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ABSTRACT Escherichia coli (E. coli) is reported to be an important pathogen associated with calf diarrhea. Antibiotic resistance genes (ARGs) and virulence factor genes (VFGs) pose a considerable threat to both animal and human health. However, little is known about the characterization of ARGs and VFGs presented in the gut microbiota of diarrheic calves caused by E. coli. In this study, we used multi-omics strategy to analyze the ARG and VFG profiles of Simmental calves with diarrhea caused by E. coli K99. We found that gut bacterial composition and their microbiome metabolic functions varied greatly in diarrheic calves compared to healthy calves. In total, 175 ARGs were identified, and diarrheal calves showed a significantly higher diversity and abundance of ARGs than healthy calves. Simmental calves with diarrhea showed higher association of VFGs with pili function, curli assembly, and ferrienterobactin transport of E. coli. Co-occurrence patterns based on Pearson correlation analysis revealed that E. coli had a highly significant (P < 0.0001) correlation coefficient (>0.8) with 16 ARGs and 7 VFGs. Metabolomics analysis showed that differentially expressed metabolites in Simmental calves with diarrhea displayed a high correlation with the aforementioned ARGs and VFGs. Phylotype analysis of E. coli genomes showed that the predominant phylogroup B1 in diarrheic Simmental calves was associated with 10 ARGs and 3 VFGs. These findings provide an overview of the diversity and abundance of the gut microbiota in diarrheic calves caused by E. coli and pave the way for further studies on the mechanisms of antibiotic resistance and virulence in the calves affected with diarrhea.IMPORTANCESimmental is a well-recognized beef cattle breed worldwide. They also suffer significant economic losses due to diarrhea. In this study, fecal metagenomic analysis was applied to characterize the antibiotic resistance gene (ARG) and virulence factor gene (VFG) profiles of diarrheic Simmental calves. We identified key ARGs and VFGs correlated with Escherichia coli isolated from Simmental calves. Additionally, metabolomics analysis showed that differentially expressed metabolites in Simmental calves with diarrhea displayed a high correlation with the aforementioned ARGs and VFGs. Our findings provide an insight into the diversity and abundance of the gut microbiota in diarrheic calves caused by Escherichia coli and pave the way for further studies on the mechanisms of antibiotic resistance and virulence in the diarrheal calves from cattle hosts.

Understanding the effects of bacteria and enzyme addition on mixed crop silage is imperative for dairy producers to make informed decisions. The current study evaluated the chemical changes in silage prepared from different ramie and elephant grass ratios (30:70, 50:50, 70:30, and 100:0) in response to bacteria (0, 100, 200, and 300 mg/kg) and enzyme addition (0, 10, 20, and 30 mg/kg) in a complete randomized design. The results indicated that the proportion of ramie in silage (p < 0.01), level of bacteria (p < 0.05), and level of enzyme added (p = 0.05) affected the CP, fiber, volatile fatty acids, and lactic acid contents and pH of silage. By comprehensive analysis, low lignin content and pH of silage with high lactic acid content was observed with a 30% ramie proportion. High CP and lactic acid contents with low ADF, lignin, and pH values were observed with the addition of bacteria (p < 0.05). High lactic acid with low ADF content was observed with the addition of enzyme in silage (p ≤ 0.05). The optimum quality of silage was observed when the ramie, bacteria, and enzymes were added at the levels of 30%, 200 mg/kg, and 20 mg/kg, respectively, in ramie and elephant grass mixed silage.

Cannabinoids are a group of bioactive compounds abundantly present in Cannabis sativa plant. The active components of cannabis with therapeutic potential are known as cannabinoids. Cannabinoids are divided into three groups: plant-derived cannabinoids (phytocannabinoids), endogenous cannabinoids (endocannabinoids), and synthetic cannabinoids. These compounds play a crucial role in the regulation various physiological processes including the immune modulation by interacting with the endocannabinoid system (A complex cell-signaling system). Cannabinoid receptor type 1 (CB1) stimulates the binding of orexigenic peptides and inhibits the attachment of anorexigenic proteins to hypothalamic neurons in mammals, increasing food intake. Digestibility is unaffected by the presence of any cannabinoids in hemp stubble. Endogenous cannabinoids are also important for the peripheral control of lipid processing in adipose tissue, in addition to their role in the hypothalamus regulation of food intake. Regardless of the kind of synaptic connection or the length of the transmission, endocannabinoids play a crucial role in inhibiting synaptic transmission through a number of mechanisms. Cannabidiol (CBD) mainly influences redox equilibrium through intrinsic mechanisms. Useful effects of cannabinoids in animals have been mentioned e.g., for disorders of the cardiovascular system, pain treatment, disorders of the respiratory system or metabolic disorders. Dietary supplementation of cannabinoids has shown positive effects on health, growth and production performance of small and large animals. Animal fed diet supplemented with hemp seeds (180 g/day) or hemp seed cake (143 g/kg DM) had achieved batter performance without any detrimental effects. But the higher level of hemp or cannabinoid supplementation suppress immune functions and reduce productive performance. With an emphasis on the poultry and ruminants, this review aims to highlight the properties of cannabinoids and their derivatives as well as their significance as a potential feed additive in their diets to improve the immune status and health performance of animals.

ABSTRACT Calf diarrhea is a multifactorial disease that affects the cattle industry and accounts for more than 50% of calf mortality. Although there is evidence of an association between altered gut microbiota and diarrhea, remarkably little is known about the microbial and metabolic mechanisms underlying the link between gut microbiota dysbiosis and the occurrence of calf diarrhea. Here, we performed fecal metagenomic and metabolomic studies on fecal samples from diarrheic and healthy calves of Xia-nan cattle breed. Results revealed that composition of the gut microbiome and metabolome was remarkably altered in diarrheic calves, and gut microbial alterations were associated with diarrhea and linked to the changes in metabolites. Metabolite profiles showed that diarrheic calves exhibited a marked decrease in some purines (adenosine, adenine, 2'-deoxyguanosine, allantoate, deoxyinosine, and deoxyguanosine) and arachidonic acid (prostaglandin F2α and prostaglandin E2) compared to healthy calves. Purine-producing microbial species, including Lactiplantibacillus plantarum, Campylobacter coli, Treponema porcinum, Klebsiella pneumoniae, and Phocaeicola coprophilus, were significantly reduced in diarrheic calves compared to healthy calves, whereas the arachidonic acid-producing species such as Neisseria gonorrhoeae, Staphylococcus aureus, and Clostridiales bacterium exhibited a marked increase. These microbial signatures were closely associated with the metabolic dysbiosis of purine and arachidonic acid in diarrhea calves. Our study showed that gut microbiota-driven metabolic disorders of purine or arachidonic acid were associated with calf diarrhea. The findings prove that altered gut microbiota plays a role in diarrhea pathogenesis and indicate that gut microbiota-targeted therapies could be useful for both prevention and treatment of diarrhea. IMPORTANCE Calf diarrhea is of great concern to the global dairy industry as it results in significant economic losses due to lower conception rates, reduced milk production, and early culling. Although there is evidence of an association between altered gut microbiota and diarrhea, remarkably little is known about the microbial and metabolic mechanisms underlying the link between gut microbiota dysbiosis and the occurrence of calf diarrhea. Here, we used fecal metagenomic and metabolomic analyses to demonstrate that gut microbiota-driven metabolic disorders of purine or arachidonic acid were associated with calf diarrhea. These altered gut microbiotas play vital roles in diarrhea pathogenesis and indicate that gut microbiota-targeted therapies could be useful for both prevention and treatment of diarrhea.

ABSTRACT: Runs of homozygosity (ROH) are a powerful tool to explore patterns of genomic inbreeding in animal populations and detect signatures of selection. The present study used ROH analysis to evaluate the genome-wide patterns of homozygosity, inbreeding levels, and distribution of ROH islands using the SNP data sets from 899 Mediterranean buffaloes. A total of 42,433 ROH segments were identified, with an average of 47.20 segments per individual. The ROH comprising mostly shorter segments (1–4 Mb) accounted for approximately 72.29% of all ROH. In contrast, the larger ROH (>8 Mb) class accounted for only 7.97% of all ROH segments. Estimated inbreeding coefficients from ROH (FROH) ranged from 0.0201 to 0.0371. Pearson correlations between FROH and genomic relationship matrix increased with the increase of ROH length. We identified ROH hotspots in 12 genomic regions, located on chromosomes 1, 2, 3, 5, 17, and 19, harboring a total of 122 genes. Protein-protein interaction (PPI) analysis revealed the clustering of these genes into 7 PPI networks. Many genes located in these regions were associated with different production traits. In addition, 5 ROH islands overlapped with cattle quantitative trait loci that were mainly associated with milk traits. These findings revealed the genome-wide autozygosity patterns and inbreeding levels in Mediterranean buffalo. Our study identified many candidate genes related to production traits that could be used to assist in selective breeding for genetic improvement of buffalo.

Abstract This study aimed to evaluate the effects of intravenous injection of cysteamine (CS) on body weight (BW), growth hormone (GH), thyroid hormones (TH) secretion, and antioxidant status of growing lambs grazing on mountain pastures. Fifteen lambs (3–4 months of age) were randomly allocated into three experimental groups which received different dosages of CS: 0, 20, and 50 mg/kg BW−1. The CS was injected on the 1st, 10th, and 20th days of the experiment to the lambs through the jugular vein. Assessment of plasma concentration of GH and TH hormones was carried out at days 0 (a day before the start of CS injections), 15, and 30 of the experiment. The antioxidant enzymes were measured at the end of the experiment. Lambs were weighed at days 0, 10, 20, and 30 of the experiment. The results showed that treatment and time affected the BW, GH, triiodothyronine (T3), and tetraiodothyronine (T4) secretion. The intravenous injection of CS increased the BW of growing lambs (p

Increase in drug resistance as well as ineffective immunization efforts against various pathogens (viruses, bacteria and fungi) pose a significant threat to the poultry industry. Spirulina is one of the most widely used natural ingredients which is becoming popular as a nutritional supplement in humans, animals, poultry and aquaculture. It contains protein, vitamins, minerals, fatty acids, pigments, and essential amino acids. Moreover, it also has considerable quantities of unique natural antioxidants including polyphenols, carotenoids, and phycocyanin. Dietary supplementation of Spirulina can beneficially affect gut microbial population, serum biochemical parameters, and growth performance of chicken. Additionally, it contains polyphenolic contents having antibacterial effects. Spirulina extracts might inhibit bacterial motility, invasion, biofilm formation, and quorum sensing in addition to acting directly on the bacterium by weakening and making the bacterial cell walls more porous, subsequently resulting in cytoplasmic content leakage. Additionally, Spirulina has shown antiviral activities against certain common human or animal viruses and this capability can be considered to exhibit potential benefits against avian viruses also. Spirulan, a calcium-rich internal polysaccharide of Spirulina, is potentially responsible for its antiviral effect through inhibiting the entry of several viruses into the host cells, boosting the production of nitric oxide in macrophages, and stimulating the generation of cytokines. Comparatively a greater emphasis has been given to the immune modulatory effects of Spirulina as a feed additive in chicken which might boost disease resistance and improve survival and growth rates, particularly under stress conditions. This manuscript reviews biological activities and immune-stimulating properties of Spirulina and its potential use as a dietary supplement in poultry to enhance growth, gut health and disease resistance.

Abstract Background Heat stress in tropics is generally associated with significant economic losses resulting from reduced performance, morbidity, and mortality of livestock. To avoid serious consequences of heat stress, it is imperative to better understand the physiological responses and biochemical changes under the state of altered body homeostasis during different seasons of the year. Objectives This study aimed to evaluate the seasonal dynamics of physiological, oxidative and metabolic responses of lactating Nili‐Ravi buffaloes to the tropical climate of South China. Methods Physiological responses including rectal temperature (RT), body surface temperature (BST) and respiratory rate (RR) along with serum biochemical and antioxidant parameters of 20 lactating Nili‐Ravi buffaloes were evaluated during different seasons of the year. Results Higher temperature‐humidity Index (THI) during the summer season (>80) resulted in a significant increases in RR and BST as compared to the winter season. Higher oxidative stress was observed in the summer season as revealed by significantly higher MDA while lower serum antioxidant enzyme (TAC, GSH‐Px, SOD and CAT) contents. Moreover, serum cortisol was also significantly higher in summer and autumn. The levels of growth hormone and ACTH were also significantly (P

Colonization and development of the gut microbiome is a crucial consideration for optimizing the health and performance of livestock animals. This is mainly attributed to the fact that dietary and management practices greatly influence the gut microbiota, subsequently leading to changes in nutrient utilization and immune response. A favorable microbiome can be implanted through dietary or management interventions of livestock animals, especially during early life. In this review, we explore all the possible factors (for example gestation, colostrum, and milk feeding, drinking water, starter feed, inoculation from healthy animals, prebiotics/probiotics, weaning time, essential oil and transgenesis), which can influence rumen microbiome colonization and development. We discuss the advantages and disadvantages of potential strategies used to manipulate gut development and microbial colonization to improve the production and health of newborn calves at an early age when they are most susceptible to enteric disease. Moreover, we provide insights into possible interventions and their potential effects on rumen development and microbiota establishment. Prospects of latest techniques like transgenesis and host genetics have also been discussed regarding their potential role in modulation of rumen microbiome and subsequent effects on gut development and performance in neonatal ruminants.

The Sox gene family constitutes transcription factors with a conserved high mobility group box (HMG) that regulate a variety of developmental processes, including sex differentiation, neural, cartilage, and early embryonic development. In this study, we systematically analyzed and characterized the 20 Sox genes from the whole buffalo genome, using comparative genomic and evolutionary analyses. All the buffalo Sox genes were divided into nine sub-groups, and each gene had a specific number of exons and introns, which contributed to different gene structures. Molecular phylogeny revealed more sequence similarity of buffalo Sox genes with those of cattle. Furthermore, evolutionary analysis revealed that the HMG domain remained conserved in the all members of the Sox gene family. Similarly, all the genes are under strong purifying selection pressure; seven segmental duplications occurred from 9.65 to 21.41 million years ago (MYA), and four potential recombination breakpoints were also predicted. Mutational analysis revealed twenty non-synonymous mutations with potential effects on physiological functions, including embryonic development and cell differentiation in the buffalo. The present study provides insights into the genetic architecture of the Sox gene family in buffalo, highlights the significance of mutations, and provides their potential utility for marker-assisted selection for targeted genetic improvement in buffalo.

Variation in food and diet shapes the diversity of the gut microbiota of ruminants. The present study investigated the microbial diversity in the fecal microbiota of yaks reared under natural grazing and feedlot system. A total of 48 domestic yaks with an average age of 7.5 years were selected from two different grazing habitats: one group grazed on natural pasture (grazing yaks—GY) while the other group was fed fodder and concentrate (feedlot yaks—FY). Crude protein, non-fiber carbohydrate, hemicelluloses, and digestible dry matter contents of natural pastures were higher than those in the feedlot. The lower insoluble fiber contents were found in grazing land. The 16S rRNA gene sequencing revealed 675 and 348 unique operational taxonomic units (OTUs) in the GY and FY, respectively, in addition to 1,778 common OTUs. Overall, a total of 9,891 OTUs were identified as a whole, of which 6,160 OTUs were from GY and 3,731 were from FY. Shannon index analysis revealed a higher bacterial diversity in GY than FY. At the phylum level, Firmicutes were dominant bacterial taxa in both groups. The relative abundance of Firmicutes in GY (56% ± 0.05) was higher than in FY (41% ± 0.08). At the family level, GY had a significantly higher abundance of Ruminococcaceae (p < 0.001) and Rikenellaceae (p < 0.001) than FY, but FY had a significantly higher abundance of Prevotellaceae than GY (p < 0.001). At the genus level, abundances of Faecalibacterium, Alloprevotella, and Succinivibrio were higher in FY than in GY. This study presents novel information on fecal bacterial composition and diversity in yaks reared under two different production systems.

Heat stress induced by long periods of high ambient temperature decreases animal productivity, leading to heavy economic losses. This devastating situation for livestock production is even becoming worse under the present climate change scenario. Strategies focused to breed animals with better thermo-tolerance and climatic resilience are keenly sought these days to mitigate impacts of heat stress especially in high input livestock production systems. The 70-kDa heat shock proteins (HSP70) are a protein family known for its potential role in thermo-tolerance and widely considered as cellular thermometers. HSP70 function as molecular chaperons and have major roles in cellular thermotolerance, apoptosis, immune-modulation and heat stress. Expression of HSP70 is controlled by various factors such as, intracellular pH, cyclic adenosine monophosphate (cyclic AMP), protein kinase C and intracellular free calcium, etc. Over expression of HSP70 has been observed under oxidative stress leading to scavenging of mitochondrial reactive oxygen species and protection of pulmonary endothelial barrier against bacterial toxins. Polymorphisms in flanking and promoter regions in HSP70 gene have shown association with heat tolerance, weaning weight, milk production, fertility and disease susceptibility in livestock. This review provides insight into pivotal roles of HSP70 which make it an ideal candidate genetic marker for selection of animals with better climate resilience, immune response and superior performance. Keywords: HSP70, Heat stress, Thermo-tolerance, Molecular chaperons, Immunity, Livestock

Nitrate is used as a methane inhibitor while cysteamine is considered as a growth promoter in ruminants. The present study evaluated the effect of sodium nitrate and cysteamine on methane (CH4) production, rumen fermentation, amino acid (AA) metabolism, and rumen microbiota in a low protein diet. Four treatments containing a 0.5 g of substrate were supplemented with 1 mg/mL sodium nitrate (SN), 100 ppm cysteamine hydrochloride (CS), and a combination of SN 1 mg/mL and CS 100 ppm (CS+SN), and a control (no additive) were applied in a completely randomized design. Each treatment group had five replicates. Two experimental runs using in vitro batch culture technique were performed for two consecutive weeks. Total gas and CH4 production were measured in each fermentation bottle at 3, 6, 9, 12, 24, 48, and 72 h of incubation. The results showed that SN and CS+SN reduced the production of total gas and CH4, increased the rumen pH, acetate, acetate to propionate ratio (A/P), and microbial protein (MCP) contents (p < 0.05), but decreased other volatile fatty acids (VFA) and total VFA (p = 0.001). The CS had no effect on CH4 production and rumen fermentation parameters except for increasing A/P. The CSN increased the populations of total bacteria, fungi, and methanogens but decreased the diversity and richness of rumen microorganisms. In conclusion, CS+SN exhibited a positive effect on rumen fermentation by increasing the number of fiber degrading and hydrogen-utilizing bacteria, with a desirable impact on rumen fermentation while reducing total gas and CH4 production.

Plant polyphenols have promoting health features, including anti-mutagenic, anti-inflammatory, anti-thrombotic, anti-atherogenic, and anti-allergic effects. These polyphenols improve the immune system by affecting the white blood cell proliferation, as well as by the synthesis of cytokines and other factors, which contribute to immunological resistance. Olive trees are one of the most famous trees in the world. Whereas, olive olive oil and derivatives represent a large group of feeding resource for farm animals. In recent years, remarkable studies have been carried out to show the possible use of olive oil and derivatives for improvement of both animal performance and product quality. In vivo application of olive oil and its derived products has shown to maintain oxidative balance owing to its polyphenolic content. Consumption of extra virgin olive oil reduces the inflammation, limits the risk of liver damage, and prevents the progression of steatohepatitis through its potent antioxidant activities. Also, the monounsaturated fatty acids content of olive oil (particularly oleic acid), might have positive impacts on lipid peroxidation and hepatic protection. Therefore, this review article aims to highlight the nutritional applications and beneficial health aspects of olive oil and its effect on poultry production.

Toll-like receptors (TLRs) are pathogen recognition receptors, and primitive sources of innate immune response that also play key roles in the defense mechanism against infectious diseases. About 10 different TLRs have been discovered in chicken that recognize ligands and participate in TLR signaling pathways. Research findings related to TLRs revealed new approaches to understand the fundamental mechanisms of the immune system, patterns of resistance against diseases, and the role of TLR-specific pathways in nutrient metabolism in chicken. In particular, the uses of specific feed ingredients encourage molecular biologists to exploit the relationship between nutrients (including different phytochemicals) and TLRs to modulate immunity in chicken. Phytonutrients and prebiotics are noteworthy dietary components to promote immunity and the production of disease-resistant chicken. Supplementations of yeast-derived products have also been extensively studied to enhance innate immunity during the last decade. Such interventions pave the way to explore nutrigenomic approaches for healthy and profitable chicken production. Additionally, single-nucleotide polymorphisms in TLRs have shown potential association with few disease outbreaks in chickens. This review aimed to provide insights into the key roles of TLRs in the immune response and discuss the potential applications of these TLRs for genomic and nutritional interventions to improve health, and resistance against different fatal diseases in chicken.

Buffalo is a luxurious genetic resource with multiple utilities (as a dairy, draft, and meat animal) and economic significance in the tropical and subtropical regions of the globe. The excellent potential to survive and perform on marginal resources makes buffalo an important source for nutritious products, particularly milk and meat. This study was aimed to investigate the evolutionary relationship, physiochemical properties, and comparative genomic analysis of the casein gene family (CSN1S1, CSN2, CSN1S2, and CSN3) in river and swamp buffalo. Phylogenetic, gene structure, motif, and conserved domain analysis revealed the evolutionarily conserved nature of the casein genes in buffalo and other closely related species. Results indicated that casein proteins were unstable, hydrophilic, and thermostable, although αs1-CN, β-CN, and κ-CN exhibited acidic properties except for αs2-CN, which behaved slightly basic. Comparative analysis of amino acid sequences revealed greater variation in the river buffalo breeds than the swamp buffalo indicating the possible role of these variations in the regulation of milk traits in buffalo. Furthermore, we identified lower transcription activators STATs and higher repressor site YY1 distribution in swamp buffalo, revealing its association with lower expression of casein genes that might subsequently affect milk production. The role of the main motifs in controlling the expression of casein genes necessitates the need for functional studies to evaluate the effect of these elements on the regulation of casein gene function in buffalo.

Ruminants inhabit the consortia of gut microbes that play a critical functional role in their maintenance and nourishment by enabling them to use cellulosic and non-cellulosic feed material. These gut microbes perform major physiological activities, including digestion and metabolism of dietary components, to derive energy to meet major protein (65–85%) and energy (ca 80%) requirements of the host. Owing to their contribution to digestive physiology, rumen microbes are considered one of the crucial factors affecting feed conversion efficiency in ruminants. Any change in the rumen microbiome has an imperative effect on animal physiology. Ruminal microbes are fundamentally anaerobic and produce various compounds during rumen fermentation, which are directly used by the host or other microbes. Methane (CH4) is produced by methanogens through utilizing metabolic hydrogen during rumen fermentation. Maximizing the flow of metabolic hydrogen in the rumen away from CH4 and toward volatile fatty acids (VFA) would increase the efficiency of ruminant production and decrease its environmental impact. Understanding of microbial diversity and rumen dynamics is not only crucial for the optimization of host efficiency but also required to mediate emission of greenhouse gases (GHGs) from ruminants. There are various strategies to modulate the rumen microbiome, mainly including dietary interventions and the use of different feed additives. Phytogenic feed additives, mainly plant secondary compounds, have been shown to modulate rumen microflora and change rumen fermentation dynamics leading to enhanced animal performance. Many in vitro and in vivo studies aimed to evaluate the use of plant secondary metabolites in ruminants have been conducted using different plants or their extract or essential oils. This review specifically aims to provide insights into dietary interactions of rumen microbes and their subsequent consequences on rumen fermentation. Moreover, a comprehensive overview of the modulation of rumen microbiome by using phytogenic compounds (essential oils, saponins, and tannins) for manipulating rumen dynamics to mediate CH4 emanation from livestock is presented. We have also discussed the pros and cons of each strategy along with future prospective of dietary modulation of rumen microbiome to improve the performance of ruminants while decreasing GHG emissions.

Hot and humid weather exposes animals to high temperature and relative humidity that ultimately reduce their ability to disperse body heat. To avoid serious consequences of heat stress, it is imperative to understand animal physiological responses and biochemical changes during a state of altered body homeostasis across different seasons of the year. This study evaluated seasonal dynamics of physiological, oxidative, and metabolic responses of Nili-Ravi buffaloes to hot and humid climate. Twenty non-lactating multiparous buffaloes were enrolled for this 1-year study. Meteorological data were recorded twice daily to calculate temperature humidity index (THI). Physiological parameters including rectal temperature (RT), body surface temperature (BST), and respiratory rate (RR) were measured weekly. Blood samples were collected once in each season (spring, summer, autumn, and winter) to analyze biochemical and antioxidant parameters. We also measured activities of liver enzymes including alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The results revealed a significantly higher THI value (82) during summer which resulted in a significant increase in RR and BST as compared to winter. Higher oxidative stress was observed in summer owing to significantly higher malondialdehyde (MDA) content and lower levels of serum antioxidant enzymes (GPx, SOD, and CAT) as compared to other seasons. Moreover, serum cortisol was also significantly higher while adrenocorticotropic hormone (ACTH), Triiodothyronine (T3), insulin, and growth hormone contents were significantly lower in summer. Contrarily, plasma thyroxin (T4) level was higher in summer. THI showed a positive correlation with physiological responses but a negative correlation with antioxidant parameters. Our study provides practical insights on the adaptive physiology of buffaloes and has several implications regarding the alleviation of heat stress in buffaloes to enhance the efficiency of production and reproduction under tropical climate. Our study suggests the use of appropriate cooling strategies to effectively manage the non-lactating buffaloes to avoid performance losses and animal welfare issues in summer season.

This study was aimed to evaluate the effect of mulberry leaf flavonoids (MLF) on oxidative stress, metabolic hormones, and milk production in Murrah buffaloes. Forty multiparous Murrah buffaloes (4 ± 1 lactations) with similar body weight (average 600 ± 50 Kg) and stage of lactation (90 ± 20 d) were randomly selected for this trial. Four treatment groups (10 buffaloes per group) with different doses of MLF included; control (0 g/d), MLF15 (15 g/d), MLF30 (30 g/d), and MLF45 (45 g/d). Buffaloes were fed with total mix ration consisting of grass (Pennisetum purpureum schum), brewery's grain and concentrate mixture for 5 weeks. Meteorological data including ambient temperature and relative humidity were recorded using the online dust monitoring system to calculate temperature-humidity index (THI). After 1 week of the adaptation, milk yield was recorded daily while physiological parameters (respiratory rate, rectal, and body surface temperature), and milk composition were measured weekly. At the end of the trial, blood samples were collected to analyze serum metabolic hormones including estradiol (E2), growth hormone (GH), prolactin (PRL), Tri-iodothyronine (T3), and Thyroxine (T4). Moreover, serum heat shock proteins (HSP), antioxidants enzymes including malondialdehyde (MDA), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) and blood biochemical indices were also analyzed. Results revealed a decrease (P = 0.012) in serum MDA level while increasing (P < 0.01) the HSP and serum GHS-Px contents in supplemented buffaloes. Treatment showed a linear and quadratic decrease (p = 0.001) in the serum T-AOC while reducing CAT contents linearly (p = 0.012) as compared to the control. However, no effect of treatment on serum SOD content was observed. Treatment resulted a linear increase (p = 0.001) in serum GH and PRL hormones while increasing serum E2 levels linearly (P < 0.001) and quadratically (P = 0.025). Treatment increased (p = 0.038) the daily milk yield as compared to the control. However, increase (P < 0.05) in serum T3 and T4 contents, fat corrected milk (4%) and milk protein (%) was observed only in MLF45. Moreover, we observed no change in serum biochemical indices except insulin which linearly increased (p = 0.002) in MLF45. Our findings indicated that MLF at 45 g per day is an appropriate level to enhance milk performance and alleviate heat stress in buffaloes.

This study was aimed to evaluate the effects of Camelina sativa oil (CO) on fermentation kinetics and methane (CH4) production in rations with different roughage (R) to concentrate (C) ratios. Three total mixed rations (TMRs) were used as substrates (R70:C30, R50:C50, and R30:C70) supplemented with different levels of CO (0, 2, 4, 6, and 8% on dry matter basis) in an in vitro batch culture system. The enteric CH4 production was determined at different times of incubation while fermentation parameters were measured at the end of incubation. Results revealed that CO significantly decreased (P < 0.05) CH4 production at 48 h in medium (R50:C50) and low- (R30:C70) roughage diets than control. Camelina oil at all levels significantly (P < 0.05) affected ammonia nitrogen (NH3-N) and microbial protein (MCP) in all rations. Propionate concentration was increased by supplementing 8% CO to R70:C30 TMR, but it decreased with increasing levels of CO for low- and medium-roughage diets. Acetate concentration was significantly (P < 0.05) higher at 4% CO supplementation, but it decreased with 8% CO level in R30:C70 TMR. For all rations, CO decreased (P < 0.001) total bacteria, protozoa, and methanogens. Total fungi counts were affected by CO in all rations, especially with a 6% level in two rations (R30:C70 and R50:C50) and 8% level with high-roughage ration (R70:C30). Supplementation of CO in medium-roughage ration (R50:C50) showed a linear (P < 0.05) decrease in bacterial richness and evenness indices along with Shannon diversity as compared to the control. Moreover, CO also increased Firmicutes to Bacteroidetes ratio in all TMRs more effectively at higher levels. Camelina oil also affected the relative abundance of Prevotella in both low- and medium-roughage diets while increasing the abundance of Ruminobacter and Pseudobutyrivibrio. The present study concluded that CO enhanced fermentation kinetics while decreasing enteric in vitro CH4 production from fibrous diets. Thus, it may be considered as a potentially effective and environmentally friendly way of mitigating CH4 emission from livestock.

This study was aimed to evaluate the effect of a mixed phytogenic (MP) on rumen bacteria and their potential association with rumen fermentation and milk yield parameters in water buffaloes. Twenty Murrah buffaloes were fed a basal diet (consisting of maize silage, brewers' grains, and concentrate mixture) for 6 weeks supplemented with 0 (control), 15 (MP15), 25 (MP25), and 35 (MP35) g of mixed phytogenic/buffalo per d. The mixed phytogenic contained fennel (seeds), ajwain (seeds), ginger (tubers), Swertia chirata (leaves), Citrullus colocynthis (fruit), turmeric, fenugreek (seeds), Terminalia chebula (fruit), licorice (roots), and Phyllanthus emblica (fruit) in equal quantities. After 2 weeks of adaptation, daily milk yield, and weekly milk composition were recorded. On the last day of the experiment (d 42), rumen contents were collected to determine rumen fermentation parameters and bacterial diversity through 16S rRNA sequencing. Results revealed no change in dry matter intake, milk yield and rumen fermentation parameters except pH, which increased (P = 0.029) in response to MP supplementation. The mixed phytogenic increased (P < 0.01) milk fatty acids (C4 to C14:0) in MP15 only. The milk C16:1 content and its unsaturation index were higher (P < 0.05) in MP35 as compared to the control and other treatments. Furthermore, C18:3n3 was higher (P < 0.05) in the control, MP15, and MP25, as compared to MP35. Supplementation of MP tended to increase (P = 0.095) the Shannon index of bacterial alpha diversity and a difference (P < 0.05) among treatment groups was observed in beta diversity. Feeding MP increased the Firmicutes, Proteobacteria, and Spirochaetes but decreased Bacteroidetes numerically. In addition, the dominant genus Prevotella decreased in all treatment groups while Pseudobutyrivibrio, Butyrivibrio, and Succinivibrioanceae increased numerically in MP25 and MP35. The mixed phytogenic promoted groups of rumen bacteria positively associated with milk and fat yield. Overall, our study revealed 14 positive correlations of rumen bacteria with milk yield and eight with rumen fermentation parameters. Our findings reveal substantial changes in the rumen bacteriome composition and milk fatty acid content in response to MP but these results should be interpreted carefully, as the sample size of our study was relatively small.

FN-III proteins are widely distributed in mammals and are usually involved in cellular growth, differentiation, and adhesion. The FNDC5/irisin regulates energy metabolism and is present in different tissues (liver, brain, etc.). The present study aimed to investigate the physiochemical characteristics and the evolution of FN-III proteins and FNDC5/irisin as a ligand targeting the gonadal receptors including androgen (AR), DDB1 and CUL4 associated factor 6 (DCAF6), estrogen-related receptor β (ERR-β), estrogen-related receptor γ (ERR-γ), Krüppel-like factor 15 (KLF15), and nuclear receptor subfamily 3 group C member 1 (NR3C1). Moreover, the putative role of irisin in folliculogenesis and spermatogenesis was also elucidated. We presented the molecular structure and function of 29 FN-III genes widely distributed in the buffalo genome. Phylogenetic analysis, motif, and conserved domain pattern demonstrated the evolutionary well-conserved nature of FN-III proteins with a variety of stable to unstable, hydrophobic to hydrophilic, and thermostable to thermo-unstable properties. The comparative structural configuration of FNDC5 revealed amino acid variations but still the FNDC5 structure of humans, buffalo, and cattle was quite similar to each other. For the first time, we predicted the binding scores and interface residues of FNDC5/irisin as a ligand for six representative receptors having a functional role in energy homeostasis, and a significant involvement in folliculogenesis and spermatogenesis in buffalo.

This study evaluated the effect of methionine on in vitro methane (CH4) production, rumen fermentation, amino acid (AA) metabolism, and rumen microbiota in a low protein diet. We evaluated three levels of methionine (M0, 0%; M1, 0.28%; and M2, 1.12%) of in the presence of sodium nitrate (1%) in a diet containing elephant grass (90%) and concentrate (10%). We used an in vitro batch culture technique by using rumen fluid from cannulated buffaloes. Total gas and CH4 production were measured in each fermentation bottle at 3, 6, 9, 12, 24, 48, 72 h of incubation. Results revealed that M0 decreased (p < 0.001) the total gas and CH4 production, but methionine exhibited no effect on these parameters. M0 decreased (p < 0.05) the individual and total volatile fatty acids (VFAs), while increasing (p < 0.05) the ruminal pH, acetate to propionate ratio, and microbial protein content. Methionine did not affect ruminal AA contents except asparagine, which substantially increased (p = 0.003). M2 increased the protozoa counts, but both M0 and M1 decreased (p < 0.05) the relative abundance of Firmicutes while increasing (p < 0.05) the Campilobacterota and Proteobacteria. However, Prevotella and γ-Proteobacteria were identified as biomarkers in the nitrate group. Our findings indicate that methionine can increase ruminal asparagine content and the population of Compylobactor.

Heat stress results in serious performance losses and adversely affects animal health and welfare under various production systems. This study was conducted to develop a thermal comfort model for lactating buffaloes under hot and humid climate. Twenty Nili-Ravi buffaloes were randomly enrolled for this one-year study. Physiological parameters including rectal temperature (RT), respiratory rate (RR), and body surface temperature (BST) and environmental variables such as wet bulb temperature (WBT), dew point temperature (DPT), and black globe temperature (BGT) were recorded twice a week on each Tuesday and Thursday (n = 1602 and 1560, respectively) at 8:00 am and 2:30 pm. Moreover, ambient temperature (AT, °C) and relative humidity (RH, %), at an interval of every 30 min were recorded. We used a typical correlation analysis to build the index models for thermal comfort. The results revealed that AT positively correlated with BGT, WBT, DPT, BST, RT, and RR, while RH negatively correlated with RT. Moreover, a physiological index model consisting of BST, RT and RR (P1 = 0.578 × BST + 0.047 × RT + 0.429 × RR) and an environmental index model (E1 = 0.881 × AT + 0.194 × RH + 0.455 × BGT − 0.347 × WBT + 0.032 × DPT) proved to be a more accurate index as a pair to reveal the state of thermal comfort in lactating buffaloes. Moreover, these models correlated well with physiological variables, indicating that this this pair of index models can be used to effectively evaluate the thermal comfort in buffaloes.

Curcumin (a polyphenolic compound in turmeric) is famous for its potent anti-inflammatory, anti-oxidant, and anti-cancer properties, and has a great potential to act as an epigenetic modulator. The epigenetic regulatory roles of curcumin include the inhibition of DNA methyltransferases (DNMTs), regulation of histone modifications via the regulation of histone acetyltransferases (HATs) and histone deacetylases (HDACs), regulation of microRNAs (miRNA), action as a DNA binding agent and interaction with transcription factors. These mechanisms are interconnected and play a vital role in tumor progression. The recent research has demonstrated the role of epigenetic inactivation of pivotal genes that regulate human pathologies such as cancers. Epigenetics helps to understand the mechanism of chemoprevention of cancer through different therapeutic agents. In this regard, dietary phytochemicals, such as curcumin, have emerged as a potential source to reverse epigenetic modifications and efficiently regulate the expression of genes and molecular targets that are involved in the promotion of tumorigenesis. The curcumin may also act as an epigenetic regulator in neurological disorders, inflammation, and diabetes. Moreover, curcumin can induce the modifications of histones (acetylation/deacetylation), which are among the most important epigenetic changes responsible for altered expression of genes leading to modulating the risks of cancers. Curcumin is an effective medicinal agent, as it regulates several important molecular signaling pathways that modulate survival, govern anti-oxidative properties like nuclear factor E2-related factor 2 (Nrf2) and inflammation pathways, e.g., nuclear factor kappa B (NF-κB). Curcumin is a potent proteasome inhibitor that increases p-53 level and induces apoptosis through caspase activation. Moreover, the disruption of 26S proteasome activity induced by curcumin through inhibiting DYRK2 in different cancerous cells resulting in the inhibition of cell proliferation opens up a new horizon for using curcumin as a potential preventive and treatment approach in proteasome-linked cancers. This review presents a brief summary of knowledge about the mechanism of epigenetic changes induced by curcumin and the potential effects of curcumin such as anti-oxidant activity, enhancement of wound healing, modulation of angiogenesis and its interaction with inflammatory cytokines. The development of curcumin as a clinical molecule for successful chemo-prevention and alternate therapeutic approach needs further mechanistic insights.

The success of the aquafeed industry mainly depends on the availability of raw ingredients with high nutritional value, such as fishmeal (FM). However, the increased demand for FM elevates its prices and leads to high feed costs. Thus, there is an urgent need to find suitable alternatives for FM in fish diets to achieve sustainability in aquaculture. Currently, attention is being paid to the possibility of using insect meals as FM substitutes in aquafeed because of their relatively high nutritional quality. TM is one of those insects that can be regarded as a unique candidate because of its relatively high nutritional value. TM are rich sources of essential amino acids (methionine), lipids, and fatty acids, which vary based on the developmental stage of the worms. Although TM have an abundant amount of chitin as a fiber source and other anti-nutritional factors, numerous studies have investigated the efficacy of partial or complete substitution of FM by T. molitor in fish diets. In this context, we reviewed the current research findings on the achievable inclusion levels of T. molitor versus FM substitution in the diets of several finfish and shellfish species. We discussed the potential use of T. molitor as an FM substitute in fish diets and evaluated its effects on growth, biometric indices, and body composition. Besides, the hematological parameters, immunological responses, antioxidative efficacy, intestinal health status, and sensory criteria of fish fed T. molitor-based diets were also assessed.

The buffalo was domesticated around 3000–6000 years ago and has substantial economic significance as a meat, dairy, and draught animal. The buffalo has remained underutilized in terms of the development of a well-annotated and assembled reference genome de novo. It is mandatory to explore the genetic architecture of a species to understand the biology that helps to manage its genetic variability, which is ultimately used for selective breeding and genomic selection. Morphological and molecular data have revealed that the swamp buffalo population has strong geographical genomic diversity with low gene flow but strong phenotypic consistency, while the river buffalo population has higher phenotypic diversity with a weak phylogeographic structure. The availability of recent high-quality reference genome and genotyping marker panels has invigorated many genome-based studies on evolutionary history, genetic diversity, functional elements, and performance traits. The increasing molecular knowledge syndicate with selective breeding should pave the way for genetic improvement in the climatic resilience, disease resistance, and production performance of water buffalo populations globally.

Leaf biomass from the mulberry plant (genus Morus and family Moraceae) is considered a potential resource for livestock feeding. Mulberry leaves (MLs) contain high protein (14.0–34.2%) and metabolizable energy (1130–2240 kcal/kg) with high dry matter (DM) digestibility (75–85%) and palatability. Flavonoid contents of MLs confer unique antioxidant properties and can potentially help alleviate oxidative stress in animals during stressful periods, such as neonatal, weaning, and periparturient periods. In addition, mulberry leaf flavonoids (MLFs) possess antimicrobial properties and can effectively decrease the population of ruminal methanogens and protozoa to reduce enteric methane (CH4) production. Owing to its rich flavonoid content, feeding MLs increases fiber digestion and utilization leading to enhanced milk production in ruminants. Dietary supplementation with MLFs alters ruminal fermentation kinetics by increasing total volatile fatty acids, propionate, and ammonia concentrations. Furthermore, they can substantially increase the population of specific cellulolytic bacteria in the rumen. Owing to their structural homology with steroid hormones, the MLFs can potentially modulate different metabolic pathways particularly those linked with energy homeostasis. This review aims to highlight the potential of ML and its flavonoids to modulate the ruminal microbiome, fermentation, and metabolic status to enhance productive performance and health in ruminants while reducing CH4 emission.

hnRNP A/B proteins modulate the alternative splicing of several mammalian and viral pre-mRNAs, and are typically viewed as proteins that enforce the activity of splicing silencers. Here we show that intronic hnRNP A/B-binding sites (ABS) can stimulate the in vitro splicing of pre-mRNAs containing artificially enlarged introns. Stimulation of in vitro splicing could also be obtained by providing intronic ABS in trans through the use of antisense oligonucleotides containing a non-hybridizing ABS-carrying tail. ABS-tailed oligonucleotides also improved the in vivo inclusion of an alternative exon flanked by an enlarged intron. Notably, binding sites for hnRNP F/H proteins (FBS) replicate the activity of ABS by improving the splicing of an enlarged intron and by modulating 5' splice-site selection. One hypothesis formulated to explain these effects is that bound hnRNP proteins self-interact to bring in closer proximity the external pair of splice sites. Consistent with this model, positioning FBS or ABS at both ends of an intron was required to stimulate splicing of some pre-mRNAs. In addition, a computational analysis of the configuration of putative FBS and ABS located at the ends of introns supports the view that these motifs have evolved to support cooperative interactions. Our results document a positive role for the hnRNP A/B and hnRNP F/H proteins in generic splicing, and suggest that these proteins may modulate the conformation of mammalian pre-mRNAs.

Summary: Camel milk and its products are gaining higher demand due to their attractive nutritional and therapeutic profile. However, manufacturing of camel milk cheese (CMC) is a challenging task. This study aimed to compare the quality of camel milk cheese made using different starter cultures. Four groups CM (camel milk + mesophilic cultures), CT (camel milk + thermophilic cultures), CBM (camel milk + 10% buffalo milk using mesophilic cultures), and CBT (camel milk + 10% buffalo milk using thermophilic cultures) were created. The cheeses were stored for 60 days at 4 °C to assess their physicochemical, sensory, and proteolytic properties. Results indicated highest moisture content in CM (70.19%) as opposed to the lowest moisture in CBT (55.19%). In addition, CM and CBM revealed lower acidity (0.63% and 0.66%) in contrast with CT and CBT (0.78% and 0.83%, respectively). On the other hand, CBM and CBT exhibited higher contents of protein (21.04, 21.57%) and fat (17.65, 17.70%) during storage. All quality indices of cheese samples significantly declined (P < 0.01) during storage excluding acidity. Sensory analyses revealed that panellists highly preferred CBT treatment followed by CBM and CT. Furthermore, urea‐PAGE and RP‐HPLC were performed for the proteolytic study of cheese samples. The study revealed that the presence of buffalo milk and thermophilic cultures in cheese resulted in a decrease concentration of intact caseins (αs1, β‐, and κ‐caseins) as storage days passed. The degree of proteolysis was found to be higher in CBT compared to CBM, CT, and CM. Based on the results, it was concluded that the use of pasteurisation at 65 °C for 30 min, pH of 5.5, 0.06% CaCl2, and the incorporation of 10% buffalo milk using thermophilic cultures (Lactobaccilus bulgaricus and Streptococcus thermophilus) resulted in desirable quality characteristics. [ABSTRACT FROM AUTHOR]