Assisted reproductive technologies (ARTs) in Mithun (Bos frontalis): What progress has been made so far? An overview.

Mithun, a unique bovine species, endemic to parts of North East India and plays an important role in the socioeconomic, cultural and religious fabrics of the local tribal population. To date, Mithuns are reared in a traditional free-range system by communities and increased deforestation, agricultural commercialization, disease outbreaks and indiscriminate slaughtering of elite Mithun for table purposes have significantly decreased its habitat and the elite Mithun population. Greater genetic gain is achieved with the implementation and effective use of assisted reproductive technologies (ARTs); however, presently it is limited to organized Mithun farms. At a slow pace, Mithun farmers are adopting semi-intensive rearing systems and interest in the use of ARTs is gradually escalating in Mithun husbandry. This article reviews the current status of ARTs such as semen collection and cryopreservation, estrus synchronization and timed artificial insemination (TAI), multiple ovulation and embryo transfer and in vitro embryo production and future perspectives in Mithun. Mithun semen collection and cryopreservation have been standardized, and estrus synchronization and TAI are suitable technologies that can be easily implemented under field conditions in near future. The establishment of an open nucleus-breeding system under community participatory mode along with the introduction of the ARTs is an alternative to the traditional breeding system for rapid genetic improvement of Mithun. Finally, the review considers the potential benefits of ARTs in Mithun and future research should include the use of these ARTs which will provide additional opportunities for improved breeding regimens in Mithun.

Genetic diversity and risk factor analysis of drug-resistant Escherichia coli recovered from broiler chicken farms.

A total of 38 Escherichia coli isolates were recovered from 120 samples collected from various sources of broiler chicken farms (n = 10 each) in Andhra Pradesh and Telangana states. Though the recovered E. coli isolates were found variably resistant to the tested antibiotics, all the tested isolates were susceptible to meropenem. Alarming multi-drug resistance (MDR) was observed (34/38) among the recovered isolates, wherein antibiotic-resistant genes (blaTEM, blaSHV, and tetA) were detected, except for blaCTX-M-9. The heatmap with cluster analysis exhibited that majority of the E. coli isolates recovered from different sources and regions clustered together based on their phenotypic resistance suggesting co-sharing of resistance. However, the pulsed-field gel electrophoresis (PFGE) typing revealed an extremely diverse genotypic profile. Further, a significant statistical association was not observed between hypothesized risk factors and recovered MDR- E. coli isolates from various sources, although a significant statistical association between antibiotic resistance with large flock size, poor biosecurity practices, poor workers' hygiene, and poor disinfection practices was noticed. Since the study highlighted an alarming level of drug resistance among the recovered E. coli isolates, further in-depth research in similar veins is required to ensure the prudent use of antimicrobials in the poultry sector and the implementation of an antimicrobial surveillance system.

Identification of sheep (Ovis aries) meat by alkaline lysis-loop mediated isothermal amplification technique targeting mitochondrial D-loop region.

Identification of meat species origin using reliable techniques is a critical requirement for ensuring label compliance, protection of consumer preference and prevention of fraudulence in the meat trade. Although a plethora of protein and DNA based meat species identification techniques are in vogue, need for rapid test suitable for under-resourced laboratories catering point-of-care (PoC) services was construed. Present study deals with development of rapid sheep (Ovis aries) meat identification technique using DNA extraction by alkaline lysis (AL) and loop-mediated isothermal amplification (LAMP) technique. The AL-LAMP specifically amplifies sheep-specific signal of mitochondrial D loop region under an isothermal temperature of 60 °C with an analytical sensitivity of 0.5 ng sheep DNA. The test was highly specific to sheep and performed well even in the presence of DNA of closely related meat animal species such as goat, cattle, buffalo and chicken. The novel primers designed for the AL-LAMP successfully detected sheep meat in raw and cooked meat samples heated up to 121 °C for 30 min. Sheep-specific AL-LAMP assay could detect 0.1% mutton-in-beef adulteration. Novel AL-LAMP assay being simple, rapid and reliable for sheep meat authentication in just 120 min; hence, it could be conveniently used by terminal laboratories engaged in rendering on-site or PoC services.

Investigation on Temperature Dependent Inductance (TDI) of a planar Multi-Layer Inductor (MLI) down to 4.2 K.

This paper presents an investigation of the temperature dependence characteristics specific to cryogenic planar Multi-Layer Inductors (MLIs). This paper establishes that the inductance of a planar MLI at a specific frequency varies with temperature when the sensor is cooled down to 4.2 K while providing a detailed analysis of various possible factors that might contribute to the variation in the sensor performance, such as the thermal deformation and the variation in the properties of sensor materials, using a combination of experiments and simulations. By calculating the interlayer capacitance, we have attempted to adopt a novel approach in the investigation of the effects of thermal deformation on the sensor. In order to arrive at that, the relative permittivity of the base material (G10CR-FR4) at cryogenic temperatures was obtained through experiments. The ANSYS static structural package was used for modeling thermally induced deformations, after which the deformed capacitance and inductance were obtained using Ansoft MAXWELL. From the analysis, we have concluded that the variation in the inductance of the sensor has a direct correlation with the electrical resistivity (hence the residual resistivity ratio) of the coil material. The number of inductor layers and the area of the component layer will also determine the temperature dependence phenomenon. These conclusions are not obvious from the established inductance models.

Breed traceability of buffalo meat using microsatellite genotyping technique.

Although buffalo has emerged as a major meat producing animal in Asia, major research on breed traceability has so far been focused on cattle (beef). This research gap on buffalo breed traceability has impelled development and validation of buffalo breed traceability using a set of eight microsatellite (STR) markers in seven Indian buffalo breeds (Bhadawari, Jaffaarabadi, Murrah, Mehsana, Nagpuri, Pandharpuri and Surti). Probability of sharing same profile by two individuals at a specific locus was computed considering different STR numbers, allele pooling in breed and population. Match probabilities per breed were considered and six most polymorphic loci were genotyped. Out of eight microsatellite markers studied, markers CSSMO47, DRB3 and CSSM060 were found most polymorphic. Developed technique was validated with known and unknown, blood and meat samples; wherein, samples were genetically traced in 24 out of 25 samples tested. Results of this study showed potential applications of the methodology and encourage other researchers to address the problem of buffalo traceability so as to create a world-wide archive of breed specific genotypes. This work is the first report of breed traceability of buffalo meat utilizing microsatellite genotyping technique.

Carcass characteristics, composition, physico-chemical, microbial and sensory quality of emu meat.

1. In order to investigate whether emu meat is a potential red meat alternative, this work was carried out with the objective of studying the carcass characteristics, proximate composition, physico-chemical and microbial characteristics and sensory attributes of emu meat. 2. Carcass characteristics clearly indicate that emus are a significant source of lean meat, fat, skin and edible by-products and these findings confirm earlier reports. 3. Proximate composition of emu meat indicated higher protein and ash content and lower fat, total lipids and cholesterol content than meat from other meat animals. 4. The pH, water holding capacity, collagen content and solubility, protein extractability, muscle fibre diameter and Warner-Bratzler shear force values of emu meat are similar to the earlier reports for meats from other food animals. 5. Emu meat is dark, cherry red in colour with significantly higher myoglobin content and the myoglobin is more prone to oxidation as evidenced by higher initial metmyoglobin percentage. The initial thiobarbituric acid reactive substances (TBARS) values and free fatty acids percentage in emu meat were higher than those in meats from other species. 6. Sensory evaluation of cooked emu meat curry revealed highly acceptable scores relative to goat meat curry, the most preferred meat in India. 7. The study shows the potential of emu meat as a new source of low fat, quality meat proteins. However, more studies are required to elucidate the effect of age, sex, muscles, pre-slaughter and post-slaughter factors on different carcass and meat quality characteristics.

A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction.

Buffalo (Bubalus bubalis) meat is a major item of export from India but export of beef i.e. meat from cattle (Bos indicus) is prohibited. Also, adulteration of buffalo meat with that of beef (meat from cattle) is a common fraudulent practice because of prohibition on cow slaughter in most states of India. Food analysts require precise identification techniques to implement such regulations. In the present study, a method of DNA extraction by Alkaline lysis from meat samples and speciation of buffalo meat using species specific Polymerase Chain Reaction (PCR) has been reported. Alkaline lysis technique is a rapid method which involves triturating meat with four volumes of 0.2N NaOH, dilution of resultant liquid extract with eight volumes of 0.2N NaOH, heating the mix 75 °C for 20 min followed by neutralization with eight volumes of 0.04N Tris HCl. Entire procedure of DNA extraction takes less than 30 min and it is economical as it involves less expensive chemicals. Method was successfully applied in animal byproducts also viz., liver, heart and kidney. For authentication of buffalo meat, pair of primers was designed based on mitochondrial D loop gene nucleotide sequence. PCR amplification using the designed primers gave amplicon of size 482 bp in buffalo and no amplification was detected in closely related species viz., cattle, sheep and goat meat samples. Results of the assay were highly repetitive and reliable. An export sample referred by export regulation authorities was also analyzed by using the Alkaline lysis method of DNA extraction and species specific PCR which enabled authentication of meat within 5 h.

Effect of dipping in pomegranate (Punica granatum) fruit juice phenolic solution on the shelf life of chicken meat under refrigerated storage (4°C).

An experiment was conducted to evaluate the effect of dipping in pomegranate fruit juice phenolics (PFJP) solution on the shelf life of chicken meat held under refrigerated storage at 4°C. Breast muscle obtained from spent hens was dipped (1:2w/v; muscle: liquid) in sterile water or in sterile water with 0.02% (v/v) PFJP, packed, stored at 4°C for 28 days and samples were analyzed on 2 days of intervals. Thiobarbituric acid reactive substance values were lower in samples treated with PFJP. Total sulfhydryl and protein bound sulfhydryl content values were higher in samples treated with PFJP. Microbial quality evaluation showed that aerobic and psychrotrophic counts were higher in samples treated without PFJP. Sensory evaluation revealed that acceptability level of samples treated without PFJP decreased on 12th day of storage. It is concluded that spent hen breast meat samples dipped in 0.02% PFJP reduced protein oxidation and inhibited microbial growth and sensorily acceptable up to 12 days of refrigerated storage at 4°C.

Identification of chicken, duck, pigeon and pig meat by species-specific markers of mitochondrial origin.

In the present study, PCR based method for meat species identification of chicken, duck, pigeon and pig was achieved by developing species-specific markers. Using mitochondrial sequences species-specific primers were designed and the sizes of them were 256bp, 292bp, 401bp and 835bp for chicken, duck, pigeon and pig, respectively. The species-specific PCR products were sequenced to confirm the specificity of the product amplified. These markers were subsequently tested for cross amplification by checking them with beef, mutton, chevon, pork, rabbit, chicken, duck, turkey and pigeon meat. DNA markers developed in this study can help identify the species of fresh, cooked and autoclaved meat of chicken, duck and pigeon and fresh and cooked meat of pig. The process of identification is simple, economical and quick as compared to other methods such as RAPD, PCR-RFLP and sequencing method of species identification.

Biochemical and physicochemical changes in spent hen breast meat during postmortem aging.

An experiment was conducted to study the biochemical and physicochemical changes with respect to improvement in tenderness of spent hen breast meat. Breast muscle obtained from freshly slaughtered spent hens (72 wk old) was divided into 5 equal lots and dipped in 1 mM NaN(3) before being packed in low-density poly-ethylene pouches under aerobic conditions and stored at refrigeration temperature (4 degrees C). Lots were removed on 0, 7, 14, 21, and 28 d of storage and analyzed for pH, TBA reactive substances (TBARS), total sulfhydryl content, protein-bound sulfhydryl content, nonprotein-bound sulfhydryl content, perimysial fraction, collagen content, free OH-proline, N, nonprotein N, and proteolysis rate. Shear force value and penetrometer readings were also determined after making patties from the stored muscle samples. Results showed that pH values were gradually decreasing over the storage period. The TBARS values were increasing (P < 0.001), whereas the sulfhydryl content was decreasing (P < 0.001) over the storage period. The TBARS values were negatively (P < 0.05) correlated with total sulfhydryl content. This suggests that sulfhydryl content may prevent further higher oxidation of lipids. The soluble collagen content, collagen solubility, free OH-proline, and proteolysis rate were increasing (P < 0.001) during postmortem aging. These results suggest that collagen degradation into free amino acids occurs postmortem. A gradual decrease (P < 0.001) in shear force value and a gradual increase (P < 0.001) in penetrometer readings were recorded in the patties made from matured breast meat. Therefore, postmortem aging of spent hen breast meat resulted in 23% improvement in tenderness of minced patties on 14 d and 39% on 28 d as evidenced by biochemical and physicochemical changes.

Polymerase chain reaction-restriction fragment length polymorphism of mitochondrial 12S rRNA gene: a simple method for identification of poultry meat species.

Chicken (Gallus gallus), duck (Anas platyrhynchos), turkey (Meleagris gallopavo), guinea fowl (Numida meleagris) and quail (Coturnix japonica) are the common poultry species consumed as meat throughout the world. In this work, a molecular technique has been developed for identification and differentiation of meat originating from these species. This tool helps in detection of misrepresentation of different poultry meats. The technique involves the extraction of DNA from the given sample, polymerase chain reaction (PCR) amplification of mitochondrial 12S rRNA gene using universal primers, restriction analysis with selected restriction enzymes, followed by identification of meat species based on restriction fragment length polymorphism (RFLP) pattern. In this study, we used HinfI, Mph1 103I, MvaI, and Eco47I to identify and differentiate to poultry species referred to above. This species identification technique has also been applied successfully to processed meat products including those cooked at 120 degrees C for 30 min. Simplicity of interpretation of results combined with versatility makes this a convenient and appropriate technique in the hands of meat analysts for identifying poultry meat species.

Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene.

Adulteration of high quality meat and meat products with their inferior/cheaper counterparts is a problem in the meat industry. The present study investigated the use of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the mitochondrial 12S rRNA gene for identification of the origin of meats. PCR-RFLP was applied for species identification of beef, buffalo meat, mutton and chevon. PCR amplification yielded a 456-bp fragment in each of these species. The amplicons were digested with AluI, HhaI, ApoI and BspTI restriction enzymes resulting in a pattern that could identify and differentiate each of the above species. This technique did not yield satisfactory results with meat mixtures/meats. However, consistent results were obtained with both fresh and processed meat samples.

Sequence analysis of mitochondrial 12S rRNA gene can identify meat species.

In this study, sequence analysis of mitochondrial 12S rRNA has been applied for meat species identification. The procedure involves polymerase chain reaction (PCR) amplification of a fragment of mitochondrial (mt) 12S rRNA gene and sequencing of amplicons. Amplified product of mt 12S rRNA gene was 456 bp in size. Species sequenced include cattle (Bos indicus), buffalo (Bubalus bubalis), sheep (Ovis aries), goat (Capra hircus) and mithun (Bos frontalis). Sequences were compared with the reported sequences of low land anoa (Bubalus depressicornis), yak (Bos grunniens) and pig (Sus scrofa). There was no effect of routinely used additives or cooking temperature (72, 90, 120 and 180 °C) on the efficacy of PCR amplification. The closely related species like cattle and buffalo, sheep and goat could also be differentiated decisively by sequence analysis. Sequencing and analysis of mt 12S rRNA gene was, hence, found to be an ideal, authentic and unambiguous qualitative method for meat species identification.