Meat Species Research Articles

Electronic Tongue as a Correlative Technique for Modeling Cattle Meat Quality and Classification of Breeds.

Discrimination and species identification of meat has always been of paramount importance in the European meat market. This is often achieved using different conventional analytical methods but advanced sensor-based methods, such as the electronic tongue (e-tongue), are also gaining attention for rapid and reliable analysis. The aim of this study was to discriminate Angus, domestic buffalo, Hungarian Grey, Hungarian Spotted cattle, and Holstein beef meat samples from the chuck steak part of the animals, which mostly contained longissimus dorsi muscles, using e-tongue as a correlative technique with conventional methods for analysis of pH, color, texture, water activity, water-holding capacity, cooking yield, water binding activity, and descriptive sensory analysis. Analysis of variance (ANOVA) was used to determine significant differences between the measured quality traits of the five-meat species after analysis with conventional analytical methods. E-tongue data were visualized with principal component analysis (PCA) before classifying the five-meat species with linear discriminant analysis (LDA). Significant differences were observed among some of the investigated quality parameter. In most cases, Hungarian Grey was most different from the other species. Using e-tongue, separation patterns could be observed in the PCA that were confirmed with 100% recognition and 97.5% prediction of all the different meat species in LDA.

Chemometrics in Tandem with Hyperspectral Imaging for Detecting Authentication of Raw and Cooked Mutton Rolls.

Authentication assurance of meat or meat products is critical in the meat industry. Various methods including DNA- or protein-based techniques are accurate for assessing meat authenticity, however, they are destructive, expensive, or laborious. This study explores the feasibility of chemometrics in tandem with hyperspectral imaging (HSI) for identifying raw and cooked mutton rolls substitution by pork and duck rolls. Raw or cooked samples (n = 180) of three meat species were prepared to collect hyperspectral images in range of 400–1000 nm. Spectra were extracted from representative regions of interest (ROIs), and spectral principal component analysis (PCA) revealed that PC1 and PC2 were effective for the identification. Different methods including standard normal variable (SNV), first and second derivatives, and normalization were individually employed for spectral preprocessing, and modeling methods of partial least squares-discriminant analysis (PLS-DA) and support vector machines (SVM) were also individually applied to develop classification models for both the raw and the cooked. Results showed that PLS-DA model developed by raw spectra presented the highest 100% correct classification rate (CCR) of success in all sets. After that, effective wavelengths selected by successive projections algorithm (SPA) built optimal simplified models which didn’t influence the modeling results compared with full spectra regardless of the meat roll states. Therefore, SPA-PLS-DA models were subsequently used to visualize the raw and cooked meat rolls classification. As a consequence, the general meat species of both raw and cooked meat rolls were readily discernible in pixel-wise manner by generating classification maps. The results showed that HSI combined with chemometrics can be used to identify the authentication of raw and cooked mutton rolls substituted by pork and duck rolls accurately. This promising methodology provides a reference which can be extended to the classification or grading of other meat rolls.

Rapid LC-MS/MS method for the detection of seven animal species in meat products

The adulteration of meat products has been reported worldwide, and detection of specific peptides through mass spectrometry (MS) is a reliable method for meat species identification. However, the practical application of this method is limited by complicated steps and long reaction time of the traditional sample preparation. Therefore, this paper introduced a convenient and time-saving sample preparation by optimizing the steps of reduction, alkylation, digestion, and purification. With the rapid sample preparation, 35 species-specific peptides for seven species (pig, cattle, sheep, deer, chicken, duck, and turkey) were screened using high-resolution MS, and a rapid LC-MS/MS method was established. The method only takes 3 h from sample receipt to results. The meat species of 20 processed meat products were detected, and three samples were found potentially adulterated. The method is proved to have high sensitivity, specificity, practicability with respect to rapid identification of meat species in meat products.

Occurrence, Risk Factors, and Antimicrobial Susceptibility Test of Thermophilic Campylobacter Species of Bovine Carcass at Municipal Abattoir and Butcher Shops of Jimma Town, Southwest Ethiopia.

BackgroundAlthough Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) cause acute diarrheal diseases in people all over the world, they are most commonly seen in other mammalian species and are a seemingly healthy carrier condition. Humans in Ethiopia, on the other hand, are largely unaware of the existence of Campylobacter in food animals as potential sources of infection.ObjectiveTo determine the occurrence, risk factors, and antimicrobial susceptibility pattern of thermophilic Campylobacter species in bovine raw meat from the abattoir and butcher shops.MethodsSwab samples were collected from 177 cattle carcasses (from abattoir 93 and butcher shops 84) and cultured using standard methods. An antimicrobial susceptibility test was performed using the disk diffusion method towards eleven antimicrobial agents.ResultsThe overall prevalence of thermophilic Campylobacter species was 14 (7.9%). From these, 11 (78.6%) were identified to be C. jejuni and 3 (21.4%) were C. coli. Lack of handwashing before meat processing and after visiting the toilet, meat contact with floors, walls, or soiling during preparation, and lack of training were the most important factors independently associated with (p<0.05) the prevalence of Campylobacter species contamination. The highest level of antimicrobial resistance of the Campylobacter isolates was recorded to ampicillin (10μg) (100%), followed by amoxicillin (30μg) (78.6%) and sulphamethoxazole-trimethoprim (57.1%) while the least resisted antimicrobials were streptomycin (25μg), erythromycin (15μg), oxytetracycline (30μg) (each 28.6%), kanamycin (30μg) 14.3%, chloramphenicol (30μg) and gentamycin (10μg) (each 7.1%).ConclusionDespite the low prevalence of thermophilic Campylobacter in the current investigation, it may pose a significant public health threat. As a result, it is vital to give retailers and customers extensive education, training, and knowledge about the correct handling and cooking of animal-derived goods. Furthermore, antimicrobials should be used with caution in both veterinary and human treatment regimens as well as a wider examination of antimicrobial resistance patterns for the use of well-targeted antimicrobials.

Application of Next-Generation Sequencing Technology Based on Single Gene Locus in Species Identification of Mixed Meat Products

Polymerase chain reaction (PCR) detection is a commonly used method for species identification of meat products. However, this method is not suitable for the analysis of meat products containing multiple mixtures. This study aimed to test whether next-generation sequencing (NGS) technology could be used as a method for the certification of mixed meat products. In this study, five kinds of common meat (pigs, cattle, sheep, chickens, and ducks) were mixed as samples with different proportions. The primers designed from mitochondrial 16S rRNA and nuclear genome gene (growth hormone receptor, GHR), respectively, were used to detect these meats. The sequencing results of NGS were analyzed using a self-designed bioinformatics program. The fragments with similar sequences were classified and compared with the database to determine their species. The results showed that all five kinds of meat components could be correctly identified using these two primers. The meat composition could be detected as low as 0.5% in the mixed samples using the NGS technology targeting GHR gene fragments, which was superior to those targeting mitochondrial 16S rRNA. However, the quantitative detection of species in the mixture was not likely to be quite accurate due to the amplification bias of PCR amplification. These results showed that the NGS technology could be applied to identify meat species in mixtures.

Prevalence and Antibiograms of Salmonella in Commercially Produced Crocodile meat in Zimbabwe

The presence of Salmonella in food products and emergence of antibiotic resistance are the major challenges facing public health policies. A total of 2749 crocodile meat samples obtained from the Central Veterinary Laboratories in Zimbabwe were screened for Salmonella specieswere collected from three Zimbabwean commercial farms between the year 2012 and 2019 for a retrospective observational study to determine the prevalence and magnitude of antibiotics resistant Salmonella species in crocodile meat. The isolation of Salmonella was in accordance with the ISO 6579:2002 and the antibiotic susceptibility testing was carried out based on Clinical and Laboratory Standard Institute’s recommendations by means of the Kirby-Bauer disk diffusion method. SILAB Database was used to determine the prevalence of Salmonella species. Prevalence was stratified by year and farms. Twenty Salmonella isolates were identified using biochemical tests, and 15 were confirmed by polymerase chain reaction (PCR). Antimicrobial susceptibility profiles of the confirmed Salmonella isolates were examined using 14 antibiotics. The overall prevalence of Salmonella species in crocodile meat samples was 0.5%. The prevalence of Salmonella species ranged from 0.04% to 0.44% in the crocodile meat samples and annual prevalence ranged from 0.01% to 1%. The highest prevalence of Salmonella (4.4%) was recorded in the year 2012. Salmonella isolates from one of the three tested farms were resistant to Erythromycin (73.33%), Ampicillin (80%), and Penicillin G (100%). Generally, Salmonella isolates displayed lower resistance to Cefepime, Ceftriaxone, Amikacin, Tetracycline, Ertapenem, Florfenicol, and Erythromycin (0-53.33%) whereas all Salmonella isolates showed susceptibility to Cefepime, Ceftriaxone, Ertapenem, and Florfenicol. Although the study indicates low prevalence of Salmonella species in crocodile meat, there is a need for strict implementation of Hazard Analysis Critical Control Point (HACCP) to reduce contamination rates in meat and its products

Wild meat trade over the last 45 years in the Peruvian Amazon.

The trade in wild meat is an important economic component of rural people's livelihoods, but it has been perceived to be among the main causes of the decline of wildlife species. Recently, the COVID-19 pandemic has brought to light an additional concern of wildlife markets as a major human-health challenge. We analyzed data from the largest longitudinal monitoring (1973-2018) of the most important urban wild-meat markets in Iquitos, Peru, to examine the trends in and impacts of these markets on people's livelihoods. Over the last 45 years, wild meat sales increased at a rate of 6.4 t/year (SD 2.17), paralleling urban population growth. Wild meat sales were highest in 2018 (442 t), contributing US$2.6 million (0.76%) to the regional gross domestic product. Five species of ungulates and rodents accounted for 88.5% of the amount of biomass traded. Vulnerable and Endangered species represented 7.0% and 0.4% of individuals sold, respectively. Despite growth in sales, the contribution of wild meat to overall urban diet was constant: 1-2%/year of total meat consumed. This result was due to greater availability and higher consumption of cheaper meats (e.g., in 2018, poultry was 45.8% cheaper and was the most consumed meat) coupled with the lack of economic incentives to harvest wild meat species in rural areas. Most wild meat was sold salted or smoked, reducing the likelihood of foodborne diseases. Community-based wildlife management plans and the continued trade bans on primates and threatened taxa may avoid biodiversity loss. Considering the recent COVID-19 pandemic, future management plans should include potential viral hosts and regulation and enforcement of hygiene practices in wild-meat markets.

Single nucleotide polymorphism-based methodology for authentication of bovine, caprine, ovine, camel, and donkey meat cuts.

To guarantee food safetyand sustainability, it is necessary to verify meat authenticity. This study focused on the development of single nucleotide polymorphism-based polymerase chain reaction-restriction fragment length polymorphism (SNP-based PCR-RFLP) and forensically informative nucleotide sequence (FINS) methodologies based on PCR amplification of the mitochondrial 12S rRNA gene for discrimination of six red meat species, that is, cattle, buffalo, goat, sheep, camel, and donkey. FINS allowed the unambiguous identification of all species analyzed. In addition, six SNPs, where a restriction site for TasI could be localized using a preliminary in silico analysis, gave a unique RFLP pattern for each species. The results revealed a low level of species substitution (8%) in the tested meat samples. In particular, one buffalo and goat samples have been substituted with cow and sheep, respectively. Finally, the developed techniques herein showed high potentials to be routinely used as reliable and fast tools to avoid meat species substitutions. PRACTICAL APPLICATION: This research deals with genetic techniques to trace meats. This kind of research helps the concerned agencies to build capacity to safeguard consumer sentiments as well as providing better market access and better food price and quality for the consumer.

Evaluation of High Resolution Melting (HRM) Analysis for Meat Species Identification of Raw and Cooked Meat

The current study aimed to examine a real-time PCR assay with high-resolution melting (HRM) analysis for the species identification of minced meat samples. Meat samples from several animal species were purchased and minced separately or as a mixture of two species. DNA was extracted from all meat samples and subjected to real-time PCR assay by amplifying species-specific mitochondrial cytochrome b regions. Regarding the meat mixtures, two separate melting curves with specific melt peak temperatures (Tm) were detected. Additionally, DNA from each species was quantified, based on the calibration curves. The results showed that a real-time PCR assay with HRM analysis is suitable for the species identification of meat products, and could be used for the detection of meat frauds.

1H-NMR-Based Metabolomics: An Integrated Approach for the Detection of the Adulteration in Chicken, Chevon, Beef and Donkey Meat.

Meat is a rich source of energy that provides high-value animal protein, fats, vitamins, minerals and trace amounts of carbohydrates. Globally, different types of meats are consumed to fulfill nutritional requirements. However, the increasing burden on the livestock industry has triggered the mixing of high-price meat species with low-quality/-price meat. This work aimed to differentiate different meat samples on the basis of metabolites. The metabolic difference between various meat samples was investigated through Nuclear Magnetic Resonance spectroscopy coupled with multivariate data analysis approaches like principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). In total, 37 metabolites were identified in the gluteal muscle tissues of cow, goat, donkey and chicken using 1H-NMR spectroscopy. PCA was found unable to completely differentiate between meat types, whereas OPLS-DA showed an apparent separation and successfully differentiated samples from all four types of meat. Lactate, creatine, choline, acetate, leucine, isoleucine, valine, formate, carnitine, glutamate, 3-hydroxybutyrate and α-mannose were found as the major discriminating metabolites between white (chicken) and red meat (chevon, beef and donkey). However, inosine, lactate, uracil, carnosine, format, pyruvate, carnitine, creatine and acetate were found responsible for differentiating chevon, beef and donkey meat. The relative quantification of differentiating metabolites was performed using one-way ANOVA and Tukey test. Our results showed that NMR-based metabolomics is a powerful tool for the identification of novel signatures (potential biomarkers) to characterize meats from different sources and could potentially be used for quality control purposes in order to differentiate different meat types.

A Practical Approach to Identifying Processed White Meat of Guinea Fowl, Rabbit, and Selected Fish Species Using End-Point PCR.

Among the foodstuff, most often adulterated are white meat and meat products as well as fish and fish products. For this reason, we evaluated in practice the possibilities of identifying selected species of white meat, i.e., guinea fowl and rabbit as well as four fish species, namely, pollock, hake, sole, and panga, in thermally treated samples. The aim was to check whether the previously published in the scientific literature species-specific primers allows for the identification of processed meat using the end-point PCR technique. To identify the six species, the short sequence fragments (from 130 to 255 bp) of 12S rRNA, COX3, mitochondrial ATP synthase Fo subunit 6 (ATP6) gene, pantophysin (Pan I) gene, 5S rRNA gene, and microsatellite markers (locus: Phy01-KUL) were selected. Stability and specificity of the six pair primers were evaluated on cooked and autoclaved meat, and commercially processed food samples such as rabbit and guinea pâtés, ready-made baby food, and breaded, fried, and deep-frozen fish products. The method proved to be useful for the authentication of severely processed food products against fraudulent species substitution and mislabelling and this approach may be an alternative to more advanced and more expensive PCR techniques.

Meat species identification accuracy improvement using sample set portioning based on joint x-y distance and laser-induced breakdown spectroscopy.

Laser-induced breakdown spectroscopy (LIBS) was suitable for the identification of meat species due to fast and less sample preparation. However, the problem of low accuracy rate of the recognition model caused by improper selection of training set samples by random split has severely restricted the development of LIBS in meat detection. Sample set portioning based on the joint x-y distance (SPXY) method was applied for dividing the meat spectra into a training set and a test set. Then, the five kinds of meat samples (shrimp, chicken, beef, scallop, and pig liver) were classified by the support vector machine (SVM). With the random split method, Kennard-Stone method, and SPXY method, the recognition accuracies of the SVM model were 90.44%, 91.95%, and 94.35%, respectively. The multidimensional scaling method was used to visualize the results of the sample split for the interpretation of the classification. The results showed that the identification performance of the SPXY method combined with the SVM model was best, and the accuracy rates of shrimp, chicken, beef, scallop, and pig liver were 100.00%, 100.00%, 100.00%, 78.57%, and 92.00%, respectively. Moreover, to verify the broad adaptability of the SPXY method, the linear discriminant analysis model, the K-nearest neighbor model, and the ensemble learning model were applied as the meat species identification model. The results demonstrated that the accuracy rate of the classification model can be improved with the SPXY method. In light of the findings, the proposed sample portioning method can improve the accuracy rate of the recognition model using LIBS.

A comparison of ELISA and real-time PCR kits for meat species identification analysis

A comparison of ELISA and real-time PCR kits for meat species identification analysis

Real-time PCR High-resolution Melting Analysis for the Species Identification of Meat Products: Focusing on Food Safety and Detection of Meat Adulterations

Background: Real-time polymerase chain reaction (PCR) and high-resolution melting (HRM) analysis are currently considered as reliable techniques for the species identification of meat-based products and widely used to detect meat adulteration. Objectives: To examine the validity of real-time PCR and HRM analysis to identify meat species in meat-based products. Methods: Meat samples from five species (i.e., cattle, sheep, chicken, turkey, and wild pig) were purchased. Minced meat from the animal species of interest was prepared at the purities of 10%, and 20% and also were prepared as single and mixtures of two species. For molecular assessments, DNA samples were extracted from all the meat samples and subjected to real-time PCR by amplifying a mitochondrial cytochrome b specific for each species. Results: All the meat species studied in this research were successfully detected in the mixed meat samples when separately examined by real-time PCR. High-resolution melting analysis showed that all the meat species of interest were efficiently distinguished when examined simultaneously. Conclusions: The data presented here shows that the real-time PCR and HRM analysis are reliable methods for the identification of meat species used in meat products.

Strand-displacement DNA polymerase induced isothermal circular amplification fluorescence sensor for identification of pork component

Proper labeling of meat products is critical for consumers who want to avoid specific meat species for cultural or religious reasons. Herein, we developed a convenient strand-displacement DNA polymerase induced isothermal circular amplification (SDPICA) fluorescence sensor for detection of undeclared pork component in meat products. The proposed assay was performed at 37 °C for 20 min with no cross-reactivity, and could detect 1 pg of pork DNA per reaction. The limit of detection of the SDPICA assay was as low as 0.1% pork in 30 mg multi-component meat mixtures. This assay would contribute to the quality control of meat products in food industry.

PCR–RFLP and species-specific PCR efficiency for the identification of adulteries in meat and meat products

The traceability of meat origin has become a necessity to safeguard consumer's confidence in commercial meat products. Our study recommends a meat species detection using qualitative approach based on PCR–RFLP and species-specific primers PCR. Here, we targeted a 359 bp fragment of the mitochondrial cytochrome b gene amplified by PCR using universal primers followed by three enzymatic digestions. Seven animal species including dromedary, rabbit, goat, turkey, rat, donkey and pork, have been efficiently detected in pure and mixed samples. The combination of PCR–RFLP and triplex PCR assays offers, in addition, the identification of chicken, dog and cat species in meat. In conclusion, by the mean of PCR-based techniques using universal primers followed by enzymatic digestion and multiplex primer-specific approach, we developed an extensible protocol by which we identified 10 animal species that could be integrated in meat analysis daily routine.

Studies on the Nutritive Value and Chemical Quality of Different Commercial Grades of Oriental Sausage

Egyptian Oriental sausage is one of the widely popular meat products. It is processed from a combination of different types of meat cuts or variant meat species minced with a suitable amount of fat. Finally, cased into natural sheep gut casing. In order to evaluate the quality of Egyptian oriental sausage, 40 samples of fresh Egyptian Oriental sausage were collected from different sources according to their price and manufacture origin at Ismailia governorate. The proximate chemical analysis and histological examination were carried out. The percentages of moisture, protein, fat, ash, total solids, pH, and red meat content were 58.82, 22.05, 6.83, 1.27,41.74, 5.6, and 78.0 respectively. All the samples from different sources were agreed with the ES: 2005 for proximate chemical composition with exception of 65 % of moisture results were not matched with the ES. While the histological examination showed wide variations among the different grades categorized according to their origin and price. This examination declared that 50% of the products contain inflamed, edematous, low-quality meat cuts, and 10% of the samples had infestation with cysticercoids.

A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species.

Multiplex PCR methods have been frequently used for authentication of meat product adulteration. Through screening of new species-specific primers designed based on the mitochondrial DNA sequences, a septuple PCR method is ultimately developed and optimized to simultaneously detect seven species including turkey (110 bp), goose (194 bp), pig (254 bp), sheep (329 bp), beef (473 bp), chicken (612 bp) and duck (718 bp) in one reaction. The proposed method has been validated to be specific, sensitive, robust and inexpensive. Taken together, the developed septuple PCR assay is reliable and efficient, not only to authenticate animal species in commercial meat products, but also easily feasible in a general laboratory without special infrastructures.

Presence and quantification of pathogenic Arcobacter and Campylobacter species in retail meats available in Japan.

We present estimations for the amounts of Arcobacter (A. butzleri, A. cryaerophilus and A. skirrowii) and Campylobacter (C. jejuni, C. coli and C. fetus) species in retail chicken, pork and beef meat using PCR-MPN. Arcobacter butzleri, A. cryaerophilus and C. jejuni were found in 100, 60 and 55% of chicken samples, respectively. No other Arcobacter or Campylobacter species were found in chicken. The MPNs of A. butzleri, A. cryaerophilus and C. jejuni were greater than 103 per 100g in 50, 0 and 5% of samples, respectively. The MPN of A. butzleri was higher than that of C. jejuni in 95% of samples. In pork, A. butzleri and A. cryaerophilus were detected in 10 and 11 (50 and 55%) of 20 samples, respectively. No other Arcobacter or Campylobacter species were found in pork. Only one pork sample had more than 103 MPN per 100g of A. cryaerophilus. For beef, only two samples tested positive for A. cryaerophilus, at 4600 and 92 MPN per 100g. Overall, we found that the presence and MPNs of Arcobacter species are very high in chicken. In contrast, the positive ratios of Arcobacter in pork were high as chicken samples, but MPNs were lower than in chicken.

Detection and discrimination of seven highly consumed meat species simultaneously in food products using heptaplex PCR-RFLP assay

Food fraud is a global problem raising increased concerns during the past decades. Beef, buffalo, chicken, duck, goat, sheep, and pork are the heavily consumed meats bearing nutritional, economic and cultural/religious importance and are often found to be mutually adulterated in raw and processed states. To authenticate these species, we developed heptaplex polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay using species-specific primer sets which amplified short-length amplicons (73–263 bp) targeting mitochondrial cytochrome b (cytb) and NADH dehydrogenase subunit 5 (ND5) genes. Target specificity was confirmed through cross-amplification reaction with 25 non-target species and PCR products were authenticated by restriction digestion with FatI, BfaI, and HPY188I enzymes followed by separation and visualization of fragments in an automated electrophoretic system. The detection limit was 0.5 % (w/w) meat in mixed matrices and food products. The assay was sufficiently stable under thermal treatments of boiling and autoclaving. A market survey on meat products revealed rampant substitution of beef with buffalo and integrity in chicken and porcine products. Given some advantageous features including short amplicons, exceptional stability and superior sensitivity, the developed assay could be conveniently used for discriminatory detection of target species in raw meat as well as processed meat products.