Entamoeba histolytica infection is the third-greatest parasitic disease responsible for death in the world. Wild rats harbouring E. histolytica can be the possible reservoir hosts for human amoebiasis. There were numerous studies on prevalence of intestinal parasites among wild rats in Malaysia but none has reported E. histolytica. Rats were captured from Sentul and Chow Kit areas, Kuala Lumpur, Malaysia. The preserved stool samples were used for microscopy examination and molecular analysis. Out of 137 samples collected, 12 were positive for E. histolytica / E. dispar / E. moshkovskii microscopically. Two E. histolytica (1.4%), 1 E. dispar (0.7%) and 6 mixed infections of E. histolytica and E. dispar (4.3%) were detected using PCR. This is the first report of molecular detection of E. histolytica/dispar infection among wild rats in Malaysia. This study provides useful information about the potential risks of zoonotic agents and the importance of developing control measures to prevent zoonotic transmission.
Dengue serotype surveillance is important as any changes in serotype distribution may result in an outbreak or increase in severe dengue cases. This study aimed to determine circulating dengue serotypes in two hospitals in Selangor. Serum samples were collected from patients admitted for dengue at these two major public hospitals i.e. Hospital Sungai Buloh (HSB) and Hospital Tunku Ampuan Rahimah (HTAR) between November 2010 and August 2011 and subjected to real-time RT-PCR using SYBR® Green. All four dengue serotypes were detected in samples from both hospitals. The predominating serotype was dengue 1 in samples from both hospitals (HSB, DENV-1; 25.53 % and HTAR, DENV-1; 32.1 %).
Malaria causes high global mortality and morbidity annually. Plasmodium knowlesi has been recognised as the fifth human Plasmodium sp. and its infection is widely distributed in Southeast Asia. Merozoite surface protein-119 (MSP-119) appears as a potential candidate for malaria blood stage vaccine as it could induce protective immunity. In this study, codon optimized P. knowlesi MSP-119 (pkMSP-119) was expressed and purified in yeast Pichia pastoris expression system. The purified recombinant protein was further evaluated using Western blot assay using knowlesi malaria, non-knowlesi human malaria, non-malarial parasitic infections and healthy serum samples (n = 50). The sensitivity of purified pkMSP-119 towards detection of knowlesi infection was as 28.6% (2/7). pkMSP-119 did not react with all non-malarial parasitic infections and healthy donor sera, yet reacted with some non-knowlesi human malaria sera, therefore lead to a specificity of 86.0% (37/43).
This study was aimed at determining the phospholipase and haemolysin activity of Candida isolates in Malaysia. A total of 37 Candida clinical isolates representing seven species, Candida albicans (12), Candida tropicalis (8), Candida glabrata (4), Candida parapsilosis (1), Candida krusei (4), Candida orthopsilosis (1) and Candida rugosa (7) were tested. In vitro phospholipase activity was determined by using egg yolk plate assay whereas in vitro haemolysin activity was tested by using blood plate assay on sheep blood Sabouraud's dextrose agar (SDA) enriched with glucose. Phospholipase activity was detected in 75% (9 out of 12) of the C. albicans isolates. Among the 25 non- C. albicans Candida isolates, phospholipase activity was detected in only 24% of these isolates. The phospholipase activity of C. albicans was significantly higher than that of the non- C. albicans Candida isolates (P=0.002). Haemolysin activity was detected in 100% of the C. albicans, C. tropicalis, C. glabrata, C. krusei, C. parapsilosis, and C. orthopsilosis isolates while 75% of the C. krusei isolates and 12.3% of the C. rugosa isolates showed haemolysin activity. The haemolytic activity of C. albicans was significantly higher than that of the non- C. albicans Candida isolates (P=0.0001).The findings in this study indicate that C. albicans isolates in Malaysia may possess greater virulence potential than the non-albicans species.
Intestinal parasitic infections are among the most common diseases affecting mankind causing major public health problems to billions of people living in developing countries. The aim of this study is to determine the prevalence of intestinal parasites in various communities residing in different habitats in Malaysia and compare the findings with 101 studies conducted over the past 42 years (1970-2013). A cross-sectional study design was conducted with the aid of a questionnaire to collect relevant information about the study population. Faecal samples were examined using the direct smear and formal ether sedimentation techniques. A total of 342 children were examined amongst whom 24.6% were positive for intestinal parasitic infections. Results showed that 32.3% of rural children, 20.6% of urban squatters and 5.4% of children from flats were positive for one or more parasites. The most common parasite encountered was Trichuris trichiura (20.2%) followed by Ascaris lumbricoides (10.5%) and hookworm (6.7%). No case of hookworm was reported in urban children whereas 12.2% of rural children were positive. The most common protozoan parasite detected was Entamoeba coli (3.2%) followed by Giardia intestinalis (1.8%), Entamoeba histolytica (1.8%) and Blastocystis hominis (1.2%). Nearly one-fifth (18.4%) of the children had single infection followed by double (12.0%) and triple infections (1.2%). Orang Asli (indigenous) children (44.3%) had the highest infection rate followed by Indians (20.2%), Malays (14.0%) and Chinese (11.9%). Twenty-eight studies carried out on plantation communities with regards to intestinal parasitic infections in Malaysia from 1970 to 2013 showed a steady decline in the prevalence rate ranging from 95.0% in the seventies to 37.0 % in 2012. Intestinal parasitic infections were more common in Orang Asli communities with prevalence ranging from over 90% in the seventies and fluctuating below 70% in most studies between 2000 to 2013 except for two studies that showed a prevalence of 98.2% and 100%. The prevalence rate among urban squatters, urban residents and those living in flats showed dramatic decrease in prevalence rate.
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.
Determination of Streptococcus pneumoniae serotypes is essential for epidemiological surveillance. Therefore accurate, reliable and cost effective serotyping method is crucial. In this study, we determined the serotypes of 41 pneumococcal isolates recovered from human anterior nares by multiplex Polymerase Chain Reaction (PCR) utilizing published primers. The data was then compared with conventional serology using latex agglutination (LA) and the Quellung reaction. Based on the PCR-approach, 8 different serogroups/serotypes were detected with one isolate classified as non-typeable (cpsA-negative). In reference to the serology-based data, the results were in agreement except for one isolate. For the latter isolate, the LA and Quellung tests failed to show a reaction but the PCR-approach and sequencing identified the isolate as serogroup 15B/C. Based on this experimental setting, we found that the PCR-approach for pneumococcal serotypes determination is reliable to serve as the alternative for determining the pneumococcal serotyping.
A survey was undertaken to investigate the prevalence of intestinal and blood parasites among wild rats in urban area of Kuala Lumpur, Malaysia. A total of 137 stool and blood samples were collected from wild rats from Sentul and Chow Kit areas. Five species of rats were captured and supplied by Kuala Lumpur City Hall. The most common was Rattus rattus diardii (Malayan Black rat), 67%, followed by Rattus norvegicus (Norway rat), 10%, Rattus argentiventer (rice-field rat), 10%, Rattus tiomanicus (Malaysian field rat), 9% and Rattus exulans (Polynesian rat), 4%. Rattus rattus diardii is commonly known to live in human environment and they are normally identified as pests to human community. More male rats were captured (61%) compared to female (39%). Out of 137 samples, 81.8% samples were positive with intestinal parasites, with 86.2% from Sentul area and 78.5% from Chow Kit area. Six different parasites were detected. The most common intestinal helminth parasite detected was Nippostrongylus brasiliensis (80.3%), followed by Hymenolepis nana (23.4%), Capillaria hepatica (13.9%) and Hymenolepis diminuta (2.9%). Intestinal protozoan detected was Entamoeba histolytica/E. dispar (8.8%). Trypanosoma lewisi (1.5%) was the only blood parasite detected.
DNA identification of blow fly species can be a very useful tool in forensic entomology. One of the potential benefits that mitochondrial DNA (mtDNA) has offered in the field of forensic entomology is species determination. Conventional identification methods have limitations for sibling and closely related species of blow fly and stage and quality of the specimen used. This could be overcome by DNA-based identification methods using mitochondrial DNA which does not demand intact or undamaged specimens. Mitochondrial DNA is usually isolated from whole blow fly and legs. Alternate sources for mitochondrial DNA isolation namely, egg, larva, puparium and empty puparium were explored in this study. The sequence of DNA obtained for each sample for every life cycle stage was 100% identical for a particular species, indicating that the egg, 1st instar, 2nd instar, 3rd instar, pupa, empty puparium and adult from the same species and obtained from same generation will exhibit similar DNA sequences. The present study also highlighted the usefulness of collecting all life cycle stages of blow fly during crime scene investigation with proper preservation and subsequent molecular analysis. Molecular identification provides a strong basis for species identification and will prove an invaluable contribution to forensic entomology as an investigative tool in Malaysia.
A study to determine the diversity and distribution of ectoparasites and endoparasites infesting wild rat population of Carey Island was carried out from June to December 2010. A total of 81 rats were captured from various locations on Carey Island. Four rat species were identified namely, Rattus tiomanicus (45.7%), Rattus rattus diardii (25.9%), Rattus argentiventer (16%) and Rattus norvegicus (12.3%). Low diversity of ecto and endoparasites were observed infecting the rodent population with 8 ecto and 8 endoparasites species recorded. The ectoparasites recovered fell under 3 broad groups, namely mites (Laelaps nuttali, Laelaps echidninus, Laelaps sculpturatus, Listrophoroides sp. and Ornithonyssus bacoti), lice (Polyplax spinulosa and Hoplopleura pacifica) and tick (Ixodes granulatus) while endoparasites recovered were cestodes (Taenia taeniaformis and Hymenolepis diminuta) and nematodes (Nippostrongylus brasiliensis, Angiostrongylus malaysiensis, Mastophorus muris, Heterakis spumosa, Hepatojarakus malayae and Syphacia muris). The rat population was observed harbouring more than one parasite species. Analysis of data also showed neither intrinsic (host age, host sex) nor extrinsic (season) factors influenced the macroparasites community structure.
This study was conducted to investigate the low prevalence of Dirofilaria immitis in dogs in Johor Bahru as reported by veterinary practitioners, using wet blood mount, Knott's Concentration Test and two heartworm antigen test kits (IDEXX Canine SNAP® 4Dx and RapiGEN®). This study also compared the two test kits used and determined the microfilaria species. Blood were collected from 100 owned dogs and 50 stray dogs in Johor Bahru via cephalic venipuncture. A thick blood smear was done and examined for samples that were positive for microfilaria species identification. The overall prevalence of D. immitis in dogs in Johor Bahru was 1.33% (2/150) and the microfilaria identified was D. immitis. The prevalence of heartworm in owned and stray dogs in this study was 1% and 2% respectively. With only one false negative result from RapiGEN® test kit, comparing the sensitivity between the two test kits could not be achieved. The low prevalence of D. immitis found in this study confirmed anecdotal evidence that prevalence of dirofilariasis is indeed low in Johor Bahru. Additionally, we speculate that dirofilariasis in dogs might be considered as an indicator of vector availability.
We report a case of Hymenolepis diminuta infection in a 2-year-old Malaysian child. This case was initially reported as 'normal' after the examination of proglottids shed from the anus of the child at a private laboratory on two occasions. The putative proglottids shed was then referred to the Parasite Southeast Asia Diagnostic (Para:SEAD) Laboratory, Department of Parasitology, Faculty of Medicine, University of Malaya for further examination. Microscopic examination confirmed that the child was infected with H. diminuta based on the characteristic eggs found in the proglottids. She was treated with a single dose praziquantel (20 mg/kg of body weight) and recovered well.
Simulium (Gomphostilbia) sofiani sp. nov. is described on the basis of reared adult female, male, pupal and larval specimens collected from Cameron Highlands, Pahang state, Malaysia. This new species is placed in the ceylonicum species-group within the subgenus Gomphostilbia and is easily distinguished from all the related known species by the combination of the following characteristics: an elongate sensory vesicle and yellow hair tuft on the stem vein of the wing in the female, the greater number of large upper-eye facets (15 or 16 vertical columns and 15 or 16 horizontal rows) and almost entirely darkened hind basitarsus in the male, and the gill bearing a long common basal stalk and 8 filaments arranged as [(1+2)+(1+2)] +2 filaments from dorsal to ventral in the pupa.
Fungal osteomyelitis is a rare opportunistic infection. It exhibits some clinical and radiological similarities to several other bone pathologies. A diagnostic delay may result in significant increase in morbidity. We report a case of a 37-year-old man with underlying hypogammaglobulinaemia presented with isolated cryptococcal osteomyelitis of the femur.
The increased frequency of antibiotic resistance is known to be associated with the dissemination of integrons in the Enterobacteriaceae. This study determined the prevalence and type of integrons amongst 160 extended-spectrum beta-lactamase producing enterobacterial isolates kept in our culture collection. Integrons were detected in 98(61.3%) isolates, including 28(62.2%) Escherichia coli, 34(64.2%) Klebsiella spp., 27(61.4%), Enterobacter spp. and 9(50.0%) Citrobacter spp. investigated in this study. Restriction analysis of the integron gene fragments revealed that class I integron was the principal integron detected in 92(57.5%) of our isolates. Class II integron was detected in 6(3.8%) of our isolates, while no class III integron was detected in this study. The high rates of integron prevalence particularly of the class I integron in the E. coli and Klebsiella spp. concur with previous studies in other geographical regions. The higher (≥50%) integron prevalence of Citrobacter and Enterobacter isolates comparing to previous studies suggests the potential of these isolates as sources for dissemination of resistance determinants. The finding in this study serves as a basis for further study on the antibiotic resistance mechanisms of enterobacterial species in this teaching hospital.
The use of ice cubes in beverages is common among patrons of food outlets in Malaysia although its safety for human consumption remains unclear. Hence, this study was designed to determine the presence of faecal coliforms and several useful water physicochemical parameters viz. free residual chlorine concentration, turbidity and pH in ice cubes from 30 randomly selected food outlets in Kubang Kerian, Kelantan. Faecal coliforms were found in ice cubes in 16 (53%) food outlets ranging between 1 CFU/100mL to >50 CFU/ 100mL, while in the remaining 14 (47%) food outlets, in samples of tap water as well as in commercially bottled drinking water, faecal coliforms were not detected. The highest faecal coliform counts of >50 CFU/100mL were observed in 3 (10%) food outlets followed by 11-50 CFU/100mL and 1-10 CFU/100mL in 7 (23%) and 6 (20%) food outlets, respectively. All samples recorded low free residual chlorine concentration (<0.10mg/L) with the pH ranging between 5.5 and 7.3 and turbidity between 0.14-1.76 NTU. Since contamination by faecal coliforms was not detected in 47% of the samples, tap water and commercially bottled drinking water, it was concluded that (1) contamination by faecal coliforms may occur due to improper handling of ice cubes at the food outlets or (2) they may not be the water sources used for making ice cubes. Since low free residual chlorine concentrations were observed (<0.10mg/ L) in all samples as well as in both tap water and commercially bottled drinking water, with the pH ranged between 5.5-7.3, ineffective disinfection of water source as a contributing factor to such high counts of faecal coliforms in ice cubes also could not be ruled out. Therefore, a periodical, yet comprehensive check on the food outlets, including that of ice cube is crucial in ensuring better food and water for human consumption.
Leptospirosis is recognized as one of the important zoonotic diseases in the world including Malaysia. A total of 145 soil and water samples were collected from selected National Service Training Centres (NSTC) in Kelantan and Terengganu. The samples were inoculated into modified semisolid Ellinghausen McCullough Johnson Harris (EMJH) medium, incubated at room temperature for 1 month and examined under the dark-field microscope. Positive growth of the leptospiral isolates were then confirmed with 8-Azaguanine Test, Polymerase Chain Reaction (PCR) assay and Microscopic Agglutination Test (MAT). Fifteen cultures (10.34%) exhibited positive growths which were seen under dark field microscope whilst only 20% (3/15) were confirmed as pathogenic species. based on 8-Azaguanine Test and PCR. Serological identification of the isolates with MAT showed that hebdomadis was the dominant serovar in Terengganu. Pathogenic leptospires can be detected in Malaysian environment and this has the potential to cause an outbreak. Therefore, precautionary steps against leptospirosis should be taken by camp authorities to ensure the safety of trainees.
The objective of our study was to study the effectiveness of CHROMagar Candida™ as the primary identification method for various clinical Candida isolates, other than the three suggested species by the manufacturer. We studied 34 clinical isolates which were isolated from patients in a local teaching hospital and 7 ATCC strains. These strains were first cultured in Sabouraud dextrose broth (SDB) for 36 hours at 35ºC, then on CHROMagar plates at 30ºC, 35ºC and 37ºC. The sensitivity of this agar to identify Candida albicans, Candida dubliniensis, Candida tropicalis, Candida glabrata, Candida rugosa, Candida krusei and Candida parapsilosis ranged between 25 and 100% at 30ºC, 14% and 100% at 35ºC, 56% and 100% at 37ºC. The specificity of this agar was 100% at 30ºC, between 97% and 100% at 35ºC, 92% and 100% at 37ºC. The efficiency of this agar ranged between 88 and 100% at 30ºC, 83% and 100% at 35ºC, 88% and 100% at 37ºC. Each species also gave rise to a variety of colony colours ranging from pink to green to blue of different colony characteristics. Therefore, the chromogenic agar was found to be useful in our study for identifying clinical Candida isolates.
Entamoeba histolytica, the causative agent of human amoebiasis remains a significant cause of morbidity and mortality in developing countries and is responsible for up to 100,000 deaths worldwide each year. Entamoeba dispar, morphologically indistinguishable from E. histolytica is more common in humans in many parts of the world. Similarly Entamoeba moshkovskii, which was long considered to be a free-living amoeba is also morphologically identical to E. histolytica and E. dispar, and is highly prevalent in some E. histolytica endemic countries. Humans are the host of infection and there would not appear to be other meaningful animal reservoirs of E. histolytica. Entamoeba. histolytica can be present in sewage and contaminated water. The infection is mainly transmitted via ingestion of water or food contaminated by faeces containing E. histolytica cysts. Clinical features of amoebiasis range from asymptomatic colonization to amoebic dysentery and invasive extraintestinal amoebiasis, which is manifested most commonly in the form of abscesses in liver and lungs. The epidemiology of amoebiasis has dramatically changed since the separation of E. histolytica and E. dispar species and the worldwide prevalence of these species has not been estimated until recently. Morever, E. moshkovskii, another morphologically indistinguishable human parasitic Entamoeba was not mentioned or considered as a contributor to the prevalence figures in endemic areas. Amoebiasis is still a major health problem especially in aboriginal settlements and amongst people living in remote area in Malaysia. However, until now there is only one data currently available to indicate the true prevalence and incidence of E. histolytica and E. dispar. Further studies are needed to determine the burden of E. histolytica, E. dispar and E. moshkovskii infections in Malaysia. In the present review, we briefly summarize all methods use in diagnosing Entamoeba species, ranging from microscopic identification to molecular detection such as culture and isoenzyme analysis, antibody detection tests, antigen detection tests, immunochromatographic assays, conventional PCR, real-time PCR and loop-mediated isothermal amplification (LAMP).
Transmission of soil-transmitted helminthes infection is by faecal oral route, and is influenced by food preference. Kelantanese love to consume ulam which are raw vegetables and herbs. Some of the herbs grow on grounds with high humidity and are abundant near drainage areas, these are also places with higher likelihood of harbouring viable parasite ova. The aim of this study was to determine the prevalence of soiltransmitted helminthes in vegetables, herbs and fruits found in our local setting. The results by microscopy showed that there was no helminthes ovum or protozoan parasite in the samples. However, Strongyloides stercoralis rhabdatiform larvae were identified in water samples used to wash pegaga, kesum and water spinach, and the number of larvae observed were 152, 9 and 16 respectively. Analysis by real-time PCR confirmed the microscopic observation of this helminth. This study highlighted that vegetables and herbs are likely sources of Strongyloides stercoralis infection in Kota Bharu, Kelantan. Thus vegetable sellers as well as the food handlers are the two important groups who are at high risk of acquiring the infection.