Mixed Meat Research Articles

A multiplex real-time PCR method for differentiation of beef, buffalo meat and pork

The objective of this study was to optimize a multiplex real-time PCR protocol for detection of DNA of beef, buffalo meat and pork, serving for food authenticity. The optimized concentrations were 200 nM primer and 100 nM specific probe for beef/buffalo meat, and 300 nM primer and 150 nM probe for pork. The amplification was performed using initial denaturation at 50oC for 2 min, 95oC for 2 min, followed by 45 cycles of denaturation at 95oC for 15 sec, and annealing and extension at 60oC for 40 sec. This protocol had high sensitivity and specificity. The detection limit of this method was found to be 0.1% in raw and heat-treated meat mix (80 – 121oC/15 min) or 0.005 ng DNA/reaction. The protocol of testing was applied for the commercial products both fresh and processed meats. The results demonstrated that 50% of raw beef sample (6/12) weren't found beef DNA. Eight of twelve beef sausage samples (66.67%) contained buffalo DNA. Beef DNA were found in all 12 samples of beef meatballs, but eight out of the 12 meatball samples were confirmed to have buffalo DNA (66.67%) and two out of the 12 meatball samples (16.67%) also contained porcine DNA.

Dietary patterns and the risk of pregnancy-associated hypertension in the Danish National Birth Cohort: a prospective longitudinal study.

To examine the association between mid-pregnancydietary patterns and pregnancy-associated hypertension (PAH). A prospective longitudinal cohort study. Denmark. About 55139 Danish women with single enrolments and recorded food frequency questionnaire dates with complete information on dietary intake. Women were eligible if they could speak Danish and were planning to carry to term. Diet was assessed using a validated semi-quantitative 360-item food frequency questionnaire and dietary patterns were derived using factor analysis. Gestational hypertension (GH) and pre-eclampsia (PE). Disease prevalence was 14% for GH (5491/39362); 2% for PE (1168/54778), and 0.4% for severe PE (234/55086). Seven dietary patterns were characterised in the population, of which two were associated with PAH. The Seafood diet characterised by high consumption of fish and vegetables was inversely associated with the odds of developing GH [odds ratio (OR) 0.86; 95% CI 0.77-0.95)] and PE (OR 0.79; 95% CI 0.65-0.97). The Western diet characterised by high consumption of potatoes (including French fries), mixed meat, margarine and white bread increased the odds of developing GH (OR 1.18; 95% CI 1.05-1.33) and PE (OR 1.40; 95% CI 1.11-1.76). No association was seen with severe PE. We found protective associations of Seafood diet and harmful associations of Western diet with PAH. Dietary interventions encouraging the reduction of Western diet may contribute to a decrease of PAH. Western diet increases (Seafood diet decreases) the likelihood of developing pre-eclampsia among Danish pregnant women.

A volatilomics approach for off-line discrimination of minced beef and pork meat and their admixture using HS-SPME GC/MS in tandem with multivariate data analysis

Beef, pork and mixed (70% beef and 30% pork) minced meat samples were obtained from a meat processing plant in Athens during a two-year survey and analyzed both microbiologically and by headspace solid-phase microextraction in combination with gas chromatography-mass spectrometry (HS-SPME/GC–MS). A validated method for the discrimination of minced meat was developed based on the volatile fingerprints. Unsupervised (PCA) and supervised (PLS-DA) multivariate statistical methods were applied to visualize, group and classify the samples. The data-set was divided 70% for model calibration and 30% for model prediction. During model calibration 99, 100 and 100% of the samples were correctly classified as beef, pork and mixed meat samples, respectively, while for model prediction the respective percentages were 100, 100 and 95% respectively. In both datasets, the overall correct classification rate amounted to 99% on average. Among the volatile compounds identified, heptanal, octanal, butanol, pentanol, hexanol, octanol, 1-penten-3-ol, 2-octen-1-ol, 3-hydroxy-2-butanone, 2-butanone and 2-heptanone were positively correlated with beef samples. Furthermore, pentanal, hexanal, decanal, nonanal, benzaldehyde, trans-2-hexenal, trans-2-heptenal, trans-2-octenal and 1-octen-3-one were positively correlated with pork. Lastly, the alcohols, 2-butanol and 1-octen-3-ol showed positive correlation with mixed samples. The results indicated that the volatilomics approach employed in this study could be used as an alternative method for robust and reliable discrimination and classification of meat samples in an off-line mode.

Duplex digital PCR for the determination of meat proportions of sausages containing meat from chicken, turkey, horse, cow, pig and sheep

The probability of accidental or intentional addition of another species of meat in meat products is high. Meat is expensive and in the context of food waste, it is reasonable to reduce loss of this precious material during the production. However, the line between fraud and waste reduction is thin. Minor variations, e.g., up to 1% of unexpected meat content, may be tolerated. But to distinguish between minor variations and severe deviations to the list of ingredients, analytical methods with a low measurement uncertainty are required. The recent introduction of digital PCR indicated that this new method may lead to a reduction of the measurement uncertainty. We present the measurement of proportions of beef, pork, chicken, turkey, sheep and horse meat in a cooked sausage matrix and a procedure to calculate the proportions (w/w) based on target DNA concentrations measured using droplet digital PCR. Six laboratories applied these methods and determined the w/w proportions of 20 sausage samples. It was shown that these methods in conjunction with conversion factors can be used to estimate meat proportions in mixed meat products with superior accuracy and precision compared to results generated by real-time PCR.

Detection of Mice DNA in Meatballs Using Real Time – PCR

Meat is one of perishable food and has the high of water content and protein content. Meatballs are processed food who are using meat as main ingredient and other ingredients, such tapioca flour, salt, pepper, and the other ingredients as food additives. Meatballs is the mostpopular because has tasty flavor, high protein content, especiallyessential amino acids.In Indonesia, until now there were still cases of counterfeiting of processed meat products. One of method is mixing the beef meat and other animalsmeat in the making process to reduce production costs, so that similar products can be obtained at a much lower cost. One common case is the mixing of beef meat with rat meat. This study aims to identify meatballs contaminated by rat meat. Meatball samples used do not have Halal certificate from LPPOM MUI. From the results of the study, obtained the green amplification curve with the highest peak showed positive control of rat DNA, followed by three other green curves from the sample that showed the occurrence of amplification, thus indicating samples positive containing rat DNA. In the positive sample, the rat DNA concentration in the sample is quite low which can be seen from the intensity of the peak produced on the amplification curve when compared with positive control. Keywords :DNA, Meat,Meatballs, Rat, RT – PCR

Macroscopic and Microscopic Studies on Sarcocystis Infection in Buffaloes - Public Health Alert

Sarcocystosis is one of the important zoonotic parasitic diseases and commonly reported in buffaloes, cattle and pigs. Meat and meat products are the main sources of infection in human beings. The present study aimed to record the prevalence of Sarcocystosis in buffaloes during the time of slaughter. Out of 61 buffaloes slaughter for meat purpose, 38 showed the presence of macroscopic sarcocysts in different organs and they were in creamy colour lying parallel in between the muscle bundles. Distribution of the sarcocysts was noticed in the muscles of oesophagus, diaphragm, heart and base of the tongue. Microscopic examination of the stained impression smears collected from the mixed meat which is ready for sale from the 61 buffaloes, 52 were found positive for microscopic bradyzoites. In conclusion, Sarcocystis is more prevalent in buffaloes and diagnosis of the disease can be done by microscopic examination of the stained slide impression smears from the suspected meat along with the macroscopic examination of the muscles.

Authentication of pork in meat mixtures using PRM mass spectrometry of myosin peptides.

Adulteration of meat products is a major concern not only for economic fraud, but also for ethical reasons. In this study, we presented a parallel reaction monitoring (PRM) mass spectrometry approach for detection of trace pork in meat mixtures (chicken, sheep, and beef). Specific peptides were identified and screened by a shotgun proteomic approach based on tryptic digests of certain protein. Five surrogate peptides from myosin were screened and then used for pork detection by PRM of Orbitrap MS. When the most sensitive peptide was selected, the LOD in mixed meat can be up to 0.5%. The RSD values between detected and designated pork levels (1%, 5% and 50%) were 4–15%. The targeted method developed can be applied to identify and quantify the pork in meat mixture.

Deteksi Daging Sapi Menggunakan Electronic Nose Berbasis Bidirectional Associative Memory

E-nose is an instrument used to detect odor. E-nose developed with Bidirectional Associative memory (BAM) algorithm has advantages in processing incomplete input data and noise. The purpose of the study was to implement the BAM algorithm to detect pure beef among samples of beef, pork, and mixed meat from aroma with e-nose.Data processing of the sample reading results begins by performing the baseline manipulation process, then do difference and integral feature extraction for the data. The characteristic extraction data will be converted into bipolar matrix patterns (1 and -1) so that the threshold data is needed to be able to determine the feature extraction data to be bipolar. Data that have become bipolar matrices will be used as test and reference data in the program with cross validation testing to obtain the percentage of truth of meat detection using BAM based e-nose.Detection of meat with BAM using integral feature extraction with bipolar the first way yields a 14,8% success percentage and the second way bipolar yields a 15,7% success rate. The extraction of characteristic difference with bipolar the first way yields a success percentage of 17,3% and the second way bipolar yields a success rate of 16,4%.

Detection and quantification of beef and pork materials in meat products by duplex droplet digital PCR.

Meat products often consist of meat from multiple animal species, and inaccurate food product adulteration and mislabeling can negatively affect consumers. Therefore, a cost-effective and reliable method for identification and quantification of animal species in meat products is required. In this study, we developed a duplex droplet digital PCR (dddPCR) detection and quantification system to simultaneously identify and quantify the source of meat in samples containing a mixture of beef (Bos taurus) and pork (Sus scrofa) in a single digital PCR reaction tube. Mixed meat samples of known composition were used to test the accuracy and applicability of this method. The limit of detection (LOD) and the limit of quantification (LOQ) of this detection and quantification system were also identified. We conclude that our dddPCR detection and quantification system is suitable for quality control and routine analyses of meat products.

Effect of transglutaminase on quality and gel properties of pork and fish mince mixtures

Pork and silver carp are two main meat in diet, and mince mixtures can improve the nutrition of meat product. TGase is usually used to enhance the texture and gel properties of meat in industry. The addition of TGase in pork/fish mince mixtures can not only improve the texture, but catalyze the synergetic interaction between proteins from different meat and further enhance gel properties of mice mixtures in a certain ratio. All of that will provide reference for meat processing and development of new meat product.

Cytochrome b conservation between six camel breeds reared in Egypt.

This study was aimed to assess cytochrome b conservation in six breeds of camels reared in Egypt and to compare its sequence with those of other livestock species. The 208-bp fragments from camel mtDNA cyto b were amplified using PCR for 54 camels belonging to 6 camel breeds reared in Egypt. The alignment of camel cyto b sequences showed the presence of two polymorphic sites resulting in four haplotypes and their nucleotide sequences were submitted to GenBank under the accession numbers: KX909894–KX909897.The genetic distances between tested camel breeds were zero between Baladi, Fallahi and Maghrabi breeds whereas they were at low value between the other three breeds: Mowaled, Sodany and Somali. Neighbor-joining showed 4 branches; one of them include most of the tested animals and another one contains 2 Somali animals which is considered a specific haplotype for this breed. The other two branches are mixed between Sodani and Mowaled breeds.Neighbor-joining tree was constructed between cyto b sequences of our tested camels and their sequences from livestock species include Camelus dromedaries, Camelus bactrianus, Ovis aries, Capra hircus, Bubalus bubalis, Bos Taurus and Sus scrofa. The result confirmed that our camel breeds belong to Camelus dromedaries and are clearly separated from other species.It is concluded that cyto b sequence is highly conserved among all camel breeds reared in Egypt which belong to Camelus dromedaries in addition to the advantage of cyto b in differentiation between different livestock sources which enables it to widely use for the adulteration detection in mixed meat.

Chemical composition and occurrence of mycotoxins in traditional meat products from the households of Bosnia and Herzegovina

The aim of this study was to investigate chemical composition and mycotoxins occurrence in traditional meat products (TMP) from the households of Bosnia and Herzegovina. During the period 2015-2016, in total 55 traditional meat products were sampled, of which 25 pork sausages, 15 mixed sausages produced from pork and beef, 7 pancetta and 8 samples of prosciutto. Basic chemical and fatty acids composition was determined by use of ISO methods, whereas mycotoxins aflatoxin B1 (AFB1) and ochratoxin A (OTA) levels were determined using immunoassay ELISA methods. The results showed variations in chemical parameters among the TMPs with significantly higher (p < 0.05) content of stearic acid and lower content of linoleic acid in the mixed meat sausages than in TMPs produced completely of pork. OTA was determined in 7 samples of different products with maximal level of 6.20 µg/kg, whereas AFB1 was determined in only one pork sausage with concentration slightly higher than the method limit of quantification (1.91 µg/kg). Mycotoxin contamination of TMP, especially by OTA, suggesting that to avoid such contamination, meat and meat products on households should be produced and processed under standardized and well-controlled conditions.

The Effect of Using Some Aromatic Medical Herbs in Broiler Diet on Biochemical Characteristics of Blood, Immune Organs and Bacteria Count in Broiler Carcasses

The objective of this study is to determine the effect of diet supplementation with coriander seed and rosemary leaves or a mixture for them on biochemical characteristics of blood, immune organs and bacteria count in meat of broiler carcass (colony forming unit/g). A total of 1080 broiler chicks , Ross-308 at one day old, were randomly allocated for nine treatment groups . Chicks in each treatment group were subdivided for three replicates with 40 chicks for each replicate. Chicks in the first and second treatment are fed on diet supplemented with coriander seed (0.25 % and 0.50 % ) , respectively . While, chicks in the third and fourth treatment are fed diet with rosemary leaves (0.25 % and 0.50 %) , respectively . Also, Mixed of the two medical herbs are added to fifth, sixth, seventh and eighth treatment .Whereas, chicks in the ninth treatment are fed diet without medical herbs supplementation and used as a control group. At marketing age (42d) 12 blood samples are taken to measure the total protein and glucose in serum. Also 9 birds from each treatment group are culled to determine Bursa and spleen weight. Also, the relative weight of Spleen and Bursa Index are calculated. Total bacteria count (colony forming unit(cfu)/ g )and coliform and Salmonella in each one gram of mixed meat from each treatment are recorded. Data show that dietary treatment doesn't significantly affect the serum protein contain, relative weight of bursa and bursa index. However the dietary treatment is significantly ( p < 0.05) increases the serum glucose and spleen relative weight . Total bacteria count, coliform and salmonella count are not affected by dietary treatment. In conclusion, diet supplementation with Rosemary or coriander seed has an effect on biochemical characteristics of blood, immune organs and bacterial count.

Molecular assay using PCR based technology to identify fraud and adulteration of some meat products

Meat species specification is a vital field to ensure the food safety to the consumers which corresponds to the laws relatedto meat and meat products. The adulteration of inferior quality meat into superior quality one is a common practice allover the world. In this study, 30 beef product samples including minced beef, luncheon and sausage (10 samples each)were collected from different markets in Cairo and Giza Governorate, transferred to the laboratory for detection ofadulteration of beef with pig, equine and dog meat using Simple and reliable multiplex-polymerase chain reaction(multiplex-PCR) for the partial-length of cytochrome b (cyt b) gene of mitochondrial DNA (mtDNA) which indicated thesuccessful detection of little as 0.05 ng % adulteration of meat products. Multiplex PCR can also be applied to detectauthentication with equal efficiency to fresh, cooked or putrefied mixed meat samples. The obtained results revealed thatone examined sample (10%) of minced beef was intermixed with equine meat only, while two samples (20%) of beefsausage were adulterated with addition of equine meat only. Moreover, the examined luncheon samples were free fromadulteration with either pork or equine meat. All examined samples were found to be free from adulteration with dogmeat. The bad effects of fraud and adulteration of meat products was discussed

Free amino acid profile of Bubalus bubalis L. meat from the Campania region

In this study, we determined the amount of carnosine and anserine in water buffalo meat without hanging treatment and the free amino acid profile by using amino acid analyser with post-column ninhydrin derivatization procedure. The main free amino acids present in samples were glutamic acid (>60 mg/100 g), followed by alanine, glycine, and arginine. Other protein amino acids were detected in minor amounts (less than 2 mg/100 g). Among the non-protein amine-containing compounds, taurine and urea were the most abundant. The analysis showed that 50% of the total free amino acids was represented by dipeptides carnosine (average ~130.3 mg/100 g) and anserine (average ~17.9 mg/100 g). Thus, this study for the first time reports the free amino acids profile of water buffalo meat and the content of carnosine and anserine, potentially involved in the darkening meat process and their ratio, that could be used to estimate the water buffalo meat portion in mixed meat products.

Accounting Information Systems of Kuwaiti Companies: Obstacles and Barriers

Moringa oleifera is mainly used as herbal medicine in most homes in Northern Nigeria. The plant is easy to grow and thrives very well regardless the type of soil. Use of moringa leaves in food production can yield attractive varieties on menu. This paper evaluates the acceptability of dishes produced with fresh moringa leaves with a view to promoting it in popular restaurants. A three course menu consisting of cream of moringa soup as the starter, mixed meat moringa sauce with semovita as the main dish and moringa roll as sweet was produced and served to a 60-member taste panel made of three groups of 20 each. Respondents were asked to rate the organoleptic qualities of the samples on a 10-point bipolar scale ranging from 1 (Dislike extremely) – 10 (Like extremely). Data collected were treated to one sample t-test and One Way ANOVA. Results show that the panelists extremely like the moringa products. It is recommended that Moringa oleifera should be incorporated into meals which is more readily acceptable than medicine.

Species-specific identification of adulteration in cooked mutton Rista (a Kashmiri Wazwan cuisine product) with beef and buffalo meat through multiplex polymerase chain reaction.

Aim:Meat adulteration is a serious problem in the meat industry and needs to be tackled to ensure the authenticity of meat products and protect the consumers from being the victims. In view of such likely problem in indigenous meat products of Kashmiri cuisine (Wazwan), the present work was performed to study the detection of beef and buffalo meat in cooked mutton Rista by mitochondrial DNA (mtDNA) based multiplex polymerase chain reaction (PCR) method under laboratory conditions.Materials and Methods:Three experimental trials were conducted wherein the products were prepared from pure mutton, beef and buffalo meat, and their admixtures in the ratios of 60:20:20, 80:10:10, 90:05:05 and 98:01:01, respectively.Results:The primers used in the study amplified the cyt b gene fragments of sizes 124 bp, 472 bp and 585 bp for buffalo, cattle and sheep, respectively. It was possible to detect cattle and buffalo meat at the level of 1% in the mixed meat cooked Rista. The multiplex PCR successfully amplified cyt b gene fragments of mtDNA of the target species and thus produced characteristic band pattern for each species. The band intensities of cattle and buffalo in the mixed meat Rista progressively decreased corresponding to their decreasing level from 20% to 1%. Processing, cooking (moist heating) and non-meat formulation ingredients had no effect on detection of meat species adulteration.Conclusion:The multiplex PCR procedure standardized and developed in this study is simple, efficient, sensitive, reliable and highly specific for detecting falsification of cooked mutton product with beef and buffalo meat up to 1% level.

Lab-on-a-chip-based PCR-RFLP assay for the confirmed detection of short-length feline DNA in food

Wider availability but lack of legal market trades has given feline meat a high potential for use as an adulterant in common meat and meat products. However, mixing of feline meat or its derivatives in food is a sensitive issue, since it is a taboo in most countries and prohibited in certain religions such as Islam and Judaism. Cat meat also has potential for contamination with of severe acute respiratory syndrome, anthrax and hepatitis, and its consumption might lead to an allergic reaction. We developed a very short-amplicon-length (69 bp) PCR assay, authenticated the amplified PCR products by AluI-restriction digestion followed by its separation and detection on a lab-on-a-chip-based automated electrophoretic system, and proved its superiority over the existing long-amplicon-based assays. Although it has been assumed that longer DNA targets are susceptible to breakdown under compromised states, scientific evidence for this hypothesis has been rarely documented. Strong evidence showed that shorter targets are more stable than the longer ones. We confirmed feline-specificity by cross-challenging the primers against 10 different species of terrestrial, aquatic and plant origins in the presence of a 141-bp site of an 18S rRNA gene as a universal eukaryotic control. RFLP analysis separated 43- and 26-bp fragments of AluI-digest in both the gel-image and electropherograms, confirming the original products. The tested detection limit was 0.01% (w/w) feline meat in binary and ternary admixed as well as meatball matrices. Shorter target, better stability and higher sensitivity mean such an assay would be valid for feline identification even in degraded specimens.

Targeted Beverage Taxes Influence Food and Beverage Purchases among Households with Preschool Children1–3

Background: How beverage taxes might influence purchases of foods and beverages among households with preschool children is unclear. Thus, we examined the relation between beverage taxes and food and beverage purchases among US households with a child 2–5 y of age.Objectives: We examined how a potential tax on sugar-sweetened beverages (SSBs), or SSBs and >1% fat and/or high-sugar milk, would influence household food and beverage purchases among US households with a preschool child. We aimed to identify the lowest tax rate associated with meaningful changes in purchases.Methods: We used household food and beverage purchase data from households with a single child who participated in the 2009–2012 Nielsen Homescan Panel. A 2-part, multilevel panel model was used to examine the relation between beverage prices and food and beverage purchases. Logistic regression was used in the first part of the model to estimate the probability of a food/beverage being purchased, whereas the second part of the model used log-linear regression to estimate predicted changes in purchases among reporting households. Estimates from both parts were combined, and bootstrapping was performed to obtain corrected SEs. In separate models, prices of SSBs, or SSBs and >1% and/or high-sugar milk, were perturbed by +10%, +15%, and +20%. Predicted changes in food and beverage purchases were compared across models.Results: Price increases of 10%, 15%, and 20% on SSBs were associated with fewer purchases of juice drinks, whereas price increases of 10%, 15%, and 20% simulated on both SSBs plus >1% fat and/or high-sugar milk (combined tax) were associated with fewer kilocalories purchased from >1% fat, low-sugar milk, and meat, poultry, fish, and mixed meat dishes.Conclusions: Our study provides further evidence that a tax on beverages high in sugar and/or fat may be associated with favorable changes in beverage purchases among US households with a preschool child.

Development of a PCR/RLB Test for Meat Source Authentication in Processed Meat and Meat Products

Concern over food authenticity has increased as a result of an increase in the consumption of processed foods containing meat or animal products. This raises a number of issues where the presence of pork in such foods is considered unacceptable in most Muslim and Jewish communities around the world. It also applied to the prohibition of beef consumption among Hindus. In order to ensure the absence of unwished meat products or mixing of meats from different sources in processed foods, a specific and sensitive test is essential. For this purpose we developed a molecular test based on DNA amplification by polymerase chain reaction (PCR) of the cytochrome b gene followed by reverse line blot analysis (RLB). Using this method many samples may be treated simultaneously and meat origins can easily be detected from processed foods or foods containing mixed meat sources; also, added pork components such as fat may be identified by this methodology. The PCR/RLB method is considered to be a sensitive and specific technique; it can detect one nucleotide change within the PCR-amplified DNA segment.