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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/drug therapy; Blastocystis Infections/parasitology*
Subtype distribution of Blastocystis isolated from humans and associated animals in an indigenous community with poor hygiene in Peninsular Malaysia

Mohammad NA, Al-Mekhlafi HM, Anuar TS

Trop Biomed, 2018 Dec 01;35(4):849-860.
PMID: 33601835

Blastocystis is one of the most common parasites inhabiting the intestinal tract of human and animals. Currently, human Blastocystis isolates are classified into nine subtypes (STs) based on the phylogeny of their small subunit ribosomal RNA (SSU rRNA) gene. Although its pathogenicity remains controversial, the possibility of zoonotic transmission was recognized since eight of the nine STs (except for ST9) have been reported in both humans and animals. A cross-sectional study was conducted to determine the prevalence and subtype distribution of Blastocystis isolated from humans and associated animals in an indigenous community with poor hygiene in Malaysia, where the risk of parasitic infection is high. A total of 275 stool samples were collected, subjected to DNA extraction and amplified by PCR assay. The Blastocystis-positive amplicons were then purified and sequenced. Phylogenetic tree of positive isolates, reference strains and outgroup were constructed using maximum likelihood method based on Hasegawa-KishinoYano+G+I model. The prevalence of Blastocystis infection among humans and domestic animals by PCR assay were 18.5% (45/243) and 6.3% (2/32), respectively. Through molecular phylogeny, 47 isolates were separated into five clusters containing isolates from both hosts. Among human isolates, ST3 (53.3%) was the predominant subtype, followed by ST1 (31.1%) and ST2 (15.6%). Chicken and cattle had lower proportions of ST6 (50%) and ST10 (50%), that were barely seen in humans. The distinct distributions of the most important STs among the host animals as well as humans examined demonstrate that there is various host-specific subtypes in the lifecycle of Blastocystis.

Matched MeSH terms: Blastocystis Infections
A case report of Blastocystis infection and Steven Johnson's syndrome

Singh A, Priyadarshi K, Raj T, Banerjee T

Trop Biomed, 2019 Dec 01;36(4):987-992.
PMID: 33597468

Blastocystis species (spp.) is an emerging pathogen. There are several unsolved issues linked to this parasite ranging from its nomenclature, commensal status, standardization of laboratory diagnostic methods, genotypes and treatment. Recently, there has been an increase in reports of Blastocystis spp. from symptomatic cases which provide enough evidence of its pathogenic potential. A range of signs and symptoms, from gastro-intestinal to cutaneous manifestations have been attributed to Blastocystis infection. Few reports have established an association between intestinal infection with Blastocystis spp. and skin manifestations in form of urticaria, palmoplantar pruritus and allergy with complete resolution of cutaneous lesions with eradication of the parasite. In this report, we describe a case of Steven Johnson's syndrome (SJS) in a 6 years old girl along with infection with Blastocystis spp. marked by diarrhea and abdominal pain. Stool examination revealed the presence of all forms of the parasite with subsequent decrease in parasite burden and diarrhea over a period of time. Interestingly, the clearance of Blastocystis spp. from stool was followed by recovery from skin lesions and other symptoms. In this case, the course of SJS was clearly associated with Blastocystis infection. Though skin manifestation with Blastocystis infection has been previously reported, this is the first report of its association with SJS. This report indicates newer insights of the parasite that are less well studied.

Matched MeSH terms: Blastocystis Infections/diagnosis*
Epitope variances demonstrated by Blastocystis sp. ST3 symptomatic and asymptomatic isolates

Sheela DS, Chandramathi S, Suresh K

Trop Biomed, 2020 Mar 01;37(1):210-217.
PMID: 33612732

Blastocystis sp. is an enteric protozoan parasite of humans and many animals. Blastocystis sp. subtype 3 (ST3) proves to be the highest frequency case in most populations around the world and it is further distinguished into symptomatic and asymptomatic isolates based on the clinical symptoms exhibited by infected individuals. Phenotypic and genotypic studies implicate the distinctiveness of this parasite which may describe its pathogenesis. However, the antigenic distinctiveness which describes the antibody mediated cell lysis of this parasite has not been explored. This study was aimed to identify the cross-reactivity and cytotoxicity effect between three isolates of symptomatic and asymptomatic Blastocystis sp. ST3 respectively. Antigen specificity and diversity of this parasite was performed by coculturing sera (10-fold dilution) obtained from mice immunised with Blastocystis sp. symptomatic and asymptomatic antigens and the respective Blastocystis sp. ST3 live cells through complement dependant cell cytotoxicity (CDC) assay. The results obtained has shown that, the sera (at 10-fold diluted concentration) from symptomatic and asymptomatic solubilised antigen immunised mice were able to specifically lyse the respective live parasites with an average percentage of 82% and 86% respectively. There were almost 50% crossreactivity observed between the three isolates of Blastocystis sp. ST3 from symptomatic and asymptomatic group proving high antigen diversity or rather low antigen specificity within the same group. However, there was only 17% cross-reactivity observed between the mice sera and parasitic cells of different groups (symptomatic vs asymptomatic isolates) suggesting high specificity between these two groups. We, for the first time have proven that through CDC analysis there were epitopes dissimilarities between Blastocystis sp. ST3 symptomatic and asymptomatic isolates which may allow the parasite to set up diverse immune modulations such as imbalanced Th1/Th2 responses in an infected host.

Matched MeSH terms: Blastocystis Infections/immunology; Blastocystis Infections/parasitology*
Prevalence and subtype distribution of Blastocystis sp. in cattle from Pahang, Malaysia

Kamaruddin SK, Mat Yusof A, Mohammad M

Trop Biomed, 2020 Mar 01;37(1):127-141.
PMID: 33612724

Blastocystis sp. is a common enteric protozoan parasite found in humans and various type of animal worldwide. Recently, genotypic distribution of Blastocystis sp. was revealed in insects, rodents, avian and mammals, which exposed its potential of transmiting the infections to human. However, very little information on current level of Blastocystis sp. infection were reported in cattle from Malaysia. Herein, a total of 120 stool samples of cattles were collected. While the potential risk of infection such as age, gender, body score, diarrheic condition of the cattle were noted, the management of the farms was also recorded. All stool sample were cultured, but 80 samples were selected for PCR sequencing analysis. The cultivation and microscopic examination revealed only 25% of the cattle (30/120) were infected with Blastocystis sp.. But, 43.8% of the cattle (35/80) were found positive upon PCR sequencing. The study also found that age, body score condition, diarrheic condition and certain farm were associated with the infection (p<0.05). Six subtypes (STs) that were discovered during the study were ST10 (21.3%;17/35), ST5 (8.8%;7/35), ST3 (7.5%;6/35), ST1 (2.5%;2/35), ST4 (2.5%;2/35) and ST14 (1.3%;1/35). Thus, moderate infections of Blastocystis sp. and variants in the genotypic distributions of the cattle suggest its potential for zoonotic transmission. Therefore, this findings could be helpful for further understanding the parasite, which assist studies of its pathogenicity.

Matched MeSH terms: Blastocystis Infections/epidemiology; Blastocystis Infections/parasitology; Blastocystis Infections/veterinary*
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/drug therapy; Blastocystis Infections/parasitology*
Blastocystis hominis in high-rise flat dwellers in Kuala Lumpur, Malaysia

Suresh K, Salim HR, Jamaiah I, Anuar AK

Trans R Soc Trop Med Hyg, 2001 10 3;95(4):377-8.
PMID: 11579877
Matched MeSH terms: Blastocystis Infections/epidemiology*
Restriction enzyme digestion analysis of PCR-amplified DNA of Blastocystis hominis isolates

Init I, Foead AL, Fong MY, Yamazaki H, Rohela M, Yong HS, et al.

Southeast Asian J. Trop. Med. Public Health, 2007 Nov;38(6):991-7.
PMID: 18613539

Genomic DNA of Blastocystis isolates released into 0.1% Triton X-100 was suitable for amplification and yielded similar results as the genomic DNA extracted with standard kit. The specific B. hominis primers (BH1: GCT TAT CTG GTT GAT CCT GCC AGT and BH2: TGA TCC TTC CGC AGG TTC ACC TAC A) successfully produced the PCR product of about 1,770 bp with all the 7 Blastocystis isolates tested. The restriction fragment length polymorphism (RFLP) patterns yielded by 13 out of 25 restriction endonucleases showed that the 7 isolates could be grouped into 4 subgroups: subgroup-1 consisted of isolate C; subgroup-2 of isolates H4 and H7; subgroup-3 of isolates KP1, Y51 and M12; and subgroup-4 of isolate 27805. The differences between subgroups manifested as clear-cut RFLP patterns. A common band of 230 bp was revealed by Eco R1 in all the Blastocystis isolates tested. The band of about 180 bp was revealed by Alu I, differentiated symptomatic from asymptomatic isolates of this parasite, and might indicate the pathogenicity of this parasite.

Matched MeSH terms: Blastocystis Infections/parasitology
Fulltext Prevalence of Blastocystis in animals from domesticated surroundings

Chuong LS, Suresh K, Mak JW, Init I, Kathijah O

Southeast Asian J. Trop. Med. Public Health, 1996 Dec;27(4):850-2.
PMID: 9253897
Matched MeSH terms: Blastocystis Infections/epidemiology*; Blastocystis Infections/transmission
Fulltext Blastocystis hominis infection, a cause of human diarrhea

Sinniah B, Rajeswari B

Southeast Asian J. Trop. Med. Public Health, 1994 Sep;25(3):490-3.
PMID: 7777913

Blastocystis hominis has long been described as a non pathogenic protozoan parasite until recently when claims have been made that it can result in pathogenic conditions. Of the 729 stool samples (614 from survey and 115 from pediatric wards) examined, 18.1% of them were found to be positive for one or more intestinal protozoan cyst. The commonest was Giardia intestinalis (8.4%) Followed by Entamoeba coli (7.1%) and Entamoeba histolytica (5.1%) in the normal children without symptoms of diarrhea. When diarrheic stools were examined, the commonest parasite encountered was Giardia (20.4%), followed by E. coli (15.9%) and E. histolytica (9.7%). Blastocystis was observed in 4.4% of the children who had diarrhea and 1.1% among the children taken from the normal population in the rural areas.

Matched MeSH terms: Blastocystis Infections/epidemiology*
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/drug therapy; 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 Stress exacerbates infectivity and pathogenicity of Blastocystis hominis: in vitro and in vivo evidences

Chandramathi S, Suresh K, Sivanandam S, Kuppusamy UR

PLoS One, 2014;9(5):e94567.
PMID: 24788756 DOI: 10.1371/journal.pone.0094567

Stress alters the oxidant-antioxidant state and immune cell responses which disrupts its function to combat infection. Blastocystis hominis, a common intestinal protozoan has been reported to be opportunistic in immunocompromised patients namely cancer. B. hominis infectivity in other altered immune system conditions especially stress is unknown. We aimed to demonstrate the stress effects towards the susceptibility and pathogenicity of B. hominis infection.

Matched MeSH terms: Blastocystis Infections/blood; Blastocystis Infections/immunology; Blastocystis Infections/physiopathology; Blastocystis Infections/psychology
Fulltext Subtype distribution of Blastocystis isolates in Sebha, Libya

Abdulsalam AM, Ithoi I, Al-Mekhlafi HM, Al-Mekhlafi AM, Ahmed A, Surin J

PLoS One, 2013;8(12):e84372.
PMID: 24376805 DOI: 10.1371/journal.pone.0084372

BACKGROUND: Blastocystis is a genetically diverse and a common intestinal parasite of humans with a controversial pathogenic potential. This study was carried out to identify the Blastocystis subtypes and their association with demographic and socioeconomic factors among outpatients living in Sebha city, Libya.
METHODS/FINDINGS: Blastocystis in stool samples were cultured followed by isolation, PCR amplification of a partial SSU rDNA gene, cloning, and sequencing. The DNA sequences of isolated clones showed 98.3% to 100% identity with the reference Blastocystis isolates from the Genbank. Multiple sequence alignment showed polymorphism from one to seven base substitution and/or insertion/deletion in several groups of non-identical nucleotides clones. Phylogenetic analysis revealed three assemblage subtypes (ST) with ST1 as the most prevalent (51.1%) followed by ST2 (24.4%), ST3 (17.8%) and mixed infections of two concurrent subtypes (6.7%).
BLASTOCYSTIS: ST1 infection was significantly associated with female (P = 0.009) and low educational level (P = 0.034). ST2 was also significantly associated with low educational level (P= 0.008) and ST3 with diarrhoea (P = 0.008).
CONCLUSION: Phylogenetic analysis of Libyan Blastocystis isolates identified three different subtypes; with ST1 being the predominant subtype and its infection was significantly associated with female gender and low educational level. More extensive studies are needed in order to relate each Blastocystis subtype with clinical symptoms and potential transmission sources in this community.

Matched MeSH terms: Blastocystis Infections/epidemiology*
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/epidemiology*; Blastocystis Infections/parasitology; Blastocystis Infections/prevention & control
Fulltext Resistance towards metronidazole in Blastocystis sp.: A pathogenic consequence

Rajamanikam A, Hooi HS, Kudva M, Samudi C, Kumar S

PLoS One, 2019;14(2):e0212542.
PMID: 30794628 DOI: 10.1371/journal.pone.0212542

Blastocsytis sp. is a protozoan parasite that has been linked to common gastrointestinal illnesses. Metronidazole, the first line therapy, was reported to show frequent inefficacy. Previously, Blastocystis sp. isolated from different population showed varying metronidazole resistance. However, the effect of metronidazole treatment on pathogenic potentials of Blastocystis sp. isolated from different populations, which is known to have different gut environment, is unclear. This study investigates the in vitro effect of metronidazole on the pathogenic potentials of Blastocystis sp. isolated from urban and orang asli individuals. Blastocystis sp. ST 3 isolated from symptomatic and asymptomatic individuals were treated with a range of metronidazole concentration. The parasites' growth characteristics, apoptotic rate, specific protease activity and the ability to proliferate cancer cells were analyzed upon treatment with 0.001 mg/l metronidazole. The study demonstrates that Blastocystis sp. isolates showed increase in the parasite numbers especially the amoebic forms (only in urban isolates) after treating with metronidazole at the concentration of 0.001 mg/ml. High number of cells in post-treated isolates coincided with increase of apoptosis. There was a significant increase in cysteine protease of Blastocystis sp. isolates upon treatment despite the initial predominance of serine protease in asymptomatic isolates. Metronidazole resistant Blastocystis sp. also showed significant increase in cancer cell proliferation. Resistance to metronidazole did not show significant different influence on the pathogenicity between Blastocystis sp. isolated from urban and orang asli individual. However, an increase in parasite numbers, higher amoebic forms, cysteine protease and ability to proliferate cancer cells implicates a pathogenic role. The study provides evidence for the first time, the effect of metronidazole towards enhancing pathogenic potentials in Blastocystis sp. when isolated from different gut environment. This necessitates the need for reassessment of metronidazole treatment modalities.

Matched MeSH terms: Blastocystis Infections/drug therapy*; Blastocystis Infections/pathology
Fulltext Effect of Blastocystis sp. in dengue patients-Increase in the treatment cost and exacerbation of symptoms

Thergarajan G, Kumar S, Bhassu S, Omar SFBS, Rampal S

PLoS One, 2019;14(3):e0211034.
PMID: 30893309 DOI: 10.1371/journal.pone.0211034

Increasing incidences of dengue have become a global health threat with major clinical manifestation including high fever and gastrointestinal symptoms. These symptoms were also expressed among Blastocystis sp. infected individuals, a parasite commonly seen in human stools. This parasite has been previously reported to replicate faster upon exposure to high temperature. The present study is a hospitalized-based cross-sectional study involved the collection of faecal sample from dengue patients. Stool examination was done by in vitro cultivation to isolate Blastocystis sp. Growth pattern of all the positive isolates were analyzed to identify the multiplication rate of Blastocystis sp. isolated from dengue patients. Distribution of Blastocystis sp. among dengue patients was 23.6%. Dengue patients who were positive for Blastocystis sp. infection denoted a significantly higher fever rate reaching 38.73°C (p<0.05) compared to the non-Blastocystis sp. infected patients (38.44°C). It was also found that Blastocystis sp. infected patients complained of frequenting the toilet more than five times a day (p<0.05) compared to those who were non-Blastocystis sp. infected. At the same time, the duration of hospitalization was significantly longer (p<0.05) for Blastocystis sp. infected dengue patients compared to the non-Blastocystis sp. infected patients. Besides, Blastocystis sp. isolated from dengue patients (in vivo thermal stress) showed a higher growth rate compared to the non-dengue isolated which was exposed to high temperature (in vitro thermal stress). Our findings suggest that presence of Blastocystis sp. during dengue infection could trigger the increase of temperature which could be due to highly elevated pro inflammatory cytokines by both parasitic and virus infection. This could justify why the temperature in Blastocystis sp. infected dengue patients is higher compared to the non-Blastocystis sp. infected patients. Higher temperature could have triggered a greater parasite multiplication rate that contributed to the aggravation of the gastrointestinal symptoms.

Matched MeSH terms: Blastocystis Infections/metabolism*; Blastocystis Infections/parasitology
Occurrence of Blastocystis sp. in water catchments at Malay villages and Aboriginal settlement during wet and dry seasons in Peninsular Malaysia

Noradilah SA, Lee IL, Anuar TS, Salleh FM, Abdul Manap SN, Mohd Mohtar NS, et al.

PeerJ, 2016;4:e2541.
PMID: 27761331

In the tropics, there are too few studies on isolation of Blastocystis sp. subtypes from water sources; in addition, there is also an absence of reported studies on the occurrence of Blastocystis sp. subtypes in water during different seasons. Therefore, this study was aimed to determine the occurrence of Blastocystis sp. subtypes in river water and other water sources that drained aboriginal vicinity of highly endemic intestinal parasitic infections during wet and dry seasons. Water samples were collected from six sampling points of Sungai Krau (K1-K6) and a point at Sungai Lompat (K7) and other water sources around the aboriginal villages. The water samples were collected during both seasons, wet and dry seasons. Filtration of the water samples were carried out using a flatbed membrane filtration system. The extracted DNA from concentrated water sediment was subjected to single round polymerase chain reaction and positive PCR products were subjected to sequencing. All samples were also subjected to filtration and cultured on membrane lactose glucuronide agar for the detection of faecal coliforms. During wet season, Blastocystis sp. ST1, ST2 and ST3 were detected in river water samples. Blastocystis sp. ST3 occurrence was sustained in the river water samples during dry season. However Blastocystis sp. ST1 and ST2 were absent during dry season. Water samples collected from various water sources showed contaminations of Blastocystis sp. ST1, ST2, ST3 and ST4, during wet season and Blastocystis sp. ST1, ST3, ST8 and ST10 during dry season. Water collected from all river sampling points during both seasons showed growth of Escherichia coli and Enterobacter aerogenes, indicating faecal contamination. In this study, Blastocystis sp. ST3 is suggested as the most robust and resistant subtype able to survive in any adverse environmental condition. Restriction and control of human and animal faecal contaminations to the river and other water sources shall prevent the transmission of Blastocystis sp. to humans and animals in this aboriginal community.

Matched MeSH terms: Blastocystis Infections
Modified fields' stain: ideal to differentiate Dientamoeba fragilis and Blastocystis sp

Ragavan AD, Govind SK

Parasitol Res, 2015 Mar;114(3):1163-6.
PMID: 25614298 DOI: 10.1007/s00436-014-4296-8

Dientamoeba fragilis, a trichomonad parasite is usually found in the gastrointestinal tract of human, and it is known to be the cause for gastrointestinal disease. The parasite is globally distributed and mostly found in rural and urban areas. The parasite is found in humans and nonhuman primates such as the macaques, baboons, and gorillas. Often, the parasite is confused with another largely found organism in stools called Blastocystis sp. especially when seen directly under light microscopy on culture samples containing both parasites. Both sometimes are seen with two nuclei with sizes tending to be similar which complicates identification. Stools were collected fresh from nine previously diagnosed persons infected with D. fragilis who also were found to be positive for Blastocystis sp. Samples were then cultured in Loeffler's medium and were stained with Giemsa, iron hematoxylin, and modified Fields' (MF) stain, respectively. D. fragilis was differentiated from Blastocystis sp. when stained with MF stain by the presence of a thinner outer membrane with clearly demarcated nuclei in the center of the cell whilst Blastocystis sp. had a darker and thicker stained outer membrane with the presence of two nuclei. The staining contrast was more evident with modified Fields' stain when compared with the other two. The simplicity in preparing the stain as well as the speed of the staining procedure make MF stain an ideal alternate. The modified Fields' stain is faster and easier to prepare when compared to the other two stains. MF stain provides a better contrast differentiating the two organisms and therefore provides a more reliable diagnostic method to precisely identify one from the other especially when cultures show mixed infections.

Matched MeSH terms: Blastocystis Infections/diagnosis*; Blastocystis Infections/parasitology
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/epidemiology; Blastocystis Infections/parasitology; Blastocystis Infections/veterinary*

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