Development of superoxide dismutase based visual and spectrophotometric method for rapid differentiation of fresh and frozen-thawed buffalo meat.

Study aimed to develop biomarker-based assay for rapid detection of fresh and frozen-thawed buffalo meat in the supply chain. The method is based on development of a solvent system and identification of suitable substrate and developer for screening of biomarkers. For the confirmation column chromatography, gel electrophoresis and Western Blotting were carried out. Validation was done by intra- and inter-day validation, storability study, and determination of thermal history. Best results were shown with pH 8.0 Tris-HCl; extraction buffer, 205 µM nicotinamide adenine dinucleotide hydrogen; substrate, 184 µM Nitroblue tetrazolium, and 1.9 µM phenazine methosulfate; developer. The thermal history ranged from 0.14 to 0.17 during storage at -20 °C. The intra- and inter-day assay precision (CV %) ranged from 5.3 to 6.5 %; in chilled and 14.1 - 9.2 % in frozen-thawed samples. The study confirmed SOD as a viable biomarker. Developed method using SOD has significant potential for rapidly differentiating chilled or frozen-thawed meat.

Effects of Slaughter Positions on Catecholamine, Blood Biochemical and Electroencephalogram Changes in Cattle Restrained Using a Modified Mark IV Box.

The proper slaughter positioning of animals is among the most crucial factors in animal welfare. The lateral position in Halal slaughter is a technique used around the world by Muslims, with a few practicing the upright position. The literature on the effects of slaughter in upright versus lateral positions on pain and stress is scarce. Thus, this study was designed to evaluate the effects of slaughter positions on blood biochemical parameters, plasma catecholamines, and electroencephalographic (EEG) responses. Twenty Brahman crossbred steers were subjected to slaughter in either lateral recumbency (LP) (n = 10) or an upright position (UP) (n = 10). There was a significant increase in adrenaline (p < 0.0001) and noradrenaline (p < 0.05) at T2 compared to T1 in the animals of both groups. A significant difference (p < 0.0001) was observed in the median frequency (MF) and total power (Ptot) of EEG, parameters for pain and stress, between the animals slaughtered in the upright and the lateral position. However, MF and delta waves were significantly higher (p < 0.05) after slaughter in the UP group than in the LP group. The results demonstrate a lesser amount of stress and pain responses among the LP group.

Recovery of Gelatin from Bovine Skin with the Aid of Pepsin and Its Effects on the Characteristics of the Extracted Gelatin.

Pepsin enzyme was used to pretreat the bovine skin at the rate of 5, 15, and 25 units of enzyme/g of skin to recover gelatin, and the recovered gelatins were referred to as Pe5, Pe15, and Pe25, respectively. The gelatin yield increased significantly (p < 0.05) from 18.17% for Pe5 to 24.67% for Pe25 as the level of pepsin increased, but the corresponding gel strength and viscosity decreased significantly (p < 0.05) from 215.49 to 56.06 g and 9.17 to 8.17 mPa·s for Pe5 and Pe25, respectively. ß- and α1- and α2-chains were degraded entirely in all the gelatins samples as observed in protein pattern elaborated by gel electrophoresis. 1H nuclear magnetic resonance (1H NMR) analysis indicated the coiled structure of gelatin protein chains. The lowest amide III amplitude of Pe25 as found by Fourier transform infrared (FTIR) spectroscopy indicated that α-helix structure of protein chains were lost to more irregular coiled structure. Thus, it could be summarized that pepsin might be used at the lower level (5 units/g of wet skin) to extract gelatin from bovine skin with good functional properties and at higher level (15/25 units/g of wet skin) to obtain gelatin of industrial grade with high yield.

Extraction, characterization and molecular structure of bovine skin gelatin extracted with plant enzymes bromelain and zingibain.

Bovine skin was incubated with plant enzymes bromelain (B) and zingibain (Z) at the level of 0, 5, 10, 15, 20 and 25 unit/g of skin and gelatin was extracted at 60 °C for 6 h. Control gelatin was extracted without enzymatic pretreatment. The yield and gel strength were 17.90% and 283.35 g for the control samples and 22.26% and 160.88 g for B20 samples. The zingibain extracted gelatin (GEZ) samples failed to form gel. Viscosities of GEZ gelatins were significantly (P < 0.05) lower than the gelatins extracted using bromelain (GEB). ß and α chains were absolutely degraded in all GEB and GEZ samples. Only smear bands were observed in GEZ gelatins whereas GEB samples revealed presence of low molecular weight polypeptides. Loss of molecular order was noticed in Z5 as elaborated by Fourier transform infrared (FTIR) spectroscopy. Larger particle size, denser and inter-connected irregular network was observed in B20 under scanning electron microscopy. Based on the results obtained, bromelain, particularly at level 20, could be used to obtain a better quality gelatin with higher yield compared to zingibain.

Effects of Slaughter Knife Sharpness on Blood Biochemical and Electroencephalogram Changes in Cattle.

The sharpness of the knife used for slaughter is of the utmost importance from an animal welfare perspective. The quantification of knife sharpness is almost impossible in abattoirs. The sharpness of the knife blade used to slaughter an animal, as well as its effects on animals' pain and stress levels, is an important area of investigation that needs to be addressed. The objective of this study was to evaluate the effects of knife sharpness on blood biochemical parameters, plasma catecholamines, and electroencephalographic (EEG) responses. Twenty Brahman crossbred steers were either subjected to slaughter with a sharp knife (n = 10) or a commercial sharp knife (n = 10); knife sharpness was measured with the ANAGO® sharpness tester. There was significant increase in adrenaline (p < 0.0001), glucose (p = 0.0167), creatinine kinase (p = 0.0123) and lactate dehydrogenase (p = 0.0151) at post-slaughter compared to pre-slaughter in commercial sharp knife group than in thesharp knife group. A significant increase was observed in the median frequency (p < 0.0001) and total power (p < 0.0001) of the EEG, the parameters for pain and stress, in the animals slaughtered with the commercial sharp knife than those slaughtered with the sharp knife. Thus, EEG results also supported the hormonal and biochemical results. From the results, it is concluded that animals slaughtered with a sharp knife experienced the least amount of pain and stress compared to those slaughtered with a commercial sharp knife.

Autolysis of bovine skin, its endogenous proteases, protease inhibitors and their effects on quality characteristics of extracted gelatin.

The autolysis of pretreated bovine skin (PBS) (treated with 0.1 M NaOH and 1% HCl), its endogenous proteases, inhibitors and their effects on quality attributes of gelatin were examined. PBS was subjected to different temperatures (20-90 °C) and pH (2-9) and treated with different protease inhibitors. Maximum autolytic activity of PBS was observed at 40 °C and pH 5. Ethylene-bis (oxyethylenenitrilo) tetraacetic acid (EGTA) was the most effective in impeding the degradation of γ-, ß- and α- chains of PBS protein indicating that metallocollagenases were the predominant endogenous proteases in bovine skin. Gelatin was extracted in the absence (GAE) and presence (GPE) of EGTA, and EGTA with papain enzyme (GPEP). GPEP had a higher yield and lower gel strength than GEA and GPE. Metallocollagenases partook in the degradation of gelatin thereby affecting its functional properties. Pretreating PBS with or without EGTA, and papain influenced the quality attributes of gelatin.

Effects of Ultrasound Assisted Extraction in Conjugation with Aid of Actinidin on the Molecular and Physicochemical Properties of Bovine Hide Gelatin.

Actinidin was used to pretreat the bovine hide and ultrasonic wave (53 kHz and 500 W) was used for the time durations of 2, 4 and 6 h at 60 °C to extract gelatin samples (UA2, UA4 and UA6, respectively). Control (UAC) gelatin was extracted using ultrasound for 6 h at 60 °C without enzyme pretreatment. There was significant (p < 0.05) increase in gelatin yield as the time duration of ultrasound treatment increased with UA6 giving the highest yield of 19.65%. Gel strength and viscosity of UAC and UA6 extracted gelatin samples were 627.53 and 502.16 g and 16.33 and 15.60 mPa.s, respectively. Longer duration of ultrasound treatment increased amino acids content of the extracted gelatin and UAC exhibited the highest content of amino acids. Progressive degradation of polypeptide chains was observed in the protein pattern of the extracted gelatin as the time duration of ultrasound extraction increased. Fourier transform infrared (FTIR) spectroscopy depicted loss of molecular order and degradation in UA6. Scanning electron microscopy (SEM) revealed protein aggregation and network formation in the gelatin samples with increasing time of ultrasound treatment. The study indicated that ultrasound assisted gelatin extraction using actinidin exhibited high yield with good quality gelatin.

Antioxidant potential of aqueous extract of some food grain powder in meat model system.

In-vitro antioxidant activity of some food grains [sprouted mung bean (Vigna radiata), mung bean, sprouted chana (Cicer arietinum), chana, corn (Zea mays), methi (Trigonella foenum-graecum) and rajma (Phaseolus vulgaris)] powder extracts (FGE) was estimated by DPPH free radical scavenging activity (SA) method. Total phenolics and reducing power were also estimated in these extracts. The antioxidant potential of these extracts was also estimated in a meat model system. Total phenolics in FGE ranged from 66.9 ± 3.4 to 248.6 ± 11.1 mg TAE/gdw respectively which differed significantly (P < 0.01) among various groups. These extracts also showed remarkable DPPH radical scavenging activity (43.9 ± 1.1 % to 69.9 ± 2.9 %). The reducing powerOD700 was observed maximum (P < 0.01) in sprouted mung bean extract as compared to corn powder extract. The correlation between DPPH free radical SA v/s total phenolics was significant (P < 0.01) with R = 0.775. correlation between DPPH free radical SA v/s reducing power and total phenolics v/s reducing power was also significant (P < 0.01) with R value 0.907 and 0.682 respectively. FGE treated raw chicken breast meat showed better (P < 0.01) oxidative stability than control groups.