Bovine Meat Research Articles

Detection of replication-competent circular single stranded DNA from Deoni cattle

Bovine meat and milk factors (BMMF) having structural similarity to plasmids of Acinetobacter baumanni and some single stranded (ss) DNA viruses are free circulating replication-competent circular ssDNA molecules which were first isolated from serum of Bos taurus animals. Isolation of BMMF from tissues of multiple sclerosis as well as some cancers led researchers to conclude that the consumption of these agents through milk and meat have a positive correlation with these diseases and that the absence of such molecules in B. indicus animals could account to the low occurrence of cancer in those areas consuming products of indigenous animals. Hence, the present study was designed to investigate its presence in Deoni, one of the indigenous cattle breeds of India. The DNA isolated from milk samples were subjected to rolling circle amplification (RCA) using exo-resistant random primers. The products of RCA were then used as template for PCR for amplification of circular DNA. The amplicons obtained of size around 2000bp were cloned in E. coli and sequenced. The BLASTn, multiple sequence alignment and phylogenetic analysis of the obtained sequences were carried out to identify the similarities of these isolates. The sequence showed maximum homology with either BMMF2 or plasmids of Acinetobactor spp. Replication initiation protein (Rep) which showed similarity to bacterial plasmids was present in the sequence. This is the pioneer report on the presence of BMMF in milk of indigenous cattle of India. Keywords: BMMF, replication-competent, circular ssDNA, Deoni

Zilpaterol in bovine liver, meat, heart, and kidney, determined by liquid chromatography–quadrupole-orbitrap high-resolution mass spectrometry

ABSTRACT An analytical method was developed for identification and quantification of zilpaterol in bovine liver, meat, heart, and kidney, using liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). It was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at six different concentrations, ranging from 0.1 to 5 μg/kg. The mean recoveries ranged from 71% to 99%, while the decision limit (CCα) and detection capability (CCβ) ranges were 0.11–0.12 μg/kg and 0.13–0.15 μg/kg, respectively. The method demonstrated good linearity (R2 > 0.9996) and the limits of detection (LODs) and of quantification (LOQs) were in the range of 0.015–0.061 μg/kg and 0.025–0.091 μg/kg, respectively. Out of 200 samples collected from local markets in Egypt, 17 contained zilpaterol residues. Liver samples revealed the highest detection frequency (26%), followed by meat (6%), at mean concentrations of 2.64 and 1.93 μg/kg, respectively.

Non destructive qualification of bovine meat tenderness

Tenderness is one of the most important criteria in bovine meat, as it determines whether the meat is used for grilling, long-time cooking or post-processing. It is therefore of great interest for the industry to measure it. Common systems require the extraction and destruction of samples, inducing time and material expenses. In this article, a new nondestructive characterization apparatus, based on monitored indentation, relaxation, and recovery, is proposed. Samples from two cuts known for their difference in tenderness were used from the same carcass. Their tenderness were assessed via compression tests and then compared to indicators from the force–displacement measurements. Results showed that the indentation and recovery speed, and the force relaxation enable the differentiation of the two cuts (p<0.05). These results were obtained on samples from a single carcass. Measurements on other carcasses should be performed to ensure that the results presented in this article can be generalized to bovine meat.

Outbreak detection in Harar town and Kersa district, Ethiopia using phylogenetic analysis and source attribution

BackgroundFoodborne diseases (FBDs) represent a significant risk to public health, with nearly one in ten people falling ill every year globally. The large incidence of foodborne diseases in African low- and middle-income countries (LMIC) shows the immediate need for action, but there is still far to a robust and efficient outbreak detection system. The detection of outbreak heavily relies on clinical diagnosis, which are often delayed or ignored due to resource limitations and inadequate surveillance systems.MethodsIn total, 68 samples of non-typhoidal Salmonella isolates from human, animal and environmental sources collected between November 2021 and January 2023 were analyzed using sequencing methods to infer phylogenetic relationships between the samples. A source attribution model using a machine-learning logit-boost that predicted the likely source of infection for 20 cases of human salmonellosis was also run and compared with the results of the cluster detection.ResultsThree clusters of samples with close relation (SNP difference < 30) were identified as non-typhoidal Salmonella in Harar town and Kersa district, Ethiopia. These three clusters were comprised of isolates from different sources, including at least two human isolates. The isolates within each cluster showed identical serovar and sequence type (ST), with few exceptions in cluster 3. The close proximity of the samples suggested the occurrence of three potential outbreaks of non-typhoidal Salmonella in the region. The results of the source attribution model found that human cases of salmonellosis could primarily be attributed to bovine meat, which the results of the phylogenetic analysis corroborated.ConclusionsThe findings of this study suggested the occurrence of three possible outbreaks of non-typhoidal Salmonella in eastern Ethiopia, emphasizing the importance of targeted intervention of food safety protocols in LMICs. It also highlighted the potential of integrated surveillance for detecting outbreak and identifying the most probable source. Source attribution models in combination with other epidemiological methods is recommended as part of a more robust and integrated surveillance system for foodborne diseases.

Optimisation and validation of liquid-liquid extraction of aminoglycosides from foods of animal origin

ABSTRACT This study focused on evaluating and optimising the liquid-liquid extraction (LLE) method for the multi-residue analysis of seven AGs in bovine liver, meat, milk, and kidney in conjunction with liquid chromatography coupled to a quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS) for quantification. Of all the hydrophilic interaction liquid chromatography (HILIC) stationary phases tested, the ZIC-HILIC column yielded the best performance. The method was validated according to Commission Implementing Regulation (CIR) EU 2021/808, which encompassed various concentration ranges from 25–7500 µg/kg. The mean recoveries ranged from 79% for liver matrix to 87% for meat matrix, with coefficients of variation (CV) below 18%. The method exhibited excellent linearity, as indicated by correlation coefficients (R2) greater than 0.9903 across all tested matrices. The decision limit (CCα) and detection capability (CCβ) varied from 62 to 5984 µg/kg and 71 to 6471 µg/kg, respectively. The limits of detection (LOD) and quantification (LOQ) were established between 1 and 45 µg/kg, and 2 and 75 µg/kg, respectively. No AG residues were detected in the 120 samples that were collected from different local markets in Egypt using the developed analytical method.

Explainable Feature Engineering for Multi-Modal Tissue State Monitoring Based on Impedance Spectroscopy.

One of the most promising approaches to food quality assessments is the use of impedance spectroscopy combined with machine learning. Thereby, feature selection is decisive for a high classification accuracy. Physically based features have particularly significant advantages because they are able to consider prior knowledge and to concentrate the data into pertinent understandable information, building a solid basis for classification. In this study, we aim to identify physically based measurable features for muscle type and freshness classifications of bovine meat based on impedance spectroscopy measurements. We carry out a combined study where features are ranked based on their F1-score, cumulative feature selection, and t-distributed Stochastic Neighbor Embedding (t-SNE). In terms of features, we analyze the characteristic points (CPs) of the impedance spectrum and the model parameters (MPs) obtained by fitting a physical model to the measurements. The results show that either MPs or CPs alone are sufficient for detecting muscle type. Combining capacitance (C) and extracellular resistance (Rex) or the modulus of the characteristic point Z1 and the phase at the characteristic frequency of the beta dispersion (Phi2) leads to accurate separation. In contrast, the detection of freshness is more challenging. It requires more distinct features. We achieved a 90% freshness separation using the MPs describing intracellular resistance (Rin) and capacitance (C). A 95.5% freshness separation was achieved by considering the phase at the end of the beta dispersion (Phi3) and Rin. Including additional features related to muscle type improves the separability of samples; ultimately, a 99.6% separation can be achieved by selecting the appropriate features.

Prevalence of perfluoroalkyl substances in paired batches of precooked and canned bovine meat and their implication on consumer safety

Meat is a highly nutritious food but there is a lot of significant evidence of negative health outcomes related to its excessive consumption, especially for processed one. Among the variety of emerging contaminants of concern for human health, a key role is played by poly- and per-fluoroalkyl substances (PFASs), which show adverse effects in humans who are exposed to them through diet. In the present study, for the first time, 70 paired batches of pre-cooked and canned bovine meat were analysed by Liquid Chromatography coupled to High Resolution Mass Spectrometry to evaluate the presence and concentration of 18 PFASs. These data were used to assess Italian consumers’ health risks by performing the PFAS intake evaluation. PFBA and PFOS were detected in the precooked and canned meat samples, with PFBA mean concentration of 0.22 ± 0.36 ng g−1, and <LOQ, respectively, and PFOS mean concentration of <LOQ in both. No PFASs were found in the jelly. The comparison between the PFBA levels in precooked and canned meat showed a significant difference. The PFAS intake evaluation showed an Estimated Daily Intake by far lower than the Tolerable Daily Intake for the average Italian consumer suggested by the European Food Safety Authority.

Super Hydrophobic Pectin-Anthocyanin Colorimetric Sensor to Indicate Freshness in Bovine Meat

Super Hydrophobic Pectin-Anthocyanin Colorimetric Sensor to Indicate Freshness in Bovine Meat

Outer membrane vesicles of Acinetobacter baumannii DS002 carry circular DNA similar to bovine meat and milk factors (BMMFs) and SPHINX 2.36 and probably play a role in interdomain lateral gene transfer.

Several independent studies have demonstrated the existence of replication competent circular DNA molecules of bacterial and viral origin in mammalian cells and tissues. However, studies about their origin and lateral mobility to mammalian cells are scarce. Our work describes the existence of circular DNA, similar to that of DNA molecules identified in mammalian cells, OMVs derived from soil isolate of A. baumannii DS002. Furthermore, the work also provides visual evidence that demonstrates the passage of labeled OMVs to different organs of experimental mice within hours after intravenously administering OMVs into experimental mice. Some of the labeled OMVs have even crossed the membrane of Neuro2A, suggesting the existence of interkingdom horizontal mobility between bacteria and mammals.

Evaluating the Effect of Temperature and Pooling in Detection and Isolation of the Major Non-O157 Shiga Toxin–Producing Escherichia coli Serogroups from Meat Samples with the Use of Alternative and Standard Methods

Pathogenic Shiga toxin–producing Escherichia coli (STEC) are an important cause of foodborne illness. The detection of STEC in finished products and during the manufacturing process has an important role as part of verification plans, to confirm that practices and procedures described in the food safety program are successfully applied to control STEC. The aim of this study is to evaluate the effect of temperature and pooling in detection and isolation of the major non-O157 STEC serogroups from meat samples with the use of alternative and standard methods. Bovine meat was experimentally inoculated with one of the “Top 6” non-O157 STEC strains (O26, O103, O111, O145, O45, and O121). Both ISO TS 13136:2012 and a novel alternative method were implemented to evaluate the impact of temperature and pooling. An increase of the enrichment temperature to 41.5 °C allowed the detection of the spiked strain in 10% more samples compared to enrichment at 37 °C. The use of 25- and 375-g sample tests demonstrated no statistically differences between both methods. And finally, this alternative method appears easy to use and time-saving for routine laboratory use.

Exploration of Microencapsulation of Arginine in Carnauba Wax (Copernicia prunifera) and Its Dietary Effect on the Quality of Beef.

The objective of this exploratory study was to assess if microencapsulated arginine influences the physicochemical quality of beef. The study included three genetic groups: Angus, Hereford, and Angus × Hereford crossbreed. Two encapsulation systems were used with carnauba wax, at ratios of 3:1 and 2:1, carnauba wax:core (arginine), respectively. A control treatment was also included with no arginine addition. Encapsulated arginine with a 3:1 ratio increased redness by 19.66 at 28 d aged beef compared to the control and 2:1 ratio with values of 18.55 and 16.77, respectively (p = 0.01). Encapsulated arginine at a 3:1 ratio showed the lowest meat shear force values with 24.32 N at 28 d of ageing (p < 0.001). The Angus breed also had a low value of 24.02 N (p < 0.001). Finally, the highest values of intramuscular fat were observed with the inclusion of arginine in a 3:1 ratio. The fat value reached 2.12% with a 3:1 ratio (p = 0.002), while in the Angus breed it was 1.59%. The addition of carnauba wax-encapsulated arginine can improve meat quality. It enhances red color, tenderness, and marbling in bovine meat.

Quick and high-throughput quantification of β-agonist residues in bovine liver, meat, milk, kidney, poultry, and egg using dispersive solid phase extraction

A reliable liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS) method was developed for the simultaneous identification and quantification of 13 β-agonist residues in bovine liver, meat, milk, kidney, poultry, and egg. Dispersive-solid phase extraction (d-SPE) using acetonitrile (ACN) was used to prepare the samples. The analyte in the extracts was separated on a reversed-phase Accucore aQ (50 mm × 2.1 mm, 2.6 μm) using a mobile phase of an aqueous solution containing 2 mM ammonium acetate and acetonitrile (ACN) 0.1 % formic acid. The method was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at six different concentrations ranging from 0.1 to 5 μg/kg. The mean recoveries ranged from 65 to 94 %, while repeatability and reproducibility values were all below 13 %. The linearity, as correlation coefficients (R2) ranged from 0.9955 to 0.9999. The decision limit (CCα) and detection capability (CCβ) ranges were 0.11–0.13 µg/kg and 0.12–0.15 µg/kg, respectively. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.004–0.048 μg/kg and 0.010–0.075 μg/kg, respectively. Of the 180 samples that were collected from local markets in Egypt, 21.11 % had β-agonist residues. The mean concentration (µg/kg) and detection frequency (%) of the most frequently found β-agonist in the samples were as follows: terbutaline (2.63 µg/kg and 90 %), ractopamine (5.14 µg/kg and 23.3 %). The method's applicability was verified by successfully completing two rounds of proficiency testing (PT).

Role of whole linseed and sunflower seed on the nutritional and organoleptic properties of Podolian × Limousine meat

The fatty acids profile of meat can be influenced by the amount and type of the lipid supplemented in animal feeding with a modulation of the rumen biohydrogenation of dietary polyunsaturated fatty acids (P UFA). Therefore, the aim of the present study was to assess the nutritional and sensory quality of bovine meat as affected by different oilseeds supplementations. Thirty two Podolian x Limousine young bulls were randomly allocated into four experimental treatments: control diet (CO) with no supplemental fat, linseed diet (LS) with a supplementation of 10% whole linseed; sunflower diet (SS) with supplementation of 10% of sunflower seeds; LS+SS diet: with supplementation of linseeds (5%) and sunflowers (5%). After finishing period (140 days), all animals were slaughtered and fatty acids profile and sensory proprieties were evaluated on meat. The results showed that dietary supplementation with linseed or/and sunflower seeds in the diet decreased the percentage of saturated fatty acids (P<0.001) and increased monounsatured (P<0.05) and polyunsaturated fatty acids (P<0.05). Feeding diets containing linseeds as respect with sunflower seeds increased the amount of total n-3 P UFA (P<0.01) and, in particular, of linolenic (P<0.01), eicosapentaenoic (P<0.001), docosapentaenoic (P<0.01) and docosaeasoenoic (P<0.05) acids. An improvement in tenderness, juiciness, chewiness and flavour intensity was observed in beef from animals supplemented with oilseeds. This study suggests that linseed alone or in combination with sunflower seeds is an effective strategy to enrich meat with beneficial fatty acids and to improve some sensory attributes appreciated by consumers.

Performance and Export Competitiveness of Meat Exports from India

The study analyses the export performance of meat products between 2000-2021. Revealed symmetric comparative advantage (RSCA) and Revealed Competitiveness (RC) were used for analysis. India consistently had RSCA for meat exports between 2010 and 2021. For frozen bovine meat, India had RSCA during the entire duration. RSCA was witnessed for eight and six years for sheep-goat meat and edible offal, respectively. For chilled/fresh meat of bovine animals, India had RSCA only for one year. In the remaining categories, there was rarely any RSCA. India had strong Revealed Competitiveness in exporting meat products. The study recommended stringent quality measures to be followed for buffalo meat exports. Society should be educated regarding the difference between beef and carabeef. The exportable surplus of goat meat should be increased.

Bovine Meat and Milk Factor-like Sequences Are Frequently Detected in Renal Cell Carcinoma Tissues.

Previous studies have indicated a potential role of diet in the pathogenesis of renal cell carcinoma (RCC). Recently, circular bovine meat and milk factor (BMMF) DNAs have been identified in peritumoral tissues of human colon and breast cancers. Here, we investigated the prevalence of the DNA of these novel human pathogenic infectious agents in RCC and adjacent peritumoral renal tissues. DNA was extracted from formalin-fixed and paraffin-embedded (FFPE) RCC and peritumoral kidney tissues, including a test (n = 11) and a validation (n = 152) collection. BMMF1 and BMMF2 consensus primers were designed to screen for the presence of BMMF1- and BMMF2-like DNA. In addition, BMMF-specific PCR was performed on selected cases to test for the presence of additional regions of BMMF1 and BMMF2 genomes. A reference collection of hepatocellular carcinomas (HCCs; n = 60) and adjacent peritumoral liver tissues (n = 50) was also included. Our results demonstrated that BMMF1 and BMMF2 DNAs are frequently found in human RCC tissues and are particularly more prevalent in peritumoral kidney tissues. Of note, BMMF1 and BMMF2 genotype heterogeneity was higher in peritumoral kidney tissues compared to RCC tissues. This is the first study to directly test human FFPE tissues for BMMF1- and BMMF2-like DNA using consensus PCR and demonstrate BMMF DNA in neoplastic and peritumoral kidney tissues. The findings are in line with the recently proposed indirect etiopathogenetic role of BMMFs in, e.g., colorectal carcinogenesis. Follow-up studies are needed to explore the potential role of BMMFs in the etiopathogenesis of RCC.

Detection of Escherichia coli O157H7 and Campylobacter jejuni in Bovine Carcasses in Two Slaughterhouses in Bio-Bío District, Chile.

Escherichia coli O157:H7 (E. coli O157:H7) and Campylobacter jejuni (C. jejuni) are pathogenic microorganisms that can cause severe clinical symptoms in humans and are associated with bovine meat consumption. Specific monitoring for E. coli O157: H7 or C. jejuni in meat is not mandatory under Chilean regulations. In this study, we analyzed 544 samples for the detection of both microorganisms, obtained from 272 bovine carcasses (280 kg average) at two slaughterhouses in the Bio-Bío District, Chile. Sampling was carried out at post-shower of carcasses and after channel passage through the cold chamber. Eleven samples were found to be positive for E. coli O157:H7 (4.0%) using microbiological and biochemical detection techniques and were subjected to a multiplex PCR to detect fliC and rfbE genes. Six samples (2.2%) were also found to be positive for the pathogenicity genes stx1, stx2, and eaeA. Twenty-two carcasses (8.0%) were found to be positive for C. jejuni using microbiological and biochemical detection techniques, but no sample with amplified mapA gene was found.

Microbiological evaluation of bovine carcasses decontaminated with lactic acid solution

Bovine meat is a nutritious food product that is highly susceptible to bacterial degradation. Organic acid spray washing can be used to reduce microbial populations in carcasses. This study aimed to examine the efficacy of lactic acid in reducing total coliforms and Escherichia coli and assess the effects of different lactic acid concentrations on meat. For this, bovine carcasses were classified into cleanliness categories and subjected to swab sampling before and after manual and automated washing with lactic acid solution at different concentrations (0, 0.5%, 1%, 2%, 3%, and 4%). Colorimetric analysis and lactic acid quantification of carcasses were performed at 0 and 24 h post mortem. The experiment followed a factorial design. Lactic acid inhibited microbial growth, particularly automated application of 3% lactic acid. Although minor changes in color were observed, lactic acid washing was beneficial from a microbial standpoint, effectively decontaminating bovine carcasses and leaving only minimum amounts of residual acid in meat.

Optimization of an HPLC-MS/MS method to analyze chlordecone in bovine serum and correlations with levels in liver, muscle and fat

Chlordecone is an organochlorine pesticide used from 1972 to 1993 in the French West Indies. Its extensive use and high persistence in soils induced massive contamination of the environment and of the food chain, especially in cattle through contaminated soil ingestion. To ensure suitability for consumption of bovine meat, monitoring plans are set up based on perirenal fat concentrations after slaughtering. In the present study, we have investigated an in-vivo monitoring approach by measuring chlordecone levels in serum samples. For this purpose, a sensitive high-performance liquid-chromatography-tandem mass spectrometry (HPLC-MS/MS) method following a QuEChERS extraction method was successfully optimized and validated, reaching a limit of quantification of 0.05 ng g−1 fresh weight. This method was applied to 121 serum samples collected from bovines originating from contaminated areas of Martinique and Guadeloupe. Chlordecone was detected in 88% of the samples, and quantified in 77% of the samples, with concentrations ranging from 0.05 to 22 ng g−1. Perirenal fat, liver, and muscle were also sampled on the same animals and the measured concentrations of chlordecone were statistically correlated to the levels determined in serum. Mean concentration ratios of 6.5 for fat/serum, 27.5 for liver/serum, and 3.3 for muscle/serum were calculated, meaning that chlordecone was not only distribute in fat (as expected), muscle and liver, but also in serum. Good correlations were found to allow prediction of chlordecone concentrations in muscle based on concentrations measured in serum. This study opens the door to possible pre-control of bovines before slaughter. In cases of probable non-compliance with maximum residue levels (MRLs), farm management could proceed to allow for depuration under controlled conditions. This would have a strong impact on both economic and food safety management measures.

Socioeconomic differences in dietary habits in Italy before and during COVID-19 pandemic: secondary analysis of a nationwide cross-sectional study

BackgroundSeveral socioeconomic conditions may influence subjects’ adherence to healthy eating habits. Food consumption may be influenced by external stress during crisis periods; however, the effects of these events on food habits are difficult to predict. Also, a pandemic crisis like the recent COVID-19 pandemic may have influenced dietary habits and food consumption. The objective of this study was to compare the dietary habits of Italian people before the COVID-19 pandemic with those belonging to the year 2020 in a nationwide population sample.Materials and methodsInformation on dietary habits has been obtained from the multi-purpose survey on families’ “Aspects of daily life”, carried out in Italy by the Italian National Statistics Institute (ISTAT). We analyzed data coming from 2016 and 2020 editions of the survey (43,000 subjects each year). We used population attributable fraction (PAF) adjusted for age, defined as the proportional reduction in unhealthy diet that would occur if all participants had had a higher education, assuming higher educated individuals as more socially advantaged. Prevalence association for each dietary exposition has been calculated through logistic regression.ResultsLooking at aggregated data from 2016 and 2020 both men and women showed a high prevalence of unhealthy dietary habits. Regarding men, excessive consumption of eggs, pork meat, and bovine meat was characterized by a PAF attributable to socioeconomic conditions to an extent greater than 30%. Women showed the same trend. Focusing on different years of investigation, in 2020, during the COVID-19 pandemic, men and women increased their consumption of eggs, cooked fats, snacks, and sweets, and reduced consumption of fruits and vegetables. Additionally, women increased the assumption of dietary products and meat. Both sexes registered an increase in overweight and obese subjects in 2020.ConclusionsTo our knowledge, this study was the first in our country to use a yearly, nationwide sample to analyze dietary habits by examining specific types of various foods on a nationwide scale and establishing a correlation between these habits and the COVID-19 pandemic. Our results showed unbalanced dietary habits of the Italian population with an excess of consumption of several foods like eggs, cooked fats snacks, and sweets with low consumption of fruits and vegetables. Socioeconomic differences influence food choices but in a complex way since they seemed to affect some wrong dietary habits but not others, especially regarding fruits and vegetables assumption where differences were less evident among social classes. Outside stressors like a crisis period such as the COVID-19 pandemic seem to have an important role in both men and women regarding the assumption of so-called “junk food”.

Assessment of Bio-Safety, Bio-Security and Microbial Ecology of Bovine Meat at the Masina Slaughterhouse

Assessment of Bio-Safety, Bio-Security and Microbial Ecology of Bovine Meat at the Masina Slaughterhouse