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Fulltext Helminth communities from two urban rat populations in Kuala Lumpur, Malaysia

Mohd Zain SN, Behnke JM, Lewis JW

Parasit Vectors, 2012 Mar 07;5:47.
PMID: 22397763 DOI: 10.1186/1756-3305-5-47

BACKGROUND: The prevalence of parasitic infections among commensal animals such as black and brown rats in many tropical countries is high and in comparison with studies on rodents in temperate climates, little is known about the community structure of their parasites. Rodent borne parasites pose threats to human health since people living in close proximity to rodent populations can be exposed to infection.
METHODS: The helminth community structures of two urban rat populations in Kuala Lumpur, Malaysia were investigated. The rats were from two contrasting sites in the city caught over a period of 21 months in 2000-2002.
RESULTS: Eleven species of helminth parasites comprising seven nematodes (Heterakis spumosum, Mastophorus muris, Nippostrongylus brasiliensis, Syphacia muris, Pterygodermatites tani/whartoni, Gongylonema neoplasticum, Angiostrongylus malaysiensis), three cestodes (Hymenolepis (Rodentolepis) nana, H. diminuta and Taenia taeniaeformis) and one acanthocephalan (Moniliformis moniliformis) were recovered from 346 Rattus rattus and 104 R. norvegicus from two urban sites, Bangsar and Chow Kit, during 2000-2002. Rattus rattus harboured over 60% of all helminths compared with R. norvegicus, although both host species played a dominant role in the different sites with, for example R. norvegicus at Bangsar and R. rattus at Chow Kit accounting for most of the nematodes. Overall 80% of rats carried at least one species of helminth, with the highest prevalences being shown by H. diminuta (35%), H. spumosum (29.8%) and H. nana (28.4%). Nevertheless, there were marked differences in prevalence rates between sites and hosts. The influence of extrinsic (year, season and site) and intrinsic (species, sex and age) factors affecting infracommunity structure (abundance and prevalence of infection) and measures of component community structure were analyzed.
CONCLUSIONS: Since at least two species of rat borne helminths in Kuala Lumpur have the potential to infect humans, and these showed high prevalences in the rats, the assessment and regular monitoring of infections carried by wild rodents have important roles to play in public health.
The anthelmintic efficacy of natural plant cysteine proteinases against two rodent cestodes Hymenolepis diminuta and Hymenolepis microstoma in vitro

Mansur F, Luoga W, Buttle DJ, Duce IR, Lowe A, Behnke JM

Vet Parasitol, 2014 Mar 17;201(1-2):48-58.
PMID: 24462509 DOI: 10.1016/j.vetpar.2013.12.018

Little is known about the efficacy of cysteine proteinases (CP) as anthelmintics for cestode infections. We examined the effects of CPs on two rodent cestodes, Hymenolepis diminuta and H. microstoma in vitro. Our data showed that naturally occurring mixtures of CPs, such as those found in papaya latex, and relatively pure preparations of fruit bromelain, papain and stem bromelain, were active in vitro against both juvenile, artificially excysted scoleces, as well as against adult worms of both rodent cestodes. Significant dose-dependent reduction in motility, ultimately leading to death of the worms, was observed with both species, and against both freshly excysted scoleces and 14-day old pre-adult worms. The most effective was fruit bromelain (after 30 min of incubation of juvenile H. diminuta and H. microstoma IC50=63 and 74 μM, respectively, and for pre-adult worms=199 and 260 μM, respectively). The least effective was stem bromelain (after 30 min of incubation of juvenile H. diminuta and H. microstoma IC50=2855 and 2772 μM, respectively, and for pre-adult worms=1374 and 1332 μM, respectively) and the efficacies of papaya latex supernatant and papain were between these extremes. In all cases these values are higher than those reported previously for efficacy of CPs against intestinal nematodes, and in contrast to nematodes, all CPs were effective against cestodes in the absence of exogenous cysteine in incubation media. The CPs appeared to attack the tegument resulting in generalised erosion mainly on the strobila. The scolex was more resistant to CP attack but nevertheless some damage to the tegument on the scolex was detected.
Seroprevalence and Associated Risk Factors for Toxoplasma gondii Infections Among Urban Poor Communities in Peninsular Malaysia

Sahimin N, Mohd Hanapi IR, Nurikhan ZA, Behnke JM, Mohd Zain SN

Acta Parasitol, 2021 Jun;66(2):524-534.
PMID: 33219942 DOI: 10.1007/s11686-020-00304-0

PURPOSE: Toxoplasmosis is a zoonotic infection linked to compromised hygiene and sanitation via the handling of infected cat faeces, eating undercooked contaminated meat or transplacental transmission. We conducted a study to determine seroprevalence and risk factors associated with toxoplasmosis among the urban poor communities in Malaysia.
METHODS: The demographic profiles for each participant were obtained through a questionnaire survey prior to blood collection. A total of 389 participants were recruited and blood samples screened for the presence of anti-Toxoplasma IgG and IgM antibody using an ELISA commercial kit, SERION ELISA classic Toxoplasma gondii IgG and IgM.
RESULTS: The overall T. gondii seroprevalence was 69.6% with 56.8% seropositive for anti-Toxoplasma IgG, 7.7% seropositive for anti-Toxoplasma IgM and 5.1% seropositive for both IgG and IgM antibodies. The presence of both antibody classes in blood samples indicated high avidity, suggesting latent infection. Univariate analysis revealed significant associations that included; age, ethnicity, location and employment status while, significant lifestyle factors included source of drinking water and eating style. A multifactorial statistical model that incorporated all the significant effects from the first-stage univariate analyses listed above revealed that age and ethnicity were the two dominant and independent effects on IgG seroprevalence. For seroprevalence of IgM, the multifactorial model revealed a significant interaction between work and accommodation. IgM seroprevalence was higher among the unemployed inhabitants of PPR (Program Perumahan Rakyat) than those living in non-PPR accommodation, and higher than among the employed irrespective of their accommodation.
CONCLUSION: High seroprevalence of Toxoplasmosis in the community calls for increased awareness of disease transmission and improvements in hygiene and sanitation.
Entamoeba infections and associated risk factors among migrant workers in Peninsular Malaysia

Sahimin N, Yunus MH, Douadi B, Yvonne Lim AL, Noordin R, Behnke JM, et al.

Trop Biomed, 2019 Dec 01;36(4):1014-1026.
PMID: 33597471

The influx of low skilled migrant workers to Malaysia from low socio-economic countries where gastrointestinal parasitic infections are prevalent has raised concerns about transmission to the local population. Three methods for detection (serology, microscopy and molecular techniques) were utilized to identify Entamoeba infections amongst the targeted cohort and determine risk factors associated with infection. Serological screening of 484 migrant workers from five working sectors in Peninsular Malaysia using IgG4 ELISA based on the rPPDK antigen showed an overall seroprevalence of 7.4% (n = 36; CL95 = 5.3-10.1%) with only one factor statistically associated with seropositivity of anti-amoebic antibodies, i.e. years of residence in Malaysia (χ2 1 = 4.007, p = 0.045). Microscopic examination of 388 faecal samples for protozoan cysts and trophozoites showed a slightly higher prevalence (11.6%; n=45; CL95: 8.4-14.8%). Meanwhile, amplification of the 16S rDNA gene detected two species i.e. Entamoeba dispar (23/388; 5.9%; CL95: 3.6-8.3%) and E. histolytica (11/388; 2.8%; CL95: 1.2-4.5%) and mixed infections with both parasites in only three samples (3/388; 0.8%; CL95: 0.2-2.2%). Entamoeba dispar infection was significantly associated with those employed in food and domestic services (χ2 4 = 12.879, p = 0.012). However, none of the factors affected the prevalence of E. histolytica infection. Despite the low prevalence of E. histolytica in faecal samples of the study cohort, the presence of this pathogenic parasite still poses potential public health risks and calls for tighter control strategies based on better availability of chemotherapeutic treatment and accessibility to appropriate health education.
Fulltext Developing novel anthelmintics: the stability of cysteine proteinase activity in a supernatant extract of papaya latex

Mansur FAF, Luoga W, Behnke JM, Buttle DJ, Duce IR, Garnett MC

Heliyon, 2021 Oct;7(10):e08125.
PMID: 34693054 DOI: 10.1016/j.heliyon.2021.e08125

Plant derived cysteine proteinases (CPs) have long been known to possess anthelmintic properties but have attracted renewed attention recently because of the acute need to discover novel methods for controlling helminth infections as a result of increasing drug resistance. However, surprisingly little is known about the stability of these proteins under typical storage and in vivo exposure conditions. We found that CPs in a supernatant preparation from papaya latex (PLS) were stable during the initial refinement process and when stored under low temperatures, but lost activity during dialysis and within 7 days of storage when kept at ambient temperature (18-20 °C). The enzyme activity in PLS was not affected by repeated freeze-thaw cycles and was also stable under typical in vitro assay conditions at 37 °C used for quantifying effects on helminths. Active enzyme activity was still detectable in the colon 3-4 h after oral administration in rodent models.
Microorganisms as an Indicator of Hygiene Status Among Migrant Food Handlers in Peninsular Malaysia

Woh PY, Thong KL, Lim YAL, Behnke JM, Lewis JW, Mohd Zain SN

Asia Pac J Public Health, 2017 Oct;29(7):599-607.
PMID: 29037053 DOI: 10.1177/1010539517735856

This study used microbial indicators to assess the hygiene status of 383 migrant food handlers from 3 urban cities in Peninsular Malaysia. Microbiological analysis revealed that all the hand swabs tested 99.5% positive for aerobic plate counts (mean [M] ± standard deviation [SD] = 3.57 ± 0.83 log10CFU [colony forming unit]), 20.8% positive for total coliform/ Escherichia coli (M ± SD = 0.30 ± 0.67 log10CFU), and 63.4% positive for Staphylococcus aureus (M ± SD = 1.38 ± 1.26 log10CFU). In addition, aerobic plate counts and Staphylococcus aureus counts exceeded the acceptable standard levels. Bacterial counts were found to be significantly associated with subjects' country of origin ( P = .019) and working responsibilities ( P = .001). Our findings indicate high probability of transmission of pathogenic bacteria from the food handlers' hands to customers during meal preparation and serving. This calls for improvements in personal hygiene and sanitation standards by the relevant health authorities among migrant food handlers.
Distribution of Giardia duodenalis (Assemblages A and B) and Cryptosporidium parvum amongst migrant workers in Peninsular Malaysia

Sahimin N, Douadi B, Yvonne Lim AL, Behnke JM, Mohd Zain SN

Acta Trop, 2018 Jun;182:178-184.
PMID: 29501402 DOI: 10.1016/j.actatropica.2018.02.033

The influx of low skilled workers from socioeconomically deprived neighbouring countries to Malaysia has raised concerns about the transmission of communicable gastrointestinal diseases such as giardiasis and cryptosporidiosis to the local population. Therefore, a cross sectional study was conducted to investigate the prevalence of both diseases and the genetic diversity of these pathogens in the migrant population. Microscopic examination of faecal samples from 388 migrant workers involved in five working sectors were screened and 10.8% (n = 42) were found to be positive with Giardia spp. and 3.1% (n = 12) with Cryptosporidium spp. infections. PCR amplicons at the triosephosphate isomerase (tpi) gene were successfully obtained for Giardia duodenalis from 30 (30/388; 7.73%) samples with assemblages AII and B in 13 (13/30; 43.3%) and 17 (17/30; 56.7%) positive samples, respectively. Nine samples (9/388; 2.3%) were identified as Cryptosporidium parvum using PCR-RFLP analysis. Country of origin, duration of residence in Malaysia and working sectors significantly influenced G. duodenalis assemblage AII infections amongst the targeted population. Meanwhile, C. parvum infection was significantly associated with those working in the food service sector. Despite the low presence of pathogenic G. duodenalis and C. parvum in the study population, the results highlight the risk of anthroponotic foodborne and waterborne transmission and therefore call for implementation of control strategies through improvements in personal hygiene and sanitation standards.
Fulltext Migrant Workers in Malaysia: Current Implications of Sociodemographic and Environmental Characteristics in the Transmission of Intestinal Parasitic Infections

Sahimin N, Lim YA, Ariffin F, Behnke JM, Lewis JW, Mohd Zain SN

PLoS Negl Trop Dis, 2016 Nov;10(11):e0005110.
PMID: 27806046 DOI: 10.1371/journal.pntd.0005110

A cross-sectional study of intestinal parasitic infections amongst migrant workers in Malaysia was conducted. A total of 388 workers were recruited from five sectors including manufacturing, construction, plantation, domestic and food services. The majority were recruited from Indonesia (n = 167, 43.3%), followed by Nepal (n = 81, 20.9%), Bangladesh (n = 70, 18%), India (n = 47, 12.1%) and Myanmar (n = 23, 5.9.2%). A total of four nematode species (Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis and hookworms), one cestode (Hymenolepis nana) and three protozoan species (Entamoeba histolytica/dispar, Giardia sp. and Cryptosporidium spp.) were identified. High prevalence of infections with A. lumbricoides (43.3%) was recorded followed by hookworms (13.1%), E. histolytica/dispar (11.6%), Giardia sp. (10.8%), T. trichura (9.5%), Cryptosporodium spp. (3.1%), H. nana (1.8%) and E. vermicularis (0.5%). Infections were significantly influenced by socio-demographic (nationality), and environmental characteristics (length of working years in the country, employment sector and educational level). Up to 84.0% of migrant workers from Nepal and 83.0% from India were infected with intestinal parasites, with the ascarid nematode A. lumbricoides occurring in 72.8% of the Nepalese and 68.1% of the Indian population. In addition, workers with an employment history of less than a year or newly arrived in Malaysia were most likely to show high levels of infection as prevalence of workers infected with A. lumbricoides was reduced from 58.2% to 35.4% following a year's residence. These findings suggest that improvement is warranted in public health and should include mandatory medical screening upon entry into the country.
Characterization of Nontyphoidal Salmonella Isolates from Asymptomatic Migrant Food Handlers in Peninsular Malaysia

Woh PY, Thong KL, Behnke JM, Lewis JW, Zain SNM

J Food Prot, 2017 Aug;80(8):1378-1383.
PMID: 28722485 DOI: 10.4315/0362-028X.JFP-16-342

Asymptomatic Salmonella carriers who work as food handlers pose food safety and public health risks, particularly during food preparation, and this has serious implications for the disease burden in society. Therefore, we conducted a study to determine the number of Salmonella carriers in a migrant cohort in several food establishments in three major cities in Peninsular Malaysia. Sociodemographic data and stool samples were collected and analyzed using standard methods of detection and isolation. Antimicrobial susceptibility tests of the positive samples were also performed. A total of 317 migrant food handlers, originating from South and Southeast Asian countries, were recruited voluntarily. Nine (2.8%) stool samples were confirmed to be Salmonella positive. PCR serotyping and pulsed-field gel electrophoresis identified four serotypes as Typhimurium (n = 3), Corvallis (n = 2), Hadar (n = 1), Agona (n = 1) and two unknown serovars. Antimicrobial susceptibility tests revealed that all nine isolates were susceptible to amoxicillin-clavulanic acid, cefotaxime, ceftazidime, ceftriaxone, and gentamycin. However, seven isolates were found to be multidrug resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, sulfonamides, streptomycin, and tetracycline. This study highlights that carriers of nontyphoidal Salmonella exist among migrant food handlers, which poses a health risk to consumers through food contamination. Our results indicate a need for authorities to enhance food safety awareness in the migrant workers and to reevaluate current health screening methods to include preventive measure such as mandatory stool screening as part of the preemployment and routine health examinations.
Hookworm infections among migrant workers in Malaysia: Molecular identification of Necator americanus and Ancylostoma duodenale

Sahimin N, Lim YAL, Douadi B, Mohd Khalid MKN, Wilson JJ, Behnke JM, et al.

Acta Trop, 2017 Sep;173:109-115.
PMID: 28610937 DOI: 10.1016/j.actatropica.2017.06.011

Ongoing urbanisation of the working population as well as cross-border migration of workers particularly into large cities has contributed to the development and growth of urban slums. These deprived areas are conducive for the transmission of intestinal pathogens including hookworm. The aim of this study was to determine both the prevalence and species identity of hookworm infections among the migrant worker community in Malaysia. A total of 388 faecal samples were collected from migrant workers between September 2014 and August 2015, representing workers from five employment sectors: construction, manufacturing, agriculture and plantations, food services and domestic services. Faecal samples were examined by microscopy and positive samples were subjected to molecular analysis. A total of 51 samples (13.1%) were positive by microscopy for hookworm infections. A two-step PCR based method amplifying a fragment of the 28S rRNA-ITS2 region was used to identify infections by Necator americanus and Ancylostoma spp. PCR products positive for Ancylostoma spp. were sequenced bidirectionally, and sequences analysed through BLAST and phylogenetic analysis. Samples containing Ancylostoma duodenale were further characterized by amplification and sequencing a fragment of cytochrome c oxidase subunit 1 (cox1) gene. PCR amplicons were successfully obtained from 42 (82.4%) of 51 samples, with 81.0% (34 of 42) identified as Necator americanus, 16.7% (7 of 42) as Ancylostoma spp. and 2.4% (1 of 42) as mixed infections of both species. All eight Ancylostoma spp. were confirmed to be Ancylostoma duodenale and this is the first time A. duodenale was reported in Malaysia. Samples containing A. duodenale from Nepalese and Indonesian workers shared high-similarity and were distinct compared to sequences from other countries. This study highlights the prevalence of hookworm infections among migrant workers living in Malaysia. Our findings underscore the necessity of screening migrant workers for hookworm infections, particularly those working in food-related services and industries.
Fulltext Socio-demographic determinants of Toxoplasma gondii seroprevalence in migrant workers of Peninsular Malaysia

Sahimin N, Lim YAL, Ariffin F, Behnke JM, Basáñez MG, Walker M, et al.

Parasit Vectors, 2017 May 15;10(1):238.
PMID: 28506241 DOI: 10.1186/s13071-017-2167-8

BACKGROUND: The number of migrants working in Malaysia has increased sharply since the 1970's and there is concern that infectious diseases endemic in other (e.g. neighbouring) countries may be inadvertently imported. Compulsory medical screening prior to entering the workforce does not include parasitic infections such as toxoplasmosis. Therefore, this study aimed to evaluate the seroprevalence of T. gondii infection among migrant workers in Peninsular Malaysia by means of serosurveys conducted on a voluntary basis among low-skilled and semi-skilled workers from five working sectors, namely, manufacturing, food service, agriculture and plantation, construction and domestic work.
METHODS: A total of 484 migrant workers originating from rural locations in neighbouring countries, namely, Indonesia (n = 247, 51.0%), Nepal (n = 99, 20.5%), Bangladesh (n = 72, 14.9%), India (n = 52, 10.7%) and Myanmar (n = 14, 2.9%) were included in this study.
RESULTS: The overall seroprevalence of T. gondii was 57.4% (n = 278; 95% CI: 52.7-61.8%) with 52.9% (n = 256; 95% CI: 48.4-57.2%) seropositive for anti-Toxoplasma IgG only, 0.8% (n = 4; 95% CI: 0.2-1.7%) seropositive for anti-Toxoplasma IgM only and 3.7% (n = 18; 95% CI: 2.1-5.4%) seropositive with both IgG and IgM antibodies. All positive samples with both IgG and IgM antibodies showed high avidity (> 40%), suggesting latent infection. Age (being older than 45 years), Nepalese nationality, manufacturing occupation, and being a newcomer in Malaysia (excepting domestic work) were positively and statistically significantly associated with seroprevalence (P
Fulltext Prevalence of anti-Leptospira antibodies and associated risk factors in the Malaysian refugee communities

Mohd Hanapi IR, Sahimin N, Maackara MJB, Annisa AS, Abdul Mutalib RNS, Lewis JW, et al.

BMC Infect Dis, 2021 Nov 01;21(1):1128.
PMID: 34724919 DOI: 10.1186/s12879-021-06830-0

BACKGROUND: Refugees in Malaysia, who are afflicted by poverty, conflict and poor health, are vulnerable to a range of zoonotic infections in the deprived environmental and social conditions under which they live. Exposure to infections such as leptospirosis, for which rodents are primary hosts, is of particular concern.
METHODS: A wellness program was conducted to determine the presence of antibodies against Leptospira (seroprevalence) in 11 refugee community schools and centers in the Klang Valley, Malaysia. A total of 433 samples were assessed for IgG and IgM antibodies against Leptospira, using enzyme-linked immunosorbent assays (ELISA).
RESULTS: Overall Leptospira seroprevalence was 24.7%, with 3.0% being seropositive for anti-Leptospira IgG and 21.7% for anti-Leptospira IgM. Factors significantly associated with overall Leptospira seroprevalence included: age, ethnicity, pet ownership, knowledge of disease and awareness of disease fatality. For IgM seroprevalence, significant risk factors included sex, ethnicity, eating habits with hands, pet ownership, the presence of rats, walking in bare feet and water recreation visits.
CONCLUSIONS: These findings highlight the need for improvements in health and well-being among the refugee community through disease awareness programs and provision of healthy behavior programs, particularly in hygiene and sanitation through community engagement activities.
Intestinal parasitic infections and risk analysis among urban refugees in the Klang Valley, Malaysia

Mohd Hanapi IR, Behnke JM, Sahimin N, Saifulazmi NF, Golam Mohammad Khan ASJ, Abdul Mutalib RNS, et al.

Trans R Soc Trop Med Hyg, 2024 Jan 02;118(1):18-32.
PMID: 37497742 DOI: 10.1093/trstmh/trad047

BACKGROUND: Global studies show intestinal parasitic infections (IPIs) have been introduced and spread with refugee inflows from low to high socio-economic countries. However, there is relatively limited information on the prevalence of infections among the community.
METHODS: A 2-year cross-sectional study was conducted to determine the prevalence and associated risk factors for infections among urban refugees in the Klang Valley, Malaysia. A total of 418 faecal samples were collected and examined by microscopy.
RESULTS: Faecal screening revealed moderate levels (32.3%) of infections in the community. Three nematode (Ascaris lumbricoides, Trichuris trichiura and hookworm) and three protozoan species (Entamoeba, Giardia and Cryptosporidium) were recorded, with the highest prevalence being A. lumbricoides (20.6%) followed by T. trichiura (10.3%), while other infections were <5%. Statistical analysis found that young males with less education were more likely to be infected with helminths. Additionally, living near waste disposal sites, the presence of stray animals, eating with bare hands, bare footedness, poor handwashing practices and no anthelmintic treatment constituted significant risk factors for helminth infections. Protozoan infections were linked to drinking tap water or from water dispensers and poor handwashing practices.
CONCLUSIONS: These findings emphasize the importance of health education in addition to introduction of biannual anthelmintic treatment to promote community health and well-being.

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