Cattle is one of an important maintenance hosts involved in the transmission of leptospirosis. Serological method is always implemented to detect current or past leptospiral infection in cattle. The results can be obtained immediately compared to isolation method which need more longer time. An in-house IgG enzyme-linked immunosorbent assay (ELISA) using heat-killed whole cells of local isolate Leptospira kmetyi serovar malaysia strain Bejo-Iso9 (ELISABejo-Iso9) was developed in this study. The perfomance of ELISA-Bejo-Iso9 was compared with in-house IgGELISA using reference strain 117123 (hardjo bovis) antigen (ELISA-117123) and CUSABIO® commercial ELISA. It was found that, the performance of ELISA-Bejo-Iso9 was promising to compete with commercial ELISA. The specificity and sensitivity was 98.75% and 53.75%, respectively. The sensitivity of ELISA-Bejo-Iso9 was lower than commercial ELISA. However, the specificity of ELISA-Bejo-Iso9 was higher than commercial ELISA. Due to economically and availability factors, this finding suggested that the ELISA-Bejo-Iso9 can be used as an alternatif method for serological diagnosis of leptospirosis in cattle.
An adult female Elephant Trunk Snake (Acrochordus javanicus) was reported to have been weak and inappetent for five days. The following morning the snake found dead, while in the process of shedding its skin. On post mortem examination, there were multiple circumscribed caseous nodules of various sizes distributed all over the liver, along the respiratory tract and on the lungs. Bacteriological analysis of the lungs and liver swab samples yielded Burkholderia pseudomallei, which was confirmed by PCR amplification of specific 16S rRNA. The condition was diagnosed as melioidosis and the organism was genotypically characterized as sequence type 51, a genotype that has been previously characterized in humans in Malaysia. Antibiotic susceptibility by both Disc diffusion or Kirby Bauer and E-test minimum inhibitory concentration (MIC) showed that the organism exhibited susceptibility to meropenem, imipenem, ceftazidime, cotrimoxazole and co-amoxyclav; the antibiotics recommended in the treatment of melioidosis.
In this study, we report the molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) among veterinary students and personnel in Malaysia. Nasal and oral swabs were collected from 103 veterinary medicine students and 28 personnel from a veterinary hospital. Antibiotic sensitivity test (AST), minimum inhibitory concentration (MIC) test, and PCR amplifications of nucA and mecA gene were performed. Molecular characterization of the isolates was conducted using multilocus sequence typing (MLST), staphylococcal protein A gene (spa) typing, and pulsed-field gel electrophoresis (PFGE). Results from MLST show the presence of the pandemic and widespread MRSA clones, ST5 and ST59. Spa gene typing revealed spa type t267 which has a wide geographical distribution. A new spa type, t5697 was found in this study. Fingerprint analysis by using PFGE show heterogeneity of the isolates. These findings affirm the importance of MRSA in veterinary settings and underscore the need for further extensive research to devise contextual control and prevention strategies.
Methicillin-resistant Staphylococcus aureus (MRSA) is known to cause nosocomial infections and is now becoming an emerging problem in veterinary medicine. The objective of the study was to determine the presence of MRSA in 100 cats and dogs sampled between November 2007 and April 2008 at the University Veterinary Hospital, Universiti Putra Malaysia. MRSA was detected in 8% of pets sampled. Ten percent (5/50) and 6% (3/50) of the isolates were from dogs and cats, respectively. All MRSA isolates possessed the mecA gene and were found to be resistant to at least three antimicrobials with a minimum of Oxacillin MIC of 8 μg/mL. One isolate (CT04) had an extremely high MIC of >256 μg/mL. The MLST type ST59 found in this study have been reported earlier from Singapore and other countries as a strain from animal and community-associated MRSA respectively. Pulsed-field gel electrophoresis revealed five pulsotypes. Two isolates from cats (CT27 and CT33) and three isolates from dogs (DG16, DG20, and DG49) were respectively assigned to pulsotypes B and D. The study suggests that cats and dogs in Malaysia are potential reservoirs for MRSA.
A nested PCR assay was used to determine the viral RNA and proviral DNA status of naturally infected cats. Selected samples that were FeLV-positive by PCR were subjected to sequencing, phylogenetic analysis, and motifs search. Of the 39 samples that were positive for FeLV p27 antigen, 87.2% (34/39) were confirmed positive with nested PCR. FeLV proviral DNA was detected in 38 (97.3%) of p27-antigen negative samples. Malaysian FeLV isolates are found to be highly similar with a homology of 91% to 100%. Phylogenetic analysis revealed that Malaysian FeLV isolates divided into two clusters, with a majority (86.2%) sharing similarity with FeLV-K01803 and fewer isolates (13.8%) with FeLV-GM1 strain. Different enhancer motifs including NF-GMa, Krox-20/WT1I-del2, BAF1, AP-2, TBP, TFIIF-beta, TRF, and TFIID are found to occur either in single, duplicate, triplicate, or sets of 5 in different positions within the U3-LTR-gag region. The present result confirms the occurrence of FeLV viral RNA and provirus DNA in naturally infected cats. Malaysian FeLV isolates are highly similar, and a majority of them are closely related to a UK isolate. This study provides the first molecular based information on FeLV in Malaysia. Additionally, different enhancer motifs likely associated with FeLV related pathogenesis have been identified.
Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a problem in veterinary medicine and is no longer considered as a mere nosocomial pathogen. We studied the occurrence of MRSA in veterinary personnel, cats and dogs and the environmental premises in University Veterinary Hospital (UVH). We found the prevalence of MRSA as follows: UVH 2/28 (7.1%) staff, 8/100 (8%) of the pets [5/50 (10%) of the dogs and 3/50 (6%) of the cats)], and 9/28 (4.5%) of the environmental samples. Antibiotic sensitivity tests (AST) show multi-resistance characteristics of the MRSA and the minimum inhibitory concentration (MIC) values for the isolates ranged from 1.5 µg to >256 µg/ml. Molecular typing by using multi-locus sequence typing (MLST), staphylococcal protein A typing (spa typing) and pulsed-field gel electrophoresis (PFGE) was conducted and the results from MLST indicated that an isolate from a veterinary personnel (PG21), typed as ST1241 belonged to the same clonal complex (CC) as the two isolates from two dogs (DG16 and DG20), both being typed as ST59. The PFGE results revealed that the two isolates from two veterinary personnel, PG21 and PG16 belonged to closely related MRSA strains with isolates from dog (DG36) and from environmental surface (EV100) respectively. The fact that PFGE revealed close similarity between isolates from humans, a dog and environmental surfaces indicates the possibility for either of them to be the source of MRSA and the potential routes and risks of spread.
The emergence of multidrug resistance (MDR), including colistin resistance, among Enterobacteriaceae recovered from food animals poses a serious public health threat because of the potential transmission of these resistant variants to humans along the food chain. Village chickens or Ayam Kampung are free-range birds and are preferred by a growing number of consumers who consider these chickens to be organic and more wholesome. The current study investigates the antibiogram profiles of Salmonella isolates recovered from village chicken flocks in South-central Peninsular Malaysia. A total of 34 isolates belonging to eight serotypes isolated from village chickens were screened for resistance towards antimicrobials including colistin according to the WHO and OIE recommendations of critical antibiotics. S. Weltevreden accounted for 20.6% of total isolates, followed by serovars Typhimurium and Agona (17.6%). The majority of isolates (73.5%) demonstrated resistance to one or more antimicrobials. Eight isolates (23.5%) were resistant to ≥3 antimicrobial classes. Colistin resistance (minimum inhibitory concentrations: 4-16 mg/L) was detected among five isolates (14.7%), including S. Weltevreden, S. Albany, S. Typhimurium, and Salmonella spp. Univariable analysis of risk factors likely to influence the occurrence of MDR Salmonella revealed that the flock size, poultry production system, and use of antibiotics in the farm were not significantly (p > 0.05) associated with MDR Salmonella. The current study highlights that MDR Salmonella occur at a lower level in village chickens compared to that found in live commercial chickens. However, MDR remains a problem even among free-range chickens with minimal exposure to antibiotics.
Apart from occasional reports of clinical disease affecting horses, there is no information about Trypanosoma evansi in horses in Peninsula Malaysia. Thus, a cross-sectional study was conducted in eight states in Peninsula Malaysia to determine the active presence of T. evansi in horses. A total of 527 blood samples were obtained and examined by haematocrit centrifugation technique (HCT), Giemsa-stained thin blood smear (GSS), morphometric measurements, polymerase chain reaction (PCR) and cloning of PCR products. The results showed an overall parasitological prevalence of 0.57% (3/527, CI: 1.6-0.19%) with both HCT and GSS. Morphometric study revealed the mean total length of the trypanosomes including the free flagellum was 27.94 ± 2.63 μm. PCR successfully amplified a trypanosome specific 257 bp in 1.14% of samples (6/527, CI: 2.4-0.52%) and was confirmed by nucleotide sequences. The mean packed cell volume (PCV) for the positive cases detected by HCT was lower (23% ± 7.00) compared to the positive cases detected by PCR alone in the state of Terengganu (35% ± 4.73). In conclusion, this study showed T. evansi infection occurred in low frequency in horses in Peninsula Malaysia, and anaemia coincided with parasitaemic animals. PCR is considered as a sensitive diagnostic tool when parasitaemia is undetectable. The slight lengthier mean of parasite and anaemia may indicate a virulent strain of T. evansi circulating throughout the country. Thus, it's highly recommended to shed light on host-parasite relationship for better epidemiological understanding.
This paper presents investigation of lungworm disease outbreaks that is based on retrospective examination of cases recorded between 1994 and 2000 on a government beef cattle breeding centre in the state of Pahang, peninsular Malaysia. The breed of cattle on the centre was Nelore and the mean population over a 7-year period (from 1994 to 2000) was 1612. All animals were allowed to graze on pasture and mixed grazing was practiced on the farm. The routine de-worming programme was performed using levamisole and ivermectin from 1994 to 1998 and abamectin in 1999 and 2000 on 1 to 3-month-old calves and an annual dose given to the adult cattle. Nelore was introduced into the farm in 1991, three years before the first outbreak from Brazil where Dictyocaulus viviparus infection had been reported. No lungworm infection had been observed in the farm prior to the animal introduction. Within the 7-year period, 36 fatalities occurred and the annual mortality rate due to lungworm infection was 0.31%. The highest rate was recorded in 1997. Among the total 36 deaths, about 75% of deaths occurred in calves aged between 6 months and 12 months, 67% were males and 33% were female cattle. The highest number of deaths (19%) occurred in the month of November. In conclusion, D. viviparus infection may have been introduced into a tropical climate along with consignments of cattle from lungworm endemic areas resulting in fatal disease outbreaks for a few years following the animal's initial introduction.
A study on causes of lung condemnation in 25 abattoirs from peninsular Malaysia for a period of seven years (1998-2004) was conducted by examining the records at the Department of Veterinary Services headquarters in Kuala Lumpur. A total of 5.3% of lungs from 233,417 cattle and buffaloes were condemned from 1998 to 2004. The main cause of condemnation was congestion (2.98%). The percentage of lungs that were condemned due to parasitic infection among the total population slaughtered was low (0.11%). Parasitic infection contributed to 2.1% of all lungs condemned. It was also found that the prevalence of parasitic infection in the lungs was generally much higher in buffaloes than in cattle.
Toxoplasmosis is a worldwide zoonosis caused by the protozoa Toxoplasma gondii which affects human and animals. Village chickens (Gallus domesticus) most commonly known as Ayam Kampung or free-range chickens, have been suggested to play a role in the epidemiology of toxoplasmosis. This study determines the presence of T. gondii in the village chicken populations in two states of Malaysia. A total of 50 serum samples from the chickens from Selangor (n=20) and Melaka (n=30) were collected and analysed using commercial serological kits. T. gondii antigen was detected in 20% (Selangor 30%; Melaka 13%) samples using ELISA test and anti-T. gondii antibody was detected in all positive ELISA samples using the indirect haemagglutination test (IHAT). Histopathological examination revealed tissue changes such as inflammation and degeneration in brain and liver of seropositive chickens. This is the first report of T. gondii infection in the village chickens in Malaysia.
A cross-sectional study was designed to assess the seroprevalence and risk factors associated with Trypanosoma evansi infection among horses, using a total of 527 blood samples obtained from eight states in Peninsular Malaysia. A structured questionnaire was used to collect data on risk factors associated with T. evansi seroprevalence. The overall seroprevalence detected by card agglutination test for T. evansi (CATT/T. evansi) was 13.90% (73/527, CI: 11.2-17.1%). Female and exogenous horses showed a higher risk in association with the disease seroprevalence compared to other groups. The majority of the horse owners were not familiar with surra (85.30%). However, most of them were very cautious with the health of their animals. In conclusion, this study showed that T. evansi occurred in low frequency among horses in Peninsular Malaysia, and the good management system adopted by horse owners was probably responsible for the low T. evansi occurrence.
Vancomycin-resistant enterococci (VRE) have been reported to be present in humans, chickens, and pigs in Malaysia. In the present study, representative samples of VRE isolated from these populations were examined for similarities and differences by using the multilocus sequence typing (MLST) method. Housekeeping genes of Enterococcus faecium (n = 14) and Enterococcus faecalis (n = 11) isolates were sequenced and analyzed using the MLST databases eBURST and goeBURST. We found five sequence types (STs) of E. faecium and six STs of E. faecalis existing in Malaysia. Enterococcus faecium isolates belonging to ST203, ST17, ST55, ST79, and ST29 were identified, and E. faecium ST203 was the most common among humans. The MLST profiles of E. faecium from humans in this study were similar to the globally reported nosocomial-related strain lineage belonging to clonal complex 17 (CC17). Isolates from chickens and pigs have few similarities to those from humans, except for one isolate from a chicken, which was identified as ST203. E. faecalis isolates were more diverse and were identified as ST4, ST6, ST87, ST108, ST274, and ST244, which were grouped as specific to the three hosts. E. faecalis, belonging to the high-risk CC2 and CC87, were detected among isolates from humans. In conclusion, even though one isolate from a chicken was found clonal to that of humans, the MLST analysis of E. faecium and E. faecalis supports the findings of others who suggest VRE to be predominantly host specific and that clinically important strains are found mainly among humans. The infrequent detection of a human VRE clone in a chicken may in fact suggest a reverse transmission of VRE from humans to animals.
A study was conducted to describe the prevalence and distribution of zoonotic Brucella melitensis in goats in Peninsular Malaysia. Using serosurveillance data of the last decade (2000-2009) involving 119,799 goats and 3555 farms, the seroprevalence of brucellosis among goats was 0.91% (95% CI=0.86-0.96) and among farms was 7.09% (95% CI=6.27-7.98). The odds of brucellosis was significantly (P<0.05) higher in the later part of the decade, in larger herd size and among the states located in the peninsula as compared to eastern Malaysia. The infection was detected throughout Malaysia but at generally low seroprevalences with states like Perlis that border neighbouring countries having higher seroprevalence of brucellosis than other non-border states.
Bovine brucellosis was first reported in Peninsular Malaysia in 1950. A subsequent survey conducted in the country revealed that the disease was widespread. Current knowledge on the potential risk factors for brucellosis occurrence on cattle farms in Malaysia is lacking. Therefore, we conducted a case-control study to identify the potential herd-level risk factors for bovine brucellosis occurrence in four states in the country, namely Kelantan, Pahang, Selangor and Negeri Sembilan. Thirty-five cases and 36 controls of herds were selected where data on farm management, biosecurity, medical history and public health were collected. Multivariable logistic regression identified that Brucella seropositive herds were more likely to; have some interaction with wildlife (OR 8.9, 95% CI = 1.59-50.05); originated from farms where multiple species such as buffalo/others (OR 41.8, 95% CI = 3.94-443.19) and goat/sheep (OR 8.9, 95%Cl = 1.10-71.83) were reared, practice extensive production system (OR 13.6, 95% CI 1.31-140.24) and have had episodes of abortion in the past (OR 51.8, 95% CI = 4.54-590.90) when compared to seronegative herds. Considering the lack of information on the epidemiology of bovine brucellosis in peninsular Malaysia and absence of information on preventing the inception or spread of the disease, this report could contribute to the on-going area-wise national brucellosis eradication program.
Bovine brucellosis is an important disease affecting cattle characterised by abortion, still birth, reduced milk production, weak foetus and infertility in both males and females. There is wide distribution of the disease among cattle and several wildlife species. Bovine brucellosis is commonly caused by B. abortus and very occasionally B. melitensis and B. suis. The distribution of bovine brucellosis in cattle has not been described in Malaysia. In this paper we describe the distribution, pattern and trend of bovine brucellosis in Peninsular Malaysia between 2000 and 2008 based on serological data obtained from nationwide B. abortus serosurveillance activities in cattle populations.
This is a retrospective study of the outbreaks of foot-and-mouth disease (FMD) in Peninsular Malaysia between 2001 and May 2007. In total, 270 outbreaks of FMD were recorded. Serotype O virus (89.95 %) and serotype A (7.7 %) had caused the outbreaks. Significant differences on the occurrence of FMD were found between the years (t = 5.73, P = 0.000, df = 11), months (t = 4.7, P = 0.000, df = 11), monsoon season (t = 2.63, P = 0.025, df = 10) and states (t = 4.84, P = 0.001, df = 10). A peak of outbreaks observed in 2003 could be due to increased animal movement and the other peak in 2006 could be due to a compromised FMD control activities due to activities on the eradication of highly pathogenic avian influenza. Cattle (86 % of outbreaks) suffered the most. However, no difference in disease occurrence between species was observed. The populations of cattle (r = 0.672, P = 0.023) and sheep (r = 0.678, P = 0.022) were significantly correlated with occurrence of FMD. Movement of animals (66 % of outbreaks) was the main source for outbreaks. A combination of control measures were implemented during outbreaks. In conclusion, the findings of this study show that FMD is endemic in Peninsular Malaysia, and information gained could be used to improve the existing control strategy.
This study determined the risk factors and characteristics of vancomycin-resistant Enterococci (VRE) among individuals working with animals in Malaysia.
Following a series of H5N1 cases in chickens and birds in a few states in Malaysia, there was much interest in the influenza A viruses subtypes that circulate among the local pig populations. Pigs may act as a mixing vessel for avian and mammal influenza viruses, resulting in new reassorted viruses. This study investigated the presence of antibodies against influenza H1N1 and H3N2 viruses in pigs from Peninsular Malaysia using Herdcheck Swine Influenza H1N1 and H3N2 Antibody Test Kits. At the same time, the presence of influenza virus was examined from the nasal swabs of seropositive pigs by virus isolation and real time RT-PCR. The list of pig farms was obtained from the headquarters of the Department of Veterinary Services, Malaysia, and pig herds were selected randomly from six of 11 states in Peninsular Malaysia. A total of 727 serum and nasal swab samples were collected from 4- to 6-month-old pigs between May and August 2005. By ELISA, the seroprevalences of swine influenza H1N1 and H3N2 among pigs were 12.2% and 12.1% respectively. Seropositivity for either of the virus subtypes was detected in less than half of the 41 sampled farms (41.4%). Combination of both subtypes was detected in 4% of all pigs and in 22% of sampled farms. However, no virus or viral nucleic acid was detected from nasal samples. This study identified that the seropositivity of pigs to H1N1 and H3N2 based on ELISA was significantly associated with factors such as size of farm, importation or purchase of pigs, proximity of farm to other pig farms and the presence of mammalian pets within the farm.