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Interaction of low-level dietary supplementation of Chlorella vulgaris Beijerinck, 1890, and feeding duration on growth hormone, growth performance and serum biochemistry of red hybrid tilapia (Oreochromis mossambicus × Oreochromis niloticus)

Ahmad MT, Shariff M, Goh YM, Banerjee S, Yusoff FM

J Fish Biol, 2023 Sep;103(3):715-726.
PMID: 37249562 DOI: 10.1111/jfb.15469

Chlorella is one of the most widely accepted Chlorophyta used by many as livestock and aquaculture feed. Nonetheless, different studies on the overall performances of fish reported the unfavourable effect of high-level supplementations of Chlorella vulgaris. The current study determined the impact of low-level dietary supplementation of C. vulgaris alongside the different feeding durations and their interactions on the growth hormone (GH), growth performances, serum-biochemical indices, hepatic function and some immunological parameters of red hybrid tilapia. The fingerlings (mean weight: 14.25 ± 0.01 g, length: 13.5 ± 0.49 cm) were fed diets containing 0, 0.99%, 2.91% and 4.76% of C. vulgaris powder per kilogram dry diet for 90 days. GH, growth performance, serum-biochemical indices (total serum protein, albumin, globulin, glucose, aspartate aminotransferase and alanine aminotransferase) and some immunological (respiratory burst and lysozyme activities) parameters of the fish were examined after 30, 60 and 90 days of feeding. The results demonstrated that tilapia fed C. vulgaris-supplemented diets showed increased levels of respiratory burst, lysozyme, albumin and total protein, GH and growth performances (P
Fulltext Clinical students' reflections on the preclinical anatomy learning experience

Syed Abd Halim SA, Yusoff MSB, Yaman MN, Razali SA, Tengku Muda TFM, Ramli RR, et al.

J Taibah Univ Med Sci, 2023 Aug;18(4):757-770.
PMID: 36852241 DOI: 10.1016/j.jtumed.2022.12.007

OBJECTIVES: Anatomy is a fundamental pillar of medical knowledge that bridges basic medical science knowledge and clinical practice. However, integrated modern medical curricula have reduced the anatomy teaching content, and cadaveric dissection is no longer conducted. Medical graduates who lack anatomy knowledge are anticipated to be inadequately equipped for safe clinical practice. This study was aimed at exploring clinical year students' experiences regarding their anatomy learning during the preclinical phase in Malaysian medical schools. The findings reflect how the students' preclinical anatomy training prepared them for their clinical years of study.
METHODS: A qualitative phenomenology study using the focus group discussion method was conducted on 30 final-year students from four public universities. Four focus group discussion sessions were conducted, and students' responses were transcribed and converted to electronic formats. The transcripts were analyzed thematically with ATLAS.ti software.
RESULTS: The first-cycle coding of the text analysis generated 157 open codes based on the phrases used by the participants. The subsequent coding cycle produced 16 axial codes-groups of open codes with similar features. During the final coding cycle, the content and interrelations between the axial codes were categorized into six codes: (1) preclinical anatomy learning experience, (2) anatomy content and teaching, (3) anatomy-related competency, (4) the importance of anatomy knowledge in clinical practice, (5) the importance of early exposure to applied clinical anatomy, and (6) suggestions for future anatomy education.
CONCLUSIONS: The six identified themes reflected students' perceptions of their anatomy learning experience, the challenges that they faced during their preclinical years, and their opinions regarding the anatomy knowledge and skills that are functionally relevant during the clinical years. Their responses also echoed the need to improve anatomy teaching and learning, thereby emphasizing the importance of early clinical integration and application.
Estimation of Total Body Surface Area Burned: A Comparison Between Burn Unit and Referring Facilities

Ho HL, Halim AS, Sulaiman WAW, Fatimah MJ

Ann Burns Fire Disasters, 2023 Mar;36(1):19-28.
PMID: 38680901

Accuracy of burn size estimation is critical in acute burn management because it directly affects the patient's outcome and prognosis. This study aims to quantify the discrepancies of total body surface area (TBSA) burned between the burn unit (TBSAb) and the referring facilities (TBSAr). Data of all referred adult and paediatric patients admitted to the Hospital Universiti Sains Malaysia Burn Unit within 24 hours post burn were retrospectively reviewed from 2015 to 2019. %TBSA discrepancies were calculated by the differences between TBSAb and TBSAr. A total of 208 patients (111 adults and 97 paediatric patients) were recruited in this study. Of these, the TBSA was overestimated in 60.58% cases, underestimated in 13.46% cases, accurate in 7.69% cases, and in 18.27% cases the referrals had no TBSAr stated. The %TBSA discrepancy was the highest in severe burns (mean 10.80% in adults and 7.59 in paediatric patients; P<0.001). The time interval between referral and reassessment and patients' body mass index (BMI) were not statistically significant for the magnitude of TBSA discrepancy. The number of burn areas involved correlated with the %TBSA discrepancies, with the highest recorded discrepancy being 21.50% in whole body involvement. There were significant discrepancies in TBSA estimations between the referring facilities and those of the Hospital Universiti Sains Malaysia (USM) burn unit, especially among the paediatric patients and those with severe burns. Implementation of educational programs by burn care experts and agreement on a universal method of TBSA assessment are necessary in reducing the discrepancies.
Influence of Symbiotic Probiont Strains on the Growth of Amphora and Chlorella and Its Potential Protections Against Vibrio spp. in Artemia

Rosland NA, Ikhsan N, Min CC, Yusoff FM, Karim M

Curr Microbiol, 2021 Nov;78(11):3901-3912.
PMID: 34522979 DOI: 10.1007/s00284-021-02642-2

The emerging aquaculture industry is in need of non-antibiotic-based disease control approaches to minimize the risk of antibiotic-resistant bacteria. Bacterial infections mainly caused by Vibrio spp. have caused mass mortalities of fish especially during the larval stages. The objectives of this study were to verify the potential of symbiotic probiont strains, isolated from microalgae (Amphora, Chlorella, and Spirulina) for suppressing the growth of Vibrio spp. and at the same time ascertain their abilities to enhance microalgal biomass by mutualistic interactions through microalgae-bacteria symbiosis. In addition, in vivo studies on Artemia bioencapsulated with probiont strains (single strain and mix strains) and microalgae were evaluated. The selected potential probionts were identified as Lysinibacillus fusiformis strain A-1 (LFA-1), Bacillus sp. strain A-2 (BA-2), Lysinibacillus fusiformis strain Cl-3 (LFCl-3), and Bacillus pocheonensis strain S-2 (BPS-2) using 16s rRNA. The cell densities of Amphora culture supplemented with BA-2 and Chlorella culture supplemented with LFCl-3 were higher than those of the controls. Artemia bioencapsulated with mix strains (LFA-1 + BA-2 + LFCl-3 + BPS-2) and Amphora demonstrated the highest survival rate compared to the controls, after being challenged with V. harveyi (60 ± 4%) and V. parahaemolyticus (78 ± 2%). Our study postulated that BA-2 and LFCl-3 were found to be good promoting bacteria for microalgal growth and microalgae serve as a vector to transport probiotic into Artemia. Moreover, mixture of potential probionts is beneficial for Artemia supplementation in conferring protection to Artemia nauplii against pathogenic Vibrios.
Massive penile lipogranuloma following olive oil injections

Fadya Nabiha AS, Fam XI, Fatimah MN, Md Hud MZ

Med J Malaysia, 2021 Sep;76(5):774-776.
PMID: 34508395

Penile self-injections are performed with the purpose of increasing the size of the penis. Commonly, mineral oils or mineral oil-like substances are used for this purpose. However, there are very few publications describing on the complications from vegetable oil injections especially olive oil. Following the injection, the resulting deformity of the male genitalia is described as penile paraffinomas lipogranuloma of the penis. We would like to report a case of penis augmentation by a 50-year-old gentleman using olive oil injections that caused a massive, severely deformed of the penis and scrotum. Surgical excision and reconstruction was done. The patient was discharged after five days postoperatively, with full graft take. Injecting any oily substances into the human body is hazardous. The differential diagnosis of nodules following injections should be kept in mind and in doubtful cases, biopsy should be done. Regular follow-up is essential to prevent the further inflammatory event. Safe and legal procedures should be implemented for penile enlargement so as to avoid self-injections and in the prevention of such debilitating problems.
Fulltext De novo transcriptome analysis of Chlorella sorokiniana: effect of glucose assimilation, and moderate light intensity

Azaman SNA, Wong DCJ, Tan SW, Yusoff FM, Nagao N, Yeap SK

Sci Rep, 2020 Oct 15;10(1):17331.
PMID: 33060668 DOI: 10.1038/s41598-020-74410-4

Chlorella can produce an unusually wide range of metabolites under various nutrient availability, carbon source, and light availability. Glucose, an essential molecule for the growth of microorganisms, also contributes significantly to the metabolism of various metabolic compounds produced by Chlorella. In addition, manipulation of light intensity also induces the formation of secondary metabolites such as pigments, and carotenoids in Chlorella. This study will focus on the effect of glucose addition, and moderate light on the regulation of carotenoid, lipid, starch, and other key metabolic pathways in Chlorella sorokiniana. To gain knowledge about this, we performed transcriptome profiling on C. sorokiniana strain NIES-2168 in response to moderate light stress supplemented with glucose under mixotrophic conditions. A total of 60,982,352 raw paired-end (PE) reads 100 bp in length was obtained from both normal, and mixotrophic samples of C. sorokiniana. After pre-processing, 93.63% high-quality PE reads were obtained, and 18,310 predicted full-length transcripts were assembled. Differential gene expression showed that a total of 937, and 1124 genes were upregulated, and downregulated in mixotrophic samples, respectively. Transcriptome analysis revealed that the mixotrophic condition caused upregulation of genes involved in carotenoids production (specifically lutein biosynthesis), fatty acid biosynthesis, TAG accumulation, and the majority of the carbon fixation pathways. Conversely, starch biosynthesis, sucrose biosynthesis, and isoprenoid biosynthesis were downregulated. Novel insights into the pathways that link the enhanced production of valuable metabolites (such as carotenoids in C. sorokiniana) grown under mixotrophic conditions is presented.
Fulltext Identification and Analysis of microRNAs in Chlorella sorokiniana Using High-Throughput Sequencing

Azaman SNA, Satharasinghe DA, Tan SW, Nagao N, Yusoff FM, Yeap SK

Genes (Basel), 2020 09 25;11(10).
PMID: 32992970 DOI: 10.3390/genes11101131

Chlorella is a popular microalga with robust physiological and biochemical characteristics, which can be cultured under various conditions. The exploration of the small RNA content of Chlorella could improve strategies for the enhancement of metabolite production from this microalga. In this study, stress was introduced to the Chlorella sorokiniana culture to produce high-value metabolites such as carotenoids and phenolic content. The small RNA transcriptome of C. sorokiniana was sequenced, focusing on microRNA (miRNA) content. From the analysis, 98 miRNAs were identified in cultures subjected to normal and stress conditions. The functional analysis result showed that the miRNA targets found were most often involved in the biosynthesis of secondary metabolites, followed by protein metabolism, cell cycle, and porphyrin and chlorophyll metabolism. Furthermore, the biosynthesis of secondary metabolites such as carotenoids, terpenoids, and lipids was found mostly in stress conditions. These results may help to improve our understanding of regulatory mechanisms of miRNA in the biological and metabolic process of Chlorella species. It is important and timely to determine the true potential of this microalga species and to support the potential for genetic engineering of microalgae as they receive increasing focus for their development as an alternative source of biofuel, food, and health supplements.
Fulltext Integration of Algae to Improve Nitrogenous Waste Management in Recirculating Aquaculture Systems: A Review

Ramli NM, Verreth JAJ, Yusoff FM, Nurulhuda K, Nagao N, Verdegem MCJ

Front Bioeng Biotechnol, 2020;8:1004.
PMID: 33015002 DOI: 10.3389/fbioe.2020.01004

This review investigates the performance and the feasibility of the integration of an algal reactor in recirculating aquaculture systems (RAS). The number of studies related to this topic is limited, despite the apparent benefit of algae that can assimilate part of the inorganic waste in RAS. We identified two major challenges related to algal integration in RAS: first, the practical feasibility for improving nitrogen removal performance by algae in RAS; second, the economic feasibility of integrating an algal reactor in RAS. The main factors that determine high algal nitrogen removal rates are light and hydraulic retention time (HRT). Besides these factors, nitrogen-loading rates and RAS configuration could be important to ensure algal performance in nitrogen removal. Since nitrogen removal rate by algae is determined by HRT, this will affect the size (area or volume) of the algal reactor due to the time required for nutrient uptake by algae and large surface area needed to capture enough light. Constraints related to design, space, light capture, and reactor management could incur additional cost for aquaculture production. However, the increased purification of RAS wastewater could reduce the cost of water discharge in places where this is subject to levees. We believe that an improved understanding of how to manage the algal reactor and technological advancement of culturing algae, such as improved algal reactor design and low-cost artificial light, will increase the practical and economic feasibility of algal integration in RAS, thus improving the potential of mass cultivation of algae in RAS.
Risk assessment of pharmaceutically active compounds (PhACs) in the Klang River estuary, Malaysia

Omar TFT, Aris AZ, Yusoff FM, Mustafa S

Environ Geochem Health, 2019 Feb;41(1):211-223.
PMID: 30051257 DOI: 10.1007/s10653-018-0157-1

The concentration profile, distribution and risk assessment of pharmaceutically active compounds (PhACs) in the coastal surface water from the Klang River estuary were measured. Surface coastal water samples were extracted using offline solid phase, applying polymeric C18 cartridges as extraction sorbent and measuring with liquid chromatography mass spectrometry-mass spectrometry (LC MS-MS) technique. Extraction method was optimized for its recovery, sensitivity and linearity. Excellent recoveries were obtained from the optimized method with percentage of recoveries ranging from 73 to 126%. The optimized analytical method achieved good sensitivity with limit of detection ranging from 0.05 to 0.15 ng L-1, while linearity of targeted compounds in the LC MS-MS system was more than 0.990. The results showed that amoxicillin has the highest concentration (102.31 ng L-1) followed by diclofenac (10.80 ng L-1) and primidone (7.74 ng L-1). The percentage of contribution (% of total concentration) for the targeted PhACs is in the following order; amoxicillin (92.90%) > diclofenac (3.95%) > primidone (1.23%) > dexamethasone (0.75%) > testosterone (0.70%) > sulfamethoxazole (0.33%) > progesterone (0.14%). Environmental risk assessment calculated based on deterministic approach (the RQ method), showed no present risk from the presence of PhACs in the coastal water of Klang River estuary. Nonetheless, this baseline assessment can be used for better understanding on PhACs pollution profile and distribution in the tropical coastal and estuarine ecosystem as well as for future comparative studies.
Fulltext Efficacy of chitosan derivative films versus hydrocolloid dressing on superficial wounds

Halim AS, Nor FM, Mat Saad AZ, Mohd Nasir NA, Norsa'adah B, Ujang Z

J Taibah Univ Med Sci, 2018 Dec;13(6):512-520.
PMID: 31435371 DOI: 10.1016/j.jtumed.2018.10.004

Objectives: Chitosan, the N-deacetylated derivative of chitin, has useful biological properties that promote haemostasis, analgesia, wound healing, and scar reduction; chitosan is bacteriostatic, biocompatible, and biodegradable. This study determined the efficacy of chitosan derivative film as a superficial wound dressing.
Methods: This multicentre randomised controlled trial included 244 patients, of whom 86 were treated with chitosan derivative film and 84 with hydrocolloid. The percentage of epithelisation, as well as patient comfort, clinical signs, and patient convenience in application and removal of the dressings were assessed.
Results: The primary outcome of this study was the percentage of epithelisation. Except for race (p = 0.04), there were no significant differences between groups in sex, age, antibiotic usage, or initial wound size (p > 0.05). There was no significant difference in the mean epithelisation percentage between groups (p = 0.29). Patients using chitosan derivative film experienced more pain during removal of dressing than those in the hydrocolloid group (p = 0.007). The chitosan derivative film group showed less exudate (p = 0.036) and less odour (p = 0.024) than the control group. Furthermore, there were no significant differences between groups in terms of adherence, ease of removal, wound drainage, erythema, itchiness, pain, and tenderness. No oedema or localised warmth was observed during the study.
Conclusion: This study concluded that chitosan derivative film is equivalent to hydrocolloid dressing and can be an option in the management of superficial and abrasion wounds.
Clinical trial No: NMRR-11-948-10565.
Fulltext Draft Genome Sequence of the Shrimp Pathogen Vibrio parahaemolyticus ST17.P5-S1, Isolated in Peninsular Malaysia

Devadas S, Bhassu S, Christie Soo TC, Yusoff FM, Shariff M

Microbiol Resour Announc, 2018 Sep;7(11).
PMID: 30533648 DOI: 10.1128/MRA.01053-18

We sequenced the genome of Vibrio parahaemolyticus strain ST17.P5-S1, isolated from Penaeus vannamei cultured in the east coast of Peninsular Malaysia. The strain contains several antibiotic resistance genes and a plasmid encoding the Photorhabdus insect-related (Pir) toxin-like genes, pirAvp and pirBvp, associated with acute hepatopancreatic necrosis disease (AHPND).
Fulltext Draft Genome Sequence of a Vibrio parahaemolyticus Strain, KS17.S5-1, with Multiple Antibiotic Resistance Genes, Which Causes Acute Hepatopancreatic Necrosis Disease in Penaeus monodon in the West Coast of Peninsular Malaysia

Devadas S, Bhassu S, Christie Soo TC, Mohamed Iqbal SN, Yusoff FM, Shariff M

Microbiol Resour Announc, 2018 Jul;7(2).
PMID: 30533806 DOI: 10.1128/MRA.00829-18

We report the first draft genome sequence of a Vibrio parahaemolyticus strain (VpAHPND), which causes acute hepatopancreatic necrosis disease (AHPND) in Penaeus monodon. The strain has a pVA1-like plasmid carrying pirAvp and pirBvp genes. Whole-genome comparisons revealed >98% similarity to VpAHPND isolates from Thailand, Mexico, and Vietnam.
New data on Cladocera (Crustacea: Branchiopoda) of Sabah State, Borneo Island, Malaysia

Sinev AY, Yusoff FM

Zootaxa, 2018 Jun 21;4438(2):362-372.
PMID: 30313150 DOI: 10.11646/zootaxa.4438.2.10

Unlike those in the mainland of Southeast Asia, the Cladocera of the Malay Archipelago has not been intensively studied, except for the state of Sabah in the north-eastern part of the Borneo island. This study aimed to complete the inventory of the Cladocera in Sabah by looking at different types of water bodies including oxbow lakes, small lakes, reservoirs, ponds, ditches and paddy fields. From 32 sites examined, 35 species of cladocerans, nine of which were new records to Sabah, were found from 25 localities. With this new finding, the total number of cladoceran species in Sabah increased to 39 species, including five species of Sididae, four species of Daphniidae, one species of Moinidae, five species of Macrothricidae, two species of Ilyocryptidae, and 22 species of Chydoridae. Only 8 % ( three species) of Sabah cladocerans are true planktonic. This study illustrated that most cladocerans were associated with substrates in the littoral zone and thus appropriate sampling methods should be employed in different microhabitats for comprehensive biodiversity assessment.
A comparison of the morphological and biochemical characteristics of Chlorella sorokiniana and Chlorella zofingiensis cultured under photoautotrophic and mixotrophic conditions

Azaman SNA, Nagao N, Yusoff FM, Tan SW, Yeap SK

PeerJ, 2017;5:e3473.
PMID: 28929006 DOI: 10.7717/peerj.3473

The responses of two species of microalgae, Chlorella sorokiniana and Chlorella zofingiensis, were compared regarding their morphological and biochemical properties under photoautotrophic and mixotrophic conditions. These microalgae were cultured under both conditions, and their crude ethanolic extracts were examined for their pigment and total phenolic contents. In addition, the microalgae's antioxidant activities were determined using a DPPH radical scavenging assay and a ferric reducing antioxidant power (FRAP) assay. Both strains showed increases in cell size due to the accumulation of lipid bodies and other cell contents, especially carotenoids, under the mixotrophic condition. Notably, reductions in phenolic and chlorophyll contents were observed to be associated with lower antioxidant activity. C. zofingiensis compared with C. sorokiniana, demonstrated higher antioxidant activity and carotenoid content. This study showed that different species of microalgae responded differently to varying conditions by producing different types of metabolites, as evidenced by the production of higher levels of phenolic compounds under the photoautotrophic condition and the production of the same levels of carotenoids under both photoautotrophic and mixotrophic conditions.
A new species of genus Ephemeroporus Frey, 1982 (Cladocera: Anomopoda: Chydoridae) from Peninsular Malaysia

Sinev AY, Yusoff FM

Zootaxa, 2016 Jun 01;4117(3):399-410.
PMID: 27395182 DOI: 10.11646/zootaxa.4117.3.7

Study of Ephemeroporus Frey, 1982 populations from Peninsular Malaysia revealed a new species. Ephemeroporus malaysiaensis sp. nov. is characterized by the presence of four-five large denticles on anal margin of postabdomen instead of two-three, and five distinctive pigmented spots on each valve. Pigmented spots on the valves were never recorded for any species of family Chydoridae. E. malaysiaensis sp. nov. seems to be another endemic Chydoridae species of South-East Asia. E. malaysiaensis sp. nov. is a rare species, associated with emergent macrophytes in the littoral zone of shallow lake. Most of Ephemeroporus populations from Peninsular Malaysia belong to eurybiotic Paleotropical species Ephemeroporus barroisi (Richard, 1984).
Identification of immune response-related genes and signalling pathways in spleen of Vibrio parahaemolyticus-infected Epinephelus fuscoguttatus (Forskal) by next-generation sequencing

Low CF, Mariana NS, Maha A, Chee HY, Fatimah MY

J Fish Dis, 2016 Mar;39(3):389-94.
PMID: 25786532 DOI: 10.1111/jfd.12359
Non-immune-related genes and signalling pathways in spleen of Vibrio parahaemolyticus-infected Epinephelus fuscoguttatus (Forskal)

Low CF, Mariana NS, Maha A, Chee HY, Fatimah MY

J Fish Dis, 2015 Aug;38(8):761-4.
PMID: 25073481 DOI: 10.1111/jfd.12283
Cladocera (Crustacea: Branchiopoda) of Sabah state in Borneo Island, Malaysia

Sinev AY, Yusoff FM

Zootaxa, 2015;4000(5):581-91.
PMID: 26623748 DOI: 10.11646/zootaxa.4000.5.7

Fauna of Cladocera (Crustacea: Branchiopoda) of Sabah state of Malaysia, Borneo Island, was evaluated for the first time. Samples from 40 locations were studied, and 31 species of Cladocera were revealed, including three species of Sididae, one species of Daphnidae, one species of Moinidae, four species of Macrothricidae, two species of Ilyocryptidae, and 20 species of Chydoridae. One species of Ilyocryptidae, Ilyocryptus yooni Jeong, Kotov and Lee, 2012, is recorded for Malaysia for the first time, and one more, Anthalona sp., is probably new for science. Of 31 species recorded for Sabah, only three are true planktonic species and 28 are substrate-associated species. Absence of large natural lakes, habitats with most rich cladoceran fauna, can be an important factor limiting diversity of Cladocera in Sabah.
Fulltext Market potential analysis for tengas (Neolissochilus sp.) in the Malaysian market

Khai, N. X., Kusairi, M. N., Ahmad, S., Syahaneem, M. Z., Fatimah, M. A.

International Food Research Journal, 2015;22(4):1429-1432.
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There are some signals that Malaysian marine landings are decreasing and that wild stocks have been over-fished. However, it has extensive inland water bodies which have the potentials to be developed into freshwater aquaculture sites. Tengas (Neolissochilus sp.), a freshwater type of fish, are found in abundance in Malaysia; but has not reached the commercial level in the market. This paper examines the potential of developing tengas as a commercial freshwater species. The data used in this study were obtained through primary data search from key informants and a survey. A semi-structured open-ended questionnaire with Malay and Chinese languages was designed for the interviews. The study areas covered Peninsular Malaysia and East Malaysia. From the survey, we found that tengas has potential to be taken as a food item and an ornamental fish. The wild stock of tengas in fact is also worthwhile to be conserved for agro-tourism development. Many restaurant operators are willing to invent cooking method on tengas.
Fulltext Prevalence and zoonotic potential of canine hookworms in Malaysia

Mahdy MA, Lim YA, Ngui R, Siti Fatimah MR, Choy SH, Yap NJ, et al.

Parasit Vectors, 2012;5:88.
PMID: 22564445 DOI: 10.1186/1756-3305-5-88

Canine hookworm infection is endemic in Southeast Asian countries with a prevalence ranging from 70% to 100%, with zoonotic transmission representing a potentially significant public health concern. However, there are limited data available on the prevalence of canine hookworms in Malaysia. This study was conducted to determine the prevalence of hookworm and Ancylostoma species among dogs in Malaysia.

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