Occurrence, Morphometric, and Molecular Investigation of Cattle and Buffalo Liver Adult Fluke in Peninsular Malaysia Main Abattoirs.

Fascioliasis is a parasitic disease of human and animal caused by Fasciola gigantica (F. gigantica) and Fasciola hepatica (F. hepatica). More than 700 million of grazing animals and over 180 million human population are at the risk of fascioliasis. This study was conducted in Banting, Ipoh, Shah Alam, and Taiping abattoirs within Peninsular Malaysia to determine the occurrence and identify the species of liver flukes, causing liver condemnation in cattle and buffaloes. Within a study period from January to December 2018, a total of 25 condemned bovine livers were collected from Banting, Ipoh, Shah Alam, and Taiping abattoirs and analyzed. Taiping abattoir had the highest occurrence of fascioliasis [14/1014 (1.38%)], and Shah Alam abattoir had the least occurrence [1/3377 (0.03%)]. From all the sampled livers, the average number of adult flukes recorded ranged from 1 to 83. A total of 440 adult flukes were studied morphometrically, using parameters such as body length (BL), body width (BW), cone length (CL), cone width (CL), body area (BA), body perimeter (BP), the distance between the ventral sucker and posterior end of body (VS-P), BL/BW ratio, BL/V-SP, and body roundness (BR). Furthermore, molecular analysis was conducted using PCR-RFLP methods to distinguish between F. gigantica and F. hepatica involving ITS1 primer and RsaI restriction enzyme. RFLP pattern with RsaI produced a consistent pattern of 360,100 and 60 bp fragments in F. hepatica, whereas F. gigantica worms had a profile of 360,170, and 60 bp in size. The morphometric and molecular analysis results indicated that cattle and buffaloes slaughtered at Banting, Shah Alam, Taiping, and Ipoh abattoirs were infected with F. gigantica.

A Retrospective Study on Bovine Fascioliasis in Veterinary Regional Laboratories in Peninsular Malaysia.

This is a retrospective study on bovine fascioliasis infection on cattle in Peninsular Malaysia, spanning from January 2007 to December 2017. Cattle were diagnosed with Fasciola based on the fecal examination and the results were reported to the Veterinary Regional Laboratories in Peninsular Malaysia. These records were analysed for the occurrence of bovine fascioliasis within that 11-year period. Records of annual diagnostic cases from five major Veterinary Regional Laboratories were examined: Bukit Tengah, Pulau Pinang (north); Kuantan, Pahang (east); Johor Bahru, Johor (south); Sepang, Selangor (west); and Kota Bharu, Kelantan (east). A positive fascioliasis infestation was calculated based on a number of positively infected cattle with Fasciola from a number of cattle examined. A total of 1988 cattle were examined during this period and 35 (1.76%) cattle were reported to be positive for bovine fascioliasis. Parasite infection was the highest at Bukit Tengah region (5.55%) where 19 cases were positive from 342 reported cases while, at Kuantan (4.96%), 15 positive cases were reported, unlike Johor Bahru (0.09%), with only 1 positive case from 1136 cattle examined. Sepang and Kelantan had no positive cases. These results showed that bovine fascioliasis was generally prevalent in the northern and southeast parts of the Peninsular Malaysia; however, there was no significant relationship between the region and the occurrence of fascioliasis.

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.

Therapeutic use of Lucilia sericata maggot in controlling bacterial bio-burden in Rat wound model.

Delayed wound healing due to extraneous bacterial contamination, antibacterial resistance and other associated factors are of great concern in dealing patients having chronically infected wound. Medicinal properties of certain maggots of Calliphoridae family are known for its effective wound debridement therapy. The objective of the study was to evaluate the wound healing potential of maggots of Lucilia sericata in an experimentally infected cutaneous wound model in Wistar rat. The study was carried out by using male Wistar rats (n=48) by creating excisional wounds and later contaminated with mixed population of gram positive and gram-negative bacteria. Animals were divided randomly in to four groups with 12 individuals each, being denominated as control, antibiotic treated, maggot treated, and antibiotic plus maggot combination treated group. Ten pre-sterilized maggots were applied per centimetre square wound bed for 24 hours. Different wound kinetics in L. sericata maggot treated wounds revealed significant reduction in wound area with maximum contraction, early elimination of bacterial bioburden as compared to group of infected control and group of rats receiving only antibiotic treatment. The histopathological examination of wounded tissue of maggot treated groups showed early and better epithelialization, collagenation and neovascularization with complete healing of wound in two weeks. The maggot effects on healing when used singly or in combination with antibiotic were recorded to be similar. The results of the present study clearly demonstrate that the maggots of L. sericata possesses a definite antibacterial action along with removal of dead tissues and effectively reduced the bacterial bio-burden in infected wound and induced wound healing quickly.

Impact of pH on the viability and morphology of Blastocystis isolates.

Blastocystis sp. is ubiquitous in avian, mammalian and human hosts and propagates in either neutral or slightly alkaline conditions within the host's gastro-intestinal tract. Of the few previous studies on this enteric protozoan parasite in feline and canine hosts, prevalence values have been shown to range between 0 to 70.8%. In view of the close association between humans, and canine and feline hosts as companion animals, faecal samples of 180 Felis catus and 82 Canis lupus, collected from Penang and Kuala Lumpur, Malaysia, were initially screened by in vitro cultivation followed by molecular characterization. No positive isolates were identified in culture but in 12 feline samples DNA barcoding detected a zoonotic subtype Blastocystis ST1 for the first time. Consequently, avian and human isolates, which had previously been successfully cultured, were used to investigate the impact of pH on the viability and morphology of Blastocystis sp. The use of Trypan blue showed that the number of viable cells increased when exposed to pH 4 and a significant increase in viability occurred in pH values of 5 to 7. Development of Blastocystis cells in both isolates was suppressed in media less than pH 5 followed by the disappearance of viable cells from avian isolates in more acidic media below pH 4. Morphologically at pH 4 cells from avian isolates were less rounded, and with wrinkled / shrunken surfaces, than the more normal rounded cells from human isolates. On the other hand, at values below pH 3, no viable cells in human isolates were visible. The present findings therefore confirm that gastro-intestinal pH is an important determinant of Blastocystis viability and consequently influences the epidemiology of infection within avian, mammalian and human hosts.

Genetic diversity of rodent Blastocystis sp. from Peninsular Malaysia.

Rodents are ubiquitous zoonotic vectors for many human pathogens including Blastocystis sp. In this study, we examined the prevalence and subtypes of Blastocystis sp. in rodents captured from Peninsular Malaysia. A total of 293 rodents predominantly brown rat (Rattus norvegicus) (290 of 293, 99.0%) and house shrew (Suncus murinus) (3 of 293, 1.0 %), were captured in the vicinity of popular eateries in two cities (Kuala Lumpur and Ipoh) in Peninsular Malaysia. In vitro cultivation method showed presence of Blastocystis sp. in approximately half (133 of 290, 45.9%) of the brown rats tested. Among the 47 Blastocystis isolates subtyped using partial small subunit ribosomal RNA gene analysis, ST4 was the most abundant (43 of 47, 91.5%) followed by ST1 (2 of 47, 4.3%), ST5 (1 of 47, 2.1%) and ST7 (1 of 47, 2.1%). Our findings highlighted the importance of rodents as a source of Blastocystis sp. infection in Malaysia and showed the high prevalence of ST4 within the rodent population infected with Blastocystis sp.

Molecular Characterization of Avian Infectious Bronchitis Virus Isolated in Malaysia during 2014-2016.

Avian Infectious Bronchitis (IB) is a highly contagious disease which can cause huge economic losses to the poultry industry. Forty five IB viruses (IBV) were isolated from poultry in Malaysia during 2014-2016. Phylogenetic analysis of the spike glycoprotein 1 (S1) gene revealed that all isolates were clustered into five distinct groups. The predominant type of IBV isolated was QX strains (47%), second was 4/91 type (27%), followed by Malaysian strain MH5365/95 (13%), Massachusetts type (11%) and finally Taiwanese strains (2%). Four types of S1 protein cleavage recognition motifs were found among the isolates which includes HRRRR, RRSRR, RRFRR and RRVRR. To our knowledge, this is the first report describing the motif RRVRR and are unique to Malaysian strains. Six IBVs were grouped in Malaysian MH5365/95 strains. Among these, one isolate was different from others where it only shared 82% identity with MH5365/95 and to others. It formed its own branch in the Malaysian cluster suggesting it may be a variant unique to Malaysia. Alignment analysis of the S1 amino acid sequences indicated that point mutations, insertions and deletions contribute to the divergence of IB variants. This study indicated at least five groups of IBV are circulating in Malaysia with most of the isolates belonged to QX strains. As new IBV variants continue to emerge, further study need to be carried out to determine whether the current available vaccine is able to give protection against the circulating virus.

Molecular Characterization of Avian Infectious Bronchitis Virus Isolated in Malaysia during 2014-2016

@#Avian Infectious Bronchitis (IB) is a highly contagious disease which can cause huge economic losses to the poultry industry. Forty five IB viruses (IBV) were isolated from poultry in Malaysia during 2014-2016. Phylogenetic analysis of the spike glycoprotein 1 (S1) gene revealed that all isolates were clustered into five distinct groups. The predominant type of IBV isolated was QX strains (47%), second was 4/91 type (27%), followed by Malaysian strain MH5365/95 (13%), Massachusetts type (11%) and finally Taiwanese strains (2%). Four types of S1 protein cleavage recognition motifs were found among the isolates which includes HRRRR, RRSRR, RRFRR and RRVRR. To our knowledge, this is the first report describing the motif RRVRR and are unique to Malaysian strains. Six IBVs were grouped in Malaysian MH5365/95 strains. Among these, one isolate was different from others where it only shared 82% identity with MH5365/95 and to others. It formed its own branch in the Malaysian cluster suggesting it may be a variant unique to Malaysia. Alignment analysis of the S1 amino acid sequences indicated that point mutations, insertions and deletions contribute to the divergence of IB variants. This study indicated at least five groups of IBV are circulating in Malaysia with most of the isolates belonged to QX strains. As new IBV variants continue to emerge, further study need to be carried out to determine whether the current available vaccine is able to give protection against the circulating virus.

Therapeutic use of Lucilia sericata maggot in controlling bacterial bio-burden in Rat wound model

@#Delayed wound healing due to extraneous bacterial contamination, antibacterial resistance and other associated factors are of great concern in dealing patients having chronically infected wound. Medicinal properties of certain maggots of Calliphoridae family are known for its effective wound debridement therapy. The objective of the study was to evaluate the wound healing potential of maggots of Lucilia sericata in an experimentally infected cutaneous wound model in Wistar rat. The study was carried out by using male Wistar rats (n=48) by creating excisional wounds and later contaminated with mixed population of gram positive and gram-negative bacteria. Animals were divided randomly in to four groups with 12 individuals each, being denominated as control, antibiotic treated, maggot treated, and antibiotic plus maggot combination treated group. Ten pre-sterilized maggots were applied per centimetre square wound bed for 24 hours. Different wound kinetics in L. sericata maggot treated wounds revealed significant reduction in wound area with maximum contraction, early elimination of bacterial bioburden as compared to group of infected control and group of rats receiving only antibiotic treatment. The histopathological examination of wounded tissue of maggot treated groups showed early and better epithelialization, collagenation and neovascularization with complete healing of wound in two weeks. The maggot effects on healing when used singly or in combination with antibiotic were recorded to be similar. The results of the present study clearly demonstrate that the maggots of L. sericata possesses a definite antibacterial action along with removal of dead tissues and effectively reduced the bacterial bio-burden in infected wound and induced wound healing quickly.

Current status of Blastocystis in cockroaches.

There are few reports on Blastocystis spp. infections in invertebrate hosts namely, cockroaches. Due to their close proximity to humans especially to their dwellings prompted this study as these organisms could possibly play a role in human transmission. A total of 151 cockroaches consisted predominantly of nymph and adult stages were captured from several types of dwellings in the state of Perak and Selangor, Malaysia. Approximately half (40.4%) of the cockroach intestinal contents screened were positive and were found associated to two main factors, host-stage and types of dwellings. The granular and vacuolated forms were the most common cell form found in the in vitro cultures and were morphologically similar to B. hominis. However, the surface coat observed was thick with an electron lucent area observed in the central vacuole. The isolates grew in room temperature but optimal growth was observed at a 24ºC similar to the reptilian Blastocystis with a high number of cells were recovered. Using the DNA barcoding method, two isolates were identified as ST3 (allele 56), one isolate was consider as the new subtype with close relation to allele 114.

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.

Evaluation of enhanced virgin coconut oil and senduduk (Melastoma malabathricum) as anthelmintics against caprine strongyle nematodes.

A study was conducted to evaluate the anthelmintic properties of enhanced virgin coconut oil (EVCO) and senduduk (Melastoma malabathricum) plant against strongyle nematodes in goats. Two preparations of 10% EVCO dissolved in 90% virgin coconut oil and 10% EVCO dissolved in 90% palm oil, were given orally to two groups of mixed breeds goats. The efficacy test indicated that EVCO was insufficiently active as an anthelmintic. Four concentrations of senduduk solution (1.25, 2.5, 5.0 and 10 mg ml(-1)) were compared with a control and albendazole in an in vitro test for larvicidal effect. There was no significant larval mortality using senduduk solution. An in vivo test of senduduk was conducted by comparing three groups of goats, namely control, levamisole and treatment groups that were given a daily oral dose of senduduk crude extract with 1g kg(-1) from Day 0 to Day 12 and 2 g kg(-1) from Day 13 to Day 30. This efficacy test with senduduk also gave negative results. The findings obtained indicated that EVCO and senduduk were ineffective as anthelmintics against caprinestrongyle nematodes at the concentrations used.

Zoonotic parasites from exotic meat in Malaysia.

Four zoonotic parasites, Sarcocystis spp., Toxoplasma gondii, Trichinella spp. and Taenia spp were screened in exotic meats. A total of forty-six (n=46) meat samples from various species of exotic animals were received from all the 14 states in Malaysia from January 2012 to April 2012. All exotic meat samples were examined macroscopically and histologically for the four zoonotic parasites. Results by histological examination of exotic meats showed the presence of Sarcocystis and Toxoplasma cysts at 8.7% (n=4) and 4.3% (n=2) respectively. No Trichinella spp. and Taenia spp. were found.

Comparative seroprevalences of bovine trypanosomiasis and anaplasmosis in five states of Malaysia.

A comparative seroprevalence study on bovine trypanosomiasis and anaplasmosis was conducted. Sera of adult cattle and buffaloes of different breeds from farms from five different states in Malaysia were collected and tested for the presence of Trypanosoma evansi antibodies by CATT and Anaplasma marginale antibodies by c-ELISA. Of the 116 samples, 14.7% tested positive for bovine trypanosomiasis and 77.6% for bovine anaplasmosis.

Comparative seroprevalences of bovine trypanosomiasis and anaplasmosis in five states of Malaysia

Abstract. A comparative seroprevalence study on bovine trypanosomiasis and anaplasmosis was conducted. Sera of adult cattle and buffaloes of different breeds from farms from five different states in Malaysia were collected and tested for the presence of Trypanosoma evansi antibodies by CATT and Anaplasma marginale antibodies by c-ELISA. Of the 116 samples, 14.7% tested positive for bovine trypanosomiasis and 77.6% for bovine anaplasmosis.

The seroprevalence of bovine babesiosis in Malaysia.

One hundred sera of Malaysian cattle were used in this seroprevalence study for bovine babesiosis. All sera were obtained from the Serological Unit of the Veterinary Research Institute (VRI), Ipoh, Perak. The sera were tested using a Veterinary Medical Research & Development (VMRD) commercial Indirect Immunofluourescent Antibody Test (IFAT) kit. The results showed that 17.0% were found to be positive for Babesia bovis, 16.0% for Babesia bigemina, and 9.0% for both B. bovis and B. bigemina infections.

Prevalence of canine ehrlichiosis in Perak State, peninsular Malaysia.

The Indirect Fluorescent Antibody Test (IFAT) and thin blood smears were conducted to establish the prevalence of Ehrlichia canis infection in dogs presented for treatment by pet owners at five private and one government veterinary clinic. Results showed that 15% of the dogs were positive for the parasite via IFAT, but none using blood smears. However, infected dogs did not show severe clinical symptoms of canine monocytic ehrlichiosis (CME).

A survey of parasitic infection on small ruminant farms in Kinta and Hilir Perak districts, Perak, Malaysia.

This paper reports the occurrence of helminth and protozoan infections on small ruminants from eight farms situated in Kinta and Perak Tengah district, Perak. The results of this survey indicate that helminthiasis and coccidiosis is rampant in sheep and goat farms. Several anthelmintics have been used for the control of helminths. The smallholders depended on health and extention services from the State Veterinary Department. This survey is part of an ongoing programme by the Department of Veterinary Services to upgrade services and report the current status of parasitic diseases in the state.

Seroprevalence of Toxoplasma gondii antibodies in pigs, goats, cattle, dogs and cats in peninsular Malaysia.

Antibodies to the protozoan parasite, Toxoplasma gondii were assayed in sera of 200 goats, 100 pigs, 126 cattle from various states of Malaysia, and 135 dogs and 55 cats around Ipoh region using an indirect fluorescence antibody test (IFAT, cut-off titer 1:200); antibodies were found in 35.5% of goats, 14.5% cats, 9.6% dogs, 7.9% local cattle and 4% yellow cattle but not in pigs. Results indicate that infection is most prevalent in goats.

Mortality of domesticated java deer attributed to Surra.

This paper reports an outbreak of trypanosomiasis due to Trypanosoma evansi in Java deer (Cervus timorensis) on a government deer farm in Lenggong, Perak. Seventeen adult female Java deer were found dead within a week. Symptoms of dullness, inappetence, anaemia, anorexia, respiratory distress and recumbency were seen prior to death in the infected Java deer. Beside trypanosomiasis, other parasitic infections such as theileriosis, helminthiasis and ectoparasite infestation were also recorded. Post mortem results showed generalized anaemia in most animals with isolated cases of jaundice. There was no significant finding with respect to bacteriological and viral investigations.