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Wintergreen oil: a novel method in Wheatley's trichrome staining technique

Salleh FM, Anuar TS, Yasin AM, Moktar N

J Microbiol Methods, 2012 Oct;91(1):174-8.
PMID: 22986100 DOI: 10.1016/j.mimet.2012.08.004

Permanent staining of faecal smears by Wheatley's trichrome technique has been used by many scientists for the detection of parasites in the past and it was found to be highly sensitive. This study was conducted to evaluate the use of Wintergreen oil in comparison with xylene in Wheatley's trichrome staining technique, as the reference technique. In a blind comparison study, 500 collected faecal samples from aboriginal communities were examined. Wintergreen oil was found to be more superior than xylene as a clearing agent in the Wheatley's trichrome staining of polyvinyl alcohol-fixed faecal smears for the identification of intestinal protozoa. Elimination of toxic, carcinogenic, and fire hazards makes Wintergreen oil the preferred choice in routine parasitology examinations.

Matched MeSH terms: Feces/parasitology*
Urinary hyaluronidase activity in rats infected with Blastocystis hominis--evidence for invasion?

Chandramathi S, Suresh KG, Mahmood AA, Kuppusamy UR

Parasitol Res, 2010 May;106(6):1459-63.
PMID: 20358228 DOI: 10.1007/s00436-010-1825-y

The fact whether Blastocystis hominis can invade has always been in question. Apart from a few sporadic studies such as that done on gnotobiotic guinea pigs which showed surface invasion and mucosal inflammation of the host's intestine caused by B. hominis infection, no real documentation of invasion has been proven. Studies have shown that hyaluronidase is secreted during the penetration into the host's skin and gut by nematode parasites. Hyaluronidase activity in protozoa namely Entamoeba histolytica has also been described previously. This study attempts to determine hyaluronidase in urine samples of B. hominis-infected rats. The presence of hyaluronidase in urine provides an indirect evidence of invasion by B. hominis into colonic epithelium causing the degradation of extracellular matrix proteins namely hyaluronic acid (HA). HA is depolymerized by hyaluronidase which may be used by organisms to invade one another. In this study, the levels of urinary hyaluronidase of Sprague-Dawley rats infected with B. hominis were monitored for 30 days. Hyaluronidase levels in the infected rats were significantly higher on days 28 and 30 compared to the day before inoculation (P < 0.01 and P < 0.05, respectively). During this stage, parasitic burden in infected stools was also at a high level. Proinflammatory cytokines, interleukin-6 and interleukin-8, were also significantly higher (P < 0.05) in the serum of infected rats. The study demonstrates that since no other pathogen was present and that amoeboid forms of the parasites have been shown to exist previously, the elevated levels of hyaluronidase in this preliminary finding suggests that the organism is capable of having invasion or penetration activity in the hosts' intestine.

Matched MeSH terms: Feces/parasitology
Fulltext Unusual faecal elements in a human diarrhoeal stool

Sinniah B, Sheikh Omar AR, Lee CC, Surin J, Subramaniam K

Med J Malaysia, 1994 Dec;49(4):419-23.
PMID: 7674981

A 16-year-old female from Rantau Panjang, Kelantan reported having diarrhoea for three months. During this period, she lost 15 lb in weight and was treated with antibiotics and anti-spasmodic tablets with no improvement. Stool examinations by private laboratories revealed "worm-like eggs". She was treated for worms with mebendazole which helped to reduce the symptoms but not completely. The patient continued passing out the abnormal "worm-like eggs" which were later identified as pollen grains.

Matched MeSH terms: Feces/parasitology*
Understanding Giardia infections among rural communities using the one health approach

Lee SC, Ngui R, Tan TK, Roslan MA, Ithoi I, Mahdy MAK, et al.

Acta Trop, 2017 Dec;176:349-354.
PMID: 28859958 DOI: 10.1016/j.actatropica.2017.08.030

The epidemiology of giardiasis in rural villages in Peninsular Malaysia was examined in the context of the One Health triad that encompasses humans, animals and environment (i.e. river water). A cross-sectional study was carried out among five rural communities in Malaysia to determine the prevalence of Giardia duodenalis in humans, animals and river water. Fecal samples collected from humans and animals were examined by light microscopy. Water was sampled from the rivers adjacent to the target communities and investigated for the occurrence of Giardia cysts. The isolated cysts were further genotyped targeting the glutamate dehydrogenase and triosephosphate isomerase genes. The overall prevalence of G. duodenalis was 6.7% (18/269) and 4.7% (8/169) among humans and animals, respectively. Giardia cysts (mean concentration range: 0.10-5.97 cysts/L) were also found in adjacent rivers at four out of the five villages examined. At Kemensah and Kuala Pangsun, Giardia cysts were isolated from humans [rate: 3.7% each (of 54 each)], animals [rates: 6.3% (of 62) and 11.3% (of 16), respectively] and river water [average concentration of 9 samples each: 0.83±0.81 and 5.97±7.00, respectively]. For both villages at Pos Piah and Paya Lebar, 12.2% (of 98) and 6.1% (of 33) of collected human samples were infected, respectively whilst none of the collected animals samples in these villages were found to be positive. The river water samples of these two villages were also contaminated (average concentration: 0.20±0.35 (of 9) and 0.10±0.19 (of 3), respectively). In conclusion, Giardia cysts were simultaneously observed in the human-animal-environment (i.e., river water) interfaces in at least two of five studied communities highlighting a vital need to improve understanding on the interplay of transmission dynamics, the role of infected humans and animals in contaminating the water sources and the role of water as a vehicle of disease transmission in these communities. Indeed, this study illustrates the One Health approach which is to recognize that the optimal health of humans are interconnected with the well-being of animals and their environment.

Matched MeSH terms: Feces/parasitology*
Two new coccidian parasites from the grand anglehead lizard, Gonocephalus grandis from Peninsular Malaysia

Maupin RS, Diong CH, McQuistion TE

J Parasitol, 1998 Dec;84(6):1210-2.
PMID: 9920316

During 3 collecting expeditions between October 1996 and December 1996, fecal samples were obtained from 43 adult Gonocephalus grandis from Tanah Rata and the Cameron Highlands in Peninsular Malaysia. Two species of coccidia (Isospora gonocephali n. sp. [9/43, 23%] and Eimeria cameronensis n. sp. [3/43, 7%]) were discovered. Sporulated oocysts of I. gonocephali are subspherical to ovoidal, 22.3 x 18.7 (19-25 x 17-23) microm with a bilayered wall composed of a thin inner wall and a striated outer wall with a pitted surface; oocyst residuum absent; 1 polar granule present; sporocysts are almond-shaped, 13.5 x 9.2 (12-15 x 8.5-10) microm, Stieda body broad, domelike, substieda body fanlike, sporocyst residuum consisting of coarse, nonuniform granules in an amorphous cluster; sporozoites sausage-shaped with 1 large terminal, refractile body and lay randomly in the sporocyst. Sporulated oocysts of E. cameronensis are bilayered, smooth-walled, ellipsoidal, 26.5 x 12.4 (25-28 x 12-13) microm; with 1, small, polar granule composed of 2-3 splinter-like structures fused together; oocyst residuum absent; sporocysts ovoidal, almost rectangular-shaped 8.8 x 6.6 (8-9 x 5-7) microm, with no Stieda or substieda bodies, containing scattered residuum and 2 sausage-shaped sporozoites with 1 terminal, ovoidal refractile body. No individual lizard was host to both coccidian species.

Matched MeSH terms: Feces/parasitology
Trematode eggs recovered from patient's faeces in Sabah

Kan SK, Hii JL, Razack KV

Southeast Asian J. Trop. Med. Public Health, 1978 Mar;9(1):113-4.
PMID: 568314
Matched MeSH terms: Feces/parasitology*
Total anthelmintic failure to control nematode parasites of small ruminants on government breeding farms in Sabah, East Malaysia

Chandrawathani P, Yusoff N, Wan LC, Ham A, Waller PJ

Vet Res Commun, 2004 Aug;28(6):479-89.
PMID: 15509022

Government-owned small-ruminant breeding farms in Malaysia provide the source of sheep and goats to smallholder farmers in the country. In the eastern Malaysian state of Sabah, high-level stock losses have been recorded on these farms for several years, frequently accompanied by clinical signs indicating pathogenic levels of infections with the nematode parasite Haemonchus contortus. This suggests that their dependence on chemotherapy to control parasite infections had failed. Accordingly, tests for anthelmintic efficacy using the faecal egg count reduction test (FECRT) on the range of drugs used to control nematode parasites were carried out on the five government small-ruminant breeding farms in Sabah. These tests showed a total failure of the benzimidazole, imidothiazole, macrocyclic lactone and salicylanilide groups of anthelmintics to control H. contortus infections of sheep and goats on all farms. Drastic changes in animal management need to be made in an attempt to deal with this situation, for which suggestions are made.

Matched MeSH terms: Feces/parasitology
The use of FAMACHA in estimation of gastrointestinal nematodes and total worm burden in Damara and Barbados Blackbelly cross sheep

Mohammed K, Abba Y, Ramli NS, Marimuthu M, Omar MA, Abdullah FF, et al.

Trop Anim Health Prod, 2016 Jun;48(5):1013-20.
PMID: 27038194 DOI: 10.1007/s11250-016-1049-y

The prevalence of gastrointestinal (GI) nematodes and total worm burden of Damara and Barbados Blackbelly cross sheep was investigated among smallholder farms in Salak Tinggi district of Selangor, Malaysia. A total of 50 sheep raised in smallholder farms comprising of 27 Damara cross and 23 Barbados Blackbelly cross were categorized based on their age into young and adults. Fecal samples were collected and examined for strongyle egg count by using modified McMaster technique. Severity of infection was categorized into mild, moderate, and heavy, based on egg per gram (EPG). Five sheep were randomly selected and slaughtered to examine the presence of adult gastrointestinal (GI) nematodes through total worm count (TWC). Faffa Malan Chart (FAMACHA) score was used for investigation of worm load based on the degree of anemia. The study revealed an overall EPG prevalence of 88 %, of which 84.1 % had mild infection. There was a significant difference (p = 0.002) in EPG among the two breeds. Based on age, significant difference (p = 0. 004) in EPG was observed among Barbados Blackbelly cross, but not for Damara cross (p = 0.941). The correlation between severity of infection and the FAMACHA score was significant (r = 0.289; p = 0.042). Haemonchus spp. were the most predominant nematode found in the gastrointestinal tract, followed by Trichostrongylus and Oesophagostomum spps. EPG and TWC for Haemonchus were positively correlated, but not significant (r = 0.85, p = 0.066). From regression analysis, 73 % of the variability in TWC for Haemonchus could be explained by EPG. Thus, it can be concluded that FAMACHA score correlates well with severity of infection of a nematode and can be used to assess the strongyle nematode burden in the different sheep crosses.

Matched MeSH terms: Feces/parasitology
The role and influence of bacteria on the growth of Blastocystis sp. subtype 3

Kaur CP, Yong CC, Rajamanikam A, Samudi C, Kumar S, Bhassu S, et al.

Parasitol Res, 2023 Jul;122(7):1463-1474.
PMID: 37162590 DOI: 10.1007/s00436-023-07842-2

Blastocystis sp. is an enteric protistan parasite that affects individuals worldwide with gastrointestinal symptoms such as abdominal discomfort, diarrhea, and flatulence. However, its pathogenicity is controversial due to its presence among asymptomatic individuals. Blastocystis sp. subtype 3 (ST3) is the most prevalent subtype among humans that have been associated with irritable bowel syndrome (IBS), Crohn's disease, ulcerative colitis, and colorectal cancer. Axenization of the parasite has been shown to impede its growth thus revealing the importance of accompanying bacteria in ensuring Blastocystis sp. survival. This study aims to identify the influence of accompanying bacteria on the growth of Blastocystis sp. ST3. Blastocystis sp. cultures were treated with Meropenem, Vancomycin, and Amoxicillin-Clavulanic acid (Augmentin). Bacteria-containing supernatant of antibiotic-treated and control cultures were isolated and identified through 16 s rRNA sequencing. Morphological changes of antibiotic-treated Blastocystis sp. ST3 were also observed. The cultures treated with meropenem and augmentin exhibited opposing effects with reduced growth of isolates from symptomatic patients and a significant increase in asymptomatic isolates. Whereas, vancomycin-treated cultures had no difference in the growth of Blastocystis sp. ST3 isolates from symptomatic and asymptomatic patients. Isolates from symptomatic and asymtomatic patients had 6 and 2 distinct bacterial species identified with Proteus mirabilis as the common bacteria among both types of isolates. Morphologically, Blastocystis sp. ST3 cultures exposed to meropenem and augmentin demonstrated an increase in pre-cystic forms. These findings demonstrate the effects of accompanying bacteria on the growth of Blastocystis sp. ST3 that could translate into clinical manifestations observed among Blastocystis sp.-infected patients.

Matched MeSH terms: Feces/parasitology
The problem of soil transmitted helminths in squatter areas around Kualalumpur

Yan CW, Ishak F, Hee GL, Devaraj JM, Ismail K, Jalleh RP, et al.

Med J Malaysia, 1978 Sep;33(1):34-43.
PMID: 571513
Matched MeSH terms: Feces/parasitology
Fulltext The prevalence and intensity of soil-transmitted helminths in some rural villages in northern peninsular Malaysia

Rahman WA

Southeast Asian J. Trop. Med. Public Health, 1994 Jun;25(2):296-9.
PMID: 7855643

The prevalence and intensity of soil-transmitted helminths in five rural villages in northern peninsular were investigated. Generally the prevalence and intensity of infection were low in the 0-10- and above 50-year-old age groups when compared with other age groups. The prevalence and intensity of infection in the five villages were quite similar, because of similar socio-economic status. The highest intensity was observed for Ascaris lumbricoides, followed by hookworms and Trichuris trichiura.

Matched MeSH terms: Feces/parasitology
The isolation, characterization and antifungal susceptibilities of Cryptococcus neoformans from bird excreta in Klang Valley, Malaysia

Tay ST, Chai HC, Na SL, Hamimah H, Rohani MY, Soo-Hoo TS

Mycopathologia, 2005 Jun;159(4):509-13.
PMID: 15983736

The occurrence of Cryptococcus neoformans in bird excreta in Klang valley, Malaysia was determined in this study. Of 544 samples of bird excreta collected from a local zoo, pet shops and public areas, 20 strains of C. neoformans were isolated. All C. neoformans strains were serotype A and thus identified as C. neoformans variety grubii. All did not produce color changes on canavanine-glycine-bromothymol blue agar. All were of alpha-mating types, as determined by a pheromone-specific PCR assay. The antifungal susceptibility testing using agar diffusion method Neo-sensitabs showed that all were susceptible to amphotericin B, fluconazole and itraconazole.

Matched MeSH terms: Feces/parasitology
The first report in Hymenolepis diminuta infection in Sabah, Malaysia

Kan SK, Kok RT, Marto S, Thomas I, Teo WW

Trans R Soc Trop Med Hyg, 1981;75(4):609.
PMID: 7324140
Matched MeSH terms: Feces/parasitology
The effect of repeated chemotherapy on age-related predisposition to Ascaris lumbricoides and Trichuris trichiura

Chan L, Kan SP, Bundy DA

Parasitology, 1992 Apr;104 ( Pt 2):371-7.
PMID: 1594301

This study examines the persistence of predisposition to Ascaris lumbricoides and Trichuris trichiura during repeated chemotherapy in an urban community in Kuala Lumpur, Malaysia. Significant predisposition was observed over 2 periods of reinfection with and without age-standardization of data. Analysis of different age groups indicated that predisposition was most strongly detectable in the younger age classes. The intensities of infection with both parasites were strongly correlated at each cycle of intervention, suggesting that individuals were similarly predisposed to both species.

Matched MeSH terms: Feces/parasitology
The control of the free-living stages of Strongyloides papillosus by the nematophagous fungus, Arthrobotrys oligospora

Chandrawathani P, Omar J, Waller PJ

Vet Parasitol, 1998 Apr 30;76(4):321-5.
PMID: 9650868

Two laboratory trials were conducted to determine the effect of the addition of spores (conidia) of the nematophagous fungus, Arthrobotrys oligospora, on the development of the ruminant parasite, Strongyloides papillosus, in cultures of bovine faeces. Both studies showed that at a concentration of 2000 conidia/g faeces virtually eliminated infective larvae (> 99% reduction), following 14 days incubation under ideal conditions (25 degrees C and saturated humidity) for free-living development of this parasite species. In one trial, a high level of control was also observed at a 10-fold decrease in conidia concentration (200 spores/g faeces). This work has demonstrated, in principle, that A. oligospora could provide a practical biological control agent against S. papillosus infecting intensively raised young ruminants in the humid tropics/subtropics.

Matched MeSH terms: Feces/parasitology
Fulltext Symptomatic chronic strongyloidiasis in children following treatment for solid organ malignancies: case reports and literature review

Norsarwany M, Abdelrahman Z, Rahmah N, Ariffin N, Norsyahida A, Madihah B, et al.

Trop Biomed, 2012 Sep;29(3):479-88.
PMID: 23018511

Strongyloidiasis is an infection caused by the intestinal nematode Strongyloides stercoralis. Infected healthy individuals are usually asymptomatic, however it is potentially fatal in immunocompromised hosts due to its capacity to cause an overwhelming hyperinfection. Strongyloidiasis could be missed during routine screening because of low and intermittent larval output in stool and variable manifestations of the symptoms. We present two cases of strongyloidiasis occurring in children with solid organ malignancies suspected to have the infection based on their clinical conditions and treatment history for cancer. Both patients were diagnosed by molecular and serological tests and were successfully treated. Thus, strongyloidiasis in patients undergoing intensive treatment for malignancies should be suspected, properly investigated and treated accordingly.

Matched MeSH terms: Feces/parasitology
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: Feces/parasitology
Studies of Angiostrongylus malaysiensis (Nematoda, Metastrongylidae) in peninsular Malaysia: natural infection in freshwater snails and rodents in ricefields and infectivity experiments

Liat LB, Fong YL, Krishnansamy M

Southeast Asian J. Trop. Med. Public Health, 1977 Mar;8(1):27-35.
PMID: 887993
Matched MeSH terms: Feces/parasitology
Fulltext Strongyloides stercoralis induced bilateral blood stained pleural effusion in patient with recurrent Non-Hodgkin lymphoma

Win TT, Sitiasma H, Zeehaida M

Trop Biomed, 2011 Apr;28(1):64-7.
PMID: 21602770

Infections and mTalignancies are common causes of pleural effusion. Among infectious causes, hyperinfection syndrome of Strongyloides stercoralis may occur in immunosuppressive patient. A 62-year-old man, known case of Non-Hodgkin lymphoma (NHL) was presented with recurrent NHL stage IV and had undergone salvage chemotherapy. Patient subsequently developed pneumonia with bilateral pleural effusion and ascites. We reported rhabditiform larvae of S. stercoralis in pleural fluid of both lungs without infiltration by lymphoma cells. Stool for microscopic examination also revealed rhabditiform larvae of S. stercoralis. This patient was a known case of NHL receiving chemotherapy resulting in immunosuppression state. Although S. stercoralis infection is not very common compared to other parasitic infections, it is common in immunosuppressive patients and may present with hyperinfection. Therefore, awareness of this parasite should be kept in mind in immunosuppressive patients.

Matched MeSH terms: Feces/parasitology
Stray animal and human defecation as sources of soil-transmitted helminth eggs in playgrounds of Peninsular Malaysia

Mohd Zain SN, Rahman R, Lewis JW

J Helminthol, 2015 Nov;89(6):740-7.
PMID: 25273274 DOI: 10.1017/S0022149X14000716

Soil contaminated with helminth eggs and protozoan cysts is a potential source of infection and poses a threat to the public, especially to young children frequenting playgrounds. The present study determines the levels of infection of helminth eggs in soil samples from urban and suburban playgrounds in five states in Peninsular Malaysia and identifies one source of contamination via faecal screening from stray animals. Three hundred soil samples from 60 playgrounds in five states in Peninsular Malaysia were screened using the centrifugal flotation technique to identify and determine egg/cyst counts per gram (EPG) for each parasite. All playgrounds, especially those in Penang, were found to be contaminated with eggs from four nematode genera, with Toxocara eggs (95.7%) the highest, followed by Ascaris (93.3%), Ancylostoma (88.3%) and Trichuris (77.0%). In addition, faeces from animal shelters were found to contain both helminth eggs and protozoan cysts, with overall infection rates being 54% and 57% for feline and canine samples, respectively. The most frequently occurring parasite in feline samples was Toxocara cati (37%; EPG, 42.47 ± 156.08), while in dog faeces it was Ancylostoma sp. (54%; EPG, 197.16 ± 383.28). Infection levels also tended to be influenced by season, type of park/playground and the texture of soil/faeces. The occurrence of Toxocara, Ancylostoma and Trichuris eggs in soil samples highlights the risk of transmission to the human population, especially children, while the presence of Ascaris eggs suggests a human source of contamination and raises the issue of hygiene standards and public health risks at sites under investigation.

Matched MeSH terms: Feces/parasitology*

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