Colibacillosis is a disease caused by avian pathogenic E. coli (APEC) and is one of the principle cause of morbidity and mortality in poultry worldwide which is represented by a complex syndrome characterized by multiple organ lesions. This study was carried out to determine phylogenetic grouping and virulenceassociated genes contained by E. coli isolates which is related in causing disease in chicken. E. coli isolates obtained from clinical cases of Veterinary ResearchInstitute were re-identified by conventional methods. Phylogenetic grouping of the isolates was determined by triplex polymerase chain reaction (PCR), and the presence of eight virulence genes were identified by multiplex PCR. A total of 125 E. coli isolates were subjected toanalysis of phylogenetic background and virulence associated genes profiling. Phylogenetic analysis demonstrated that most of the E. coli isolated from chicken in this study belonged to group B1 (36.0%),group D (28.0%), group A (27.2%) and group B2 (8.8%). Multiplex PCR analysis demonstrated that 96 (78.6%) of the E. coli isolates harbored at least one virulencegene, while 29 (23.3%) did not contain any virulence genes tested. The most prevalent virulence genes identified were iss (51.2%), followed by iucD (36.0%),tsh (32.8%), vat (16.0%), astA (13.6%), irp2 (11.2%), papC (9.6%) and the least is cva/cvi gene (0%). None of the isolates harbored more than four virulence genes.Each of phylogenetic groups presented with different combinations of virulence genes, with no specific combinations of virulence genes found to correlate withE. coli phylogroups. None of the E. coli isolates harbored more than four virulence genes, suggesting that E. coli isolates from chicken in this study appear to bederived from commensal strains and may relate to environmental predispose factors especially stress factors in the host to establish infection.

A total of thirty-eight Mafriwal cattle were selected from a localcattle herd of a government cattle farm; of which 36 animals were sub-fertile Mafriwal female dams and two bulls which were considered as control animals (one male Mafriwal and one male Jersey). Two markers were used in the detection of Y chromosome in the sub-fertile female animal which are testis specific proteins Y-encoded (TSPY) and amelogenin (AMLX/AMLY) genes. The genes were amplified using PCR. The DNA bands from a normal male for TSPY gene size was approximately 260 bp while AMLX/ AMLY gene were approximately 341 and 467 bp. The examination of all samples showed that the sub-fertile cow revealedonly 467 bp while three fragments were detected in the control group; 260 bp (testis specific protein, Y-encoded gene), 341 and 467 bp (Amelogenin gene). The results showed that the sex chromosomeanomalies associated with Y chromosome did not occur in this group. These two sex markers can be used for the diagnosis of Y chromosome abnormality in a sub-fertile cow through polymerase chain reactionwhich is a rapid and reliable method for use in breeding herds.

A comprehensive study was conducted on the clinical observationsincluding clinical history, physical examination along with haematobiochemical alteration on 41 naturally occurring cases of canine babesiosis from Punjab state, India. Examination of 964 dogs revealed 4.25 percent (41/964) prevalence of the disease including 3.84 percent (37) B. gibsoni and 0.41 percent (4) B. canis infected cases. Clinical and parasitological diagnosis was finally confirmed by polymerase chain reaction. A large variation of clinical anifestations including rare findings of paraplegia, blindness, ocular bleeding, immune mediated haemolytic anaemia (IMHA), ascites and skin lesions were observed among the affected animals. Bloodfilms showed anisocytosis and nucleated erythrocytes indicating regenerative anaemia. Blood parameters of the affected dogs revealed significant decrease in Hb, TEC, PCV and thrombocytes. Significantdecrease in lymphocytes was found in B. gibsoni affected animals. The affected dogs showed significant increase in serum bilirubin, ALT, AKP, BUN and creatinine. Haemato-biochemical observations wereindicative of severity of babesiosis in dogs.

A study was carried out to report the phylogenetic analysis of Brucella abortus and Brucella melitensisby using molecular techniques from samples submitted to the Regional Veterinary Laboratory, Bukit Tengah.In this study, identification and genetic characterization of Brucella isolated samples using molecular analysis based on IS711 sequence between localisolates and foreign countries accesses in GenBank was done successfully. A total of 31 samples were isolated for Brucella species and then were amplified byPCR, directly sequenced and compared genetically to published sequences which were obtained from GenBank. The most common Brucella species that was found in both bovine (76.5%) and caprine (85.7%) through diagnostic samples in Regional Veterinary Laboratory, Bukit Tengah, was Brucella melitensis. PCR and sequencing were confirmed positive with 76.5% for Brucella melitensis, 23.5% for Brucella abortus and 23.5% for mixed infectionfrom the total of 17 bovine samples. In caprine, the detection of Brucella melintesis and Brucella abortus showed 85.7% and 21.4% respectively meanwhile total mixedinfection showed 21.4%. These clustering between local isolates of Brucella melitensis were phylogenetically related to other Asian countries such as Singapore,Yemen and Saudi Arabia. The Neighbour Joining Analysis clustered the Brucella abortus local isolates for both bovine and caprine were most closely related to India,Iran, Italy and USA. Interestingly, all the isolates within Malaysia have a close relationship (>95%) with the low level of genetic diversity. When local isolates arecompared to GenBank data, it gives an indication on the possible sources of these infections. Eventually, it will improve the import and export policies to controlbrucellosis in Malaysia.

Goose Parvovirus (GPV) also known as Derzy’s disease is aninfectious viral disease of waterfowl which causes serious economic loss in industrial production of geese and Muscovy ducks. In year 2014, Derzy’s disease was detected in Pekin ducks from Sarawak. The affected farm recorded up to 50% mortality, affecting only young ducklings (starting at the age of 3 weeks). Polymerase chainreaction (PCR) from liver samples were performed based on partial region of VP3 gene of GPV, generated amplicon of 801 bp. Sequence analysis showed that the isolate shared 99% sequence similaritywith goose parvovirus strain YBLJ and YZYZ20130304 from China. Phylogeny based on VP3 showed that this isolate is grouped under Asian strains. This is the first report of GPV in Malaysia focusingon the molecular analysis. Notably, this study revealed that GPV not only can be detected from goose and Muscovy but also from Pekin duck.

Recently, Fowl Adenovirus (FAdV) cases have been reported in many countries worldwide. FAdV is a contagious agent associated with inclusion body hepatitis (IBH) and hydropericardium syndrome (HPS) in chickens. It belongs to the Aviadenovirus genus of the family Adenoviridae. The virus is classified into five species (A to E) and further divided into 12 serotypes. Depending on the serotypes, they have diverse characteristics of virus that can either be pathogenic or nonpathogenic strain. From the viewpoint of epidemiological as well as vaccine development, it is very important to detect FAdV strains. Previous studies have been conducted on molecular research, but the continuity of this study in Malaysia has been limited. This study aims to identify the serotype classification of five Malaysian FAdV isolates obtained from field outbreaks during 2017-2019. In this study, polymerase chain reactions (PCR) were conducted based on Hexon gene. Results from the nucleotide sequence analysis discovered that the five isolates showed high similarity with FAdV-8b strains. High bootstrap values in phylogenetic analysis supported the clustering of the Malaysian FAdVs isolates into FAdVs species E. Consequently, the result of this study contributed important information on the epidemiology and culminated in the importance of control strategies against FAdV infection in Malaysia.

Newcastle disease (ND) is an economically important, contagious poultry viral disease reported across the globe. No recent reports on ND circulating in Malaysia. Therefore, the aim of the study is to characterize 16 Newcastle disease viruses (NDVs) isolated from chickens in Malaysia in the year of 2019. All isolates were genotypically analyzed using reverse transcription-polymerase chain reaction (RT-PCR) with primers specific to the viral fusion (F) protein gene. Analysis of the F protein cleavage site’s deduced amino acid sequences revealed that from the Newcastle disease virus (NDV) isolates, three of them were virulent with two different motifs of 112RRQKRF117 and 12RRRKRF117 while other isolates were avirulent. Phylogenetic analysis demonstrated that three isolates were grouped in genotype VII, five in genotype I while eight in genotype II. All genotype VII isolates were clustered under sub-genotype VII.2 (VIIh and VIIi) which is the same strain causing previous outbreaks in Malaysia. Therefore, findings in this study demonstrated that there is no new introduction of NDV genotypes in Malaysia. However, farms should implement biosecurity measures at strict level as well as executing continuous monitoring and surveillance of the disease as these implementations would help them to conduct proper preventive measures and control of panzootic viruses in future.

Locally, venison is considered as a premium and exotic meat, as it is not commonly found in fresh food markets and grocery stores. Despite its limited availability, demand is always high in its niche market, especially during festive seasons which highly escalate the price. However, as an expensive delicacy, deer meat is highly susceptible to fraudulent substitution and adulteration. Authentic deer meat are currently only recognized by consumers based on their own experience, meat texture, and taste which can be quite subjective. To assist in authenticating local deer meat in the market and protect consumers from fraudsters, Polymerase Chain Reaction (PCR) analysis can be carried out to distinguish between venison and other animal meat and products. Farmed venison in Malaysia are mostly from the species Rusa timorensis while Rusa unicolor is bred in the wild. Here, we detailed a newly developed conventional PCR method that is able to detect R. timorensis and R. unicolor based on partial mitochondrial cytochrome c oxidase subunit I (COI) in a single run, thus providing a simple and more accurate alternative in venison authentication.