Anthropogenic particles in the muscle, gill, and gastrointestinal tract of marine fish sold for human consumption

Contamination of marine fish with the widespread distribution of anthropogenic particles (APs) becomes increasingly severe, however, related research on the assessment of the occurrence of APs in the edible tissue of commercial fish is scarce. The objective of this study was to evaluate the features of APs pollution based on seven species of commercial marine fish (n = 12 per species) and investigate the accumulation of APs in different tissues of fish namely gill and gastrointestinal tract (GIT), and muscle. The results show that a total of 62 APs were detected in 33 out of 84 (39.3%) fresh fish samples using a micro-Raman spectrometer which in particular is characterized by a blue color, shape-like fiber, and size smaller than 0.5 mm. Among them, 47 (75.8%) particles were identified as pigments such as indigo, chrome yellow-orange, disperse yellow, and pigment black. The other 11 (17.7%) particles were plastic including polypropylene (PP), polyethylene terephthalate (PET), and polyacrylonitrile (PAN). And the rest 4 (6.5%) particles were anthropogenic cellulose fibers. Muscle tissue from six species of fish was detected to contain a total of 15 APs. Based on the total mean of APs found in fish muscle (0.018 AP items/g tissue) and on the consumption of fish in Malaysia (59 kg/capita/year), the estimated human intake of APs through fish consumption was 1062 AP items/year/capita. Considering that food consumption is an important route of human exposure to APs, it is suggested to add APs testing into the guidelines of food safety management systems and adopt mitigation measures to reduce the APs pollution in food

Nanoparticle sensor for label free detection of swine DNA in mixed biological samples

We used 40 ± 5 nm gold nanoparticles (GNPs) as colorimetric sensor to visually detect swine-specific conserved sequence and nucleotide mismatch in PCR-amplified and non-amplified mitochondrial DNA mixtures to authenticate species. Colloidal GNPs changed color from pinkish-red to gray-purple in 2 mM PBS. Visually observed results were clearly reflected by the dramatic reduction of surface plasmon resonance peak at 530 nm and the appearance of new features in the 620–800 nm regions in their absorption spectra. The particles were stabilized against salt-induced aggregation upon the adsorption of single-stranded DNA. The PCR products, without any additional processing, were hybridized with a 17-base probe prior to exposure to GNPs. At a critical annealing temperature (55 °C) that differentiated matched and mismatched base pairing, the probe was hybridized to pig PCR product and dehybridized from the deer product. The dehybridized probe stuck to GNPs to prevent them from salt-induced aggregation and retained their characteristic red color. Hybridization of a 27-nucleotide probe to swine mitochondrial DNA identified them in pork–venison, pork–shad and venison–shad binary admixtures, eliminating the need of PCR amplification. Thus the assay was applied to authenticate species both in PCR-amplified and non-amplified heterogeneous biological samples. The results were determined visually and validated by absorption spectroscopy. The entire assay (hybridization plus visual detection) was performed in less than 10 min. The LOD (for genomic DNA) of the assay was 6 µg ml − 1 swine DNA in mixed meat samples. We believe the assay can be applied for species assignment in food analysis, mismatch detection in genetic screening and homology studies between closely related species

Determination of lethal (LD) and growth reduction (GR)doses on acute and chronic gamma- irradiated Bambara groundnut [Vigna subterranea (L.) Verdc.]varieties

Bambara groundnut is a highly nutritious underutilized legume with enormous potential to sustain food security in resource-poor countries. However, its potential for improvement through conventional breeding (< 2% success rate) limitation due to the nature of the flowers. Thus, the most viable method of improving this crop is by creating genetic variability through induced mutagenesis. The present study was conducted to evaluate the radiosensitivity of two Bambara groundnut varieties irradiated with acute and chronic gamma irradiations to determine the lethal dose (LD) and growth reduction dose (GR). Healthy seeds of both varieties were exposed to acute gamma irradiation using Cesium-137 at 0, 25, 50, 75, 100, 125, 150, 175, 200, 250, and 300 Gy. For chronic irradiation, two-week-old seedlings of the two genotypes were exposed to accumulated doses of 0, 8.52, 17.04, 35.56, 34.09, 42.61, 59.65, 93.74, 144.87, 255.64, and 570.94 Gy, respectively, in Gamma Green House (GGH) for 60 days. The result from the variance analysis indicated highly significant differences (P < 0.01) for all evaluated traits except for internode length. A linear regression model was developed to determine the mean LD and GR of both genotypes. The established lethal doses (LD25, 50, 75) for acute gamma irradiation on Ex-Sokoto variety were 75, 160, and 250 Gy while 68, 148, and 227 Gy were recorded for Karo variety, respectively. For chronic irradiation, the established growth reduction doses for Ex-Sokoto were 47, 250, and 444 Gy, whereas 70, 264, and 452 Gy were observed in Karo. Variations were observed between the gamma-irradiated genotypes and the methods of irradiations. Generally, the growth, development, and survival rate of Bambara groundnut increase with a decrease in gamma-irradiation doses. The established LD and GR doses from this study can be utilized in large-scale mutagenesis breeding programs for generating a wide range of mutants in Bambara groundnut

Detection of raw pork targeting porcine-specific mitochondrial cytochrome B gene by molecular beacon probe and real-time polymerase chain reaction

A real-time polymerase chain reaction (PCR) assay with molecular beacon probe was developed for the detection of pork in raw states. The method combined the swine-specific primers and molecular beacon probe to specifically amplify a 119-bp fragment of porcine mitochondrial cytochrome b (mt-cyt b) gene. Mitochondrial genes are present in multiple copies and thus ensure available targets even in degraded samples. A pair of 18-nucleotide swine-cytb-specific primers were designed using Primer 3 Plus software. On the other hand, a 34-nt molecular beacon probe was designed using Beacon Designer 4 software. The porcine specificity of the primers and probe were checked by basic local alignment search tools (BLAST) to avoid mismatches with other species. A specificity test with 10 ng DNA of nine common meat providing land and aquatic species yielded a Cq value of 18.70±0.12 to 19.08±0.06 only with the pork DNA in a 40 cycle PCR reaction, demonstrating the swine specificity of the primers and probe. The swine-specificity was further confirmed in a binary mixture of pork and beef. The method detected 0.1% pork in binary pork-beef mixture with a Cq of 25.79±0.20. A sensitivity test with 10-fold serial dilution revealed that the assay can determine 0.0001 ng of porcine DNA with a PCR efficiency of 96% with a good reproducibility, precision and high correlation coefficient (r2=0.9989). The shorter length target (119-bp) and strong sensitivity and specificity suggest the method can be used for the routine analysis of pork adulteration in raw meats

Review on halal forensic: A focus on dna-based methods for pork authentication

Food product authentication is important at every level of the food manufacturing process, starting from raw materials until finished products. Authentication also plays an important role in assuring accurate food labelling, which is required to help consumers select suitable types of food products. Food adulteration is one of the vital issues addressed by halal authentication, especially for food products that contain pig traces or porcine ingredients. Various methods that aim to guarantee the authenticity of foods have been developed over the past years. In this article, a short review of recent food analytical methods related to authenticity studies, with special regard to pork identification, is provided. The focus of this review is DNA-based methods, which have gained the interest of the scientific community. The specificity, sensitivity and fast and high throughput of the methods are highlighted. In the present case, methods that are capable of detecting pork by using DNA barcode, polymerase chain reaction (PCR)-restriction fragment length polymorphism, conventional PCR, real-time PCR and isothermal amplification are discussed. Although PCR is the most popular method, recent studies have shown that isothermal amplification is a potential alternative because it is rapid, simple and does not require the use of any complicated instruments, such as a thermal cycler and sequencer

Detection of pork in pastry products for halal verification via polymerase chain reaction (PCR) techniques

Pork identification in pastry fillings using polymerase chain reaction (PCR) was developed. Genomic DNA of the pastry filling were successfully extracted and found to be good quality and produced clear PCR products. In this study, two methods of PCR were used for detection of pork adulteration in several types of pastry fillings, including Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and species-specific PCR amplification. PCR amplification of mitochondrial cytochrome b gene (mt-cyt b) for PCR-RFLP produced DNA fragments of approximately 360 bp. This generated amplicons were then digested with restriction enzyme BsaJI, resulting in RFLP profiles of 228 bp and 131 bp in sizes which are specific for pork. Besides that, the species-specific PCR amplification of cyt b gene produced DNA fragments with approximate size of 398 bp as expected for the positive results. The findings from this study can serve as a basis of reference for the research in halal food authentication

Rapid Detection of Porcine DNA in Meatball Using Recombinase Polymerase Amplification Couple with Lateral Flow Immunoassay for Halal Authentication

Point-of-care diagnostic methods for animal species determination are critical for rapid, simple, and accurate enforcement of food labelling. PCR is the most common method for species identification. However, the requirement of using a thermal cycler created drawbacks for the PCR application, particularly in low-resource settings. Hence, in this study, a method for porcine DNA detection using recombinase polymerase amplification (RPA), coupled with nucleic acid lateral flow immunoassay (NALFIA), was developed. Porcine-specific primers targeting pig (Sus scrofa) cytochrome b gene fragments specifically amplify a 197 bp fragment of the mitochondrial gene as being visualized by 2% agarose gel and PCRD NALFIA. The reaction temperature and time were 39 &deg;C and 20 min, respectively. Herein, the specificity of the primers to porcine was confirmed after being assayed against six animal species, namely cow, goat, chicken, duck, dog, and rabbit. The porcine-specific RPA assay shows a high limit of detection of 0.01 ng/&micro;L pork DNA. Based on the preliminary performance data obtained from this study, the potential of this method as a rapid and sensitive tool for porcine DNA detection in meat-based products is foreseen

Exploration of Bambara groundnut (Vigna subterranea (L.) Verdc.), an underutilized crop, to aid global food security: varietal improvement, genetic diversity and processing

Currently, the global agricultural system is focused on a limited number of crop species, thereby presenting a threat to food security and supply, especially with predicted global climate change conditions. The importance of ‘underutilized’ crop species in meeting the world’s demand for food has been duly recognized by research communities, governments and policy makers worldwide. The development of underutilized crops, with their vast genetic resources and beneficial traits, may be a useful step towards solving food security challenges by offering a multifaceted agricultural system that includes additional important food resources. Bambara groundnut is among the beneficial underutilized crop species that may have a positive impact on global food security through organized and well-coordinated multidimensional breeding programs. The excessive degrees of allelic difference in Bambara groundnut germplasm could be exploited in breeding activities to develop new varieties. It is important to match recognized breeding objectives with documented diversity in order to significantly improve breeding. This review assesses the genetic diversity of Bambara groundnut, as well as important factors involved in realizing and harnessing the potential of this crop