcass decomposition. The female fly was allowed to oviposit on chicken liver in a container and the resulting larvae were reared to the adult stage. Along the developmental process, several individuals from each instar were collected and preserved in 70% ethanol and then processed on the slides. We recorded the duration of development for each instar and described its larval features for the first time. The third instar larvae of H. fumipennis showed accessory oral sclerite present, anterior spiracle with 13-15 papillae, intersegmental spines mostly unicuspid with pointed end, and posterior spiracles heavily sclerotized with inter-slit projections. Some larval differences between H. fumipennis and Hypopygiopsis violacea were noted.

The fallopian canal: a comprehensive review and proposal of a new classification.

INTRODUCTION: The facial nerve follows a complex course through the skull base. Understanding its anatomy is crucial during standard skull base approaches and resection of certain skull base tumors closely related to the nerve, especially, tumors at the cerebellopontine angle. METHODS: Herein, we review the fallopian canal and its implications in surgical approaches to the skull base. Furthermore, we suggest a new classification. CONCLUSIONS: Based on the anatomy and literature, we propose that the meatal segment of the facial nerve be included as a component of the fallopian canal. A comprehensive knowledge of the course of the facial nerve is important to those who treat patients with pathology of or near this cranial nerve.

Human and animal invasive muscular sarcocystosis in Malaysia--recent cases, review and hypotheses.

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.

Dermatitis caused by Paederus fuscipes Curtis, 1840 (Coleoptera: Staphilinidae) in student hostels in Selangor, Malaysia.

We report a series of dermatitis cases caused by the staphilinid beetles, Paederusfuscipes Curtis, among university students staying in the residential college in Puncak Alam, Selangor, Malaysia from 1 January to 31 December 2010. A total of 360 cases (6.0%) were recorded in the Student Health Center throughout the year; the majority of patients stayed at a hostel near an oil palm plantation. Skin symptoms included erythema, edema, vesicular papules, painful blisters, burning sensation, pruritus, hyper pigmentation and peeling of skin. The commonly involved sites were the face, neck, shoulders and arms. Most students noticed the symptoms upon awakening in the morning. The patients were treated with fusidic acid cream and the symptoms resolved within 5 days. These beetles are nocturnally active and enter the room whenever a light source is available. The unintentional crushing of these beetles during sleep causes the release of its hemolymph (paederin) which is the cause of the dermatitis.

Gabriele Fallopio (1523-1562) and his contributions to the development of medicine and anatomy.

INTRODUCTION: Gabriele Fallopio was one of the greatest anatomists of the sixteenth century. He discovered and named numerous parts of the human body. His name survives to this day as it is associated with several anatomical structures including the Fallopian canal, Fallopian hiatus, Fallopian valve, Fallopian muscle, and the Fallopian tube. CONCLUSIONS: Our current knowledge of human anatomy is based on giants such as Fallopio. His contributions to neuroanatomy laid the foundations for the development of this discipline.

High prevalence of muscular sarcocystosis in cattle and water buffaloes from Selangor, Malaysia.

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.

Seasonal variation in Sarcocystis species infections in goats in northern Iraq.

We investigated the prevalence of sarcocystosis in 826 goats slaughtered in the winter season from November to April in northern Iraq. The prevalence of macrocysts was on average 34%, with only 20% infected animals in November, but 46% in February. The infection rate in 1-, 3- and 6-year-old goats was 4%, 48%, and 83%, respectively. The highest specificity of infection was in the oesophagus (99%) and the lowest in the diaphragm (3%). Grossly, we identified 2 forms of macroscopic sarcocysts, fat and thin, with different morphological characteristics. The prevalence of microcysts was 97% and no effects of age, sex and seasonal variations were observed. Development of microcysts in the small intestine of dogs and cats has also been investigated. The pre-patent period in experimentally infected dogs was 12-14 days and the patent period lasted 64-66 days. A dog shed about 155 million sporocysts, but no sporocysts were shed by cats that had been fed the same infected tissues, thus identifying the microcysts as Sarcocystis capracanis.

Cryptosporidiosis in children in a north Jordanian paediatric hospital

We investigated the rate of infection by Cryptosporidium parvum among children from birth to 12 years attending Princess Rahma Teaching Hospital in Irbid, Jordan and evaluated various diagnostic methods. We collected single stool specimens from 300 children; 7 specimens were from children undergoing chemotherapy treatment for cancer. Diagnostic methods used for detection of infection were direct wet mount preparation, flotation concentration, cold Kinyoun Ziehl-Neelsen stain and direct immunofluorescence. We detected C. parvum oocysts in 112 samples [37.3%] using direct immunofluorescence, which showed the highest sensitivity. Source of drinking water appeared to be an important risk factor for transmission of infection. A higher incidence of infection was recorded during January-May, the rainy season

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.

Immunization of sheep against Theileria hirci infection with schizonts propagated in tissue culture.

Two experiments were carried out. In the first, three groups of lambs were inoculated subcutaneously with 3 X 10(6) schizonts of different passages (3, 30 and 63) of Theileria hirci propagated in tissue culture. Severe reactions were observed in lambs inoculated with organisms derived from the 3rd passage. In the second experiment, four groups were inoculated with 5 X 10(5), 3 X 10(6), 1 X 10(7) and 5 X 10(8) schizonts of the 63rd passage. No clinical reactions or parasites were detected in lambs inoculated with 5 X 10(5) schizonts. Mild reactions were observed in lambs inoculated with 3 X 10(6), 1 X 10(7), and 5 X 10(7) schizonts. Lambs inoculated with 3 X 10(6) schizonts were resistant to challenge with a virulent strain. The indirect fluorescent antibody (IFA) test was used to determine the antibody titre.