Displaying publications 41 - 58 of 58 in total

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  1. Gut microbiota of healthy Asians and their discriminative features revealed by metagenomics approach
    Taha SFM, Bhassu S, Omar H, Raju CS, Rajamanikam A, Govind SKP, et al.
    3 Biotech, 2023 Aug;13(8):275.
    PMID: 37457869 DOI: 10.1007/s13205-023-03671-3
    This study is conducted to identify the microbial architecture and its functional capacity in the Asian population via the whole metagenomics approach. A brief comparison of the Asian countries namely Malaysia, India, China, and Thailand, was conducted, giving a total of 916 taxa under observation. Results show a close representation of the taxonomic diversity in the gut microbiota of Malaysia, India, and China, where Bacteroidetes, Firmicutes, and Actinobacteria were more predominant compared to other phyla. Mainly due to the multi-racial population in Malaysia, which also consists of Malays, Indian, and Chinese, the population tend to share similar dietary preferences, culture, and lifestyle, which are major influences that shapes the structure of the gut microbiota. Moreover, Thailand showed a more distinct diversity in the gut microbiota which was highly dominated by Firmicutes. Meanwhile, functional profiles show 1034 gene families that are common between the four countries. The Malaysia samples are having the most unique gene families with a total of 67,517 gene families, and 51 unique KEGG Orthologs, mainly dominated by the metabolic pathways, followed by microbial metabolism in diverse environments. In conclusion, this study provides some general overview on the structure of the Asian gut microbiota, with some additional highlights on the Malaysian population.

    SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-023-03671-3.

  2. Fulltext RNA-seq analysis of Macrobrachium rosenbergii hepatopancreas in response to Vibrio parahaemolyticus infection
    Rao R, Bing Zhu Y, Alinejad T, Tiruvayipati S, Lin Thong K, Wang J, et al.
    Gut Pathog, 2015;7:6.
    PMID: 25922623 DOI: 10.1186/s13099-015-0052-6
    The Malaysian giant freshwater prawn, Macrobrachium rosenbergii, is an economically important crustacean worldwide. However, production of this prawn is facing a serious threat from Vibriosis disease caused by Vibrio species such as Vibrio parahaemolyticus. Unfortunately, the mechanisms involved in the immune response of this species to bacterial infection are not fully understood. We therefore used a high-throughput deep sequencing technology to investigate the transcriptome and comparative expression profiles of the hepatopancreas from this freshwater prawn infected with V. parahaemolyticus to gain an increased understanding of the molecular mechanisms underlying the species' immune response to this pathogenic bacteria.
  3. Fulltext Genome anatomy of the gastrointestinal pathogen, Vibrio parahaemolyticus of crustacean origin
    Tiruvayipati S, Bhassu S, Kumar N, Baddam R, Shaik S, Gurindapalli AK, et al.
    Gut Pathog, 2013 Dec 11;5(1):37.
    PMID: 24330647 DOI: 10.1186/1757-4749-5-37
    Vibrio parahaemolyticus, an important human pathogen, is associated with gastroenteritis and transmitted through partially cooked seafood. It has become a major concern in the production and trade of marine food products. The prevalence of potentially virulent and pathogenic V. parahaemolyticus in raw seafood is of public health significance. Here we describe the genome sequence of a V. parahaemolyticus isolate of crustacean origin which was cultured from prawns in 2008 in Selangor, Malaysia (isolate PCV08-7). The next generation sequencing and analysis revealed that the genome of isolate PCV08-7 has closest similarity to that of V. parahaemolyticus RIMD2210633. However, there are certain unique features of the PCV08-7 genome such as the absence of TDH-related hemolysin (TRH), and the presence of HU-alpha insertion. The genome of isolate PCV08-7 encodes a thermostable direct hemolysin (TDH), an important virulence factor that classifies PCV08-7 isolate to be a serovariant of O3:K6 strain. Apart from these, we observed that there is certain pattern of genetic rearrangements that makes V. parahaemolyticus PCV08-7 a non-pandemic clone. We present detailed genome statistics and important genetic features of this bacterium and discuss how its survival, adaptation and virulence in marine and terrestrial hosts can be understood through the genomic blueprint and that the availability of genome sequence entailing this important Malaysian isolate would likely enhance our understanding of the epidemiology, evolution and transmission of foodborne Vibrios in Malaysia and elsewhere.
  4. Deep parallel sequencing reveals conserved and novel miRNAs in gill and hepatopancreas of giant freshwater prawn
    Tan TT, Chen M, Harikrishna JA, Khairuddin N, Mohd Shamsudin MI, Zhang G, et al.
    Fish Shellfish Immunol, 2013 Oct;35(4):1061-9.
    PMID: 23816854 DOI: 10.1016/j.fsi.2013.06.017
    MicroRNAs (miRNAs) are ~20-22 nucleotides, non protein-coding RNA regulatory genes that post-transcriptionally regulate many protein-coding genes, influencing critical biological and metabolic processes. While the number of known microRNA is increasing, there is currently no published data for miRNA from giant freshwater prawns, Macrobrachium rosenbergii (M. rosenbergii), a commercially cultured and economically important food species. In this study, we identified novel miRNAs in the gill and hepatopancreas of M. rosenbergii. Through a deep parallel sequencing analysis and an in silico data analysis approach, 327 miRNA families were identified from small RNA libraries with reference to both the de novo transcriptome of M. rosenbergii obtained from RNA-Seq and to miRBase (Release 18.0, November 2012). Based on the identified mature miRNA and recovered precursor sequences that form appropriate hairpin structures, three conserved miRNA (miR125, miR750, miR993) and 27 novel miRNA candidates encoding messenger-like non-coding RNA were identified. miR-125, miR-750, G-m0002/H-m0009, G-m0005, G-m0008/H-m0016, G-m0011/H-m0027 and G-m0015 were selected for experimental validation with stem-loop quantitative RT-PCR and were found to be coherent with the expression profile of deep sequencing data as evaluated with Pearson's correlation coefficient (r = 0.835178 for miRNA in gill, r = 0.724131 for miRNA in hepatopancreas). Using a combinatorial approach of pathway enrichment analysis and inverse expression relationship of miRNA and mRNA, four co-expressed novel miRNA candidates (G-m0005, G-m0008/H-m0016, G-m0011/H-m0027, and G-m0015) were found to be associated with energy metabolism. In addition, the expression of the three novel miRNA candidates (G-m0005, G-m0008/H-m0016, and G-m0011/H-m0027) were also found to be significantly reduced at 9 and 24 h post infection in M. rosenbergii challenged with infectious hypodermal and hematopoietic necrosis virus, suggesting a functional role of these miRNAs in crustacean immune defense.
  5. A transcriptome study on Macrobrachium rosenbergii hepatopancreas experimentally challenged with white spot syndrome virus (WSSV)
    Rao R, Bhassu S, Bing RZ, Alinejad T, Hassan SS, Wang J
    J Invertebr Pathol, 2016 05;136:10-22.
    PMID: 26880158 DOI: 10.1016/j.jip.2016.01.002
    The world production of shrimp such as the Malaysian giant freshwater prawn, Macrobrachium rosenbergii is seriously affected by the white spot syndrome virus (WSSV). There is an urgent need to understand the host pathogen interaction between M. rosenbergii and WSSV which will be able to provide a solution in controlling the spread of this infectious disease and lastly save the aquaculture industry. Now, using Next Generation Sequencing (NGS), we will be able to capture the response of the M. rosenbergii to the pathogen and have a better understanding of the host defence mechanism. Two cDNA libraries, one of WSSV-challenged M. rosenbergii and a normal control one, were sequenced using the Illumina HiSeq™ 2000 platform. After de novo assembly and clustering of the unigenes from both libraries, 63,584 standard unigenes were generated with a mean size of 698bp and an N50 of 1137bp. We successfully annotated 35.31% of all unigenes by using BLASTX program (E-value <10-5) against NCBI non-redundant (Nr), Swiss-Prot, Kyoto Encyclopedia of Genes and Genome pathway (KEGG) and Orthologous Groups of proteins (COG) databases. Gene Ontology (GO) assessment was conducted using BLAST2GO software. Differentially expressed genes (DEGs) by using the FPKM method showed 8443 host genes were significantly up-regulated whereas 5973 genes were significantly down-regulated. The differentially expressed immune related genes were grouped into 15 animal immune functions. The present study showed that WSSV infection has a significant impact on the transcriptome profile of M. rosenbergii's hepatopancreas, and further enhanced the knowledge of this host-virus interaction. Furthermore, the high number of transcripts generated in this study will provide a platform for future genomic research on freshwater prawns.
  6. Molecular Identification and Genetic Variation of Rattus Species From Oil Palm Plantations of Malaysia Based on Mitochondrial Cytochrome Oxidase Subunit I (COI) Gene Sequences
    Nasir MH, Bhassu S, Mispan MS, Bakar SA, Jing KJ, Omar H
    Zoolog Sci, 2022 Dec;39(6):554-561.
    PMID: 36495490 DOI: 10.2108/zs210093
    Rats (Rattus species) are the most notorious vertebrate pests in Malaysian oil palm plantations. Although many studies have been conducted on Asian rats, little attention has been paid to their species composition and phylogenetic relationships in oil palm plantations in Peninsular Malaysia. We determined the mitochondrial cytochrome oxidase subunit I (COI) gene sequence (708 bp) for 216 individual rats collected from five oil palm plantations in Peninsular Malaysia. Phylogenetic analysis in conjunction with comparison with sequences from the nucleotide sequence database revealed five distinct lineages in the Malaysian oil plantations: Rattus tiomanicus, Rattus argentiventer, Rattus exulans, Rattus tanezumi, and a taxon corresponding to the Malayan house rat, which was most frequently observed (∼50%). The last taxon has traditionally been classified as a synonym of Rattus rattus (Rattus rattus diardii) or Rattus tanezumi, but our phylogenetic analysis placed it as an independent lineage, which is not particularly closely related to R. rattus or R. tanezumi, and which we refer to as Rattus diardii. The construction of the network showed that there is considerable genetic variation within the lineages of R. diardii and R tiomanicus, suggesting that these two species are native to the Malay Peninsula.
  7. Knowledge, attitudes and practices on antimicrobial use and antimicrobial resistance among shrimp aquaculturists in Peninsular Malaysia
    Devadas S, Zakaria Z, Din MSM, Bhassu S, Karim MMA, Ikhsan N, et al.
    Prev Vet Med, 2025 Mar 13;239:106513.
    PMID: 40117849 DOI: 10.1016/j.prevetmed.2025.106513
    In aquaculture, production intensification and the increasing occurrence of diseases are equally driving antimicrobial use (AMU) and the development of antimicrobial resistance (AMR). There is inadequate awareness and knowledge regarding AMU and AMR among the public in Malaysia. Thereby, AMR reduction action plans critically require the involvement of farmers as end users of the antimicrobials. The knowledge, attitudes and practices (KAP) survey can evaluate the farmers' awareness level about the issue. To date, no KAP survey has been conducted among shrimp aquaculture farmers in Malaysia regarding AMU and AMR. In addition, no suitable questionnaire is available to conduct the study. Therefore, this study has developed a questionnaire to evaluate the KAP of Penaeus vannamei and Penaeus monodon aquaculture farmers towards AMU and AMR in selected states in Peninsular Malaysia. In total, 113 farmers participated in this survey. The majority (88.1 %, n = 89) of 101 respondents who completed answering the knowledge- and attitude-related KAP items had inadequate knowledge of AMU and AMR. Regarding attitude, the percentages of farmers with good attitudes (49.5 %, n = 50) and poor attitudes (50.5 %, n = 51) are comparable. Nevertheless, a high percentage (95.0 %, n = 96) of them have never used antibiotics on farmed shrimp, indicating good practices in terms of AMU. The Pearson correlation revealed a large positive association between knowledge and attitudes. To the best of our knowledge, this study is the first to develop a questionnaire and assess the KAP regarding AMU and AMR among shrimp aquaculture farmers in Malaysia.
  8. Detection and phylogenetic profiling of nodavirus associated with white tail disease in Malaysian Macrobrachium rosenbergii de Man
    Saedi TA, Moeini H, Tan WS, Yusoff K, Daud HM, Chu KB, et al.
    Mol Biol Rep, 2012 May;39(5):5785-90.
    PMID: 22223294 DOI: 10.1007/s11033-011-1389-7
    White tail disease (WTD) is a serious viral disease in the hatcheries and nursery ponds of Macrobrachium rosenbergii in many parts of the world. A new disease similar to WTD was observed in larvae and post larvae of M. rosenbergii cultured in Malaysia. In the present study, RT-PCR assay was used to detect the causative agents of WTD, M. rosenbergii nodavirus (MrNV) and extra small virus (XSV) using specific primers for MrNV RNA2 and XSV. The results showed the presence of MrNV in the samples with or without signs of WTD. However, XSV was only detected in some of the MrNV-positive samples. Phylogenetic analysis showed that the RNA2 of our Malaysian isolates were significantly different from the other isolates. Histopathological studies revealed myofiber degeneration of the tail muscles and liquefactive myopathy in the infected prawns. This was the first report on the occurrence of MrNV in the Malaysian freshwater prawn.
  9. Fulltext In-depth tanscriptomic analysis on giant freshwater prawns
    Mohd-Shamsudin MI, Kang Y, Lili Z, Tan TT, Kwong QB, Liu H, et al.
    PLoS One, 2013;8(5):e60839.
    PMID: 23734171 DOI: 10.1371/journal.pone.0060839
    Gene discovery in the Malaysian giant freshwater prawn (Macrobrachium rosenbergii) has been limited to small scale data collection, despite great interest in various research fields related to the commercial significance of this species. Next generation sequencing technologies that have been developed recently and enabled whole transcriptome sequencing (RNA-seq), have allowed generation of large scale functional genomics data sets in a shorter time than was previously possible. Using this technology, transcriptome sequencing of three tissue types: hepatopancreas, gill and muscle, has been undertaken to generate functional genomics data for M. rosenbergii at a massive scale. De novo assembly of 75-bp paired end Ilumina reads has generated 102,230 unigenes. Sequence homology search and in silico prediction have identified known and novel protein coding candidate genes (∼24%), non-coding RNA, and repetitive elements in the transcriptome. Potential markers consisting of simple sequence repeats associated with known protein coding genes have been successfully identified. Using KEGG pathway enrichment, differentially expressed genes in different tissues were systematically represented. The functions of gill and hepatopancreas in the context of neuroactive regulation, metabolism, reproduction, environmental stress and disease responses are described and support relevant experimental studies conducted previously in M. rosenbergii and other crustaceans. This large scale gene discovery represents the most extensive transcriptome data for freshwater prawn. Comparison with model organisms has paved the path to address the possible conserved biological entities shared between vertebrates and crustaceans. The functional genomics resources generated from this study provide the basis for constructing hypotheses for future molecular research in the freshwater shrimp.
  10. TaqMan probe real-time polymerase chain reaction assay for the quantification of canine DNA in chicken nugget
    Rahman MM, Hamid SB, Basirun WJ, Bhassu S, Rashid NR, Mustafa S, et al.
    Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2016;33(1):10-8.
    PMID: 26458055 DOI: 10.1080/19440049.2015.1104558
    This paper describes a short-amplicon-based TaqMan probe quantitative real-time PCR (qPCR) assay for the quantitative detection of canine meat in chicken nuggets, which are very popular across the world, including Malaysia. The assay targeted a 100-bp fragment of canine cytb gene using a canine-specific primer and TaqMan probe. Specificity against 10 different animals and plants species demonstrated threshold cycles (Ct) of 16.13 ± 0.12 to 16.25 ± 0.23 for canine DNA and negative results for the others in a 40-cycle reaction. The assay was tested for the quantification of up to 0.01% canine meat in deliberately spiked chicken nuggets with 99.7% PCR efficiency and 0.995 correlation coefficient. The analysis of the actual and qPCR predicted values showed a high recovery rate (from 87% ± 28% to 112% ± 19%) with a linear regression close to unity (R(2) = 0.999). Finally, samples of three halal-branded commercial chicken nuggets collected from different Malaysian outlets were screened for canine meat, but no contamination was demonstrated.
  11. Fulltext Revolutionizing colorectal cancer detection: A breakthrough in microbiome data analysis
    Mulenga M, Rajamanikam A, Kumar S, Muhammad SB, Bhassu S, Samudid C, et al.
    PLoS One, 2025;20(1):e0316493.
    PMID: 39879257 DOI: 10.1371/journal.pone.0316493
    The emergence of Next Generation Sequencing (NGS) technology has catalyzed a paradigm shift in clinical diagnostics and personalized medicine, enabling unprecedented access to high-throughput microbiome data. However, the inherent high dimensionality, noise, and variability of microbiome data present substantial obstacles to conventional statistical methods and machine learning techniques. Even the promising deep learning (DL) methods are not immune to these challenges. This paper introduces a novel feature engineering method that circumvents these limitations by amalgamating two feature sets derived from input data to generate a new dataset, which is then subjected to feature selection. This innovative approach markedly enhances the Area Under the Curve (AUC) performance of the Deep Neural Network (DNN) algorithm in colorectal cancer (CRC) detection using gut microbiome data, elevating it from 0.800 to 0.923. The proposed method constitutes a significant advancement in the field, providing a robust solution to the intricacies of microbiome data analysis and amplifying the potential of DL methods in disease detection.
  12. The role and influence of bacteria on the growth of Blastocystis sp. subtype 3
    Kaur CP, Yong CC, Rajamanikam A, Samudi C, Kumar S, Bhassu S, et al.
    Parasitol Res, 2023 Jul;122(7):1463-1474.
    PMID: 37162590 DOI: 10.1007/s00436-023-07842-2
    Blastocystis sp. is an enteric protistan parasite that affects individuals worldwide with gastrointestinal symptoms such as abdominal discomfort, diarrhea, and flatulence. However, its pathogenicity is controversial due to its presence among asymptomatic individuals. Blastocystis sp. subtype 3 (ST3) is the most prevalent subtype among humans that have been associated with irritable bowel syndrome (IBS), Crohn's disease, ulcerative colitis, and colorectal cancer. Axenization of the parasite has been shown to impede its growth thus revealing the importance of accompanying bacteria in ensuring Blastocystis sp. survival. This study aims to identify the influence of accompanying bacteria on the growth of Blastocystis sp. ST3. Blastocystis sp. cultures were treated with Meropenem, Vancomycin, and Amoxicillin-Clavulanic acid (Augmentin). Bacteria-containing supernatant of antibiotic-treated and control cultures were isolated and identified through 16 s rRNA sequencing. Morphological changes of antibiotic-treated Blastocystis sp. ST3 were also observed. The cultures treated with meropenem and augmentin exhibited opposing effects with reduced growth of isolates from symptomatic patients and a significant increase in asymptomatic isolates. Whereas, vancomycin-treated cultures had no difference in the growth of Blastocystis sp. ST3 isolates from symptomatic and asymptomatic patients. Isolates from symptomatic and asymtomatic patients had 6 and 2 distinct bacterial species identified with Proteus mirabilis as the common bacteria among both types of isolates. Morphologically, Blastocystis sp. ST3 cultures exposed to meropenem and augmentin demonstrated an increase in pre-cystic forms. These findings demonstrate the effects of accompanying bacteria on the growth of Blastocystis sp. ST3 that could translate into clinical manifestations observed among Blastocystis sp.-infected patients.
  13. A survey on the occurrence of papaya ringspot virus-P (PRSV-P) in Malaysia and genetic diversity assessment of the coat protein region
    Mohsin Mohsen N, Aziz MA, Thangaraja K, Mohammad Nasir MA, Md Zoqratt MZH, Bhassu S, et al.
    Environ Monit Assess, 2024 Nov 04;196(12):1154.
    PMID: 39495404 DOI: 10.1007/s10661-024-13321-7
    Papaya ringspot virus (PRSV) is a plant virus transmitted by aphids that has spread throughout many countries, including Malaysia, causing yield losses and economic impacts to the papaya industry worldwide. PRSV infection in papaya-distinctive ring-shaped patterns on papaya leaves resulted in stunted growth and reduced fruit quality. Management strategies such as the use of resistant varieties, cultural practices, and vector control are employed to mitigate the spread of PRSV. However, the evolution of new virus strains and the uncertainties posed by climate change pose ongoing challenges for the management of PRSV worldwide. Therefore, in this present study, we aim to confirm the presence of PRSV in symptomatic papaya leaves, to depict the current status of PRSV in Malaysia. Using reverse-transcription PCR (RT-PCR) targeting PRSV partial nuclear inclusion b protein (NIb) and coat protein (CP), 13 out of 40 papaya leaves collected were found positive for the PRSV strain-P (PRSV-P). Nucleotide analysis revealed a high similarity with strains from Taiwan and India, showing 96.83%, 97.03%, and 97.03% identity with the Taiwan strains (DQ340771, AY027810) and the India strain (KJ755852), respectively. Compared to the CP gene of Malaysian isolates reported in 2016 (EU082207), several nonsynonymous mutations have been discovered suggesting genetic diversity within the PRSV population in Malaysia. Overall, this study confirms the current circulation of PRSV infection in Malaysia since it was first identified in Johore in 1991. The re-occurrence of PRSV-P in this study highlights the need for continuous monitoring and targeted management strategies to prevent the further spread of PRSV-P in Malaysia.
  14. Fulltext Electronic Properties of Synthetic Shrimp Pathogens-derived DNA Schottky Diodes
    Rizan N, Yew CY, Niknam MR, Krishnasamy J, Bhassu S, Hong GZ, et al.
    Sci Rep, 2018 01 17;8(1):896.
    PMID: 29343758 DOI: 10.1038/s41598-017-18825-6
    The exciting discovery of the semiconducting-like properties of deoxyribonucleic acid (DNA) and its potential applications in molecular genetics and diagnostics in recent times has resulted in a paradigm shift in biophysics research. Recent studies in our laboratory provide a platform towards detecting charge transfer mechanism and understanding the electronic properties of DNA based on the sequence-specific electronic response, which can be applied as an alternative to identify or detect DNA. In this study, we demonstrate a novel method for identification of DNA from different shrimp viruses and bacteria using electronic properties of DNA obtained from both negative and positive bias regions in current-voltage (I-V) profiles. Characteristic electronic properties were calculated and used for quantification and further understanding in the identification process. Aquaculture in shrimp industry is a fast-growing food sector throughout the world. However, shrimp culture in many Asian countries faced a huge economic loss due to disease outbreaks. Scientists have been using specific established methods for detecting shrimp infection, but those methods do have their significant drawbacks due to many inherent factors. As such, we believe that this simple, rapid, sensitive and cost-effective tool can be used for detection and identification of DNA from different shrimp viruses and bacteria.
  15. Fulltext Publisher Correction: Electronic Properties of Synthetic Shrimp Pathogens-derived DNA Schottky Diodes
    Rizan N, Yew CY, Niknam MR, Krishnasamy J, Bhassu S, Hong GZ, et al.
    Sci Rep, 2018 Apr 12;8(1):6009.
    PMID: 29651139 DOI: 10.1038/s41598-018-24116-5
    A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
  16. Zoonotic infection with Onchocerca dewittei japonica in an 11-year-old boy in Kansai Region, Western Honshu, Japan
    Uni S, Fukuda M, Ogawa K, Lim YA, Agatsuma T, Bunchom N, et al.
    Parasitol Int, 2017 Oct;66(5):593-595.
    PMID: 28648713 DOI: 10.1016/j.parint.2017.06.006
    An 11-year-old boy living in Otsu City, Shiga Prefecture, Kansai Region, Western Honshu, Japan had zoonotic onchocercosis. The patient developed a painful swelling on the little finger of his left hand. The worm detected in the excised mass had external transverse ridges but did not have inner striae in the cuticle. On the basis of the parasite's histopathological characteristics, the causative agent was identified as a female Onchocerca dewittei japonica (Spirurida: Onchocercidae). The species of the filarial parasite was confirmed by sequencing the cox1 gene of the parasite. The Japanese wild boar Sus scrofa leucomystax is a definitive host for O. dewittei japonica, which is then transmitted by blackflies as the vector to humans. The current case described occurred in the Kansai Region, Western Honshu, where such infections were previously not reported.
  17. Fulltext Description, molecular characteristics and Wolbachia endosymbionts of Onchocerca borneensis Uni, Mat Udin & Takaoka n. sp. (Nematoda: Filarioidea) from the Bornean bearded pig Sus barbatus Müller (Cetartiodactyla: Suidae) of Sarawak, Malaysia
    Uni S, Mat Udin AS, Agatsuma T, Junker K, Saijuntha W, Bunchom N, et al.
    Parasit Vectors, 2020 Feb 06;13(1):50.
    PMID: 32028994 DOI: 10.1186/s13071-020-3907-8
    BACKGROUND: The genus Onchocerca Diesing, 1841 includes species of medical importance, such as O. volvulus (Leuckart, 1893), which causes river blindness in the tropics. Recently, zoonotic onchocercosis has been reported in humans worldwide. In Japan, O. dewittei japonica Uni, Bain & Takaoka, 2001 from wild boars is a causative agent for this zoonosis. Many filarioid nematodes are infected with Wolbachia endosymbionts which exhibit various evolutionary relationships with their hosts. While investigating the filarial fauna of Borneo, we discovered an undescribed Onchocerca species in the bearded pig Sus barbatus Müller (Cetartiodactyla: Suidae).

    METHODS: We isolated Onchocerca specimens from bearded pigs and examined their morphology. For comparative material, we collected fresh specimens of O. d. dewittei Bain, Ramachandran, Petter & Mak, 1977 from banded pigs (S. scrofa vittatus Boie) in Peninsular Malaysia. Partial sequences of three different genes (two mitochondrial genes, cox1 and 12S rRNA, and one nuclear ITS region) of these filarioids were analysed. By multi-locus sequence analyses based on six genes (16S rDNA, ftsZ, dnaA, coxA, fbpA and gatB) of Wolbachia, we determined the supergroups in the specimens from bearded pigs and those of O. d. dewittei.

    RESULTS: Onchocerca borneensis Uni, Mat Udin & Takaoka n. sp. is described on the basis of morphological characteristics and its genetic divergence from congeners. Molecular characteristics of the new species revealed its close evolutionary relationship with O. d. dewittei. Calculated p-distance for the cox1 gene sequences between O. borneensis n. sp. and O. d. dewittei was 5.9%, while that between O. d. dewittei and O. d. japonica was 7.6%. No intraspecific genetic variation was found for the new species. Wolbachia strains identified in the new species and O. d. dewittei belonged to supergroup C and are closely related.

    CONCLUSIONS: Our molecular analyses of filarioids from Asian suids indicate that the new species is sister to O. d. dewittei. On the basis of its morphological and molecular characteristics, we propose to elevate O. d. japonica to species level as O. japonica Uni, Bain & Takaoka, 2001. Coevolutionary relationships exist between the Wolbachia strains and their filarial hosts in Borneo and Peninsular Malaysia.

  18. Fulltext Morphological and molecular characteristics of Malayfilaria sofiani Uni, Mat Udin & Takaoka n. g., n. sp. (Nematoda: Filarioidea) from the common treeshrew Tupaia glis Diard & Duvaucel (Mammalia: Scandentia) in Peninsular Malaysia
    Uni S, Mat Udin AS, Agatsuma T, Saijuntha W, Junker K, Ramli R, et al.
    Parasit Vectors, 2017 Apr 20;10(1):194.
    PMID: 28427478 DOI: 10.1186/s13071-017-2105-9
    BACKGROUND: The filarial nematodes Wuchereria bancrofti (Cobbold, 1877), Brugia malayi (Brug, 1927) and B. timori Partono, Purnomo, Dennis, Atmosoedjono, Oemijati & Cross, 1977 cause lymphatic diseases in humans in the tropics, while B. pahangi (Buckley & Edeson, 1956) infects carnivores and causes zoonotic diseases in humans in Malaysia. Wuchereria bancrofti, W. kalimantani Palmieri, Pulnomo, Dennis & Marwoto, 1980 and six out of ten Brugia spp. have been described from Australia, Southeast Asia, Sri Lanka and India. However, the origin and evolution of the species in the Wuchereria-Brugia clade remain unclear. While investigating the diversity of filarial parasites in Malaysia, we discovered an undescribed species in the common treeshrew Tupaia glis Diard & Duvaucel (Mammalia: Scandentia).

    METHODS: We examined 81 common treeshrews from 14 areas in nine states and the Federal Territory of Peninsular Malaysia for filarial parasites. Once any filariae that were found had been isolated, we examined their morphological characteristics and determined the partial sequences of their mitochondrial cytochrome c oxidase subunit 1 (cox1) and 12S rRNA genes. Polymerase chain reaction (PCR) products of the internal transcribed spacer 1 (ITS1) region were then cloned into the pGEM-T vector, and the recombinant plasmids were used as templates for sequencing.

    RESULTS: Malayfilaria sofiani Uni, Mat Udin & Takaoka, n. g., n. sp. is described based on the morphological characteristics of adults and microfilariae found in common treeshrews from Jeram Pasu, Kelantan, Malaysia. The Kimura 2-parameter distance between the cox1 gene sequences of the new species and W. bancrofti was 11.8%. Based on the three gene sequences, the new species forms a monophyletic clade with W. bancrofti and Brugia spp. The adult parasites were found in tissues surrounding the lymph nodes of the neck of common treeshrews.

    CONCLUSIONS: The newly described species appears most closely related to Wuchereria spp. and Brugia spp., but differs from these in several morphological characteristics. Molecular analyses based on the cox1 and 12S rRNA genes and the ITS1 region indicated that this species differs from both W. bancrofti and Brugia spp. at the genus level. We thus propose a new genus, Malayfilaria, along with the new species M. sofiani.

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