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Transmission of pathogenic microorganisms in the last decades has been considered a significant health hazard and pathogenic E. coli, particularly antibiotic-resistant strains, have long been identified as a zoonotic problem. This study aimed to investigate multidrug resistant pathogenic E. coli isolates from wild birds, chickens, and environment in selected Orang Asli and Malay villages in Peninsular Malaysia. The bacteriological culture-based technique, disc diffusion method, and multiplex Polymerase Chain Reaction (mPCR) assay was used to determine the occurrence of pathogenic E. coli strains in the several samples in the study. E. coli isolates showed a variety of multi-drug resistant (MDR) antibiotypes and Enteropathogenic E. coli (EPEC) and Enteroinvasive E. coli (EIEC) were the most predominantly identified pathogenic E. coli strains. The findings of this study demonstrated the significance of animal reservoirs and the environment as sources of pathogenic E. coli, resistant bacteria, and resistance genes. Hence, there is a need for adoption of a practical surveillance approach on MDR pathogens to control foodborne contamination.

Foodborne pathogens have become a major concern not only due to the diseases they cause, but also because of the rise of antibiotic resistant strains in human and animals. The purposes of this study were to determine the occurrence of Campylobacter jejuni and Escherichia coli and their antibiotic resistance profiles in wild birds, chickens, humans, and the environment in Malay villages in Malaysia. Three Malay villages in Kota Setar, Kedah were chosen. Three hundred nine (309) samples were collected in this study including wild birds (38), chickens (71), humans (47), and the environment (153). Subsequently, the C. jejuni and E coli isolates were tested against antibiotics using the disc diffusion method. Campylobacter jejuni was found positive in 17 (37.8%) flies and 8 (11.3%) chickens. Also, E. coli was found positive in 89.4% of human, 47.4% of bird, 44 62% of chicken and in 71.2% of the environmental samples. Ten antibiotics were used to determine the susceptibility of the isolates. Eighty four percent (84%) of C. jejuni and 100% of E. coli isolates were found to show resistance towards at least one antibiotic. The isolates showed high resistance to cefpodoxime and tetracycline.

Introduction: Proventricular dilatation disease (PDD) is caused by avian bornavirus (ABV) has been identified in psittacine, non-psittacine birds and waterfowl. Birds may show signs of gastrointestinal tract deficit or neurological dysfunction or even both. The objectives of this study were to determine the molecular prevalence, risk factors and public awareness of ABV and PDD among captive and non-captive birds in Peninsular Malaysia. Material and Methods: A total of 344 cloacal swabs or faeces were collected and subjected to detection using the RT-PCR assay. Meanwhile, KAP questionnaires were distributed by using the Google forms platform. Results: Molecular prevalence studies revealed that 4.5% (9/201) of the pet birds were ABV-positive, whereas 0% (0/143) in waterfowl. Nine positive pet birds were identified to be PaBV-2, which is closest to ABV isolates EU781967 (USA). Among the risk factors analysed, category, age and, location, were found to show an association with the ABV positivity. The KAP survey result showed: the respondents have low knowledge (32.9%), however, they showed positive attitude (60.8%) and good practice (94.9%). The association between knowledge, attitude and practice showed that there was a significant association between knowledge-attitude and also attitude-practice (P < 0.05). Conclusion: This study proved that avian bornavirus (ABV) causes proventricular dilatation disease (PDD) among a group of pet birds of Psittaciformes, but it is present in Peninsular Malaysia with a low prevalence rate. Furthermore, in addition to the useful databases obtained from this study, the level of public awareness on the importance of avian bornavirus that causes fatal disorders among a wide range of bird species is satisfactorily raised.

Orthopedic conditions are among the most frequent avian emergency cases admitted to veterinary hospitals and clinics. An external skeletal fixator is commonly used as surgical treatment in avian orthopedics. This review aimed to systematically evaluate the use of external skeletal fixator (ESF) in published cases of avian orthopedics. Four electronic databases were searched to identify all relevant case reports and case series. Altogether, 24 primary reports met the inclusion criteria. Of those, 16 reports were case reports; and eight were case series. In total, 166 avian patients with different orthopedic conditions were undergone ESF stabilization techniques; and raptor species formed the majority of the patients. The characteristics of the fractured bone were the key factor in the suitability of different ESF techniques. In addition, other factors such as type, location, and severity of the orthopedic condition should be considered. ESF implants are promising techniques for treating avian orthopedic conditions, as the complications related to the fixators were few, and the percentage of successfully released raptors was high. Unfortunately, there are so many cases of avian orthopedics that were not published. Considering this caveat, it is not easy to draw a firm conclusion on this topic. However, this paper can serve as a guiding principle for clinicians.

Over the years, development of molecular diagnostics has evolved significantly in the detection of pathogens within humans and their surroundings. Researchers have discovered new species and strains of viruses, while mitigating the viral infections that occur, owing to the accessibility of nucleic acid screening methods such as polymerase chain reaction (PCR), quantitative (real-time) polymerase chain reaction (qPCR) and reverse-transcription qPCR (RT-qPCR). While such molecular detection methods are widely utilized as the benchmark, the invention of isothermal amplifications has also emerged as a reliable tool to improvise on-field diagnosis without dependence on thermocyclers. Among the established isothermal amplification technologies are loop-mediated isothermal amplification (LAMP), recombinant polymerase amplification (RPA), strand displacement activity (SDA), nucleic acid sequence-based amplification (NASBA), helicase-dependent amplification (HDA) and rolling circle amplification (RCA). This review highlights the past research on and future prospects of LAMP, its principles and applications as a promising point-of-care diagnostic method against avian viruses.

The aptitude of Mycoplasma gallisepticum (MG) for changing its surface proteins allows the pathogen to reduce the efficacy of antimicrobial agents, especially those targeting surface proteins. Although antibiotic treatment cannot be a solution for the eradication of avian mycoplasmosis, it can be considered a good option to minimize the number of deaths. One of the challenges of antibiotic treatments is the development of antimicrobial resistance (AMR) among field isolates. Monitoring the antimicrobial susceptibility profile of field isolates can be a practical way of avoiding AMR. Thus, various tests have been developed to determine the antimicrobial susceptibility profile of field isolates. A modified broth microdilution minimum inhibitory concentration (MIC) test has been regularly used to measure the antimicrobial susceptibility profile of MG field isolates. Numerous studies have reported an increase in the number of antibiotic-resistant field isolates of MG. Therefore, this study presents a systematic review on antimicrobial MIC values of MG isolates to gather recent knowledge and investigate the prevalence and distribution of AMR among MG field isolates. A thorough search was conducted for related studies throughout 3 electronic databases. Altogether, 23 studies were identified as eligible studies and were used for further analysis. The results showed that enrofloxacin, oxytetracycline, and tylosin had the highest number of resistant isolates in most of the geographical distributions, respectively. Minimum inhibitory concentrations of these antibiotics need to be determined regularly to optimize treatment dosages.

Fowl cholera has caused significant economic losses in many poultry producing countries worldwide. In Malaysia, outbreaks of fowl cholera are frequently reported and encountered in different types of poultry productions. The objective of this study was to characterise 13 avian Pasteurella multocida, isolated from fowl cholera outbreaks in Central Peninsular Malaysia in the period between 2000 and 2018. The isolates were subjected to multiplex polymerase chain reaction (PCR) for capsular serotyping, disc diffusion method for antimicrobial susceptibility profiles, and molecular genotyping using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The capsular serotyping showed all 13 Pasteurella multocida isolates belonging to capsular serotype A. The antimicrobial susceptibility showed several multidrug resistance strains among the P. multocida isolates. All the isolates were resistant to erythromycin (100%), streptomycin (68%), tetracycline (37%), enrofloxacin (37%), florfenicol (23%), penicillin G (14%), gentamicin (14%), and amoxicillin (14%). The PFGE analysis clustered the isolates into three clones. Group A included isolates with a similarity of 87% from the year 2000, 2013, and 2018. Three sequence types were identified using MLST typing namely, ST129, ST231, and ST355. The ST355 was assigned for the first time in the Rural Industries Research and Development Corporation (RIRDC) database. Besides, ST129 has been reported in India, China, and Sri Lanka, which highlights the possibility of transmission between Asian countries. This study provides an insight into epidemiological information of Pasteurella multocida that causes fowl cholera outbreaks in the central region of Peninsular Malaysia.

Aims: Psittacine birds such as parrots, macaws, cockatoos, lovebirds and parakeets, are widely reared as household pets or at aviary due to their attractive features. However, the status of virus-causing diseases of psittacine species in Malaysia is fairly under-documented. Therefore, this study was aimed to detect the presence of three common avian viruses that infect psittacine birds, i.e. beak and feather disease virus (BFDV), avian polyomavirus and avian papillomavirus. Methodology and results: Faecal samples from twelve asymptomatic captive psittacine birds of different species were collected from an undisclosed animal garden in Serdang, Selangor, Malaysia. Briefly, the sample was homogenised and resuspended with SM buffer with the ratio 1:1 (weight of sample/g: volume of SM buffer/mL) before centrifugation at 1,000 × g for 20 min. The supernatant was collected and filtered before subjected to genomic DNA extraction using a commercialised kit. Polymerase chain reaction (PCR) technique was used to screen the V1, VP1 and L1 genes of beak and feather disease virus (BFDV), avian polyomavirus and avian papillomavirus, respectively. Findings revealed that the samples were negative for BFDV and avian polyomavirus. However, positive results of 1.5 kbp PCR amplicon were detected for avian papillomavirus in four out of the 12 samples (33.33%), which was from the white-crested cockatoo, African grey parrot, yellow-collared macaw and Senegal parrot. Sequence analysis of the L1 gene from the Senegal parrot Poicephalus senegalus revealed 93% identity to a reference Psittacus erithacus timneh avian papillomavirus. Conclusion, significance and impact of study: This study added to the limited prevalence data of three important avian viruses which infect captive psittacines in Seri Kembangan, Selangor, Malaysia. Avian papillomavirus, but not BFDV and avian polyomavirus, was detected in the collected captive psittacine birds. Therefore, a routine screening can be performed to mon

The role of wildlife such as wild birds, macaques, and bats in the spreading and maintenance of deadly zoonotic pathogens in nature have been well documented in many parts of the world. One such pathogen is the mosquitoes borne virus, namely the West Nile Virus (WNV). Previous research has shown that 1:7 and 1:6 Malaysian wild birds are WNV antibody and RNA positive, respectively, and bats in North America may not be susceptible to the WNV infection. This study was conducted to determine the status of WNV in Malaysian macaques and bats found in mangrove forests and caves, respectively. Archive sera and oropharyngeal swabs from long-tailed macaques were subjected to the antibody detection using WNV competitive enzyme-linked immunosorbent assay (c-ELISA) and WNV RNA using RT-PCR, respectively, while the archive oropharyngeal and rectal swabs from bats were subjected to RT-PCR without serological analysis due to the unavailability of serum samples. The analysis revealed a WNV seropositivity of 29.63% (24/81) and none of the macaques were positive for WNV RNA. Meanwhile, 12.2% (5/41) of the bats from Pteropodidae, Emballonuridae, and Rhinolophidae families tested positive for WNV RNA. Here, we show a high WNV antibody prevalence in macaques and a moderate WNV RNA in various Malaysian bat species, suggesting that WNV circulates through Malaysian wild animals and Malaysian bat species may be susceptible to the WNV infection.

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.

West Nile virus (WNV) is a zoonotic mosquito-borne flavivirus that is harbored and amplified by wild birds via the enzootic transmission cycle. Wide range of hosts are found to be susceptible to WNV infection including mammals, amphibians and reptiles across the world. Several studies have demonstrated that WNV was present in the Malaysian Orang Asli and captive birds. However, no data are available on the WNV prevalence in wild birds found in Malaysia. Therefore this study was conducted to determine the serological and molecular prevalence of WNV in wild birds in selected areas in the West Coast of Peninsular Malaysia. Two types of wild birds were screened, namely migratory and resident birds in order to explore any possibility of WNV transmission from the migratory birds to the resident birds. Thus, a cross-sectional study was conducted at the migratory birds sanctuary located in Kuala Gula, Perak and Kapar, Selangor by catching 163 migratory birds, and 97 resident birds from Kuala Gula and Parit Buntar, Perak at different time between 2016 and 2017 (Total, n = 260). Blood and oropharyngeal swabs were collected for serological and molecular analysis, respectively. Serum were screened for WNV antibodies using a commercial competitive ELISA (c-ELISA) (ID Screen® West Nile Competition Multi-species ELISA, ID VET, Montpellier, France) and cross-reactivity towards Japanese Encephalitis virus (JEV) was also carried out using the JEV-double antigen sandwich (DAS) ELISA. Oropharyngeal swabs were subjected to one-step RT-PCR to detect WNV RNA, in which positive reactions were subsequently sequenced. WNV seropositive rate of 18.71% (29/155) at 95% CI (0.131 to 0.260) and molecular prevalence of 15.2% (16/105) at 95% CI (0.092 to 0.239) were demonstrated in migratory and resident wild birds found in West Coast Malaysia. Phylogenetic analyses of the 16 WNV isolates found in this study revealed that the local strains have 99% similarity to the strains from South Africa and wer

In 1994, Mycoplasma gallisepticum (MG) was reported to be responsible for conjunctivitis outbreak in the North American house finch population. This new course of MG infection in passerine was the result of spillover infections from the poultry strains. In severe cases of the disease, the conjunctival lesions might cause blindness and death, but in the mild form, there is a chance of recovery. The immune system of the recovered birds develops a resistance to the previous strains. However, the incomplete immune responses and the ability of MG to rapidly alter its surface antigens allow the pathogen to evolve new strains that can infect the birds that have already developed immune resistance. Although the rate of mortality decreases as a result of developing resistance, the persistence of the disease continues due to the increase in both virulence and the replication rate of the new strains. Therefore, the morbidity rate has remained steady, and new species of birds become infected as a result of evolutionary adaptation of the new strains. In this regard, the objective of this study is to provide a review of the mycoplasma conjunctivitis in passerine species, notably by looking at it from the host-pathogen interaction point of view.

Aim: This study aimed to investigate the occurrence of Helicobacter pullorum in broiler chickens and their farm environment. Materials and Methods: The ceca from 100 broiler chickens from ten farms were sampled from processing sites or markets. The cecal contents were aseptically collected from each cecum and cultured. The farms were visited, and environmental samples were collected which included water, house flies, floor swabs and soils in chicken houses. Results: H. pullorum was present in 51% of the broilers; 17.5% of the flies were found to carry H. pullorum and Campylobacter spp., 30% of house floors were positive, while all water samples were negative. Conclusion: Flies could have picked up the organisms from the chickens' feces and/or the environment of the chicken houses or they could be one of the sources in the spread of the organisms. This study also showed that broiler chickens are potential reservoirs for H. pullorum and may serve as a source of infection for humans through the food chain.

JALILA BINTI ABU/// MOHAMED YOUSIF IBRAHIM MOHAMED,SALEHA BINTI ABDUL AZIZ,ZUNITA BINTI ZAKARIA/// Abdul Rashid,Elmutaz Atta Awad In developing countries, Escherichia coli and Campylobacter jejuni were found to be the prominent bacterial causes of gastroenteritis in human gastroenteritis, and they also showed an increased trend towards antibiotic resistance. Thus, the study aimed to determine the occurrence of E. coli and C. jejuni in wild birds, chickens, human and environment in villages in Perak and to determine the antibiotic resistance amongst E. coli and C. jejuni isolates. Three Orang Asli villages in Perak were chosen. Wild birds were trapped from nearby areas of the villages. We collected 52 human stool samples, 40 cloacal swabs from wild birds and 42 from chickens. For the environment, samples were collected from water (40 samples), flies (40 samples) and soil (40 samples). Two (5%) flies and 16 (38.1%) chickens were found positive for C. jejuni. Also, a total of 50 (96.2%) humans, 40 (100%) birds, 40 (95.2%) chickens were positive for E. coli and 111 (92.5%) of the 120 environmental samples tested positive for E. coli. The disk diffusion method was employed to determine the susceptibility of C. jejuni and E. coli isolates against ten antibiotics. All of the E. coli (100%) and C. jejuni (66%) isolates were found resistance towards at least one antibiotic. Hundred percent (100%) of the E. coli isolates obtained from wild birds were found to show multidrug resistance (MDR). Campylobacter jejuni isolates from flies and chickens showed high resistance towards nalidixic acid.

Aims: Campylobacter is a major cause of gastroenteritis in humans worldwide, particularly in developed countries and is reported to show an increased trend in antibiotic resistance. The purpose of this study was to determine the occurrence of Campylobacter in wild birds, poultry and in poultry environments in Selangor, Malaysia as well as to determine the rate of antibiotic resistance among Campylobacter isolates from poultry and wild birds. Methodology and results: The wild birds were trapped near poultry farm areas and in open areas which were more than 5 km away from poultry farms (referred to as open environment). Of 57 birds trapped near the farm's environment, 17.5% were positive for Campylobacter and out of these, 90% were C. jejuni. Of a total of 77 birds in the open environment, 22.1% were positive for Campylobacter and 88.7% were C. jejuni. The poultry farms consisted of 3 chicken and 2 duck farms. About 60% of the chickens and 44.8% of the ducks were positive for Campylobacter of which 80% were C. jejuni, while 20% were C. coli. The Campylobacter isolates were subjected to antibiotic susceptibility test using disk diffusion method against 12 antibiotics. All the isolates (100%) from wild birds around poultry houses were resistant to at least one antibiotic. Conclusion, significance and impact of study: The findings showed 93% of the isolates from wild birds were resistant to at least two antibiotics. Campylobacter isolates from poultry in the farms were resistant to at least one antibiotic. The antibiotic resistant Campylobacter is of public health importance.

The objective of this study was to evaluate the effect of shackling and halal slaughter with head-only electrical stunning (ES) using water bath or no stunning (NS) on plasma levels of adrenaline (AD), noradrenaline (NAD) and corticosterone (CORT) levels of broiler chickens. Male and female broiler chickens (Cobb 500) 38 days old from a commercial farm in Johore, Malaysia, were placed in plastic crates (0.80 × 0.60 × 0.31 m) at 10 birds per crate, and transported for 2.5 hours in an open truck. The ambient temperature during transportation was 25 to 27 °C. Upon arrival at a commercial processing plant, a total of 50 male broiler chickens (2.2 - 2.5 kg) were randomly selected from the flocks that were delivered, transferred to different five crates (10 birds per crate) and held in a lairage for 2 to 3 h. Blood samples were collected at various points, namely after lairaging (P1) (first sampling point for both ES and NS chickens, regarded as basal values), shackling (P2) (second sampling point for both ES and NS chickens), stunning (P3) (third sampling point for ES chickens), and ventral neck incision in stunned (P4) (fourth sampling point for ES chickens) and no stun (P5) (third sampling point for NS chickens). Ten birds were sampled at each sampling point (50 birds in total). Both stunned and no stun birds were shackled. Results show that shackling (P2) did not significantly affect the AD, NAD and CORT levels. However, electrical stunning elevated both the AD and CORT levels significantly, but not NAD. Neck cut had negligible effect on the AD and NAD in NS and ES broilers, but significantly increased CORT compared with the basal CORT value. In conclusion, the study demonstrated that head-only electrical stunning using a water bath might elevate AD and CORT. The study concluded that, as measured by AD and NAD, slaughter with or without stunning may not induce a physiological stress response in broilers.

The growing demand of edible bird’s nest (EBN) worldwide as well as competition from neighbouring countries has made the EBN industry one of the rising industries in Malaysia with stringent exportation requirement by China. However, as majority of the EBN products in the market is in commercialised form, studies on the nutritional composition of these commercialised EBN in comparison with farmed, raw EBN are limited. The farmed EBN samples were taken from 4 different regions of Malaysia: Perak (central), Kelantan (eastern), Johor (southern) and Sarawak (west Borneo) while the commercialised sample was obtained from a local drug store. Proximate, amino acid and elemental composition were performed on these EBNs. Farmed EBNs mainly comprised protein followed by carbohydrate while the commercialised EBN had similar protein and carbohydrate composition. The total collection of essential amino acid in commercialised EBN was higher (237.9 mg/g protein) compared to the farmed EBN which was between 156.81 − 236.6 mg amino acid/g protein. Among the essential amino acids, valine was found to be highest in both commercialised and farmed EBN. The differences between the nutritional compositions of EBNs could be due to the process of commercialisation of the EBN as well as seasonal, breeding sites and diet of the swiftlets. Farmed EBN therefore can be considered to be more nutritional due to higher protein levels.