Poverty related diseases amongst Malaysia's low-income community: a review.

Poverty, as proven by several studies, is a driving force behind poor health and hygiene practices. This review attempts to outline common communicable and non-communicable diseases that disproportionately affect Malaysia's 2.91 million low-income households. The current study also looks into the government's housing and healthcare programmes for this demographic to improve their health and well-being. The initial examination yielded incredibly little research on this marginalised community, with event reporting typically generalised to the Malaysian community as a whole rather than analysing disease incidences based on household income, which would better reflect povertydriven diseases. As a result, there is an acute need for more accurate information on the epidemiology of diseases among the poor in order to address this public health issue and provide conclusions that can drive policy designs.

In silico structural modeling and quality assessment of Plasmodium knowlesi apical membrane antigen 1 using comparative protein models.

Plasmodium knowlesi is the most common zoonotic parasite associated with human malaria infection in Malaysia. Apical membrane antigen 1 (AMA1) protein in the parasite plays a critical role in parasite invasion into host cells. To date, there is no complete three-dimensional ectodomain structure of P. knowlesi AMA1 (PkAMA1) protein. The knowledge of a protein structure is important to understand the protein molecular functions. Three in silico servers with respective structure prediction methods were used in this study, i.e., SWISS-MODEL for homology modeling and Phyre2 for protein threading, which are template-based modeling, while I-TASSER for template-free ab initio modeling. Two query sequences were used in the study, i.e., native ectodomain of PkAMA1 strain H protein designated as PkAMA1-H and a modified PkAMA1 (mPkAMA1) protein sequence in adaptation for Pichia pastoris expression. The quality of each model was assessed by ProSA-web, QMEAN and SAVES v6.0 (ERRAT, Verify3D and Ramachandran plot) servers. Generated models were then superimposed with two models of Plasmodium AMA1 deposited in Protein Data Bank (PDB), i.e., PkAMA1 (4UV6.B) and Plasmodium vivax AMA1 (PvAMA1, 1W81) protein structures for similarity assessment, quantified by root-meansquare deviation (RMSD) value. SWISS-MODEL, Phyre2 and I-TASSER server generated two, one and five models, respectively. All models are of good quality according to ProSA-web assessment. Based on the average values of model quality assessment and superimposition, the models that recorded highest values for most parameters were selected as best predicted models, i.e., model 2 for both PkAMA1-H and mPkAMA1 from SWISS-MODEL as well as model 1 of PkAMA1-H and model 3 of mPkAMA1 from I-TASSER. Template-based method is useful if known template is available, but template-free method is more suitable if there is no known available template. Generated models can be used as guidance in further protein study that requires protein structural data, i.e., protein-protein interaction study.

Higher efficacy of a single dosage albendazole and different soil-transmitted helminths re-infection profiles amongst indigenous Negritos from inland jungle versus those in resettlement at town peripheries.

Demarginalization through initiation of resettlement program since 1978 is an inevitable progress faced by the indigenous Orang Asli (OA) population in Peninsular Malaysia. As Malaysian huntergatherers, the Negrito has been exposed to various environmental-cultural variations. These changes may influence the pattern of soil-transmitted helminth (STH) infections, the common malady amongst OA. This study evaluated the deworming effects of single-dosage albendazole (400 mg) and STH-reinfection rate between Negritos who are still living in the inland jungle versus those living in resettlements at town peripheries (RPS). Stool samples from the consented participants were first examined using the direct faecal smear, formalin-ether sedimentation and Kato Katz techniques. Subsequently, stool collections were carried out in three time points following treatment (i.e., 21 days, 3 months and 6 months). In brief, a total number of 54 Negritos (inland: 24; RPS: 30) with a complete set of stool collection was included in this longitudinal study. This study revealed 72.2% cure rate against T. trichiura in the inland but only 15.0% in the RPS. Although the efficacy of albendazole against T. trichiura was ultimately low in the RPS, 62.6% egg reduction rate (ERR) (arithmetic mean) was noted (p = 0.001). For A. lumbricoides and hookworm, high cure rates were found in both communities (85.7-100.0%). Reinfection for T. trichiura was seen in less than 1 month with higher rate in the RPS (90.0%) as opposed to the inland (44.4%) at 21 days following treatment. This study found that the inland OA had better tolerability to single-dosage albendazole and experienced slower STH reinfection rates versus the RPS. Hence, the selection of albendazole dosage should be targeted and the use of single- dosage albendazole (biannually) would be more suitable for the inland OA. Conversely, we propose the use of 3-days albendazole regimens in the resettled RPS population.

Imported case ofLeishmania tropica cutaneous leishmaniasis in a 10-year-old child in Malaysia.

The present paper reported a first imported case of cutaneous leishmaniasis in a 10-year- old child who returned from Saudi Arabia to Malaysia. Six weeks after his travel to Malaysia, two erythematous dermal nodules were developed over his right cheek and chin. Occurrence of intracellular amastigote of Leishmania was observed through examination of skin biopsy with hematoxylin and eosin stain. Furthermore, molecular analysis of ribosomal internal transcribed spacer 1 (ITS1) of Leishmania spp. confirmed the child was infected with Leishmania tropica. The child was given oral fluconazole and he had a 80% recovery before he went back to Saudi Arabia.

A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces: development and validation.

Soil-transmitted helminth (STH) infections, mainly caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms, are among the most common intestinal parasites that infect humans. The infections are widely distributed throughout tropical and subtropical countries, including Malaysia, particularly in underprivileged communities. Microscopic and culture techniques have been used as a gold standard for diagnostic techniques. However, these methods yield low sensitivity and specificity, laborious and time-consuming. Therefore, simple, rapid, and accurate alternative methods are needed for the simultaneous detection of STH infections. Although advanced technologies such as real-time multiplex PCR have been established, the use of this technique as a routine diagnostic is limited due to the high cost of the instrument. Therefore, a single-round multiplex conventional PCR assay for rapid detection of four STH species in the fecal sample was developed in this study. To perform the single-round multiplex PCR, each pair of species-specific primers was selected from target genes, including Ancylostoma duodenale (Internal Transcribed Spacer 2; accession No. AJ001594; 156 base pair), Necator americanus (ITS 2; accession No. AJ001599; 225 base pair), Ascaris lumbricoides (Internal Transcribed Spacer 1; accession No. AJ000895; 334 base pair) and Trichuris triciura (partial ITS 1, 5.8s rRNA and partial ITS 2; accession No. AM992981; 518 base pair). The results showed that the newly designed primers could detect the DNA of STH at low concentrations (0.001 ng/ µl) with no cross-amplification with other species. This assay enables the differentiation of single infections as well as mixed infections. It could be used as an alternative and is a convenient method for the detection of STHs, especially for the differentiation of N. americanus and A. duodenale.

Bioefficacy of mosquito mat vaporizers and associated metabolic detoxication mechanisms in Aedes aegypti (Linnaeus) in Selangor, Malaysia: A statewide assessment.

This study aims to examine the efficacy of mosquito mat vaporizers on Aedes aegypti and their associated metabolic detoxication mechanisms. For this purpose, Aedes aegypti (Linnaeus) was collected from nine districts in Selangor, Malaysia and tested with mosquito vaporizing mat bioassays. The same populations were also subjected to biochemical assays to investigate activities of detoxifying enzymes, namely non-specific esterase (EST), glutathione-S-transferase (GST) and mixed function oxidase (MFO). The efficacy of Ae. aegypti on the active ingredients tested in decreasing order were d- allethrin > dimefluthrin > prallethrin with PBO > prallethrin. The results further indicated significant enhancement mean levels of EST, GST and MFO in pyrethroid-resistant populations. The mortality rate of Ae. aegypti in response to pyrethroid active ingredients was associated with MFO activity, suggesting it is an important detoxification enzyme for the populations tested. In view of the presence of resistance against household insecticide products, pyrethroid efficacy on Ae. aegypti populations needs to be monitored closely to ensure the implementation of an effective vector control program in Malaysia.

Epidemiological study of human intestinal parasites in Sarawak, East Malaysia: A review.

Intestinal parasitic infections are endemic in rural settings and may account for asymptomatic infections to various health complications. These infections are a cause of concern for communities of lower economic status, especially in developing countries. In Sarawak, indigenous populations residing in geographically inaccessible areas are socially and economically disadvantaged. Through close association with nature, these populations are prone to intestinal parasitism. Currently, scattered information has led to a continual state of neglect at each level of parasitic infection control. This urges for a review of their distribution and transmission based on previous reports to understand the pattern of the diseases in the state which can further address the improvement of mass controlling programs. A literature search was conducted to collect previous reports on human intestinal parasites in Sarawak, East Malaysia from PubMed (Medline), SCOPUS, ScienceDirect and Web of Science from January 2019 to March 2021. Extrapolating the current data in Sarawak which is still considered limited, further interdisciplinary strategies are demanded to give insights in the epidemiology and true prevalence of intestinal parasites in Sarawak. This review addresses for redirection of attitude towards intestinal parasitic infections where it should be given with ample attention by rural populations. In tandem to that, improvement of rural livelihood such as standard of living and sanitation in Sarawak should be accredited as part of the efforts to reduce the number of intestinal parasitic infections in the state. As a control measure, mass deworming should be reconsidered especially to the rural populations.

Bioinformatics characterization of Plasmodium knowlesi apical membrane antigen 1 (PkAMA1) for multi-epitope vaccine design.

Malaria caused by Plasmodium knowlesi species has become a public health concern, especially in Malaysia. Plasmodium knowlesi parasite which originates from the macaque species, infects human through the bite of the Anopheles mosquitoes. Research on malaria vaccine has been a continuous effort to eradicate the malaria infection, yet there is no vaccine against P. knowlesi malaria to date. Apical membrane antigen 1 (AMA1) is a unique surface protein of all apicomplexan parasites that plays a crucial role in parasite-host cell invasion and thus has been a long-standing malaria vaccine candidate. The selection of protective epitopes in silico has led to significant advances in the design of the vaccine. The present study aimed to employ bioinformatics tools to predict the potential immunogenic B- and T-cell epitopes in designing malaria vaccine targeting P. knowlesi AMA1 (PkAMA1). B-cell epitopes were predicted using four bioinformatics tools, i.e., BepiPred, ABCpred, BcePred, and IEDB servers whereas T-cell epitopes were predicted using two bioinformatics servers, i.e., NetMHCpan4.1 and NetMHCIIpan-4.0 targeting human major histocompatibility complex (MHC) class I and class II molecules, respectively. The antigenicity of the selected epitopes computed by both B- and T-cell predictors were further analyzed using the VaxiJen server. The results demonstrated that PkAMA1 protein encompasses multi antigenic regions that have the potential for the development of multi-epitope vaccine. Two B- and T-cell epitopes consensus regions, i.e., NSGIRIDLGEDAEVGNSKYRIPAGKCP (codons 28-54) and KTHAASFVIAEDQNTSY RHPAVYDEKNKT (codons 122-150) at domain I (DI) of PkAMA1 were reported. Advancement of bioinformatics in characterization of the target protein may facilitate vaccine development especially in vaccine design which is costly and cumbersome process. Thus, comprehensive B-cell and T-cell epitope prediction of PkAMA1 offers a promising pipeline for the development and design of multi-epitope vaccine against P. knowlesi.

Oral bacteria detection among children with cancer in a tertiary teaching hospital in Kuala Lumpur, Malaysia.

This study sought to determine the prevalence of pathogenic and non-pathogenic bacteria in the oral cavities of children with cancer. There were 68 paediatric patients with cancer who were included in this study. Oral swab samples from the dorsum of tongues and mouth floors of these patients were subjected to culture, staining, and molecular methods to detect the bacteria. The overall prevalence of gram-positive and gram-negative bacteria was 79.4% (54/68; 95% CI = 68.4 - 87.3) and 25% (17/68; 95% CI = 16.2 - 36.4), respectively. Streptococcus salivarius and Streptococcus parasanguinis were the predominant pathogenic grampositive bacteria, while Neisseria subflava and Neisseria perflava were the most common pathogenic gram-negative bacteria. The results revealed that the number of bacteria isolates recovered in patients receiving cancer treatment was higher (55.9%) than those who had not received treatment (16.2%). Therefore, more isolated pathogenic bacteria were observed post-therapy (54.4%). Pathogenic organisms can have significant implications on patient health. Awareness of the types of bacteria inhabiting the oral cavity is essential to predict and prevent dental problems, and their associated systemic complications. Findings on the diversity of oral microflora can also provide a better understanding of the aetiology of oral diseases in paediatric patients receiving cancer treatment.

Serological survey of canine vector-borne diseases in two animal shelters in central Peninsular Malaysia.

Canine vector-borne diseases (CVBDs) are increasingly becoming a cause for global concern because of their high morbidity and mortality rates in dogs. However, information on their occurrence in Malaysia is still scanty. In this study, a total of 103 dog blood samples were collected from two animal shelters in central Peninsular Malaysia and tested for the antibodies against Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and the antigen of Dirofilaria immitis. Of the 103 tested dogs, 44.7% (46) were found to be seropositive for Ehrlichia spp., 30.1% (31) for Anaplasma spp. and 13.6% (14) for D. immitis. Co-infections of Anaplasma spp. + Ehrlichia spp. (18.5%, 19) were most prevalent, followed by Anaplasma spp. + D. immitis (1.9%; two) and D. immitis + Ehrlichia spp. (1.0%; one). Furthermore, three dogs (2.9%) were also found to have triple infection, testing seropositive for Ehrlichia spp., Anaplasma spp. and D. immitis. The dogs which were found to be seropositive with at least one pathogen were 66.7% (32/51) at shelter A, and 55.8% (29/52) at shelter B. Serological evidence showed that the exposure of major vector-borne diseases in dogs in shelters was relatively high in the surveyed areas. Routine detection and control of vector-borne diseases are of paramount importance for reducing the risk of CVBDs transmission in dogs and humans.

Molecular detection of porcine Enterocytozoon bieneusi infection in Peninsular Malaysia and epidemiological risk factors associated with potentially zoonotic genotypes.

Enterocytozoon bieneusi is an emerging opportunistic pathogen infecting humans, and both domestic and wild pigs are known to harbour zoonotic genotypes. There remains a paucity of information on the prevalence and epidemiology of this enteropathogen in Southeast Asia. The present study was undertaken to determine the molecular prevalence and risk factors associated with E. bieneusi infection among commercially farmed pigs in Malaysia. Faecal samples were collected from 450 pigs from 15 different farms and subjected to nested PCR amplification of the ribosomal internal transcribed spacer (ITS) gene of E. bieneusi. Phylogenetic analysis involved 28 nucleotide sequences of the ITS region of E. bieneusi. An interviewer-administered questionnaire provided information on the animal hosts, farm management systems and environmental factors and was statistically analysed to determine the risk factors for infection. The prevalence of E. bieneusi infection was relatively high (40.7%). The highest prevalence (51.3%) was recorded among the piglets, while the adults showed the lowest level of infection (31.3%). Multivariate analysis indicated that age of the pigs, distance of the farm from human settlement and farm management system were significant risk factors of infection. Three genotypes (EbpA, EbpC and Henan-III) detected among the pigs are potentially zoonotic. The high prevalence of E. bieneusi among locally reared pigs, the presence of zoonotic genotypes and the spatial distribution of pig farms and human settlements warrant further investigation on the possibility of zoonotic transmission.

Toxoplasma gondii infection among selected indigenous community in Sarawak, East Malaysia.

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis in humans. To date, little is known about T. gondii infection among the indigenous community, particularly in East Malaysia. This study was conducted to determine the status of T. gondii infection and to investigate associated risk factors among the indigenous community of Sarawak, East Malaysia. The sociodemographic data was obtained using a pretested questionnaire. A serological test was done to detect the presence of specific IgM and IgG antibodies against T. gondii in serum samples. A nested polymerase chain reaction (PCR) was used to determine acute infection among seropositive individuals. The overall seroprevalence of T. gondii infection was 50% (95% CI = 43.3 - 56.7). From this subset, 40.1%, 5.7%, and 4.2% were positive for anti-T. Gondii IgG antibodies, IgM, and both IgG and IgM, respectively. Four seropositive samples were amplified through PCR. None of the pregnant women tested positive for T. gondii infection based on the serological and PCR assays. A significant association was found between age, low monthly household income, unemployment, usage of untreated water and close contact with T. gondii seropositive cats. These results provide basic information on T. gondii infection and may be useful for policymakers to initiate prevention and control programs, especially amongst pregnant women and women of childbearing age in the indigenous community.

Detection of human Sarcocystosis using dried blood on filter papers: An Immunofluorescent Antibody Test.

Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats' muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis.

Genetic diversity of Merozoite Surface Protein-1 gene block 2 allelic types in Plasmodium falciparum isolates from Malaysia and Thailand.

Malaria is the most common vector-borne parasitic disease in Malaysia and Thailand, especially in Malayan Borneo and along the Thailand border areas, but little is known about the genetic diversity of the parasite. Present study aims to investigate the genetic diversity of Plasmodium falciparum isolates in these two countries and eventually contributes to more effective malaria control strategies, particularly in vaccine and antimalarial treatment. One hundred and seventy three P. falciparum isolates were collected from Malaysia (n = 67) and Thailand (n = 106) and genotyped using nested PCR targeting the polymorphic region of MSP-1, block 2. Sequence analysis was conducted to investigate the allele diversity of the isolates. Three allelic families were identified in Malaysian and Thailand P. falciparum isolates, MAD20, K1 and RO33. Sequence analysis revealed that there were 5 different MAD20, 1 K1 and 2 different RO33 for Malaysian isolates. Thailand isolates exhibited greater polymorphism because there were 13 different MAD20, 6 different K1 and 2 different RO33 identified in this study. Multiclonal infections were observed for the isolates in both countries, however, low multiplicity of infection (MOI) was observed for Malaysian (1.1) and Thailand (1.2) isolates. Phylogenetic analysis showed that P. falciparum isolates of Malaysia and Thailand were clustered in the same group for all the allelic families. Population structure of P. falciparum isolates in Malaysia and Thailand exhibit extensive genetic polymorphism but showed high similarities as well as comparable MOI.

Observations of scrotal mass, liver mass, haemolytic jaundice, and central vestibular disorder in Brugia pahangi-infected dogs.

Brugia pahangi is known to infect humans and dogs. Its associated symptoms and complications, however, have not been fully understood in dogs. Herein, we reported the observations of B. pahangi infections in dogs with scrotal mass, liver mass, haemolytic jaundice, and central vestibular disorder.

Molecular detection and epidemiological risk factors associated with Cryptosporidium infection among cattle in Peninsular Malaysia.

Enteric protozoa infection among cattle may pose a threat to productivity and survival leading to negative impacts on the livestock industry. A number of these pathogens are also known to be zoonotic and are of public health concern. Despite the importance of these enteric protozoa to both animal and human health, there remains a paucity of published information on the epidemiological risk factors that may be associated with bovine cryptosporidiosis in Southeast Asia. The present study was undertaken to determine the molecular prevalence and associated risk factors for Cryptosporidium infection among beef and dairy cattle in Peninsular Malaysia. Faecal samples were collected from 824 cattle in 39 farms (526 beef and 298 dairy) situated in 33 locations throughout the country, and subjected to PCR detection for Cryptosporidium using primers targeting the 18S SSUrRNA gene. Epidemiological variables including host, environment and management factors were subjected to univariate and multivariate logistic regression analyses to determine the potential risk factors for infection. The prevalence of Cryptosporidium among the cattle was 12.5%, with significant difference in the infection rate among the various breeds. There was no significant effect of gender, and both the beef and dairy cattle were at similar odds for infection. The younger cattle had a significantly higher infection rate compared to the older animals. Multivariate analysis revealed that deworming practice, distance to human settlement, geographical location (zone) and farm management system were significant risk factors associated with Cryptosporidium infection. The cattle that were reared on farms located in the northeast of the country, closest (≤200 m) to human settlements, reared extensively, and dewormed every four months were at highest risk of infection. The present study constitutes the first attempt to analyze the multivariable epidemiological risk factors involved in bovine cryptosporidiosis in Malaysia and in Southeast Asia. It is envisaged that the data obtained will facilitate better control and prevention measures for Cryptosporidium infection among cattle in the region. Due to the potential zoonotic nature of the infection, serious steps should be instituted for animal treatment and biohazard waste management on local cattle farms.

Soil-transmitted helminthiasis among indigenous communities in Malaysia: Is this the endless malady with no solution?

Soil-transmitted helminths (STHs) are the most common intestinal parasitic infections of medical importance in human. The infections are widely distributed throughout the tropical and subtropical countries including Malaysia particularly among disadvantaged and underprivileged communities. This study was conducted to determine the prevalence and pattern of STH infections among Temuan indigenous subgroup. A cross sectional study was conducted among five villages in Peninsular Malaysia. Faecal samples and socioeconomic data were collected from each consented participant. Faecal samples were processed using formalin-ether sedimentation and examined under microscope. Data analysis was carried out using SPSS software programme for Windows version 24. A total of 411 participants voluntarily participated in this study. The overall prevalence of STH infections was 72.7% (95% CI = 68.2 - 77%). The most common STH species recorded was Trichuris trichiura (58.4%, 95% CI = 53.5 - 63.2%) followed by Ascaris lumbricoides (45.5%, 95% CI = 40.6 - 50.5%) and hookworm (23.1%, 95% CI = 19.1 - 27.5%). Multivariate analysis demonstrated that using untreated water was a significant predictor of STH infections in these communities. Our findings demonstrated that STH infections are still prevalent and co-exist with the low SES among this subgroup. Poverty and poor sanitation are the leading factors contributing to this malady. Hence, the reassessments of the existing control measures are needed.

Microsporidia infection among various groups of the immunocompromised patients.

While information with regards to the bacterial and viral infections are commonly available among clinicians, data on parasitic infection, particularly Microsporidia among immunocompromised patient is currently lacking in Malaysia. This study was conducted to determine the prevalence of Microsporidia among a various group of immunocompromised patient. Two hundred and eighty-eight archived stool samples were examined for the presence of Microsporidia with Gram-Chromotrope Kinyoun staining method. The overall prevalence of Microsporidia was 29.2 % (84/288; 95% CI=24.2-34.5). The end-stage renal failure (ESRF) patients (32.1%) recorded the highest infection rate, followed by cancer (26.2%), human immunodeficiency virus (HIV/AIDS) (22.6%) and acute gastroenteritis (AGE) (7.1%). Meanwhile, organ transplant recipients and autoimmune disease patients recorded the lowest prevalence rate (6.0%). Other intestinal parasites were Strongyloides stercoralis, Trichuris trichiura, Ascaris lumbricoides and Cryptosporidium species. Diarrhoea was the most common symptoms among patients with microsporidiosis. The present study showed that the prevalence of Microsporidia infection was relatively high among immunocompromised patients. This finding highlighted the importance to include detection of microsporidia infection as a routine differential diagnosis in immunocompromised patients, which serves the benefit of treatment to the patients.

Nematode control failure due to anthelmintic resistance in a sheep farm in Malaysia: First identification of the F200Y mutation in the isotype 1 ß-tubulin gene.

This paper reports total nematode anthelmintic resistance towards albendazole, fenbendazole, levamisole and ivermectin in a commercial sheep farm located in Terengganu, Malaysia. Faecal Egg Count Reduction Test (FECRT) was conducted on 25 sheep, where five sheep in each group were treated with the respective four anthelmintics based on live bodyweight. The balance of five sheep placed in the control group were not treated with any anthelmintics. At day 13 post-treatment, faecal egg count was conducted and nematode worm egg count reduction percentage was calculated to determine the resistance status towards the respective anthelmintics tested. Results showed that nematodes were resistant to all the anthelmintics tested, namely albendazole, fenbendazole, levamisole and ivermectin with reduction percentage of 87%, 46%, 94% and 68%, respectively. Subsequently, the third stage larvae of Haemonchus contortus and Trichostrongylus colubriformis recovered from post-treatment faecal cultures were subjected to allele-specific polymerase chain reaction (AS-PCR) assay to determine the presence of the benzimidazole resistance gene. This study reports the occurrence of the classical F200Y mutation in the isotype 1 ßtubulin gene, for the first time in Malaysia.

Occurrence of gastro-intestinal parasites among small ruminants in Malaysia: highlighting Dicrocoelium infection in goats.

The aim of the present study was to determine the gastro-intestinal (GI) parasitic infections among small ruminants (i.e., goats, sheep, deer) in Malaysia through formalin-ether concentration technique. Overall, 70.9% or 302 out of 426 small ruminants (79.4% or 251/316 goats; 87.5% or 35/40 sheep; 22.9% or 16/70 deer) were infected with at least one species of GI parasites. Overall, ten types of GI parasites [Helminth: strongyle (57.7%), Moniezia spp. (5.4%), Paramphistomum spp. (4.5%), Strongyloides spp. (4.2%), Dicrocoelium spp. (2.3%), Trichuris spp. (2.3%); Protozoa: Eimeria spp. (23.7%), Entamoeba spp. (18.8%), Giardia spp. (1.9%), Cryptosporidium spp. (0.2%)] were detected in this study. Among the studied animals, goats harboured the highest diversity of GI parasites (ten types), followed by sheep (six types) and deer (two types). Polyparasitism was observed in goats (43.7% or 138 of 316) and sheep (15.0% or 6 of 40). Cumulatively, a total of 32 combinations of coinfections (Helminth+Helminth: 8 combinations; Helminth+Protozoa: 20 combinations; Protozoa+Protozoa: 4 combinations) between detected parasites with up to quintuple infections were reported. Among these parasites, "strongyle + Eimeria spp." and "Moniezia spp. + strongyle" were the commonest infections in goats (13.5% or 34 of 251) and sheep (5.7% or 2 of 6), respectively. This study is a comprehensive documentation on multiple GI parasitisms among small ruminant in Malaysia, and the findings are crucial for effective farm management, especially for the formulation of parasitic control and elimination strategies.