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Genetic diversity of caprine Blastocystis from Peninsular Malaysia

Tan TC, Tan PC, Sharma R, Sugnaseelan S, Suresh KG

Parasitol Res, 2013 Jan;112(1):85-9.
PMID: 22961236 DOI: 10.1007/s00436-012-3107-3

Blastocystis sp. is a common intestinal parasite found in humans and animals. The possibility of zoonotic transmission to humans from livestock especially goats led us to investigate the genetic diversity of caprine Blastocystis sp. obtained from five different farms in Peninsular Malaysia. Moreover, there is a lack of information on the prevalence as well as genetic diversity of Blastocystis sp. in goat worldwide. Results showed that 73/236 (30.9 %) of the goats were found to be positive for Blastocystis infection. The most predominant Blastocystis sp. subtype was ST1 (60.3 %) followed by ST7 (41.1 %), ST6 (41.1 %), and ST3 (11.0 %) when amplified by PCR using sequenced-tagged site (STS) primers. Four farms had goats infected only with ST1 whereas the fifth showed mixed infections with multiple STs. The proximity of the fifth farm to human dwellings, nearby domesticated animals and grass land as opposed to a sterile captive environment in the first four farms may account for the multiple STs seen in the fifth farm. Since ST1, ST3, ST6 and ST 7 were previously reported in human infection worldwide in particular Malaysia, the potential of the zoonotic transmission of blastocystosis should not be disregarded. The implications of different farm management systems on the distribution of Blastocystis sp. STs are discussed.

Matched MeSH terms: Blastocystis Infections/parasitology
Effects of symptomatic and asymptomatic isolates of Blastocystis hominis on colorectal cancer cell line, HCT116

Chan KH, Chandramathi S, Suresh K, Chua KH, Kuppusamy UR

Parasitol Res, 2012 Jun;110(6):2475-80.
PMID: 22278727 DOI: 10.1007/s00436-011-2788-3

The pathogenesis of Blastocystis hominis in human hosts has always been a matter of debate as it is present in both symptomatic and asymptomatic individuals. A recent report showed that B. hominis isolated from an asymptomatic individual could facilitate the proliferation and growth of existing cancer cells while having the potential to downregulate the host immune response. The present study investigated the differences between the effects of symptomatic and asymptomatic derived solubilized antigen of B. hominis (Blasto-Ag) on the cell viability and proliferation of colorectal cancer cells. Besides that, the gene expression of cytokine and nuclear transcriptional factors in response to the symptomatic and asymptomatic B. hominis antigen in HCT116 was also compared. In the current study, an increase in cell proliferation was observed in HCT116 cells which led to the speculation that B. hominis infection could facilitate the growth of colorectal cancer cells. In addition, a more significant upregulation of Th2 cytokines observed in HCT116 may lead to the postulation that symptomatic Blasto-Ag may have the potential in weakening the cellular immune response, allowing the progression of existing tumor cells. The upregulation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB) was observed in HCT116 exposed to symptomatic Blasto-Ag, while asymptomatic Blasto-Ag exhibited an insignificant effect on NF-κB gene expression in HCT116. HCT116 cells exposed to symptomatic and asymptomatic Blasto-Ag caused a significant upregulation of CTSB which lead to the postulation that the Blasto-Ag may enhance the invasive and metastasis properties of colorectal cancer. In conclusion, antigen isolated from a symptomatic individual is more pathogenic as compared to asymptomatic isolates as it caused a more extensive inflammatory reaction as well as more enhanced proliferation of cancer cells.

Matched MeSH terms: Blastocystis Infections/parasitology*
Genetic variability of Blastocystis sp. isolates obtained from cancer and HIV/AIDS patients

Tan TC, Ong SC, Suresh KG

Parasitol Res, 2009 Oct;105(5):1283-6.
PMID: 19603182 DOI: 10.1007/s00436-009-1551-5

This represents the first study to determine the genetic diversity of Blastocystis sp. among cancer and HIV/AIDS patients. Forty Blastocystis sp. isolates obtained from 20 cancer and 20 HIV/AIDS patients were genotyped by PCR using seven pairs of known sequenced-tagged site primers. Out of the 40 isolates, 38 were identified as one of the known genotypes and two isolates were negative with all the STS primers. Blastocystis sp. subtype 3 which is reported to be associated with disease was found to be predominant among the study subjects.

Matched MeSH terms: Blastocystis Infections/parasitology*
Phenotypic and genotypic characterisation of Blastocystis hominis isolates implicates subtype 3 as a subtype with pathogenic potential

Tan TC, Suresh KG, Smith HV

Parasitol Res, 2008 Dec;104(1):85-93.
PMID: 18795333 DOI: 10.1007/s00436-008-1163-5

Despite frequent reports on the presence of Blastocystis hominis in human intestinal tract, its pathogenicity remains a matter of intense debate. These discrepancies may be due to the varying pathogenic potential or virulence of the isolates studied. The present study represents the first to investigate both phenotypic and genotypic characteristics of B. hominis obtained from symptomatic and asymptomatic individuals. Symptomatic isolates had a significantly greater size range and lower growth rate in Jones' medium than asymptomatic isolates. The parasite cells of symptomatic isolates exhibited rougher surface topography and greater binding affinity to Canavalia ensiformis (ConA) and Helix pomatia (HPA). The present study also identifies further phenotypic characteristics, which aided in differentiating the pathogenic forms from the non-pathogenic forms of B. hominis. Blastocystis subtype 3 was found to be correlated well with the disease.

Matched MeSH terms: Blastocystis Infections/parasitology*
Apoptosis in Blastocystis spp. is related to subtype

Dhurga DB, Suresh KG, Tan TC, Chandramathi S

Trans R Soc Trop Med Hyg, 2012 Dec;106(12):725-30.
PMID: 23141370 DOI: 10.1016/j.trstmh.2012.08.005

Previous studies have shown that apoptosis-like features are observed in Blastocystis spp., an intestinal protozoan parasite, when exposed to the cytotoxic drug metronidazole (MTZ). This study reports that among the four subtypes of Blastocystis spp. investigated for rate of apoptosis when treated with MTZ, subtype 3 showed the highest significant increase after 72h of in vitro culture when treated with MTZ at 0.1mg/ml (79%; p<0.01) and 0.0001mg/ml (89%; p<0.001). The close correlation between viable cells and apoptotic cells for both dosages implies that the pathogenic potential of these isolates has been enhanced when treated with MTZ. This suggests that there is a mechanism in Blastocystis spp. that actually regulates the apoptotic process to produce higher number of viable cells when treated. Apoptosis may not just be programmed cell death but instead a mechanism to increase the number of viable cells to ensure survival during stressed conditions. The findings of the present study have an important contribution to influence chemotherapeutic approaches when developing drugs against the emerging Blastocystis spp. infections.

Matched MeSH terms: Blastocystis Infections/parasitology*
Fulltext Predominance of Blastocystis sp. Infection among School Children in Peninsular Malaysia

Nithyamathi K, Chandramathi S, Kumar S

PLoS One, 2016;11(2):e0136709.
PMID: 26914483 DOI: 10.1371/journal.pone.0136709

BACKGROUND: One of the largest cross-sectional study in recent years was carried out to investigate the prevalence of intestinal parasitic infections among urban and rural school children from five states namely Selangor, Perak, Pahang, Kedah and Johor in Peninsula Malaysia. This information would be vital for school authorities to influence strategies for providing better health especially in terms of reducing intestinal parasitism.
METHODS AND PRINCIPAL FINDINGS: A total of 3776 stool cups was distributed to 26 schools throughout the country. 1760 (46.61%) responded. The overall prevalence of intestinal parasitic infection in both rural and urban areas was 13.3%, with Blastocystis sp (10.6%) being the most predominant, followed by Trichuris trichiura (3.4%), Ascaris lumbricoides (1.5%) and hook worm infection (0.9%). Only rural school children had helminthic infection. In general Perak had the highest infection (37.2%, total, n = 317), followed by Selangor (10.4%, total, n = 729), Pahang (8.6%, total, n = 221), Kedah (6.2%, total, n = 195) and Johor (3.4%, total, n = 298). School children from rural schools had higher infection (13.7%, total, n = 922) than urban school children (7.2%, total, n = 838). Subtype (ST) 3 (54.3%) is the most predominant ST with persons infected with only ST1 and ST3 showing symptoms. Blastocystis sp infection significantly associated with low household income, low parent's education and presence of symptoms (p<0.05).
CONCLUSION: It is critical that we institute deworming and treatment to eradicate the parasite especially in rural school children.

Matched MeSH terms: Blastocystis Infections/parasitology
Polypeptides associated with in vitro cyst formation of Blastocystis hominis

Init I, Mak JW, Top S, Zulhainan Z, Prummongkol S, Nissapatorn V, et al.

Southeast Asian J. Trop. Med. Public Health, 2003 Dec;34(4):727-32.
PMID: 15115079

The objective of this study was to characterize the polypeptides associated with cysts of Blastocystis hominis. This form is believed to be infective and plays a role in parasite resistance to anti-B. hominis drugs currently used for treatment of Blastocystis associated diarrhea. Cysts were induced through in vitro culture of the parasite in complete medium supplemented with bacterial extract with trypticase, metronidazole or doxycycline. SDS-PAGE analysis showed almost similar polypeptide patterns of parasite extracts obtained from in vitro cultured parasites before and after exposure with the three supplements. Polypeptide bands at 76, 58.5, 48, 45, 40, 38, 32, 25 and 22 kDa were constantly seen in all antigenic preparations and no specific cyst-associated polypeptide was present. However, on immunoblot analysis, 3 out of 16 blastocystosis human sera identified a cyst-associated polypeptide at 60 kDa in all parasite extracts prepared from cultures with the three supplements. In addition, there were associated morphological changes detected in these parasites stained with acridine orange and observed under fluorescence microscopy. Metronidazole induced cyst forms (reddish cells) as early as 12 hours post-exposure; more cyst production (with stronger immunoblot bands) occurred after 24 hours exposure. However, cysts rupture with release and destruction of B. hominis daughters cells occurred after 48 hours exposure. Doxycycline induced less cyst-like forms at 24 hours (weaker 60 kDa band) and less destruction of the cysts (60 kDa band still present at 72 hours post exposure). Bacterial extract and trypticase also induced cysts at 12 hours with increasing numbers up to 72 hours exposure (corresponding increase in intensity of 60 kDa band from samples harvested at 12 to 72 hours post exposure) without any sign of deleterious effect on the parasite.

Matched MeSH terms: Blastocystis Infections/parasitology*
Prevalence and subtypes of Blastocystis among migrant workers from different working sectors in Peninsular Malaysia

Sahimin N, Meor Termizi FH, Rajamanikam A, Mohd Nazri NA, Govind SK, Mohd Zain SN

Parasitol Res, 2020 Oct;119(10):3555-3558.
PMID: 32875397 DOI: 10.1007/s00436-020-06865-3

Blastocystis sp. is a common enteric parasite of humans and animals associated with inadequate sanitation and poor personal hygiene. Over the years, the Malaysian thriving economy has been facilitated largely by migrant workers from developing countries, and there is concern that diseases endemic to their countries may be imported. Therefore, this study aimed to determine the current status of Blastocystis infection as well as subtypes (STs) from fecal samples among migrant workers in Selangor and Kuala Lumpur, Malaysia. Overall, almost a third of the study cohort (30.9%; n = 68/220) screened were infected with Blastocystis sp. predominantly with ST3 (54.5%; n = 12), followed by ST1 (36.4%; n = 8) and ST2 (9.1%; n = 2). Infection levels was almost similar among the different sectors; manufacturing (32.8%), domestic service (32.3%), and food service (27.3%) with common symptoms for infection included stomach and abdominal pain or discomfort and diarrhea (48.5%; n = 33). None of the socio-demographic risk factors evaluated were significant. Therefore, this study warrants continuous monitoring as well as understanding the impact of transmission among the migrant community with the local population especially those involved in food service sector.

Matched MeSH terms: Blastocystis Infections/parasitology*
Isolate resistance of Blastocystis hominis to metronidazole

Haresh K, Suresh K, Khairul Anus A, Saminathan S

Trop Med Int Health, 1999 Apr;4(4):274-7.
PMID: 10357863

Isolates of Blastocystis hominis from infected immigrant workers from Indonesia, Bangladesh and infected individuals from Singapore and Malaysia were assessed for growth pattern and degree of resistance to different concentrations of metronidazole. Viability of the cells was assessed using eosin-brillian cresyl blue which stained viable cells green and nonviable cells red. The Bangladeshi and Singaporean isolates were nonviable even at the lowest concentration of 0.01 mg/ml, whereas 40% of the initial inoculum of parasites from the Indonesian isolate at day one were still viable in cultures with 1.0 mg/ml metronidazole. The study shows that isolates of B. hominis of different geographical origin have different levels of resistance to metronidazole. The search for more effective drugs to eliminate th parasite appears inevitable, especially since surviving parasites from metronidazole cultures show greater ability to multiply in subcultures than controls.

Matched MeSH terms: Blastocystis Infections/parasitology*
Fulltext Molecular epidemiology of blastocystosis in Malaysia: does seasonal variation play an important role in determining the distribution and risk factors of Blastocystis subtype infections in the Aboriginal community?

Noradilah SA, Moktar N, Anuar TS, Lee IL, Salleh FM, Manap SNAA, et al.

Parasit Vectors, 2017 Jul 31;10(1):360.
PMID: 28760145 DOI: 10.1186/s13071-017-2294-2

BACKGROUND: Alternating wet and dry seasons may play an important role in the acquisition and distribution of Blastocystis subtype infection in the tropics. This cross-sectional study was therefore conducted to provide the prevalence of Blastocystis and to determine the potential risk factors associated with each subtype during the wet and dry seasons in the Aboriginal community, Pahang, Malaysia.
METHODS: A total of 473 faecal samples were collected: 256 (54.1%) and 217 (45.9%) samples were obtained during the wet (October-November 2014) and the dry season (June 2015), respectively. All fresh faecal samples were subjected to molecular analysis for subtype and allele identification.
RESULTS: Of the 473 samples, 42.6% and 37.8% were positive for Blastocystis ST1, ST2, ST3 and ST4 during wet and dry seasons, respectively. Prevalence of Blastocystis ST1 was significantly higher during the wet season compared to the dry season (Z = 2.146, P

Matched MeSH terms: Blastocystis Infections/parasitology
Fulltext Comparison of apoptotic responses in Blastocystis sp. upon treatment with Tongkat Ali and Metronidazole

Girish S, Kumar S, Aminudin N, Hashim NM

Sci Rep, 2021 04 09;11(1):7833.
PMID: 33837230 DOI: 10.1038/s41598-021-81418-x

Blastocystis sp. infection, although many remain asymptomatic, there is growing data in recent studies that suggests it is a frequent cause of gastrointestinal symptoms in children and adults. This proposes that treatment against this infection is necessary however metronidazole (MTZ), which is the current choice of treatment, has expressed non-uniformity in its efficacy in combating this infection which has led to the study of alternative treatment. In our previous study, it was established that Tongkat Ali fractions exhibited promising anti-protozoal properties which leads to the current aim of the study, to further narrow down the purification process in order to identify the specific active compound promoting the anti-protozoal effect through HPLC analysis. Based on the data analysis and in-vitro susceptibility assay, the collected Tongkat Ali fraction that demonstrated anti-blastocystis property was shown to contain eurycomanone. Previous studies have suggested that there is a mechanism in Blastocystis sp. that regulates the apoptotic process to produce higher number of viable cells when treated. In reference to this, our current study also aims to investigate the apoptotic response of Tongkat Ali extract and eurycomanone across different subtype groups with comparison to MTZ. Based on our investigation, both Tongkat Ali extract and eurycomanone induced the high apoptotic rate however exhibited a reduction in viable cell count (p

Matched MeSH terms: Blastocystis Infections/parasitology*
Fulltext Prevalence, predictors and clinical significance of Blastocystis sp. in Sebha, Libya

Abdulsalam AM, Ithoi I, Al-Mekhlafi HM, Khan AH, Ahmed A, Surin J, et al.

Parasit Vectors, 2013;6:86.
PMID: 23566585 DOI: 10.1186/1756-3305-6-86

BACKGROUND: Blastocystis sp. has a worldwide distribution and is often the most common human intestinal protozoan reported in children and adults in developing countries. The clinical relevance of Blastocystis sp. remains controversial. This study was undertaken to determine the prevalence of Blastocystis infection and its association with gastrointestinal symptoms among outpatients in Sebha city, Libya.
METHODS: A total of 380 stool samples were collected from outpatients attending the Central Laboratory in Sebha, Libya for routine stool examination. The presence of Blastocystis sp. was screened comparing light microscopy of direct smears against in vitro cultivation. Demographic and socioeconomic information were collected with a standardized questionnaire.
RESULTS: The us (OR = 2.2; 95% CI = 1.02, 4.70) as significant predictors of Blastocystis infection among this population. In those who had only Blastocystis infection but no other gastrointestinal parasitic infections, the prevalence of gastrointestinal symptoms was higher compared to those without Blastocystis infection (35.3% vs 13.2%; x² = 25.8; P < 0.001). The most common symptoms among these patients were abdominal pain (76.4%), flatulence (41.1%) and diarrhoea (21.5%).overall prevalence of Blastocystis infection was 22.1%. The prevalence was significantly higher among patients aged ≥18 years compared to those aged < 18 years (29.4% vs 9.9%; x² = 19.746; P < 0.001), and in males compared to females (26.4% vs 17.5%; x² = 4.374; P = 0.036). Univariate analysis showed significant associations between Blastocystis infection and the occupational status (P = 0.017), family size (P = 0.023) and educational level (P = 0.042) of the participants. Multiple logistic regression analysis confirmed that the age of ≥ 18 years (OR = 5.7; 95% CI = 2.21; 9.86) and occupational stat
CONCLUSIONS: Blastocystis sp. is prevalent and associated with gastrointestinal symptoms among communities in Sebha city, Libya. Age and occupational status were the significant predictors of infection. However, more studies from different areas in Libya are needed in order to delineate the epidemiology and clinical significance of this infection.

Matched MeSH terms: Blastocystis Infections/parasitology
Predominance of Blastocystis sp. subtype 4 in rural communities, Nepal

Lee IL, Tan TC, Tan PC, Nanthiney DR, Biraj MK, Surendra KM, et al.

Parasitol Res, 2012 Apr;110(4):1553-62.
PMID: 22076050 DOI: 10.1007/s00436-011-2665-0

Blastocystis sp. is a common intestinal parasite. To date, there have been sporadic and scanty studies on Blastocystis sp. carried out in rural communities in Nepal. We surveyed the prevalence of Blastocystis sp. and its possible associated risk factors, and reported the predominant Blastocystis sp. subtype in two rural communities, Bolde Phediche and Bahunipati, in Nepal. Human faecal samples were collected from 241 participants, cultured using in vitro cultivation and examined for Blastocystis sp. The presence of Blastocystis sp. in faecal samples was further confirmed by polymerase chain reaction (PCR) and subsequently genotyped using subtype-specific sequence tagged site (STS) primers. There were 26.1% (63/241) of the participants that were infected by Blastocystis sp. We detected 84.1% (53/63) of Blastocystis sp. subtype 4 infections in these rural communities. The unusually high prevalence of Blastocystis sp. subtype 4 can be attributed to the rearing of family-owned animals in barns built close to their houses. Eighty one percent (51/63) of the Blastocystis sp. infected participants drank not boiled or unfiltered water. The present study revealed that Blastocystis sp. could pose a health concern to the communities and travellers to the hilly area in Nepal. Infection may be transmitted through human-to-human, zoonotic and waterborne transmissions. We provide recommendations to ensure good public health practices.

Matched MeSH terms: Blastocystis Infections/parasitology*
Increase number of mitochondrion-like organelle in symptomatic Blastocystis subtype 3 due to metronidazole treatment

Raman K, Kumar S, Chye TT

Parasitol Res, 2016 Jan;115(1):391-6.
PMID: 26481491 DOI: 10.1007/s00436-015-4760-0

Blastocystis sp., an intestinal organism is known to cause diarrhea with metronidazole regarded as the first line of treatment despite reports of its resistance. The conflicting reports of variation in drug treatment have been ascribed to subtype differences. The present study evaluated in vitro responses due to metronidazole on ST3 isolated from three symptomatic and asymptomatic patients, respectively. Symptomatic isolates were obtained from clinical patients who showed symptoms such as diarrhea and abdominal bloating. Asymptomatic isolates from a stool survey carried out in a rural area. These patients had no other pathogens other than Blastocystis. Ultrastructural studies using transmission electron microscopy (TEM) and scanning electron microscopy (SEM) revealed drug-treated ST3 from symptomatic patients were irregular and amoebic with surface showing high-convoluted folding when treated with metronidazole. These organisms had higher number of mitochondrion-like organelle (MLO) with prominent cristae. However, the drug-treated ST3 from asymptomatic persons remained spherical in shape. Asymptomatic ST3 showed increase in the size of its central body with the MLO located at the periphery.

Matched MeSH terms: Blastocystis Infections/parasitology*

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