Presence of vancomycin resistance among enterococcus isolates from stray cats in Universiti Putra Malaysia and selected neighbourhood in Sri Serdang, Selangor, Malaysia

The present study was undertaken to determine the distribution of Enterococcus species from stray cats in Universiti Putra Malaysia and Seri Serdang and the presence of vancomycin resistance among the isolates. Fifty-five rectal swabs were collected from stray cats found in UPM and around the area of Sri Serdang. The Enterococcus species isolated were inoculated onto vancomycin resistant enterococci (VRE) agar supplemented with 8 μg/mL of vancomycin. Biochemical tests such as catalase, bile-aesculin and 6.5% NaCI were conducted to further confirm VRE isolates. Multiplex polymerase chain reaction assay (PCR) were performed for Enterococcus genus and species identification and vancomycin-resistant gene detection. Presence of Enterococcus spp. were demonstrated in every rectal sample tested. Two species were identified: Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium). Among 55 isolates of Enterococcus tested, none was resistant to vancomycin at 8 μg/mL

Molecular study of Babesia in canine blood and comparison between conventional and molecular diagnostic methods.

A molecular study was conducted to detect the presence and determine the prevalence of Babesia species in stray and pet dogs in Kuala Lumpur using the Polymerase Chain Reaction (PCR) method. Seventy dogs, 35 from pet dogs presented to clinics around Kuala Lumpur and 35 from stray dogs from the Dewan Bandaraya Kuala Lumpur (DBKL) dog pound, were included in this study. Thin blood films were made, stained with Giemsa and examined under a light microscope for the detection of Babesia organisms. Two out of 70 dogs (2.8%) were positive for canine Babesia. One was identified as Babesiacanis positive and the other Babesiagibsoni positive. Genus-specific screening PCR was performed on DNA extracted from all 70 samples followed by Babesia canis-specific and Babesia gibsoni-specific PCR. Nine out of 70 dogs (12.8%) were positive following genus-specific screening PCR but of the 9, only one was positive for Babesia canis and one for Babesia gibsoni. The two positive samples were the same as those detected using light microscopy. Both of the positive samples were from the stray group. Haematological abnormalities in the two Babesia positive dogs included anemia and thrombocytopaenia. The prevalence rate of canine babesiosis was 5.8% for the stray group and 0% for the pet group. The overall prevalence of canine babesiosis in Kuala Lumpur was found to be 2.85%.This is the first molecular study of canine Babesia in Malaysia

Evaluation of the survival of vancomycin-resistant enterococci (VRE) isolated from chickens and possible inactivation by in-use concentration of Lindores®, Ecos Timsen® and Omnicide®

Vancomycin-resistant enterococci (VRE) are well-known ascendant nosocomial pathogens. The recent detection of epidemiologic strain carrying vanA gene in the community of people working with animals and in chickens has brought to the forefront the potential public health danger posed by these organism. The farm environment is a major source of VRE persistence in poultry farms. We carried out survival test to test the survival of the VRE isolates on dry condition and surface test to evaluate the inactivation of the isolates by in-use concentration of commonly used disinfectants. In the survival test, all isolates survived for at least 4 weeks in colony counts of (1.00 × 103 – 3.86 × 103 CFU/ml) under clean condition and (1.00 × 103 – 2.02 × 104) for soiled condition. Those that were suspended in 5% BSA solution to mimic organic matter load as obtainable on farms survived for at least 8 weeks at (1.54 × 102 – 1.34 × 103 CFU/ml). In the surface test, inactivation of VRE isolates by in-use concentration of Lindores, Omnicide and Ecos Timsen was tested using the European surface test (EST). All the tested disinfectants were active against the VRE isolates on both the standard test surface (stainless steel) and our test surface (wooden). The results shows microbiocidal effects (ME)for test disinfectants, i.e. the log10 CFU of micro-organisms compared between test biocide and control treated with distilled water, after 7 min of exposure as follows; Lindores® active on both surfaces 5.24 and 3.17, Ecos Timsen® active significantly on steel 4.90 than wood 2.98 and Omnicide® significantly less active on stainless steel 2.40 than on wood 3.50

Toxoplasma gondii infection in native village chickens (Gallus domesticus) in Selangor and Melaka, Malaysia

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 villagechicken 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%) samplesusing 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

Species distribution and resistance phenotypes of vancomycin-resistant enterococcus isolated from pigs in Pulau Pinang, Malaysia

Vancomycin-resistant enterococci (VRE) are important nosocomial pathogens. The extensive use of avoparcin as a growth promoter in poultry and pigs is the hypothesized factor for the emergence of vancomycin resistance in enterococci in animals. As pork is one of the major protein sources for 30% of Malaysians, the present study was conducted to elucidate the role of pigs in the epidemiology of VRE. In this study, 220 rectal swabs were collected from pigs at 12 pig farms in Pulau Pinang. The study found 10 of 12 farms (83.3%) and 92 (41.8%) of the sampled pigs were positive for VRE. Of the 92 isolates examined by PCR, E. faecium (14%), E. casseliflavus (21.7%), E. gallinarum (1.1%) and other Enterococcus species (63.0%) were identified. VanA was detected in E. faecium and E. gallinarum. Questionnaire survey indicated that none of the sampled farms had used glycopeptides, either for growth promotion or for therapy. Tylosin, which has also been associated with vancomycin cross-resistance, was used in 41.8% of the sampled farms; however, there was no significant difference (P>0.05) between the proportion of VRE detected in the farms which used tylosin to those farms which did not. E-test on selected 49 isolates showed 16.0% of the isolates had MIC≤8 and 22.0% had MIC≥32. Single isolates of E. faecium and E. gallinarum, both possessed the resistance gene vanA, showed very high resistance (MIC>256). About 10.0% of the isolates, in which van genes was not detected, had MIC>32. In conclusion E. faecium and E. faecalis were found to be present at a low rate in the pigs sampled in this study. However, detection of vanA with high level of vancomycin resistance (MIC>256) highlights the potential public health threat associated with the pigs

Awareness of operators on the requirements and procedures for animal quarantine and the distribution of temporary animal quarantine stations in Peninsular Malaysia

Temporary quarantine stations (TQS) are transitory premises that have been approved to facilitate the quarantine of imported live animals in Malaysia. These stations must abide to the standard operating procedures (SOP) for animal quarantine as outlined by the veterinary authority in Malaysia. However, the level of awareness for the quarantine procedures among the TQS operators and managers has not been assessed. This study was conducted to describe the distribution of the TQS in 2012-2013 and the level of awareness among its operators on the quarantine procedures and the fundamental requirements for quarantine establishments. Eight TQS from 25 were selected and operators or managers were interviewed using a questionnaire and the facility was visited. The study found that majority (82.5%) of the TQS operators were aware of the quarantine procedures but the auditors from the veterinary authority revealed vice versa

Prevalence of methicillin-resistant Staphylococcus aureus in stray cats around colleges of Universiti Putra Malaysia and selected neighbourhoods of Seri Serdang, Selangor, Malaysia

The existence of Methicillin-resistant Staphylococcus aureus (MRSA) is due to the widespread utilization of antimicrobial agents. Besides humans, MRSA has not only been reported in livestock animals but also in pet animals such as the dogs and cats. This study was designed to estimate the prevalence of MRSA in stray cats around colleges of Universiti Putra Malaysia (UPM) and in selected neighbourhood of Sri Serdang. The samples for this study were taken from the nostrils of the stray cats using small sterile cotton swabs. The swabs were then inoculated onto blood agar (BA) and suspected colonies were gram stained. Gram positive cocci bacteria colonies were then inoculated onto mannitol salt agar (MSA) and yellowish colonies that grew were subjected to a series of biochemical tests such as the catalase, coagulase and Staphytect Plus latex agglutination test before they were inoculated onto the oxacillin resistance screening agar base (ORSAB). A total of seven (12.73%) MRSA from 55 samples were isolated from the stray cats. The results showed that the prevalence of MRSA in stray cats at colleges of Universiti Putra Malaysia and Seri Serdang are high (>10%). The stray cats may have contracted MRSA from contact with infected humans or via contaminated environments at the health care facilities such as the Student Medical Centre which is located in the college area of UPM and also from consuming contaminated raw meat from markets in Sri Serdang

Molecular relatedness of methicillin-resistant s. aureus isolates from staff, environment and pets at university veterinary hospital in Malaysia

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 detection and comparison of antimicrobial resistance pattern of vancomycin-resistant enterococci and Salmonella isolated from eggs of commercial layers and free-range chickens

The prevalence of vancomycin-resistant enterococci (VRE) and Salmonella in eggs produced by free-range chickens and commercial layers was compared to establish the microbial safety of the food and to determine if management, especially feeding management and the environment, playa role in the occurrence of the two organisms. The isolates were subjected to an antibiotic sensitivity test to determine the antibiotic resistance pattern. It was observed that there was a slight difference in the occurrence of the organisms and in the pattern of antibiotic resistance of isolates from free-range chickens' eggs and those from commercial layers. Enterococcus was detected in 10% and 7.7% of eggs from commercial layers and free-range chickens respectively. Salmonella spp occurred in 7.5% and 12.8% of commercial layers and free-range chicken eggs, respectively. The study suggests that free-range eggs may not be as wholesome as often believed and that the environment plays an important role in the development of antibiotic resistance

Genetic variability of vancomycin-resistant Enterococcus faecium and Enterococcus faecalis isolates from humans, chickens and pigs in Malaysia

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