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Electroencephalography (EEG) can be reliable for assessing the brain’s electrical activity of preslaughter stress and pain. The duration between the ventral neck cut and induction of a state of unconsciousness/insensibility is crucial in the slaughtering of animals, reducing pain, fear, and distress. Various EEG variables, such as median frequency (F50), the total power of EEG spectrum (Ptot), waves patterns (amplitude and frequencies), epileptiform EEG, index of consciousness, and isoelectric EEG, are used to identify a valid indicator of the state of unconsciousness. Association among various behavioral, physiological, and hematological parameters with EEG variables could provide an overall assessment and deep insights into the animal stress levels or welfare status during various managemental and preslaughter operations, such as transport, stunning, and slaughtering operations. The application of EEG could help in further refining the stunning technologies and slaughter protocols in livestock, poultry, and fish. The present review analyzed the application of EEG as a neurophysiological tool for assessing animal welfare during the critical state of preslaughter handling and slaughter, thus ensuring proper compliance with animal welfare principles.

Domestic beef production in Malaysia is mainly contributed by the indigenous Kedah-Kelantan (KK) and their crossbred cattle. This study was conducted to compare growth performance and sexual maturity of KK and KK × Brangus bulls. Ten (10) experimental bulls with similar ages were kept under the same paddock and fed with equal quality and quantity Guinea grasses and palm kernel cakes. Body weight was measured at monthly basis by using a digital animal weighing scale. Age at sexual maturity was determined by the time of semen ejaculation and quality. The results revealed significantly (p<0.05) greater body weight in KK × Brangus bulls (230.50±9.3 kg) as compared to the pure-bred KK bulls (204.5 ±13.2 kg) at 24 months of ages. KK bulls attained sexual maturity at an earlier age of 21.5±0.4 months, while KK × Brangus bulls attained at 28.8±0.5 months. The growth performance of KK × Brangus bulls was significantly (p˂0.05) higher than KK bulls. Age showed a significant positive correlation with body weight and average daily weight gains. In conclusion, KK pure-bred bulls were superior in terms of attaining age at puberty and sexual maturity earlier, whereas KK × Brangus bulls were better for growth performance.

Mammalian long bone growth occurs through endochondral ossification, majorly regulated by the controlled enlargement of chondrocytes at the growth plate (GP). This study aimed to investigate the roles of Na+/H+ (sodium hydrogen exchanger (NHE1)) and HCO3− (anion exchanger [AE2]) during longitudinal bone growth in mammals. Bones from P10 SpragueDawley rat pups were cultured exvivo in the presence or absence of NHE1 and AE2 inhibitors to determine their effect on long bone growth. Gross morphometry, histomorphometry, and immunohistochemistry were used to assess the bone growth. The results revealed that the culture of the bones in the presence of NHE1 and AE2 inhibitors reduces bone growth significantly (p < 0.05) by approximately 11%. The inhibitor significantly (p < 0.05) reduces bone growth velocity and the length of the hypertrophic chondrocyte zone without any effect on the total GP length. The total GP chondrocyte density was significantly (p < 0.05) reduced, but hypertrophic chondrocyte densities remained constant. NHE1 fluorescence signaling across the GP length was higher than AE2, and their localization was significantly (p < 0.05) inhibited at the hypertrophic chondrocytes zone. The GP lengthening was majorly driven by an increase in the overall GP chondrocyte and hypertrophic chondrocyte densities apart from the regulatory volume phenomenon. This may suggest that NHE1 and AE2 could have a regulatory role in long bone growth.

With the ever-increasing global population, it is impossible to meet the demand for animal protein by relying only on conventional methods due to the depleting natural resources. It is very challenging to ensure a sustainable supply of animal proteins from a single source or form and requires a holistic approach by using all suitable options. The present review critically reviewed various technological, sustainability, nutritional value, regulatory framework, food safety challenge, and prospect aspects of plant-based meat analogs, in vitro meat, edible insect, and single-cell proteins as suitable candidates for future food security and supply of animal protein in a sustainable way. For in vitro meat, the technological challenge in the supply of raw inputs, large-size bioreactors, and scale-up remains a major issue. Although having a lower environmental impact, the acceptance of edible insects to more comprehensive sections and associated food safety risks remains a major concern. There is a need for uniform and proper regulations of these alternatives/novel foods across the globe, covering various aspects throughout the food supply chain. Plant-based meat analogs, in vitro meat, insects, and single-cell proteins along with conventional meat can meet the demand for high-quality protein in the near future.

Various potential sources of bioactive components exist in nature which are fairly underutilized due to the lack of a scientific approach that can be sustainable as well as practically feasible. The recovery of bioactive compounds is a big challenge and its use in food industry to develop functional foods is a promising area of research. Various techniques are available for the extraction of these bioactives but due to their thermolabile nature, there is demand for nonthermal or green technologies which can lower the cost of operation and decrease operational time and energy consumption as compared to conventional methods. Ultrasound-assisted extraction (UAE) is gaining popularity due to its relative advantages over solvent extraction. Thereafter, ultrasonication as an encapsulating tool helps in protecting the core components against adverse food environmental conditions during processing and storage. The review mainly aims to discuss ultrasound technology, its applications, the fundamental principles of ultrasonic-assisted extraction and encapsulation, the parameters affecting them, and applications of ultrasound-assisted extraction and encapsulation in food systems. Additionally, future research areas are highlighted with an emphasis on the energy sustainability of the whole process.

Meat analogs have opened a new horizon of opportunities for developing a sustainable alternative for meat and meat products. Proteins are an integral part of meat analogs and their functionalities have been extensively studied to mimic meat-like appearance and texture. Proteins have a vital role in imparting texture, nutritive value, and organoleptic attributes to meat analogs. Processing of suitable proteins from vegetable, mycoproteins, algal, and single-cell protein sources remains a challenge and several technological interventions ranging from the isolation of proteins to the processing of products are required. The present paper reviews and discusses in detail various proteins (soy proteins, wheat gluten, zein, algal proteins, mycoproteins, pulses, potato, oilseeds, pseudo-cereals, and grass) and their suitability for meat analog production. The review also discusses other associated aspects such as processing interventions that can be adapted to improve the functional and textural attributes of proteins in the processing of meat analogs (extrusion, spinning, Couette shear cell, additive manufacturing/3D printing, and freeze structuring).

Stress induces various physiological and biochemical alterations in the animal body, which are used to assess the stress status of animals. Blood profiles, serum hormones, enzymes, and physiological conditions such as body temperature, heart, and breathing rate of animals are the most commonly used stress biomarkers in the livestock sector. Previous exposure, genetics, stress adaptation, intensity, duration, and rearing practices result in wide intra- and inter-animal variations in the expression of various stress biomarkers. The use of meat proteomics by adequately analyzing the expression of various muscle proteins such as heat shock proteins (HSPs), acute phase proteins (APPs), texture, and tenderness biomarkers help predict meat quality and stress in animals before slaughter. Thus, there is a need to identify non-invasive, rapid, and accurate stress biomarkers that can objectively assess stress in animals. The present manuscript critically reviews various aspects of stress biomarkers in animals and their application in mitigating preslaughter stress in meat production.

Palm kernel cake (PKC) is a by-product of oil extraction from palm fruits and has been included in poultry diets as an alternative to soybean meal and yellow corn. Due to its high content of fibre, coarse texture and gritty appearance, the use of PKC in poultry nutrition is limited. In order to increase the nutritive value of PKC, there is a tendency nowadays to create solid state fermentation (SSF) by using cellulolytic microbes. This paper reviews the impact of feeding fermented and non-fermented PKC on the performance of broiler chickens. Recent studies have reported that SSF by cellulolytic microorganisms improved the nutritive value of PKC. The nutrient digestibility has been increased significantly in PKC fermented using Paenibacillus polymyxya ATCC 842 or Weisella confusa SR-17b. The availability of valine, histidine, methionine and arginine was 70.42%, 71.50%, 71.92% and 81.15%, respectively, in PKC fermented using P. polymyxa ATCC 842. The digestibility of crude protein (CP) increased by 61.83% and 59.90% in PKC fermented using P. polymyxya ATCC 842 or W. confusa SR-17b, respectively. In addition, body weight gain (BWG) and feed conversion ratio (FCR) improved significantly in broilers fed 15% fermented PKC compared to those fed 15% non-fermented PKC (2000.43 g versus 1823.23 g and 1.75 versus 1.91, respectively). The intestinal Enterobacteriaceae decreased (4.03 CFU/g) and lactic acid bacteria increased (5.56 CFU/g) in birds fed 15% PKC fermented by P. polymyxa ATCC 842. Therefore, fermented PKC can be included in a broiler diet up to 15%, replacing part of soybean and yellow corn in the diet, leading to a decrease in the overall cost of poultry feeding.

Incorporation of the coloring agent, tartrazine and curcumin affects healthy physiological system leading to changes in hematology, biochemistry and enzymatic activity. Consequences of oral administration of tartrazine and curcumin were explored for 15, 30 and 45 days in female adult rat. Two doses were based on the admissible daily intake (ADI) of 9.6 and 96 (high) mg/kg/body weight for tartrazine, 3.85 and 38.5 mg/kg/body weight for curcumin. Phytochemicals such as saponins, glycosides, alkaloids, flavonoids, tannin, carbohydrates and phenolics were present in coloring agents. The renal function tests and lipid profile showed alterations in the values of uric acid, urea, total protein, albumin and cholesterol in all treatment groups. Additionally, the levels of liver enzymes fluctuated after 45 days of treatment with curcumin and tartrazine. Histopathological changes were also recorded in liver and kidney. In contrast, blood biochemistry depicted the significant reduction of RBCs, Hb, MCH, glucose, WBCs, MCHC and LDH at 15, 30 and 45 days. Besides, IgG, IgM and overall antibody GMT at 45 days were significantly increased. Study indicated that ADI and doses up to 10 times of the ADI of food colors intake exerted adverse effects on immune response and altered the biochemical parameters.

This preliminary trial investigated the effect of transportation and lairage periods on physiological parameters of goats subjected to slaughter. Nine male Boer cross goats aged 8–12 months were transported for 6 h and kept at lairage for 3, 6, or 16 h (n = 3). Blood samples were collected at pre- (pre-T) and post-transportation (post-T), and post-slaughter (post-S) for determination of hematological parameters, serum enzyme, protein, and cortisol concentrations. Electroencephalogram readings were taken at pre-T, post-T, pre-slaughter (pre-S), and post-S to determine the median frequency (F50) and total power (Ptot) values. At post-T, there were manifestations of stress leukogram; increase in hematocrit, total protein, and muscle enzyme concentrations; and decrease in Ptot (p < 0.05). The high pre-T cortisol concentration suggests that the goats were already under stress before transportation. Stress leukogram became less evident after lairage, indicating that the goats had recovered from the stress of transportation. Although the Ptot increased at post-S especially following 3 h of lairage, F50 values at post-S did not differ from pre-L, suggesting that the pre-slaughter stress may have affected the pain threshold. It is suggested that after 6 h of transportation, goats should ideally be placed in lairage for a minimum period of 3 h before slaughter.

A comprehensive stress assessment is vital in understanding the impact of the pre-slaughter procedure on animal welfare. The transportation and handling process was commonly reported to cause stress in animals. This research utilises electroencephalography (EEG) as an alternative stress indicator to non-painful acute stress measurement. EEG has been proved to be instantaneous and sensitive with specific results. Therefore, this study was aimed to determine the stress level of goats subjected to two different transportation duration and the effect of lairage based on their EEG activities and blood parameters changes. Eighteen adult male goats were divided into two transportation stress groups based on the transport duration: the two-hour (TS2) and six-hour (TS6) groups. Then, each group was then again divided into three smaller groups according to the lairage duration, which was three-hour (L3), six-hour (L6), and overnight (L12) groups. Blood was sampled before transport, after transport, and during slaughter while EEG was recorded before transport, after transport, after lairage, and during slaughter. Results revealed that there was a significant decrease in beta wave activity compared to baseline in TS2 goats (P < 0.05) after transportation, whereas no significant difference was detected in the TS6 goats. At the same time, goats from the TS2 group showed increase in creatine kinase (CK) and lactate dehydrogenase (LDH) compared to that in TS6 goats. Together with the observed cortisol concentration, these findings showed that the TS6 goats were fully adapted to the transportation stress while the TS2 goats were still under stress. As for the lairage duration, it was observed that the TS2L3 goats showed lower EEG activities than the values obtained after two-hour transportation, while lower EEG activities were found from the TS6L6 goats after six-hour transportation. Therefore, it can be concluded that three-hour lairage was adequate to lower the impact of two hours

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.

This study’s objective was to evaluate the effects of distance and stocking density on physicochemical properties and oxidative stability of meat and acute-phase proteins in Brahman crossbred cattle transported by road under hot and humid tropical conditions. Sixty Brahman crossbred heifers were subjected to road transport from a cattle feedlot farm located in Universiti Putra Malaysia (UPM), Serdang, to a commercial ruminant abattoir in Shah Alam, Selangor. Animals were assigned to long and short distances and high, medium, and low stocking densities. The results revealed that the intensity of response significantly increased in meat samples from animals subjected to long-distance transportation and higher stocking density. Alpha-1-acid glycoprotein and serum amyloid-A values increased considerably and were different from the baseline values recorded at preload. In conclusion, the current results revealed that the color, pH, shear force values, water holding capacity (WHC), glycogen level, and malondilaldehyde assay (MDA) concentrations in meat and acute-phase proteins (APP) were affected by both distances and stocking densities, as evidenced by the significant changes recorded from the parameters above.

Storage temperature and duration plays an important role in meat processing. Observations in poultry processing plants have shown a serious deviation in storage condition compared to the recommended procedures. Furthermore, there is still a paucity of evidence on the effects of storage temperature and duration on meat quality and microbial population. The aim of this study was to determine the effects of different temperature and duration during storage on physico-chemical properties and microbiological quality of broiler chicken Pectoralis major muscle. Eighty birds were slaughtered and processed, following which the packed boneless breast (PBB) (each bird was to provide two breast muscle samples; left breast and right breast) was divided into four groups, each consisted of 40 PBB. Each group was subsequently assigned to storage either at 4 °C, -10 °C, -18 °C or -40 °C, for 24 h before 20 PBB samples from each group were transported to the respective laboratory for meat quality and microbiological analysis. The remaining 20 PBB from each storage temperature were stored for 72 h before being transported for analysis. Results have shown significant increases in drip loss and cooking loss as the storage temperature decreases. Similarly, storage duration significantly affected cooking loss, of which, samples stored for 72 h exhibited higher cooking loss compared to those stored for 24 h. For color, significant differences were only observed in lightness (L*) and redness (a*) values. Longer duration of storage had significantly improved tenderness whereby, samples that have been stored for a shorter duration presented higher pH values. Populations of coliform and Salmonella decreased significantly with decreasing temperature and increasing storage duration.

A fracture model in rats for the study of secondary bone healing was described. Standard open midshaft transverse metatarsal fracture was produced with bone cutting forceps in 28 rats. The commonly open and close fracture models utilized for bone and mineral researches are associated with varying degree of complications ranging from a high degree of fracture comminution to severe associated soft tissue injury which interferes with the healing process. We hypothesized that fracture model in rat third metatarsal bone could be associated with low -post-surgical complications and could be a reproducible model. To test this, open mid-shaft transverse fractures were created on the metatarsals of 28 rats. The objectives of the study were to evaluate the fracture complications, to determine the nature of fracture produced, evaluate the fracture consolidation during healing periods, and to assess the histological and radiographic healing of the fracture. The fracture produced in the mid metatarsal shaft of all rats was 100% transverse, 73% located at the midshaft. Minimal fracture angulations were recorded (0.48 ± 0.09o; 0.78 ± 0.17o) for anterior-posterior and lateral views respectively. Minimal soft tissue injury was recorded immediately post-surgery, but no infection and the delayed union was observed. Varying degrees of weight-bearing lameness was also recorded but seized at day six onward post-operative. Callus index observed was peaked at week 2 and 3 (2.02 ± 0.1, 1.99 ± 0.13) respectively but declined to 1.10 ± 0.04 at week 7 during the consolidation period. The fracture line disappeared completely at week 7. The histological and radiographic healing scores were (3.5 ± 0.13 and 3.75 ± 0.25) respectively (out of the maximum healing score of 4) at week 7 post-operative. There was a positive correlation between the histological and radiographic healing scores. The metatarsal fracture model is considered to be a suitable model for in vivo study of secondary fracture heali

This research was conducted to estimate the effects of Persicaria odorata leaf meal (POLM) on haematological indices, serum biochemical attributes, and internal organs parameters, including histomorphological features of the liver, in broiler chickens. A total of 120 one-day-old male broiler chicks (Cobb-500) were randomly allocated into four experimental groups. The dietary treatments were basal diet (BD), which served as the control (C), along with BD + 2 g/kg POLM (Po2), BD + 4 g/kg POLM (Po4), BD + 8 g/kg POLM (Po8), which were the supplemented groups. The body weight gain (BWG) showed a linear increase and feed conversion ratio (FCR) showed a linear decrease with increasing POLM dosage at day 42 (p ˂ 0.05) and for the overall growth performance period (p ˂ 0.01). On day 21 and day 42, the values of red blood cells (RBCs), white blood cells (WBCs), haemoglobin (Hb), and packed cell volume (PCV) showed linear increases (p ˂0.05) as the dosage of POLM increased in the diet. On day 21, dietary supplementation of POLM linearly decreased (p ˂ 0.05) the serum activity of alkaline phosphatase (ALP), aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), and serum levels of urea and creatinine. On the other hand, serum levels of total protein (TP), albumin, and globulin showed a linear increase (p ˂ 0.05) as the POLM dosage increased. On day 42, the serum activity of AST and ALT and serum levels of glucose, cholesterol, triglycerides, urea, and creatinine showed linear decreases (p ˂ 0.05) with increased levels of POLM in the diet. However, POLM supplementation linearly increased (p ˂ 0.05) the serum levels of TP and globulin. Dietary inclusion of POLM did not influence the organ parameters and showed no adverse effects on the liver histomorphology. In conclusion, supplementation of POLM increased the growth performance, improving haematological indices and serum biochemistry profiles of broiler chickens without any deleterious effects on the liver histomor

Due to antimicrobial resistance and the public health hazard of antibiotic growth promoters, there is a grave need to find potential alternatives for sustainable poultry production. Piper betle (PB) and Persicaria odorata (PO) are herbs, which have been reported for antimicrobial, antioxidant, and anti-inflammatory properties. The present study aimed to estimate the influence of different dose supplementation of Piper betle leaf meal (PBLM) and Persicaria odorata leaf meal (POLM) on growth performance, ileal digestibility and gut morphology of broilers chickens. A total of 210 one day-old broiler chicks were randomly grouped into 7 treatments, and each treatment group has 3 replicates (n = 10) with a total number of 30 chicks. The treatments included T1 control (basal diet (BD) with no supplementation), T2 (BD + 2 g/kg PBLM); T3 (BD + 4 g/kg PBLM), T4 (BD + 8 g/kg PBLM), T5 (BD + 2 g/kg POLM), T6 (BD + 4 g/kg POLM), T7 (BD + 8 g/kg POLM). Growth performance, gut morphology and ileal digestibility were measured. Except for T4 (8 g/kg PBLM), graded dose inclusion of PBLM and POLM increased (P < 0.05) the body weight gain (BWG), positively modulated the gut architecture and enhanced nutrient digestibility in both stater and finisher growth phases of broiler chickens. Birds fed on PBLM 4 g/kg (T3), and POLM 8 g/kg (T7) had significantly higher (P < 0.05) BWG with superior (P < 0.05) feed efficiency in the overall growth period. Chickens fed on diets T3 and T7 had longer (P < 0.05) villi for duodenum as well as for jejunum. Furthermore, the birds fed on supplementations T3 and T7 showed improved (P < 0.05) digestibility of ether extract (EE), and dry matter (DM) compared to the control group. However, least (P < 0.05) crude protein (CP) digestibility was recorded for T4. In conclusion, dietary supplementations of PBLM 4 g/kg and POLM 8 g/kg were positively modulated the intestinal microarchitecture with enhanced nutrient digestibility, resulted in maximum body weight gai

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 the sharp 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.