Piper cubeba L. is traditionally recognised as flavouring ingredient in various types of foods and has been used to marinate meat. Scientifically, it has been reported to possess various valuable nutritional and pharmacological properties including antimicrobial potential. The aim of the present work was to determine the antibacterial activity of ethanolic P. cubeba L. extract against Escherichia coli and its effect on the microbiological quality of raw chicken meat during storage. Disc diffusion assay was done and resulted in 8.40 ± 0.10 mm of inhibition zone. The bacteriostatic and bactericidal effects of the extract were determined at 0.63 ± 0.00 mg/mL and 1.25 ± 0.00 mg/mL of concentration by MIC and MBC methods, respectively. The killing time was recorded at 2 × MIC (1.25 mg/mL) for 4 h. The application of the extract on chicken meat samples showed reduction in TPC and E. coli count with the observed optimum condition at 5.00% concentration stored at -18°C for 14 days based on the consistent reduction. Sensory attributes acceptability evaluation by 9-point hedonic scale showed acceptable score for colour, odour, texture and overall acceptability of the treated raw chicken meat samples. The findings implies that P. cubeba L. can be listed as one of the alternatives to reduce the bacterial load of raw chicken meat prior to cooking which is very important in ensuring food safety as well as reducing the occurrence of foodborne poisoning associated with chicken meat.
Mitochondrial DNA (mtDNA) is a useful genetic marker that can be used for species identification. The cytochrome b (Cyt b) gene is a suitable mtDNA candidate gene for use in phylogenetic analyses due to its sequence variability, which makes it appropriate for comparisons at the subspecies, species, and genus levels. This study was conducted to develop a rapid molecular method for species identification of Malayan gaur (Bos gaurus hubbacki), Kedah-Kelantan (KK) (Bos indicus), and Bali (Bos javanicus) cattle in Malaysia. DNA was extracted from blood samples of 8 Malayan gaurs, 30 KK, and 28 Bali cattle. A set of both specific and universal primers for the Cyt b gene were used in PCR amplification. DNA sequences obtained were then analyzed using BioEdit and Restriction Mapper softwares. The PCR products obtained from Cyt b gene amplification were then subjected to restriction enzyme digestion. The amplification, using both specific and universal primers, produced a 154- and a 603-bp fragment, respectively, in all three species. Two restriction enzymes, NlaIV and SspI, were used to obtain specific restriction profiles that allowed direct identification of Malayan gaur, KK, and Bali cattle. Our findings indicate that all three species can be identified separately using a combination of universal primers and the restriction enzyme SspI.
Bali cattle is a domestic cattle breed that can be found in Malaysia. It is a domestic cattle that was purely derived from a domestication event in Banteng (Bos javanicus) around 3,500 BC in Indonesia. This research was conducted to portray the phylogenetic relationships of the Bali cattle with other cattle species in Malaysia based on maternal and paternal lineage. We analyzed the cytochrome c oxidase I (COI) mitochondrial gene and SRY of Y chromosome obtained from five species of the Bos genus (B. javanicus, Bos gaurus, Bos indicus, Bos taurus, and Bos grunniens). The water buffalo (Bubalus bubalis) was used as an outgroup. The phylogenetic relationships were observed by employing several algorithms: Neighbor-Joining (PAUP version 4.0), Maximum parsimony (PAUP version 4.0) and Bayesian inference (MrBayes 3.1). Results from the maternal data showed that the Bali cattle formed a monophyletic clade, and together with the B. gaurus clade formed a wild cattle clade. Results were supported by high bootstrap and posterior probability values together with genetic distance data. For the paternal lineage, the sequence variation is low (with parsimony informative characters: 2/660) resulting an unresolved Neighbor-Joining tree. However, Bali cattle and other domestic cattle appear in two monophyletic clades distinct from yak, gaur and selembu. This study expresses the potential of the COI gene in portraying the phylogenetic relationships between several Bos species which is important for conservation efforts especially in decision making since cattle is highly bred and hybrid breeds are often formed. Genetic conservation for this high quality beef cattle breed is important by maintaining its genetic characters to prevent extinction or even decreased the genetic quality.