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Fulltext Development of indirect IgG elisa for detection of leptospiral infection in cattle using Malaysian local isolate Leptospira kmetyi serovar Malaysia strain Bejo-Iso9

Sabri, A.R., Khairani-Bejo, S., Zunita, Z., Hassan, L.

Jurnal Veterinar Malaysia, 2017;29(2):5-8.
MyJurnal

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.
Fulltext Salmonella in pet and captive reptiles with reference to some studies in Malaysia and countries worldwide

Saleha, A.A., Khor, K.H., Zunita, Z., Jalila, A.

Jurnal Veterinar Malaysia, 2015;27(1):7-11.
MyJurnal

Reptiles, which included snakes, chelonians (turtles, tortoises), iguanas and lizards, have been reported as important reservoirs
for Salmonella. In Malaysia, reports on Salmonella in reptiles are not many. A search for works in Malaysia on salmonellae in
reptiles found 28.0% chelonians, 27.5 - 61.0% snakes, 26.5 – 36.0% lizards and 31.0% house geckos carried Salmonella. The
prevalence in some countries worldwide which included the United States of America, Brazil, Japan, Nigeria and few others were
shown. The public health risk of the presence of salmonellae particularly in pet reptiles are discussed.
Fulltext Oral microbes of pet sugar gliders and detection of Salmonella in their faeces

Nur Diana, H., Saleha, A.A., Azlan, C.M., Bejo, S.K., Zunita, Z., Fauziah, N.

Jurnal Veterinar Malaysia, 2016;28(2):24-25.
MyJurnal

Clinically healthy pets may carry zoonotic pathogens and shed them, thus act as potential public health threat. Small, exotic animals are gaining popularity as pets which include iguanas, turtles and sugar gliders. Salmonellosis is an important disease affecting human and animal populations worldwide. It is reported that sugar gliders are becoming popular pets among young Malaysians, not only are they cute and adorable but are easily carried around. Of 35 pet sugar gliders and 17 from a breeder, 15% were positive for salmonellae and the serovars identified were Salmonella enterica serovar Albany (62.5%) and Salmonella enterica serovar London (37.5%). Staphylococcus spp. (41.0%) were most frequently identified from oral mucosae, followed by Enterococcus faecalis (17%), Streptococcus viridians (15.0%), Acinetobacter calcoaceticus (13.0%) and Enterococcus faecium, Pasteurella spp., Staphylococcus delphini and Escherichia coli (2.0–4.0%). The close interaction between infected pet sugar gliders and their owners poses public health risk. Breeders with infected animals present direct risk to pet shops and pet owners.
Molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) among veterinary students and personnel at a veterinary hospital in Malaysia

Aklilu E, Zunita Z, Hassan L, Cheng CH

Vet Microbiol, 2013 Jun 28;164(3-4):352-8.
PMID: 23523336 DOI: 10.1016/j.vetmic.2013.02.030

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

Annas S, Zamri-Saad M, Jesse FF, Zunita Z

Microb Pathog, 2015 Nov;88:94-102.
PMID: 26298001 DOI: 10.1016/j.micpath.2015.08.009

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

Bashir A, Zunita Z, Jesse FFA, Ramanoon SZ

Microbiol Resour Announc, 2019 Feb;8(6).
PMID: 30746526 DOI: 10.1128/MRA.01618-18

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

Aklilu E, Zunita Z, Hassan L, Chen HC

Trop Biomed, 2010 Dec;27(3):483-92.
PMID: 21399590

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

Aliyu AB, Saleha AA, Jalila A, Zunita Z

BMC Public Health, 2016 08 02;16:699.
PMID: 27484086 DOI: 10.1186/s12889-016-3377-2

BACKGROUND: The significant role of retail poultry meat as an important exposure pathway for the acquisition and transmission of extended spectrum β-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.
Fulltext New sites of localisation of Pasteurella multocida B:2 in buffalo surviving experimental haemorrhagic septicaemia

Annas S, Zamri-Saad M, Jesse FF, Zunita Z

BMC Vet Res, 2014;10:88.
PMID: 24721163 DOI: 10.1186/1746-6148-10-88

Haemorrhagic septicaemia (HS) is an acute septicaemic disease of buffalo and cattle caused by Pasteurella multocida B:2 and E:2. Field outbreaks of HS are known to result in localisation of bacteria in the tonsils of surviving buffalo, confirming that animals can become carriers and the role of respiratory tract in the transmission of the disease. This report describes additional sites of localisation of P. multocida B:2 in surviving buffalo following experimental induction of HS.
Prevalence, distribution and antibiotic resistance of emergent Arcobacter spp. from clinically healthy cattle and goats

Shah AH, Saleha AA, Zunita Z, Murugaiyah M, Aliyu AB, Jafri N

Transbound Emerg Dis, 2013 Feb;60(1):9-16.
PMID: 22280210 DOI: 10.1111/j.1865-1682.2012.01311.x

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

Bashir A, Zunita Z, Jesse FFA, Ramanoon SZ, Mohd-Azmi ML

Microbiol Resour Announc, 2019 Nov 27;8(48).
PMID: 31776215 DOI: 10.1128/MRA.01057-19

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.
Fulltext Correction for Bashir et al., "Whole-Genome Shotgun Sequence of Streptococcus agalactiae Sequence Type 176 Strain 3966RFQB from a Dairy Herd in Selangor, Malaysia"

Bashir A, Zunita Z, Jesse FFA, Ramanoon SZ, Mohd-Azmi ML

Microbiol Resour Announc, 2019 Apr 18;8(16).
PMID: 31000558 DOI: 10.1128/MRA.00331-19
Prevalence and distribution of Arcobacter spp. in raw milk and retail raw beef

Shah AH, Saleha AA, Murugaiyah M, Zunita Z, Memon AA

J Food Prot, 2012 Aug;75(8):1474-8.
PMID: 22856572 DOI: 10.4315/0362-028X.JFP-11-487

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.
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

Hasheimi SR, Zulkifli I, Somchit MN, Zunita Z, Loh TC, Soleimani AF, et al.

J Anim Physiol Anim Nutr (Berl), 2013 Aug;97(4):632-8.
PMID: 22533311 DOI: 10.1111/j.1439-0396.2012.01302.x

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.
Genetic characterization of Arcobacter isolates from various sources

Shah AH, Saleha AA, Zunita Z, Cheah YK, Murugaiyah M, Korejo NA

Vet Microbiol, 2012 Dec 7;160(3-4):355-61.
PMID: 22739058 DOI: 10.1016/j.vetmic.2012.05.037

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.
Fulltext In vitro transfer of methicillin resistance determinants mecA from methicillin resistant Staphylococcus aureus (MRSA) to methicillin susceptible Staphylococcus aureus (MSSA)

Bitrus AA, Zunita Z, Bejo SK, Othman S, Nadzir NA

BMC Microbiol, 2017 04 04;17(1):83.
PMID: 28376716 DOI: 10.1186/s12866-017-0994-6

BACKGROUND: Staphylococcus aureus more than any other human pathogen is a better model for the study of the adaptive evolution of bacterial resistance to antibiotics, as it has demonstrated a remarkable ability in its response to new antibiotics. This study was designed to investigate the in vitro transfer of mecA gene from methicillin resistant S. aureus to methicillin susceptible S. aureus.
RESULT: The recipient transconjugants were resistant to erythromycin, cefpodoxime and were mecA positive. PCR amplification of mecA after mix culture plating on Luria Bertani agar containing 100 μg/mL showed that 75% of the donor and 58.3% of the recipient transconjugants were mecA positive. Additionally, 61.5% of both the donor cells and recipient transconjugants were mecA positive, while 46.2% and 41.75% of both donor and recipient transconjugants were mecA positive on LB agar containing 50 μg/mL and 30 μg/mL respectively.
CONCLUSION: In this study, the direction of transfer of phenotypic resistance as well as mecA was observed to have occurred from the donor to the recipient strains. This study affirmed the importance of horizontal transfer events in the dissemination of antibiotics resistance among different strains of MRSA.
Staphylococcal cassette chromosome mec (SCCmec) and characterization of the attachment site (attB) of methicillin resistant Staphylococcus aureus (MRSA) and methicillin susceptible Staphylococcus aureus (MSSA) isolates

Bitrus AA, Zunita Z, Khairani-Bejo S, Othman S, Ahmad Nadzir NA

Microb Pathog, 2018 Oct;123:323-329.
PMID: 30053600 DOI: 10.1016/j.micpath.2018.07.033

This study was designed to screen for SCCmec types and to characterize the attachment site (attB) and universal insertion site (orfX) of SCCmec in a collection of 27 isolates (n = 11) methicillin resistant S. aureus and (n = 16) methicillin susceptible S. aureus isolates in Malaysia. Screening of SCCmec types and characterization of the attachment site was carried out using PCR amplification and Sanger's sequencing method. The result showed that a large proportion of the MRSA isolates carried SCCmec type III 7/11 (63%). Three isolates 3/11 (27%) and 1/11 (9.0%) carried SCCmec type II and IVd respectively. Amplification of the universal insertion site of the SCCmec (orfX) and attachment site (attB) showed that all 16 S. aureus isolates were positive for the orfX gene, while only 7 were positive for the attB gene. Phylogenetic diversity showed that the isolates clustered around strains with features similar to a community acquired MRSA. In conclusion, a high carriage rate of SCCmec type III was observed. The result also showed that all the S. aureus isolates have the orfX structure; however, not all isolates possesses the attB site on the 3' end of the orfX region.
Occurrence of Campylobacter species from broiler chickens and chicken meat in Malaysia

Sinulingga TS, Aziz SA, Bitrus AA, Zunita Z, Abu J

Trop Anim Health Prod, 2020 Jan;52(1):151-157.
PMID: 31273582 DOI: 10.1007/s11250-019-01995-y

Campylobacter is reported as a major cause of foodborne illness worldwide. Consumption of contaminated chicken meat is considered a significant risk factor of Campylobacter infection in humans. This study investigated the occurrence of non-Campylobacter jejuni-Campylobacter coli, in broiler chickens (n = 210) and chicken meat (n = 109). The samples were collected from seven broiler chicken farms (n = 210 cloacal swabs), 11 markets (n = 84 chicken meat), and 5 supermarkets (n = 25 chicken meat) located in different districts of Selangor State. Campylobacter were isolated from cloacal swabs using the Cape Town Protocol and from meat samples using the method of Duffy et al. (2007) with some modifications for Campylobacter isolations which were reported effective in the isolation of non-C. jejuni-C. coli Campylobacter species. The isolates were identified by Gram staining for cellular morphology, wet mount for motility and biochemical tests. Confirmation of presumed Campylobacter isolates was carried out using multiplex PCR (mPCR). One hundred seven (107/210) or 50.9% and twenty-nine (29/109) or 26.6% of chickens and chicken meat samples respectively were positive for Campylobacter species. Among the Campylobacter isolates from chickens, C. jejuni was the most predominantly isolated species (69.5%), followed by C. coli (16.2%). Campylobacter fetus and C. upsaliensis were the non-C. jejuni-C. coli Campylobacter species isolated in this study, at 9.3% and 2.5% respectively. Overall, the findings indicated broiler chickens were colonized not only by the common Campylobacter species but also by other Campylobacter species. We found the Cape Town Protocol useful to detect the occurrence of non-C. jejuni-C. coli isolates in chickens.

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