Detection Of Pork And Lard Adulteration In Food Products Using Molecular Biology Techniques For Halal Authentication

Adulteration of food products has become a common problem in many countries. Adulteration may take the form of substitution of one species for another whereby the food products from one species have been mixed intentionally with either similar substitute material or cheaper species. In most cases, food manufacturers often choose lard as a substitute ingredient for oil because it is cheap and easily available. However, the usage of pork and lard is a serious matter in Islam because foods containing ingredients from pig sources are haram (unlawful or prohibited) for Muslims to consume. Therefore, a reliable technique for detection of pork and lard adulteration in food products is necessary in order to protect Muslim consumers from intentional or non- intentional fraud. Polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) techniques have been developed for species identification in various types of food products such as canned fish, peanut and milk. These techniques are proven to be rapid and specific in detecting species adulteration. In this study, rapid methods using PCR and ELISA were utilized to detect pork and lard adulteration in selected food products. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR detection of the conserved region in the mitochondrial (mt) cytochrome (cyt) b gene and mt 12S rRNA gene, respectively, for species identification from raw meat, fat and three types of food products were optimized and developed. Genomic DNA of raw meats, fats and sausages were successfully extracted and were found to be of good quality. Genomic DNA was not detected from the extraction of casing samples and the yield of genomic DNA extracted from bread and biscuit samples were very low. PCR amplifications of mt cyt b gene and 12S rRNA gene produced DNA fragments of approximately 360 bp and 387 bp, respectively from the meat samples. However, no amplification product was observed from the bread and biscuit samples. The amplicons from the mt cyt b gene amplification were then digested with RE BsaJI resulting in species-specific RFLP profiles. The cyt b PCR-RFLP and species-specific PCR identification yielded excellent results for detection of pig derivatives in food products Crude protein were successfully extracted from three types of food products and subjected to ELISA. Using this technique, ten samples were shown to be contaminated with pig derivatives. Positive results were confirmed by observing the colour changes in the well of the ELISA plate. This technique highlighted an alternative in detection of pig derivatives in food products. From these studies, the utilization of PCR-RFLP and the utilization of mt cyt b and 12S rRNA gene in detecting pork and lard adulteration in selected food products were demonstrated. The use of ELISA was also shown to be fast and reliable in identification of pork and lard adulterated food products. However, PCR-RFLP and specific PCR techniques were determined to be better detection techniques for pork and lard adulteration in food products compared to ELISA. The findings from this study can serve as a basis of reference for the research in halal food authentication

下水処理UASBリアクターにおいて嫌気的硫黄酸化反応に関与する微生物群集の解明

国立大学法人長岡技術科学大

Metagenomic studies of the gut microbiota: the snake gut microbiota as a model organism

Microbes play a very important role in each individual. The microbial communities and its genetic blueprint greatly influence in many human diseases. Most of the microbe populations are grow in an individual’s gut. Therefore, metagenomics studies on gut microbes are essential to understand the microbial diversity in gut and the knowledge on microbial composition associates with terrestrial animals will be very important for further understand nutrition, diseases and physiological state. Besides, the availability of next generation sequencing technologies gives a better understanding on gut microbiotas communities compare to the first generation sequencing. This paper, we suggested snakes as a model to study microbial metagenomics due to its various compounds can help to cure various illnesses, even kill off unwanted germs from body. Therefore, this paper mainly review on snake gut microbes, secondary metabolites produce by microbes and the benefits of molecular technologies used in metagenomics which can be useful in medical industries and treatment of infectious diseases

Diversity profile of microbes associated with anaerobic sulfur oxidation in an upflow anaerobic sludge blanket reactor treating municipal sewage

We herein analyzed the diversity of microbes involved in anaerobic sulfur oxidation in an upflow anaerobic sludge blanket (UASB) reactor used for treating municipal sewage under low-temperature conditions. Anaerobic sulfur oxidation occurred in the absence of oxygen, with nitrite and nitrate as electron acceptors; however, reactor performance parameters demonstrated that anaerobic conditions were maintained. In order to gain insights into the underlying basis of anaerobic sulfur oxidation, the microbial diversity that exists in the UASB sludge was analyzed comprehensively to determine their identities and contribution to sulfur oxidation. Sludge samples were collected from the UASB reactor over a period of 2 years and used for bacterial 16S rRNA gene-based terminal restriction fragment length polymorphism (T-RFLP) and next-generation sequencing analyses. T-RFLP and sequencing results both showed that microbial community patterns changed markedly from day 537 onwards. Bacteria belonging to the genus Desulforhabdus within the phylum Proteobacteria and uncultured bacteria within the phylum Fusobacteria were the main groups observed during the period of anaerobic sulfur oxidation. Their abundance correlated with temperature, suggesting that these bacterial groups played roles in anaerobic sulfur oxidation in UASB reactors

Molecular characterization of Escherichia coli isolated from different food sources

Escherichia coli and Escherichia coli O157 were identified from "selom" (Oenanthe stolonifera), "pegaga" (Centella asiatica), beef, chicken, lamb, buffalo, "ulam Raja" (Cosmos caudatus) and "tenggek burung" (Euodia redlevi). The bacteria were recovered using chromagenic agar. Isolated Escherichia coli and Escherichia coli 0157 were further characterized by plasmid profiling and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The virulence genes of the isolates (VT1, VT2, LT, ST, eaeA, inV) that produces pathogenic Escherichia coli and 16S rRNA gene were screened by a multiplex PCR assay. The plasmid profiling analysis showed that out of 176 isolates, only 103 isolates contained plasmids. ERIC-PCR analysis generated amplified products in the range of ~150 bp to > 1000 bp categorizing isolates into a total of 52 different profiles. Multiplex PCR showed that 20 (32.3%) of the isolates carried eaeA gene, 6 (9.7%) isolates possessed inV genes, only 1 (1.6%) have VT2 genes and 1 (1.6%) as well carried VT1 genes, 2 (3.2%) of the isolates harboured LT genes, and only 1 (1.6%) isolate possessed ST genes. There were no correlation between plasmid, ERIC-PCR and virulence genes profiles

Exploring Muslim Consumers’ Acceptance of Cultured Beef Meat

The advancement in cultured meat research in cellular agriculture has greatly surged. The concerns of halalness and thayibban (cleanliness and permissibility to consume) of cultured beef meat will arise among Muslim consumers, prompting the question, “Who will consume the cultured meat, and are Muslims ready to consume it?” This study aimed to clarify how Muslims perceive cultured meat and the issues surrounding their acceptance. A chi-square test and a binary logistic regression analysis were applied to reveal the acceptance of cultured meat. The results revealed that 44.1% of the respondents accepted cultured meat as their food, while 55.9% expressed doubts due to religious concerns. Their attitudes toward cultured meat influenced their decision to accept it as food. Some consumers had high expectations for cultured meat because they believed it would be superior in taste and have nutritional value and health effects. In conclusion, those Muslims who did not doubt cultured meat accepted it as future food with expectations for better function and value

Exploring Muslim consumers' acceptance of cultured beef meat

The advancement in cultured meat research in cellular agriculture has greatly surged. The concerns of halalness and thayibban (cleanliness and permissibility to consume) of cultured beef meat will arise among Muslim consumers, prompting the question, “Who will consume the cultured meat, and are Muslims ready to consume it?” This study aimed to clarify how Muslims perceive cultured meat and the issues surrounding their acceptance. A chi-square test and a binary logistic regression analysis were applied to reveal the acceptance of cultured meat. The results revealed that 44.1% of the respondents accepted cultured meat as their food, while 55.9% expressed doubts due to religious concerns. Their attitudes toward cultured meat influenced their decision to accept it as food. Some consumers had high expectations for cultured meat because they believed it would be superior in taste and have nutritional value and health effects. In conclusion, those Muslims who did not doubt cultured meat accepted it as future food with expectations for better function and value

Specific polymerase chain reaction (PCR) analysis of raw meats and fats of pigs for Halal authentication

Species-specific polymerase chain reaction (PCR) analysis of a conserved region in the mitochondrial (mt) 12S ribosomal RNA (rRNA) gene was developed for species identification from raw pork and lard samples. Genomic DNA of pork and lard were successfully extracted and were found to be of good quality. The extracted genomic DNA was then subjected to PCR amplification targeting the specific regions of the 12S rRNA gene and produced clear PCR products on the amplification of 12S rRNA gene of 387 base pairs (bp) from pig species. The species-specific PCR identification yielded excellent results for identification of pig. This made it deal for quality control purposes and a potentially reliable technique to avoid species adulteration for Halal authentication and verification

The use of spectroscopic methods in combination with multivariate data analysis for determination of omega fatty acids: a review

Omega-3 fatty acids (ω-3 FAs), typically found in fish oils and marine-based products, are important fatty acids due to their beneficial activities toward human health, such as anti- inflammation, immune-stimulant, lowering the risk of cardiovascular disease and reducing blood pressure. Therefore, the determination of ω-3 FAs for quality control of products containing these FAs is very important. Molecular spectroscopic methods offered simple, fast, and reliable analytical methods for quality controls of food and pharmaceutical products containing ω-3 FAs since a large amount of information could be retrieved from molecular spectra. This review highlighted the employment of molecular spectroscopy such as near-infrared (NIR), Fourier transform infrared (FTIR), Raman, and nuclear magnetic resonance (NMR) spectrometer combined with multivariate data analysis or chemometrics for analysis of ω-3 FAs in fish oil-based products. From this review, it is reported that the combination of molecular spectroscopy and chemometrics could be used as effective analytical techniques for the analysis of ω-3 FAs, especially eicosapentaenoic acid (C20:5, ω-3) and docosahexaenoic acid (C22:6, ω-3), with high accuracy and high precision. The results of quantitative analysis of ω-3 FAs from NIR, FTIR, Raman, and NMR were comparable to those reference results obtained from gas chromatography-mass spectrometry measurement. In the future, collaborative studies through proficiency testing should be performed to get standardized methods based on molecular spectroscopy and chemometrics