This study was conducted to detect the presence of chicken and porcine DNA in meatballs using mitochondria DNA (mtDNA) of cytochrome b (cyt b) and nuclear DNA (nDNA) short interspersed nuclear element (SINE) species-specific primers, respectively. While, the mtDNA primers targeted transfer RNA-ATP8 (tRNA-ATP8) gene was used for 1 and 5% (w/w) chicken meatball spiked with commercial porcine blood plasm. Chicken meatballs spiked with 1% and 5% (v/w) fresh and commercial porcine blood plasma, respectively were prepared and heat-treated using five (n = 5) cooking methods: boiling, pan-frying, roasting, microwaving and autoclaving. Two pairs of mtDNA and nDNA primers used, produced 129 and 161 bp amplicons, respectively. Whereas, tRNA-ATP8 primers produced 212 bp of amplicon. Electrophoresis analysis showed positive results for porcine DNA at 1% and 5% (w/w or v/v) for all of the different cooking techniques, either for fresh or commercial blood plasma using SINE primers but not for tRNA-ATP8 primers. The present study has highlighted the useful of species-specific primers of SINE primers in PCR analysis for detecting porcine DNA blood plasma in heat-treated chicken meatballs.
In order to invent a porcine gelatine detection device using microbial resources, bacterial enzymes with a preference towards porcine gelatine and their candidate genes were evaluated. Five (n = 5) bacterial strains isolated from hot spring water and wet clay, Malaysia were screened for their gelatinase activity. The gelatinase enzyme was extracted and purified using ammonium sulphate precipitation prior to performing gelatinase assay on porcine, bovine and fish gelatine medium substrates. The G2 strain or Enterobacter aerogenes (Strain EA1) was selected for whole genome sequenced after showing a consistent trend of preference towards porcine gelatine. The gelatinase candidate gene gelEA1_9 was cloned and expressed. Based on one-way analysis of variance (ANOVA) with POST-HOC Duncan test (α = 0.05), the final product of gelEA1_9 was identified as a novel gelatinase. This gelatinase presented no significant difference in activity towards porcine gelatine. Hence, the present study demonstrated an enzyme-substrate interaction for porcine gelatine identification.
Cultured meat is meat produced from stem cell biopsies of cattle. Stem cells were cultured in a bioreactor in the presence of serum to grow the flesh to maturity. Cultured meat technology originated from regenerative medical technology; however, it has been given a new lease of life to produce cultured meat as an innovative food source in the future without involving cattle breeding. This technology can reduce the negative environmental impacts of global warming, water use, soil, and unethical handling of animals. In the excitement of accepting this new technology, the halal status of cultured meat is in question, as it can be produced from embryonic stem cells and myosatellite cells, each of which can be disputed for their halal status. Additionally, the process of culturing and maturation of stem cells involves the use of an impure medium derived from animal blood. Thus, cultured meat is acceptable to Muslims only if the stem cells, medium and scaffold biomaterials used to manufacture it are from Halal sources and shall be in line with the six principles discussed in this study. The discussion is based on Halal and haram animals; Animal slaughtering; Not derived from a source of najs (impurity); Istihalah tammah (perfect substance change); Maslahah (public interest or benefit) and mafsadah (damage); and Darurat (exigency) of cultured meat)).
Meat culturing technology goes beyond laboratory research and materialises in the market. Nonetheless, this technology has raised concerns among Muslim consumers worldwide due to its medium, especially foetal bovine serum (FBS), which originates from blood. Thus, the aim of this research was to determine the halal status of cultured meat by detecting species-specific DNA of bovine serum as one of the media used during meat production. Polymerase chain reaction (PCR) analysis was conducted by targeting mitochondrial cytochrome oxidase II (COII) gene sequences, producing a 165 bp amplicon. The sequences of the primers used were Bovine-F, 5'-CAT CAT AGC AAT TGC CAT AGT CC-3' and Bovine-R, 5'-GTA CTA GTA GTA TTA GAG CTA GAA TTA G-3'. DNA extraction was conducted using a QIAGEN Blood and Tissue™ commercial kit. The presence study also included a literature review on the Istihalah (transformation) concept in order to determine the halal status of cultured meat. The results revealed that bovine DNA was detected in all samples tested using PCR analysis. Therefore, Istihalah tammah (perfect transformation) does not occur due to the ability of PCR analysis to detect bovine DNA in FBS and is prohibited according to Shariah law.
A total of 24 strains of Vibrio alginolyticus were isolated from cockles (Anadara granosa) and identified for VibA and gyrB genes. All V. alginolyticus isolates were then tested against nine different antibiotics. In this study, the highest percentage of antibiotic resistance was obtained against penicillin (37.50%), followed by ampicillin, vancomycin (12.50%) and erythromycin (8.33%). All of V. alginolyticus isolates were susceptible against streptomycin, kanamycin, tetracycline, chloramphenicol and sulfamethoxazole. Polymerase chain reaction (PCR) assay has confirmed the presence of four antibiotic resistance genes of penicillin (pbp2a), ampicillin (blaOXA), erythromycin (ermB) and vancomycin (vanB). Out of 24 V. alginolyticus isolates, 2 isolates possessed the tdh-related hemolysin (trh) (strains VA15 and VA16) and none for the thermostable direct hemolysin (tdh) gene. Both strains of the tdh-related hemolysin (trh) were susceptible to all antibiotics tested. The multiple antibiotic resistance (MAR) index ranging between 0.2 and 0.3 with 5 antibiograms (A1-A5) was observed. Combination of enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and antibiotic resistance indicated 18 genome types which showed genetic heterogeneity of those V. alginolyticus isolates. The results demonstrated the presence of V. alginolyticus strain found in cockles can be a potential risk to consumers and can contribute to the deterioration of human health in the study area. Thus, it is essential for local authority to provide the preventive measures in ensuring the cockles are safe for consumption.