Cryptosporidium fragile sp. n. (Apicomplexa) is described from black-spined toads, Duttaphrynus melanostictus (Schneider) (Amphibia, Anura, Bufonidae) from the Malay Peninsula. The parasitized animals were directly imported from Malaysia and harboured C. fragile at the time of arrival. Oocysts were subspherical to elliptical with irregular contour in optical section, measuring 6.2 (5.5-7.0) x 5.5 (5.0-6.5) microm. Oocyst wall was smooth and colourless in light microscopy. The endogenous development of C. fragile in the stomach of black-spined toad was analysed in detail using light and electron microscopy. Cryptosporidian developmental stages were confined to the surface of gastric epithelial cells. In transmission experiments, C. fragile has not been infective for one fish species, four amphibian species, one species of reptile and SCID mice. Full length small subunit rRNA gene sequence was obtained. Phylogenetic reconstruction revealed distinct status of C. fragile within the clade of species with gastric localisation including Cryptosporidium muris Tyzzer, 1907, Cryptosporidium serpentis Levine, 1980 and Cryptosporidium andersoni Lindsay, Upton, Owens, Morgan, Mead et Blagburn, 2000. Described characteristics differentiate C. fragile from the currently recognized Cryptosporidium species. Our experience with the description of C. fragile has led us to revise the recommended criteria for an introduction of a new Cryptosporidium species name. C. fragile is the first species described and named from an amphibian host. Its prevalence of 83% (15/18) in black-spined toads within the 3 months after importation calls for strict quarantine measures and import regulation for lower vertebrates.
Data on soil-transmitted helminth (STH) infections and reinfection among Orang Asli (aborigine) schoolchildren and their nutritional and socioeconomic status were analyzed to investigate the pattern and the possible predictors of STH reinfection. In this longitudinal study, 120 (60 males and 60 females) Orang Asli primary schoolchildren aged 7-12 years and living in remote areas in Pos Betau, Kuala Lipis, Pahang were screened for the presence of STH using modified cellophane thick smear and Harada Mori techniques. The overall prevalence of ascariasis, trichuriasis and hookworm infections were 65.8, 97.5 and 10.8%, respectively. After complete deworming with a 3-day course of 400mg/daily of albendazole tablets, children were re-examined at 3 and 6 months from baseline. The reinfection rate, by one or more of STH species, at 3 months after deworming was high (49.5%) while 79.6% of the children were reinfected at 6 months after deworming. Logistic regression analyses showed that females, stunted children and those living in houses without toilets had significantly higher reinfection rates than others at 3 months (P<0.05). At 6 months, maternal employment status emerged as another predictor where children of working mothers had significantly higher reinfection rates (P=0.026). In conclusion, reinfection rate of STH is high and thus necessitates frequent and periodic deworming among children. Public health personnel need to re-look at the current control measures and identify innovative and integrated ways in order to reduce STH significantly in the rural communities.
This study was conducted to reassess the risk factors for giardiasis in communities of the Orang Asli (indigenous people) in Pahang, Malaysia. Stool samples were collected from 321 individuals (2-76 years old; 160 males, 161 females). Data were collected via laboratory analysis of faecal samples and a pre-tested standard questionnaire. River water samples were tested for Giardia cysts and Cryptosporidium oocysts. The overall prevalence of G. intestinalis infection was 23.7%. Children < or =12 years old had the highest infection rate and have been identified as a high risk group (odds ratio (OR)=6.2, 95% CI 1.5-27.0, P<0.005). The risk of getting giardiasis also appeared to be significantly associated with drinking piped water (OR=5.1, 95% CI 0.06-0.7, P<0.005) and eating raw vegetables (OR=2.4, 95% CI 0.2-0.6, P<0.005). In conclusion, sociodemographic factors have always been associated with the high prevalence of Giardia infections in Malaysia. However, the present study also highlights the need to look into the possibility of other risks such as water and food transmission routes. In future, it is necessary that these two aspects be considered in control strategies.
A large focus of human infections with Plasmodium knowlesi, a simian parasite naturally found in long-tailed and pig-tailed macaques was discovered in the Kapit Division of Sarawak, Malaysian Borneo. A study was initiated to identify the vectors of malaria, to elucidate where transmission is taking place and to understand the bionomics of the vectors in Kapit.
There have been three major rabies epidemics in China since the 1950s. To gain more insights into the molecular epidemiology of rabies viruses (RVs) for the third (the current) epidemic, we isolated RV from dogs and humans in major endemic areas, and characterized these isolates genetically by sequencing the entire glycoprotein (G) gene and the G-L non-coding region. These sequences were also compared phylogenetically with RVs isolated in China during previous epidemics and those around the world. Comparison of the entire G genes among the Chinese isolates revealed up to 21.8% divergence at the nucleotide level and 17.8% at the amino acid level. The available Chinese isolates could be divided into two distinct clades, each of which could be further divided into six lineages. Viruses in clade I include most of the Chinese viruses as well as viruses from southeast Asian countries including Indonesia, Malaysia, the Philippines, Thailand, and Vietnam. The viruses in the other clade were found infrequently in China, but are closely related to viruses distributed worldwide among terrestrial animals. Interestingly, most of the viruses isolated during the past 10 years belong to lineage A viruses within clade I whereas most of the viruses isolated before 1996 belong to other lineages within clades I and II. Our results indicated that lineages A viruses have been predominant during the past 10 years and thus are largely responsible for the third and the current epidemic in China. Our results also suggested that the Chinese RV isolates in clade I share a common recent ancestor with those circulating in southeast Asia.
In recent years, Spirulina has gained more and more attention from medical scientists as a nutraceutical and a source of potential pharmaceuticals. The present study was conducted to elucidate the immunomodulatory effect of Spirulina fusiformis (a cyanobacterium of the family Oscillatoriaceae) in vivo and in vitro. The in vivo effect of S. fusiformis (400 or 800 mg/kg body wt.) on humoral immune response, cell-mediated immune response and tumour necrosis factor alpha was investigated in mice. We also evaluated the effect of S. fusiformis (50 or 100 microg/ml) in vitro on mitogen (phytohaemagglutinin)-induced T lymphocyte proliferation in heparinized human peripheral blood. For comparison, dexamethasone was used as a standard. In mice, S. fusiformis (400 or 800 mg/kg body wt.) administration significantly inhibited the humoral immune response, cell-mediated immune response (delayed-type hypersensitivity reaction (DTH)) and tumour necrosis factor alpha in a dose-dependent manner. In vitro, S. fusiformis (50 or 100 microg/ml) decreased the mitogen (phytohaemagglutinin)-induced T lymphocyte proliferation in a concentration-dependent manner when compared with control cells. These observations clearly suggest that S. fusiformis has a remarkable immunosuppressive effect, which provides a scientific validation for the popular use of this drug, and helped us in further work on investigating its complete mechanism of action.
Typhonium flagelliforme (Lodd.) Blume (Araceae) is a Malaysian plant used locally to combat cancer. In order to evaluate its antiproliferative activity in vitro and to possibly identify the active chemical constituents, a bioactivity guided study was conducted on the extracts of this plant.
Fifty faecal samples from diarrheic calves between 1 and 6 months old were collected per rectum from 5 farms around Petaling District in Selangor, Malaysia for Cryptosporidium species detection and genotyping investigation. Oocysts were purified using sedimentation and gradient centrifugation, then examined by immunofluorescence assay (IFAT). Genomic DNA was extracted from all samples and nested PCR was performed to amplify the SSU rRNA gene. Eighteen samples (36%) were positive for Cryptosporidium species by PCR. The sequence and phylogenetic analysis of 14 isolates indicated that Cryptosporidium parvum was most common (11 isolates) followed by Cryptosporidium deer-like genotype (3 isolates). The present work reports the first data on Cryptosporidium genotyping from cattle in Malaysia.
We have defined DNA repeat variability in the 3'-terminus of the cagA gene of Helicobacter pylori strains from Malaysian patients of different ethnicities. We identified different alleles based on the EPIYA repeats. cagA types A-B-D and A-B-B-D are more similar to the sequence of Japanese strains, whereas cagA types A-B-C, A-B-C-C, A-B and A-C displayed similarity to strain 26695 sequences. A significant association was found between cagA genotypes and patients' ethnicity, with cagA type A-B-D being predominantly isolated from Chinese patients and cagA type A-B-C from Malays and Indians. Our data further corroborate the possibility that variant biological activity of CagA may affect the host specificity and/or pathogenicity of H. pylori.
The anti-pyretic activity of a standardized methanol/water (50/50) extract of Orthosiphon stamineus Benth. (SEOS) was investigated for its effect on normal body temperature and yeast-induced pyrexia in Sprague Dawley (SD) rats. The SEOS showed no effect on normal body temperature. Doses of 500 and 1000 mg/kg body weight of SEOS significantly reduced the yeast-induced elevation in body temperature. This effect persisted up to 4 h following the administration of the extract. The anti-pyretic effect of SEOS was comparable with that of paracetamol (acetaminophen in U.S) (150 mg/kg p.o.), a standard anti-pyretic agent. HPLC study revealed that rosmarinic acid, sinensetin, eupatorin and tetramethoxyflavone were present in SEOS in the amounts of 7.58%, 0.2%, 0.34% and 0.24% respectively. The LD(50) of the extract in rats was higher than 5000 mg/kg body weight. Therefore, the present study ascertained that SEOS possesses a significant anti-pyretic activity.
A minisequencing method based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) was developed for rapid identification of single nucleotide polymorphisms at bla(TEM) gene codons 104, 164 and 238 associated with extended-spectrum activity on TEM-type beta-lactamases. The method was validated by testing the Escherichia coli and Klebsiella pneumoniae strains possessing the known bla(TEM) gene sequences.
This article is a review of the latest information on the prevalence of G. lamblia in South Asia, South East Asia and Far East, characterizing the current endemic situation within these regions. Around 33 published papers from 2002-2007 were collected on G. lamblia. The included countries were Nepal, Bangladesh, India, Cambodia, Vietnam, Malaysia, Philippines, Indonesia, Thailand, Republic of Korea, and China. Only five published papers were discarded because data was extracted before 2002-2007 or they are not included within our regions, emphasizing more on G. lamblia in animals, or performed at extensive molecular level. The prevalence of G. lamblia varied markedly between studies illustrating higher levels in the urban than in the rural areas, more among poor communities, slightly higher in males than in females with age range of 2-5-year-old children, and among university students, old-aged people, HIV-positive patients, and gastric carcinoma patients. Though G. lamblia is not a life-threatening parasite, nevertheless, it is still considered as the most common water-borne diarrhea-causing disease. It is important to understand the etiology, frequency, and consequences of acute diarrhea in children. Routine surveillance such as bi-annual follow-up treatments, treating G. duodenalis cysts and other protozoa oocysts detected in ground water sources, and continuous health education are the most preventive measures.
The sensitivity and specificity of 18S rRNA polymerase chain reaction (PCR) in the detection of fungal aetiology of microbial keratitis was determined in thirty patients with clinical diagnosis of microbial keratitis.
A total of 225 samples from poultry farms and the surrounding environment were screened for vancomycin-resistant enterococci (VRE) and bifunctional aminoglycoside-resistant enterococci using conventional microbiological tests and a nanoplex polymerase chain reaction (PCR) assay. Three (1.3%) of the samples were found to contain vancomycin-resistant isolates (MIC>256 microg/mL) that had a vanA genotype. The three vanA positive VRE isolates were identified as different species. Only one isolate (Enterococcus faecium F 4/13_54) was sensitive to teicoplanin (MIC<0. 12-0.35 microg/mL); the other two VRE (E. faecalis A 21_35 and E. gallinarum F 5/10_1) were resistant to teicoplanin (MIC 3.6-->16 microg/mL). The vanC genotype was observed in nine (4%) of the samples collected. High-level gentamicin-resistant (HLGR) enterococci (with MIC ranging between 100 and 500 microg/mL) were detected in 44 samples. However, only 40 of these were found to possess the aac(6')-aph(2'') gene. The overall prevalence of VRE among the samples from the poultry farms and environment was 5.3%, but the prevalence of the clinically significant vanA VRE was 1.3%, and the prevalence of bifunctional aminoglycoside-resistant enterococci was slightly higher, at 19.5%.
Although well established in the West, stool antigen tests (SAT) are not widely used in Asia. Data on the accuracy of this test in Asia is sparse and, to date, there have been no studies looking at the more refined monoclonal SAT. The aim of this study is to validate the diagnostic accuracy of a stool antigen test, Hp STAR, for the detection of Helicobacter pylori.
Choline-binding proteins (CBP) have been associated with the pathogenesis of Streptococcus pneumoniae. We screened, using PCR, for the presence of genes (cbpA, D, E, G) encoding these proteins in 34 isolates of pneumococci of known serotypes and penicillin susceptibility from invasive and non-invasive disease. All isolates harboured cbpD and cbpE whereas cbpA and cbpG were found in 47% and 59% respectively; the latter were more frequent in vaccine-associated types and together accounted for 77% of these isolates. No association was observed with penicillin susceptibility but 85% of non-invasive isolates were positive for these genes.
Multi-resistant Enterobacteriaceae pose a serious threat of hospital acquired infections and their rapid identification is important for better clinical outcome. This study describes the rapid identification of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae of the sulphydryl variable-type by fluorescent in-situ hybridization. The method which rapidly identifies the target genes within 1 h could be a potentially rapid bacterial diagnostic tool.
Burkholderia pseudomallei, the causative agent of melioidosis, is endemic in southeast Asia and northern Australia. In recent years, the incidence of melioidosis has increased worldwide. Septic arthritis is a rare but well-recognized manifestation of melioidosis. Patients with underlying medical conditions, such as diabetes mellitus, renal impairment, cirrhosis, and malignancy are at greater risk. The presentations of melioidotic septic arthritis often mimic other disease processes and patients may not always be clinically septic. We present a case of septic arthritis due to B. pseudomallei in a 66-year-old male with diabetes mellitus presenting with a history of fever and ankle swelling. Follow-up ankle X-ray showed soft tissue swelling. Synovial fluid and blood samples grew B. pseudomallei. The patient improved gradually after parenteral administration of ceftazidime (2 g 8-hourly) and cotrimoxazole (1440 mg 8-hourly). He was discharged on oral cotrimoxazole (1440 mg 12-hourly), doxycycline (100 mg 12-hourly), and chloramphenicol (500 mg 6-hourly) for 6 months. This case highlights the possible occurrence of melioidotic septic arthritis, and the importance of prompt initiation of appropriate antimicrobials to achieve good outcomes.
Study site: Emergency department, University of Malaya Medical Centre (UMMC), Kuala Lumpur, Malaysia
Data on the carriage rate and antibiotic sensitivity pattern of Staphylococcus aureus strains prevalent in the community are not available for many developing countries including Malaysia. To estimate the extent of community S. aureus transmission, in particular methicillin-resistant S. aureus (MRSA), the prevalence of S. aureus nasal colonization in a population of healthy adults was determined. Factors associated with S. aureus nasal carriage and antibiotic sensitivity patterns of the isolates were also analyzed.