A cross-sectional survey of 365 individuals, (51.9% males, 48.1% females; ages 5-85 years), from five remote interior communities in upper Rejang River basin Sarawak, Malaysia, found 24.4% were anemic. The range and mean of Hb concentration in male and female were: 7.2-17.0 mg/ml and 13.7 mg/ml and 7.9-15.7 mg/ml and 12.9 mg/ml respectively. Amongst the five tribes surveyed, the prevalence of anemia (range: 10.6-46.7%), was higher among the Penans (46.7%), Kenyahs (31.1%), Kajangs (27.8%) and Kayans (19.3%), than amongst the Ukits (10.6%). Anemia is more common among males >40 years and among adolescents and young reproductive females, as well as elderly females > 61 years old. Of the 83 anemic individuals, 6.0% and 3.6% had Trichuris trichiura or hookworm respectively; however there is no clear association with intestinal worm infection.
A survey of Angiostrongylus malaysiensis among wild rodent and molluscan hosts was made in the Tuaran Central Agricultural Research Station and within the vicinity of Tuaran, Sabah. Three of 19 Rattus rattus diardii, one of 2 R. exulans and one R. argentiventer were found naturally infected with the parasite. In this survey 56 of 382 molluscs comprising of Pila scutata, Achatina fulica and two species of land slugs, Laevicaulis alte and Microparmarion malayanus, were found naturally infected with the parasite. Samples of larvae from each of these molluscs were experimentally transferred to laboratory albino rats and adult worms consistent with A. malaysiensis were recovered. Comparison of the rat hosts and the molluscan intermediate hosts of the parasite in Peninsular Malaysia, Sarawak and Sabah was made, and the finding of A. malaysiensis in Tuaran is the first report of the parasite from Sabah. The distribution of the parasite throughout Malaysia is discussed. Observation on the human consumption of the freshwater snail, P. scutata, was made. Although the infection rate of this snail is low compared with other molluscan hosts examined. The importance of this mollusc as a potential source of human infection should not be overlooked. Hospital records for 1974 and 1975 were examined and clinical human angiostrongyliasis was rarely recorded in Sabah.
Discrepancies in angiotensin-1 converting enzyme (ACE) allele genetic susceptibility with disease etiology have been attributed to ethnic differences. We investigated ACE gene polymorphism of the multiethnic Malaysian population by utilizing nested polymerase chain reaction. Allelic frequency of 0.65 and 0.35 for I and D allele, respectively in the pooled population was comparable with other Asian populations. A significant association was found between the Malaysian ethnic groups and ACE I/D genotype. The II genotype was found at higher frequency among the Malays but a greater frequency of DD genotype among Indians.
Studies were carried out to observe the species composition of mosquitos and to determine the vectors responsible for the transmission of filariasis in Grik, Perak, Malaysia. A total of 2,155 mosquitos belonging to 7 genera and 30 species were collected. Anopheles donaldi comprised 24.1% of the collection. Twelve out of 519 An. donaldi were infected with L3 larvae of Brugia malayi. The peak biting time was around 23.00-24.00 hours. The infective bites per month ranged from 0 to 6.3.
Collections of adult anopheline mosquitos were made from a cow-baited trap in nine coastal villages located along nearly 160km of northwest peninsular Malaysia. Two collections, separated by 1.5 to 6 months, were made at each site. Nearly 6,000 anophelines of 19 species were collected. The dominant species were Anopheles peditaeniatus. An. sinensis, An. subpictus and An. lesteri paraliae. Small numbers of the malaria vectors An. maculatus (at one site) and An. campestris (at four sites) were collected, but no An. sundaicus were recorded.
There are few small animals models for filariasis, even more so for onchocerciasis. Therefore it is difficult to test under drug screening conditions large numbers of potentially macrofilaricidal compounds. One way around this difficulty is to use mice infected with Trichinella spiralis which by reason of anatomical location in the host would show some correlation in antinematode activity between the test and target organisms. This study investigated the activity of 16 compounds against the immature larval stage of T. spiralis. All the nine benzimidazole compounds (albendazole, flubendazole, mebendazole, oxfendazole, oxibendazole 780118, 780120, 790163, and 790392) were active, the most potent being oxfendazole. The benzothiazoles (CGP21306, CGP20376, CGP21833 and CGP24588A) also indicated some anti-nematode activity together with 35vr, an imidazopyridine, but not as marked as the benzimidazole group. However, the organic arsenical compounds (Mel Ga and Mel Ni) showed little activity and this was at a rather highly toxic level. The prospects of using the Trichinella-mouse model as a primary screen to test for potential macrofilaricides are discussed.
Forty samples of Malaysian cooked foods were examined for the presence of antibiotic-resistant coliforms and R plasmids. Twenty seven (68%) of the foods had antibiotic-resistant coliforms and 5 (13%) had R plasmids. Nineteen samples (48%) had total bacterial counts over 10(6) per gm and in 5 samples, no coliforms were detected. Our findings show that cooked food may be one possible way by which R plasmids are spread. The control of the spread of R plasmids is discussed.
Enteropathogenic Escherichia coli (EPEC) is a leading cause of diarrhea among infants in developing countries. A total of 38 EPEC isolates, obtained from diarrhea patients of Hospital Miri, Sarawak, were investigated through plasmid profile, antibiotic resistance and randomly amplified polymorphic DNA (RAPD) analysis. From the 8 types of antibiotics used, all isolates were 100% resistant to furoxime, cephalothin and sulphamethoxazole and showed high multiple antibiotic resistant (MAR) indexes, ranging from 0.5 to 1.0. In plasmid profiling, 22 isolates (58%) showed the presence of one or more plasmids in the range 1.0 to 30.9 mDa. The dendrogram obtained from the results of the RAPD-PCR discriminated the isolates into 30 single isolates and 3 clusters at the level of 40% similarity. The EPEC isolates were highly diverse, as shown by their differing plasmid profiles, antibiotic resistance patterns and RAPD profiles.
Western blot analysis of infective larvae (L3) antigen of Brugia malayi were performed on 200 sera from six groups of individuals: 36 samples from B. malayi microfilaremic individuals; 10 samples from individuals with elephantiasis; 50 and 20 samples from amicrofilaremic individuals in a B. malayi endemic area with no anti-filarial IgG4 antibodies (towards microfilaria and adult worm antigens) and samples with high titres of the anti-filarial IgG4 antibodies respectively; 50 samples from non-endemic normals and 34 samples from geohelminth-infected individuals. After protein transfer, PVDF membrane strips were successively incubated with blocking solution, human sera, monoclonal anti-human IgG4 antibody-HRP and developed with luminol chemiluminescence substrate. 28/36 (78%), 1/10 (10%) and 16/20(80%) of sera from individuals with microfilariae, elephantiasis and amicrofilaremic individuals with high titers of anti-filarial IgG4 antibodies respectively recognized L3 antigenic epitopes; the dominant and consistent antigenic bands were of approximately MW 43 kDa, 14 kDa, 15 kDa and 59 kDa. The rest of the sera were unreactive. This study showed that microfilaremics may or may not mount a notable antibody response to somatic L3 antigens, thus lending evidence that antibody response to this antigen is not protective against establishment of Brugia malayi infection.
The seroprevalence of Orientia tsutsugamushi (OT), Rickettsia typhi (RT) and TT118 spotted fever group rickettsiae (SFGR) among blood donors and febrile Malaysian patients in the urban areas was determined. Of the 240 blood donors, 5.4%, 9.2% and 1.7% had either present or previous exposure to OT, RT and SFG rickettsiae, respectively. Patients admitted to an urban hospital had high seroprevalences of OT (43.5%) and RT (22.9%), as compared to SFGR (11.6%). Antibody levels suggestive of recent infections of scrub typhus, murine typhus and tick typhus were detected in 16.8%, 12.7% and 8.2% of patients respectively. No significant difference was noted in the distribution of rickettsial antibodies among urban patients from 2 geographical locations. However, the serologic patterns of rickettsial infection in the urban areas were different form those of rural areas.
The seroprevalence of various Orientia tsutsugamushi (OT) strains among Malaysian patients with suspected scrub typhus infections was determined using an indirect immunoperoxidase (IIP) assay. IgG against a single OT strain were detected in six sera (3 Karp, 1 Gilliam and 2 TC586), whereas IgM antibodies against a single OT strain (Gilliam) were noted in 3 sera (Gilliam). IgG reactive to all OT strains were present in 33 (47.1%) of the 70 sera and IgM reactive to all OT strains were present in 22 (78.6%) of the 28 sera. The fact that most sera were reactive to multiple OT strains suggests that group-specific antigens are involved in scrub typhus infections, whereas very few were due to strain-specific epitopes present on these strains. Peak IgG and IgM titers were noted more frequently against Gilliam, Karp, and TA763 strains: this suggests that these strains may be the commonest infecting strains among Malaysian patients. Two predominant OT polypeptides consistently reacted with patients' sera were the 70 kDa and 56 kDa proteins.
Fifteen independent E. coli strains of avian, bovine and porcine origin in Peninsular Malaysia were tested for antibiotic resistance and conjugative R plasmids. Eight (53%) isolates were found to be antibiotic resistant. Among them, 37.5% were mono-resistant and 62.5% were resistant to three or more antibiotics, i.e., multi-resistant. All of them were resistant to Tc and sensitive to Gm and Nx. Three of the eight antibiotic resistant strains were able to transfer all or part of their resistance to an E. coli K12 recipient by conjugation. The transfer frequencies of Km, Sm and Tc resistance of the three donors varied between 4.5 X 10(-8) to 6.8 X 10(-7). Analysis of the plasmid profiles of all the three donors and their respective transconjugants after agarose gel electrophoresis provided conclusive evidence that the transferable resistance traits were plasmid-mediated.
This study was conducted to determine the antibiotic susceptibility pattern and distribution of exoU and exoS among 44 clinical isolates of P. aeruginosa collected from different patients over a 3-month period in 2010 at a major Malaysian hospital. Susceptibility data by disk diffusion method for cefepime (30 microg), ceftazidime (30 microg), gentamicin (10 microg), piperacillin-tazobactam (100/10 microg) and ciprofloxacin (5 microg) were available for 38 isolates. Resistance to ceftazidime and piperacillin-tazobactam was the most common (74%) with five isolates not susceptible to three or more different antibiotics. PCR detection of exoU and exoS of all 44 isolates showed the former gene to be present in 18 and exoS in 41. In analyzing the two genes together, 17 isolates were detected for exoU and exoS with only two being negative for both genes. Only one isolate was detected for exoU alone whereas 24 for exoS alone. Distribution of the genes in relation to antibiotic susceptibility was inapplicable due to the majority of the isolates having similar susceptibility patterns, but the tendency of exoU-carrying isolates to be present in male patients (83%) and respiratory sites (61%) was observed (p < 0.050). The finding warrants further investigation in a larger sample of isolates.\
Burkholderia pseudomallei, the causative agent of melioidosis is an important cause of morbidity and mortality particularly among diabetics. We evaluated 228 isolates of B. pseudomallei for antimicrobial sensitivity during 2005-2010 using the disc diffusion technique, of which 144 were obtained from blood culture. More than 90% of the strains were susceptible to cefoperazone, ceftazidime, chloramphenicol and imipenem. Eighty-two percent of the isolates were susceptible to tetracycline and amoxicillin/clavulanate. The susceptibilities to ciprofloxacin was 78% and to trimethoprim-sulfamethoxezole was 47%. The susceptibilities to aminoglycoside antibiotics were low (21% to gentamicin and 6% to amikacin). The susceptibilities were similar between isolates from females and males, bacteremic and abacteremic cases, diabetics and non-diabetics, pneumonia and non-pneumonia cases and between those who died and those who survived. Our findings show antibiotic susceptibility patterns are not a major factor in determining outcomes of B. pseudomallei infection. Monitoring the drug susceptibilities among B. pseudomallei isolates needs to be conducted regularly to guide empiric therapy for melioidosis, as it causes high mortality, especially among diabetic cases.
Partial gene sequences of phenylalanyl-tRNA synthase alpha subunit (pheS) and RNA polymerase alpha subunit (rpoA) were evaluated for species delineation and detection of recombination among enterococci populations recovered from a bathing beach impacted by low tide river flow. At inter-species level, a maximum similarity of 86.5% and 94.8% was observed among the enterococci pheS and rpoA sequence, respectively. A superimposed plot of delimited pairwise similarity values obtained for 266 pair-wise observations revealed that while there was a harmony between species identity obtained from both genes, pheS was more discriminatory than rpoA. The difference was more pronounced for inter-species comparison. A number of putative recombination events between indigenous and non-indigenous strains was detected based on a library of aligned sequences. Virulence genes cyl, esp, gelE and asa were detected in 7, 22, 100 and 63%, respectively among river isolates but at lower proportion of 0, 20, 67 and 42%, respectively among beach water isolates. Random amplified polymorphic DNA profiling presented evidence suggesting low tide river as a source of fecal enterococci entering the recreation beach water. Multilocus sequence typing analysis of a number of Enterococcus faecalis isolates presented four sequence types, ST59, 117, 181 and 474. The presence of genetically diverse fecal enterococci with associated virulence traits and a background of recombination events in surface recreational water could present a potential public health risk.
A representative sample of 20 isolates of Salmonella weltevreden strains from stool cultures of patients admitted at the University Hospital, Kuala Lumpur, Malaysia were analyzed. All the strains were susceptible to ampicillin, ceftriaxone, ciprofloxacin, chloramphenicol, tetracycline, trimethoprim, gentamicin and co-trimoxazole. Ribosomal RNA gene restriction pattern analysis of PstI-digested DNA gave three ribotypes while pulsed-field gel electrophoresis (PFGE) analysis of XbaI-digested DNA gave ten distinct profiles. PFGE was more discriminative than ribotyping in distinguishing the strains. The majority of the strains analyzed were very closely related with similarity coefficient values ranging from 0.8 to 1.0. Both PFGE and ribotyping could distinguish one of the strains which was obtained from a patient following a bone marrow transplant for beta-thalassemia major, indicating that this particular strain was unrelated to the rest of the strains from patients with acute gastroenteritis.
Biotechnological tools are being used in malaria, filariasis and dengue research. The main emphasis has been on the production of reagents for immunodiagnosis and research. In this respect monoclonal antibodies (McAbs) against various species and stages of the above pathogens have been produced. It is hoped that these McAbs will be useful not only in immunodiagnosis but also for seroepidemiological applications. A DNA probe against Brugia malayi has been tested in Malaysia and was found to be sensitive and specific.