Interrogating the Whole-Genome Shotgun Sequence of Escherichia coli Sequence Type 127 Strain 1538RHQ, Which Harbors Virulent Antigenic Factors, Isolated from a Mastitic Cow.

We report the whole-genome sequence of Escherichia coli sequence type 127 (ST127) strain 1538RHQ, recovered from a mastitic cow in a dairy herd in Selangor, Malaysia. The objective of this study was to identify the antigenic and virulence properties that can be used as suitable targets for vaccine development against bovine mastitis.

Whole-Genome Shotgun Sequence of Streptococcus agalactiae Sequence Type 176 Strain 3966RFQB from a Dairy Herd in Selangor, Malaysia.

Streptococcus agalactiae, commonly known as group B streptococcus (GBS), is among the most implicated pathogens in bovine mastitis worldwide. Proper control measures can curb both economic and public health effects it may cause. Here, we report the sequenced genome of S. agalactiae sequence type 167 (ST167) strain 3966RFQB obtained from a bovine mastitis case at a dairy herd in Banting, Selangor, Malaysia (longitude 2.8121°N, latitude 101.5026°E).

Molecular detection of Haemophilus parasuis serotypes 4, 5 or 12 and 13 in Peninsular Malaysia.

Although the economic importance of Haemophilus parasuis infection causing Glasser's disease is prevalent throughout pig farms in Peninsular Malaysia, there is a dearth of knowledge on its actual nature. In this study, a multiplex PCR was performed to screen for three major predominant virulent strains of H. parasuis, which are serotypes 4, 5 or 12 and 13. A total of 175 tissues or bodily fluid samples of various parts were collected from diseased animals from October, 2016 to February, 2018; with total of 62.9% positive detection of H. parasuis. The highest detection was found to be in the pericardial sac fibrin (90.9%) followed by pleural fibrin, lung, pleural fluid, tonsil, pericardial sac, peritoneal fluid, abdominal fibrin, joint fluid, brain and pericardium. Serotype 13 was the highest (40/110) followed by serotype 4(37/110), serotype 5(31/110) and 12 samples were nontypable (12/110). The presence of untypable serotype also drives to further identification of other serotypes in Malaysia.

Molecular detection of Leptospira sp. in cattle and goats in Kelantan, Malaysia after a massive flood using multiplex polymerase chain reaction.

Flood is a potential driver in spreading waterborne diseases including leptospirosis, which is a zoonotic disease caused by pathogenic bacteria of the genus Leptospira. In the case of leptospirosis, cattle and goats can be incidental hosts and potential carriers of leptospirosis. Traditionally, serology such as microscopic agglutination test (MAT) and isolation of the organisms have been commonly used as the diagnostic approaches in diagnosing leptospirosis. However, nowadays, various molecular techniques have been developed for specific detection of Leptospira sp. such as, polymerase chain reaction (PCR), which is sensitive, specific and rapid in detecting the species. This study detected Leptospira sp. directly from the blood and urine of the animals such as, cattle, goats and sheep in Kelantan after a massive flood by using multiplex PCR (mPCR). From the results collected in the study, four blood samples (0.63%; 4/635) were found to be positive with Leptospira sp. and one urine sample (3.23%; 1/31) was detected as positive with Leptospira sp. The blood and urine samples that were detected to be positive with Leptospira sp. were collected from cattle and goats exposed to the flood. However, no Leptospira sp. was detected from the sheep in this study. Multiplex PCR (mPCR) was successfully used to detect the presence of Leptospira sp. in animals. Apart from that, it is also suggested that flood has a significant role in transmitting the disease to animals.

Molecular detection of Leptospira sp. in cattle and goats in Kelantan, Malaysia after a massive flood using multiplex polymerase chain reaction

@#Flood is a potential driver in spreading waterborne diseases including leptospirosis, which is a zoonotic disease caused by pathogenic bacteria of the genus Leptospira. In the case of leptospirosis, cattle and goats can be incidental hosts and potential carriers of leptospirosis. Traditionally, serology such as microscopic agglutination test (MAT) and isolation of the organisms have been commonly used as the diagnostic approaches in diagnosing leptospirosis. However, nowadays, various molecular techniques have been developed for specific detection of Leptospira sp. such as, polymerase chain reaction (PCR), which is sensitive, specific and rapid in detecting the species. This study detected Leptospira sp. directly from the blood and urine of the animals such as, cattle, goats and sheep in Kelantan after a massive flood by using multiplex PCR (mPCR). From the results collected in the study, four blood samples (0.63%; 4/635) were found to be positive with Leptospira sp. and one urine sample (3.23%; 1/31) was detected as positive with Leptospira sp. The blood and urine samples that were detected to be positive with Leptospira sp. were collected from cattle and goats exposed to the flood. However, no Leptospira sp. was detected from the sheep in this study. Multiplex PCR (mPCR) was successfully used to detect the presence of Leptospira sp. in animals. Apart from that, it is also suggested that flood has a significant role in transmitting the disease to animals.

Molecular detection of Haemophilus parasuis serotypes 4, 5 or 12 and 13 in Peninsular Malaysia

@#Although the economic importance of Haemophilus parasuis infection causing Glasser’s disease is prevalent throughout pig farms in Peninsular Malaysia, there is a dearth of knowledge on its actual nature. In this study, a multiplex PCR was performed to screen for three major predominant virulent strains of H. parasuis, which are serotypes 4, 5 or 12 and 13. A total of 175 tissues or bodily fluid samples of various parts were collected from diseased animals from October, 2016 to February, 2018; with total of 62.9% positive detection of H. parasuis. The highest detection was found to be in the pericardial sac fibrin (90.9%) followed by pleural fibrin, lung, pleural fluid, tonsil, pericardial sac, peritoneal fluid, abdominal fibrin, joint fluid, brain and pericardium. Serotype 13 was the highest (40/110) followed by serotype 4(37/110), serotype 5(31/110) and 12 samples were nontypable (12/110). The presence of untypable serotype also drives to further identification of other serotypes in Malaysia.

Risk factors and spatial distribution of extended spectrum ß-lactamase-producing- Escherichia coli at retail poultry meat markets in Malaysia: a cross-sectional study.

BACKGROUND: The significant role of retail poultry meat as an important exposure pathway for the acquisition and transmission of extended spectrum ß-lactamase-producing Escherichia coli (ESBL-EC) into the human population warrants understanding concerning those operational practices associated with dissemination of ESBL-EC in poultry meat retailing. Hence, the objective of this study was to determine the prevalence, spatial distribution and potential risk factors associated with the dissemination of ESBL-EC in poultry meat retail at wet-markets in Selangor, Malaysia. METHODS: Poultry meat (breast, wing, thigh, and keel) as well as the contact surfaces of weighing scales and cutting boards were sampled to detect ESBL-EC by using culture and disk combination methods and polymerase chain reaction assays. Besides, questionnaire was used to obtain data and information pertaining to those operational practices that may possibly explain the occurrence of ESBL-EC. The data were analysed using logistic regression analysis at 95 % CI. RESULTS: The overall prevalence of ESBL-EC was 48.8 % (95 % CI, 42 - 55 %). Among the risk factors that were explored, type of countertop, sanitation of the stall environment, source of cleaning water, and type of cutting board were found to be significantly associated with the presence of ESBL-EC. CONCLUSIONS: Thus, in order to prevent or reduce the presence of ESBL-EC and other contaminants at the retail-outlet, there is a need to design a process control system based on the current prevailing practices in order to reduce cross contamination, as well as to improve food safety and consumer health.

Comparative clinicopathological changes in buffalo and cattle following infection by Pasteurella multocida B:2.

Haemorrhagic septicaemia (HS) is an acute, septicaemic disease of cattle and buffalo of Asia and Africa caused by Pasteurella multocida B:2 or E:2. Buffaloes are believed to be more susceptible than cattle. In this study, 9 buffaloes of 8 months old were divided equally into 3 groups (Groups 1, 3, 5). Similarly, 9 cattle of 8 months old were equally divided into 3 groups (Groups 2, 4, 6). Animals of Groups 1 and 2 were inoculated with PBS while Groups 3 and 4 were inoculated subcutaneously with 10(5) cfu/ml of P. multocida B:2. Animals of Groups 5 and 6 were inoculated intranasally with the same inoculum. Both buffaloes and cattle that were inoculated subcutaneously succumbed to the infection at 16 h and 18 h, respectively. Two buffaloes that were inoculated intranasally (Group 5) succumbed at 68 h while the remaining cattle and buffaloes survived the 72-h study period. Endotoxin was detected in the blood of infected cattle (Group 4) and buffaloes (Groups 3 and 5) prior to the detection of P. multocida B:2 in the blood. The endotoxin was detected in the blood of buffaloes of Group 3 and cattle of Group 4 at 0.5 h post-inoculation while buffaloes of Group 5 and cattle of Group 6 at 1.5 h. On the other hand, bacteraemia was detected at 2.5 h in buffaloes of Group 3 and cattle of Group 4 and at 12 h in buffaloes of Group 5 and cattle of Group 6. Affected cattle and buffaloes showed lesions typical of haemorrhagic septicaemia. These included congestion and haemorrhages in the organs of respiratory, gastrointestinal and urinary tracts with evidence of acute inflammatory reactions. The severity of gross and histopathology lesions in cattle and buffalo calves that succumbed to the infection showed insignificant (p > 0.05) difference. However, inoculated buffalo and cattle that survived the infection showed significantly (p < 0.05) less severe gross and histopathological changes than those that succumbed. In general, cattle are more resistant to intranasal infection by P. multocida B:2 than buffaloes.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) among veterinary students and personnel at a veterinary hospital in Malaysia.

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.

Prevalence, distribution and antibiotic resistance of emergent Arcobacter spp. from clinically healthy cattle and goats.

Prevalence, distribution and antibiotic resistance of Arcobacter spp. were investigated in cattle, goats, floor and treated water samples in this study. The prevalence of Arcobacter in adult and young was recorded as 8/110 (7.27%) and 4/83 (4.81%), respectively, which showed insignificant difference (P = 0.3503) in detection rates between adult and young cattle. A total of 33.33% of the floor samples and 11.11% of the treated water samples analysed were determined as positive for Arcobacter. Among the species isolated, over all, A. butzleri (45%) was the most frequently detected species, followed by A. skirrowii (5%). A. butzleri was isolated from adult cattle, floor and water samples at the rates of 75.0%, 33.4% and 50%, respectively. Co-colonization of species was not uncommon, and 50% of the samples were carrying more than one Arcobacter species. Only 12.5% sample from cattle (adult) was detected positive for only A. skirrowii. All samples from young animals, floor and water contained mixed isolates. None of the samples from goat farm was found to be carrying Arcobacter species. On profiling of antimicrobial resistance patterns, it was found that only one A. butzleri isolate (3.7%) was sensitive to all nine antibiotics tested. A. butzleri was found highly resistant to ampicillin (55.6%), followed by cefotaxime (33.4%) and ciprofloxacin (33.4%). Overall, 20% of the isolates showed multidrug resistance (resistant ≥4 antibiotics). Gentamicin and enrofloxacin can be used as drugs of choice for the treatment for Arcobacter infections.

Dietary supplementation of Zingiber officinale and Zingiber zerumbet to heat-stressed broiler chickens and its effect on heat shock protein 70 expression, blood parameters and body temperature.

The present study was conducted to assess the effects of dietary supplementation of Zingiber officinale and Zingiber zerumbet and to heat-stressed broiler chickens on heat shock protein (HSP) 70 density, plasma corticosterone concentration (CORT), heterophil to lymphocyte ratio (HLR) and body temperature. Beginning from day 28, chicks were divided into five dietary groups: (i) basal diet (control), (ii) basal diet +1%Z. zerumbet powder (ZZ1%), (iii) basal diet +2%Z. zerumbet powder (ZZ2%), (iv) basal diet +1%Z. officinale powder (ZO1%) and (v) basal diet +2%Z. officinale powder (ZO2%). From day 35-42, heat stress was induced by exposing birds to 38±1°C and 80% RH for 2 h/day. Irrespective of diet, heat challenge elevated HSP70 expression, CORT and HLR on day 42. On day 42, following heat challenge, the ZZ1% birds showed lower body temperatures than those of control, ZO1% and ZO2%. Neither CORT nor HLR was significantly affected by diet. The ZO2% and ZZ2% diets enhanced HSP70 expression when compared to the control groups. We concluded that dietary supplementation of Z. officinale and Z. zerumbet powder may induce HSP70 reaction in broiler chickens exposed to heat stress.

Prevalence and distribution of Arcobacter spp. in raw milk and retail raw beef.

A total of 106 beef samples which consisted of local (n = 59) and imported (n = 47) beef and 180 milk samples from cows (n = 86) and goats (n = 94) were collected from Selangor, Malaysia. Overall, 30.2% (32 of 106) of beef samples were found positive for Arcobacter species. Imported beef was significantly more contaminated (46.80%) than local beef (16.9%). Arcobacter butzleri was the species isolated most frequently from imported (81.8%) and local (60%) beef, followed by Arcobacter cryaerophilus in local (33.3%) and imported (18.2%) beef samples. Only one local beef sample (10%) yielded Arcobacter skirrowii. Arcobacter species were detected from cow's milk (5.8%), with A. butzleri as the dominant species (60%), followed by A. cryaerophilus (40%), whereas none of the goat's milk samples were found positive for Arcobacter. This is the first report of the detection of Arcobacter in milk and beef in Malaysia.

Genetic characterization of Arcobacter isolates from various sources.

Arcobacter is getting more attention due to its detection from wide host-range and foods of animal origin. The objective of this study was to determine the prevalence of Arcobacter spp. in various sources at farm level and beef retailed in markets in Malaysia and to assess the genetic relatedness among them. A total of 273 samples from dairy cattle including cattle (n=120), floor (n=30), water (n=18) and milk (n=105) as well as 148 beef samples collected from retail markets were studied. The overall prevalence of Arcobacter in various sources was 15% (63/421). However, source-wise detection rate of Arcobacter spp. was recorded as 26.66% (8/30) in floor, 26.3% (39/148) in beef, 11.11% (2/18) in water, 7.6% (8/105) in milk and 6.66% (8/120) in cattle. Arcobacter butzleri was the frequently isolated species however, a total of 75%, 66.7%, 53.8%, 50% and 12.5%% samples from floor, milk, beef, water and cattle, respectively, were carrying more than one species simultaneously. One (12.5%) cattle and beef sample (2.5%) found to be carrying one Arcobacter spp., A. skirrowii, only. Typing of Arcobacter isolates was done though pulsed field gel electrophoresis (PFGE) after digested with Eag1 restriction endonuclease (RE). Digestion of genomic DNA of Arcobacter from various sources yielded 12 major clusters (≥ 50% similarity) which included 29 different band patterns. A number of closely related A. butzleri isolates were found from beef samples which indicate cross contamination of common type of Arcobacter. Fecal shedding of Arcobacter by healthy animals can contaminate water and milk which may act as source of infection in humans.

Phenotypic and genotypic characterization of methicillin-resistant Staphylococcus aureus (MRSA) isolated from dogs and cats at University Veterinary Hospital, Universiti Putra Malaysia.

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.