Yvonne Al Lim

Increased detection of Plasmodium knowlesi in Sandakan division, Sabah as revealed by PlasmoNex™

BackgroundPlasmodium knowlesi is a simian malaria parasite that is widespread in humans in Malaysian Borneo. However, little is known about the incidence and distribution of this parasite in the Sandakan division, Malaysian Borneo. Therefore, the aim of the present epidemiological study was to investigate the incidence and distribution of P. knowlesi as well as other Plasmodium species in this division based on a most recent developed hexaplex PCR system (PlasmoNex™).MethodsA total of 189 whole blood samples were collected from Telupid Health Clinic, Sabah, Malaysia, from 2008 to 2011. All patients who participated in the study were microscopically malaria positive before recruitment. Complete demographic details and haematological profiles were obtained from 85 patients (13 females and 72 males). Identification of Plasmodium species was conducted using PlasmoNex™ targeting the 18S ssu rRNA gene.ResultsA total of 178 samples were positive for Plasmodium species by using PlasmoNex™. Plasmodium falciparum was identified in 68 samples (38.2%) followed by 64 cases (36.0%) of Plasmodium vivax, 42 (23.6%) cases of P. knowlesi, two (1.1%) cases of Plasmodium malariae and two (1.1%) mixed-species infections (i e, P. vivax/P. falciparum). Thirty-five PlasmoNex™ positive P. knowlesi samples were misdiagnosed as P. malariae by microscopy. Plasmodium knowlesi was detected in all four districts of Sandakan division with the highest incidence in the Kinabatangan district. Thrombocytopaenia and anaemia showed to be the most frequent malaria-associated haematological complications in this study.ConclusionsThe discovery of P. knowlesi in Sandakan division showed that prospective studies on the epidemiological risk factors and transmission dynamics of P. knowlesi in these areas are crucial in order to develop strategies for effective malaria control. The availability of advanced diagnostic tool PlasmoNex™ enhanced the accuracy and accelerated the speed in the diagnosis of malaria.

Developing and evaluating health education learning package (HELP) to control soil-transmitted helminth infections among Orang Asli children in Malaysia

BackgroundThis study was carried out to develop a health education learning package (HELP) about soil-transmitted helminth (STH) infections, and to evaluate what impact such a package could have in terms of reducing the incidence and intensity of STH infections among Orang Asli schoolchildren in Pahang, Malaysia.MethodsTo identify the key risk factors of STH in Orang Asli communities, we applied an extensive mixed methods approach which involved an intensive literature review, as well as community-based discussions with children, their parents, teachers and health personnel, whilst also placing the children under direct observation. To evaluate the package, 317 children from two schools in Lipis, Pahang were screened for STH infections, treated by a 3-day course of albendazole and then followed up over the next 6xa0months. The knowledge of teachers, parents and children towards STH infections were assessed at baseline and after 3xa0months.ResultsThe developed package consists of a half day workshop for teachers, a teacher’s guide book to STH infections, posters, a comic book, a music video, a puppet show, drawing activities and an aid kit. The package was well-received with effective contributions being made by teachers, children and their parents. The incidence rates of hookworm infection at different assessment points were significantly lower among children in the intervention school compared to those in the control school. Similarly, the intensity of trichuriasis, ascariasis and hookworm infections were found to be significantly lower among children in the HELP group compared to those in the control group (Pu2009<u20090.05). Moreover, the package significantly improved the knowledge, attitude and practices (KAP) of Orang Asli people and the knowledge of teachers towards STH infections.ConclusionA school-based health education learning package (HELP) was developed which displayed a significant impact in terms of reducing the intensity of all three main STH infections, as well as in reducing the prevalence of hookworm infections. Moreover, the knowledge levels of both teachers and the Orang Asli population regarding STH was significantly improved, a fact which greatly helped in attracting community participation and thus raising the general level of awareness regarding these forms of infections.

Malaria in Hadhramout, a southeast province of Yemen: prevalence, risk factors, knowledge, attitude and practices (KAPs)

BackgroundYemen is a Mediterranean country where 65% of its population is at risk of malaria, with 43% at high risk. Yemen is still in the control phase without sustainable reduction in the proportion of malaria cases. A cross-sectional household survey was carried out in different districts in the southeast of the country to determine malaria prevalence and identify factors that impede progress of the elimination phase.MethodsBlood specimens were collected from 735 individuals aged 1–66 years. Plasmodium species were detected and identified by microscopic examination of Giemsa-stained thick and thin blood smears. A household-based questionnaire was used to collect demographic, socioeconomic and environmental data.ResultsThe overall prevalence of malaria was 18.8% with Plasmodium falciparum as the predominant species (99.3%), with a low rate of Plasmodium vivax detected (0.7%). The infection rate was higher in Al-Raydah and Qusyer districts (21.8%) compared to Hajer district (11.8%). Fifty-two percent of the persons positive for Plasmodium were asymptomatic with low parasite density. The adults had a higher infection rate as compared to children. Univariate analysis identified those whose household’s head are fishermen (ORu2009=u200911.3, 95% CI: 3.13 – 40.5) and farmers (ORu2009=u20094.84, 95% CI: 1.73 – 13.6) as high-risk groups. A higher number of positive smears were observed in people living in houses with uncemented brick walls (ORu2009=u20092.1, 95% CI: 1.32 – 3.30), without access to toilets (ORu2009=u20091.6, 95% CI: 1.05 – 2.32), without a fridge (ORu2009=u20091. 6, 95% CI: 1.05 – 2.30), or without TV (ORu2009=u20091. 6, (95% CI: 1.05 – 2.30). People living in houses with water collection points located less than 200 meters away were also at higher risk of acquiring malaria (ORu2009=u20091.6, 95% CI: 1.05 – 2.30). Knowledge about the importance of using insecticide-treated mosquito nets (ITNs) and indoor residual spraying (IRS) for prevention of malaria was 7% and 2%, respectively.ConclusionsSeveral environmental, socioeconomic and behavioral issues were discovered to be the contributing factors to the high prevalence of malaria in southeast Yemen. Novel strategies adapted to the local situations need to be established in order to improve the effectiveness of malaria control.

Plasmodium ovale infection in Malaysia: first imported case

BackgroundPlasmodium ovale infection is rarely reported in Malaysia. This is the first imported case of P. ovale infection in Malaysia which was initially misdiagnosed as Plasmodium vivax.MethodsPeripheral blood sample was first examined by Giemsa-stained microscopy examination and further confirmed using a patented in-house multiplex PCR followed by sequencing.Results and DiscussionInitial results from peripheral blood smear examination diagnosed P. vivax infection. However further analysis using a patented in-house multiplex PCR followed by sequencing confirmed the presence of P. ovale. Given that Anopheles maculatus and Anopheles dirus, vectors of P. ovale are found in Malaysia, this finding has significant implication on Malaysias public health sector.ConclusionsThe current finding should serve as an alert to epidemiologists, clinicians and laboratory technicians in the possibility of finding P. ovale in Malaysia. P. ovale should be considered in the differential diagnosis of imported malaria cases in Malaysia due to the exponential increase in the number of visitors from P. ovale endemic regions and the long latent period of P. ovale. It is also timely that conventional diagnosis of malaria via microscopy should be coupled with more advanced molecular tools for effective diagnosis.

Waterborne parasites: a current status from the Philippines

BackgroundDespite the amount of awareness created, waterborne disease still poses threat, especially in developing countries. Due to the scarcity of reported data on waterborne parasites, the consumption of unsafe water prolongs. Thus, the occurrences of waterborne parasites from various samples were investigated from one of the Southeast Asian country, the Philippines.MethodsA total of thirty three samples, each consisting of twelve liters, were collected and processed to obtain the sediment. Ten liters of sample each was processed to detect Cryptosporidium spp. and Giardia spp. using an immunomagnetic separation method prior to enumeration via fluorescence microscope. Meanwhile, the remaining two liters were cultured to detect Acanthamoeba and Naegleria through microscopy examination and polymerase chain reaction (PCR) analysis.ResultsTwelve samples (36.4%) from river (5), swimming pool (1), pond (3), rain tank (1), and natural lake (2) were positive for Cryptosporidium spp., 17 (45.5%) samples from river (9), pond (2), swimming pool (1), rain tank (1), and natural lake (4) were positive for Giardia spp. while, 13 (33.3%) samples from river (3), swimming pool (2), pond (2), dispenser (1), well (1), tap (2) and natural lake (2) were positive for Acanthamoeba spp. and 5 (18.2%) samples from river (1), natural lake (1), tap (1), dispenser (1) and mineral (1) were Naegleria spp. positive. Physical parameters such as temperature, conductivity, total dissolved solid (TDS), salinity, dissolved oxygen (DO), pH, and turbidity and chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite were also measured. The highest chemical contamination was observed at pond 2. A good correlation was observed between Giardia and nitrite (ru2009=u20090.736, pu2009<u20090.01) and Giardia and nitrate (ru2009=u20090.502, pu2009<u20090.01).ConclusionThis study was aimed to create greater awareness of parasitic contamination in the water environment in the Philippines and also to act as a platform of the current scenario for policymakers as water pollution is a key health issue in this region.

Aquatic biomonitoring of Giardia cysts and Cryptosporidium oocysts in peninsular Malaysia

An aquatic biomonitoring of Giardia cysts and Cryptosporidium oocysts in river water corresponding to five villages situated in three states in peninsular Malaysia was determined. There were 51.3xa0% (20/39) and 23.1xa0% (9/39) samples positive for Giardia and Cryptosporidium (oo)cysts, respectively. Overall mean concentration between villages for Giardia cysts ranged from 0.10 to 25.80 cysts/l whilst Cryptosporidium oocysts ranged from 0.10 to 0.90 oocysts/l. Detailed results of the river samples from five villages indicated that Kuala Pangsun 100xa0% (9/9), Kemensah 77.8xa0% (7/9), Pos Piah 33.3xa0% (3/9) and Paya Lebar 33.3xa0% (1/3) were contaminated with Giardia cysts whilst Cryptosporidium (oo)cysts were only detected in Kemensah (100xa0%; 9/9) and Kuala Pangsun (66.6xa0%; 6/9). However, the water samples from Bentong were all negative for these waterborne parasites. Samples were collected from lower point, midpoint and upper point. Midpoint refers to the section of the river where the studied communities are highly populated. Meanwhile, the position of the lower point is at least 2xa0km southward of the midpoint and upper point is at least 2xa0km northward of the midpoint. The highest mean concentration for (oo)cysts was found at the lower points [3.15u2009±u20096.09 (oo)cysts/l], followed by midpoints [0.66u2009±u20091.10 (oo)cysts/l] and upper points [0.66u2009±u20090.92 (oo)cysts/l]. The mean concentration of Giardia cysts was highest at Kuala Pangsun (i.e. 5.97u2009±u20097.0 cysts/l), followed by Kemensah (0.83u2009±u20090.81 cysts/l), Pos Piah (0.20u2009±u20090.35 cysts/l) and Paya Lebar (0.10u2009±u20090.19 cysts/l). On the other hand, the mean concentration of Cryptosporidium oocysts was higher at Kemensah (0.31u2009±u20090.19 cysts/l) compared to Kuala Pangsun (0.03u2009±u20090.03cysts/l). All the physical and chemical parameters did not show significant correlation with both protozoa. In future, viability status and molecular characterisation of Giardia and Cryptosporidium should be applied to identify species and genotypes/subgenotypes for better understanding of the epidemiology of these waterborne parasites.

Inferred metagenomic comparison of mucosal and fecal microbiota from individuals undergoing routine screening colonoscopy reveals similar differences observed during active inflammation.

The mucosal microbiota lives in close proximity with the intestinal epithelium and may interact more directly with the host immune system than the luminal/fecal bacteria. The availability of nutrients in the mucus layer of the epithelium is also very different from the gut lumen environment. Inferred metagenomic analysis for microbial function of the mucosal microbiota is possible by PICRUSt. We recently found that by using this approach, actively inflamed tissue of ulcerative colitis (UC) patients have mucosal communities enriched for genes involved in lipid and amino acid metabolism, and reduced for carbohydrate and nucleotide metabolism. Here, we find that the same bacterial taxa (e.g. Acinetobacter) and predicted microbial pathways enriched in actively inflamed colitis tissue are also enriched in the mucosa of subjects undergoing routine screening colonoscopies, when compared with paired samples of luminal/fecal bacteria. These results suggest that the mucosa of healthy individuals may be a reservoir of aerotolerant microbial communities expanded during colitis.

Molecular detection of human Plasmodium species in Sabah using PlasmoNex™ multiplex PCR and hydrolysis probes real-time PCR

BackgroundMalaria is a vector borne-parasitic disease transmitted through the bite of the infective female Anopheles mosquitoes. Five Plasmodium species have been recognized by World Health Organization (WHO) as the causative agents of human malaria. Generally, microscopic examination is the gold standard for routine malaria diagnosis. However, molecular PCR assays in many cases have shown improvement on the sensitivity and specificity over microscopic or other immunochromatographic assays.MethodsThe present study attempts to screen 207 suspected malaria samples from patients seeking treatment in clinics around Sabah state, Malaysia, using two panels of multiplex PCRs, conventional PCR system (PlasmoNex™) and real-time PCR based on hydrolysis probe technology. Discordance results between two PCR assays were further confirmed by sequencing using 18S ssu rRNA species-specific primers.ResultsOf the 207 malaria samples, Plasmodium knowlesi (73.4% vs 72.0%) was the most prevalent species based on two PCR assays, followed by Plasmodium falciparum (15.9% vs 17.9%), and Plasmodium vivax (9.7% vs 7.7%), respectively. Neither Plasmodium malariae nor Plasmodium ovale was detected in this study. Nine discrepant species identification based on both the PCR assays were further confirmed through DNA sequencing. Species-specific real-time PCR only accurately diagnosed 198 of 207 (95.7%) malaria samples up to species level in contrast to PlasmoNex™ assay which had 100% sensitivity and specificity based on sequencing results.ConclusionsMultiplex PCR accelerate the speed in the diagnosis of malaria. The PlasmoNex™ PCR assay seems to be more accurate than real-time PCR in the speciation of all five human malaria parasites. The present study also showed a significant increase of the potential fatal P. knowlesi infection in Sabah state as revealed by molecular PCR assays.

Determining intestinal parasitic infections (IPIs) in inmates from Kajang Prison, Selangor, Malaysia for improved prison management

BackgroundThe prison management in Malaysia is proactively seeking to improve the health status of the prison inmates. Intestinal parasitic infections (IPIs) are widely distributed throughout the world and are still gaining great concern due to their significant morbidity and mortality among infected humans. In Malaysia, there is a paucity of information on IPIs among prison inmates. In order to further enhance the current health strategies employed, the present study aims to establish firm data on the prevalence and diversity of IPIs among HIV-infected and non-HIV-infected individuals in a prison, an area in which informed knowledge is still very limited.MethodsSamples were subjected to microscopy examination and serological test (only for Strongyloides). Speciation for parasites on microscopy-positive samples and seropositive samples for Strongyloides were further determined via polymerase chain reaction. SPSS was used for statistical analysis.ResultsA total of 294 stool and blood samples each were successfully collected, involving 131 HIV positive and 163 HIV negative adult male inmates whose age ranged from 21xa0toxa069-years-old. Overall prevalence showed 26.5xa0% was positive for various IPIs. The IPIs detected included Blastocystis sp., Strongyloides stercoralis, Entamoeba spp., Cryptosporidium spp., Giardia spp., and Trichuris trichiura. Comparatively, the rate of IPIs was slightly higher among the HIV positive inmates (27.5xa0%) than HIV negative inmates (25.8xa0%). Interestingly, seropositivity for S. stercoralis was more predominant in HIV negative inmates (10.4xa0%) compared to HIV-infected inmates (6.9xa0%), however these findings were not statistically significant. Polymerase chain reaction (PCR) confirmed the presence of Blastocystis, Strongyloides, Entamoeba histolytica and E. dispar.ConclusionsThese data will enable the health care providers and prison management staff to understand the trend and epidemiological situations in HIV/parasitic co-infections in a prison. This information will further assist in providing evidence-based guidance to improve prevention, control and management strategies of IPIs co-infections among both HIV positive and HIV negative inmates in a prison environment.

Epidemiological Characteristics of Strongyloidiasis in Inhabitants of Indigenous Communities in Borneo Island, Malaysia

Epidemiological study on strongyloidiasis in humans is currently lacking in Malaysia. Thus, a cross-sectional study was carried out to determine the prevalence of Strongyloides stercoralis infection among the inhabitants of longhouse indigenous communities in Sarawak. A single stool and blood sample were collected from each participant and subjected to microscopy, serological and molecular techniques. Five species of intestinal parasites were identified by stool microscopy. None of the stool samples were positive for S. stercoralis. However, 11% of 236 serum samples were seropositive for strongyloidiasis. Further confirmation using molecular technique on stool samples of the seropositive individuals successfully amplified 5 samples, suggesting current active infections. The prevalence was significantly higher in adult males and tended to increase with age. S. stercoralis should no longer be neglected in any intestinal parasitic survey. Combination of more than 1 diagnostic technique is necessary to increase the likelihood of estimating the ‘true’ prevalence of S. stercoralis.