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Silver nanodisks (AgNDs) have been successfully synthesized by using ferric chloride as an auxiliary agent in the presence of polyvinylpyrrolidone and

This experiment was conducted to study the regularity influence in apparent digestibility, rumen fermentation parameters, and relative bacterial abundance in buffaloes with different concentrate to roughage ratios. Three adult female barren buffaloes with permanent rumen fistulas were fed five diets with concentrate to roughage ratios of 20:80, 35:65, 50:50, 65:35, and 80:20 according to an incomplete Latin square design of 3×5, respectively. The pre-feeding period of each period was 12 d. From day 13 to 15 of the experiment, the feed intake of each buffalo was accurately recorded and all feces were collected. Samples of diet and feces were collected for nutrient analysis. Rumen fluid was collected to determine rumen fermentation parameters, and rumen bacteria were analyzed by 16S rRNA sequencing. After 2 hours of feeding in the morning on the 15th day of the experiment, rumen bacteria were analyzed. The daily gain and DMI (G/kgW0.75) reached the highest at the concentrate to roughage ratio of 50:50, and the apparent digestibility of ADF NDF NDICP ADICP reached the highest at the concentrate to roughage ratio of 35:65. The weight loss of the experimental animals when the concentrate to roughage was 80:20. Rumen pH decreased with the increase of concentrate level, starch content and NFC content. The rumen ammonia nitrogen concentration increased with the increase in the concentrate to roughage ratio and protein content. The concentration of total volatile fatty acids (mmol/L) in the rumen decreased with the increase of NDF but increased with the increase of NFC and CP, and the acetic acid proportion increased with the increase of NDF. There were differences in the relative abundance of rumen bacterial microbiota with the different concentrate to roughage ratios. Moreover, the relative abundance of each bacterial microbiota changed regularly with the change of concentrate to roughage ratio. The relative abundance of Bacteroidetes, Firmicutes, and other rumen core microbiota varied linearly or nonlinearly with the change of concentrate to roughage ratio. This suggests that the relative abundance of the ratio of rumen microbiota can be used to specify or measure rumen health and subacute rumen acidosis/rumen acidosis.

Buffalo milk is nutrient-rich and contains less cholesterol than cow milk. Dairy buffaloes are widely distributed at different altitudes in the Yunnan Province, China; however, the impacts of altitude on the whey-derived functional metabolites of buffalo milk whey are not well understood. Here, we used non-targeted and targeted metabolomics to evaluate the differential metabolites in the milk whey of buffaloes raised at low altitudes (LA), medium altitudes (MA), and high altitudes (HA). ANOVA statistical test was performed to acquire differential metabolites using IBM SPSS statistics 22 software. The results showed that LA- and MA-milk whey had higher levels of amino acids (glutamine and pyroglutamic acid) and vitamin B6 than HA-milk whey. LA-milk whey had higher levels of the carbohydrates involved in galactose, amino sugar, and nucleotide sugar metabolism than MA- and HA-milk whey, but HA-milk whey showed significantly higher levels of free fatty acids. In conclusion, owing to the biological functions of their most abundant components, LA-milk is more suitable for the production of functional milk with high levels of amino acids, vitamin B6, and carbohydrates; while HA-milk is suitable as raw milk for the production of dairy products with high free fatty acid content.

Synthesis of silver nanowires (Ag NWs) has been studied for decades. However, Ag NWs with diameters below 20 nm synthesised by simple and robust approaches are still rarely reported. In this work, Ag NWs with an average diameter of ∼15 nm and an aspect ratio of over 1000 have been prepared by using a Grignard reagent 5-chloro-2-thienylmagnesium bromide as an assistant additive. In particular, the presence of 5-chloro-2-thienylmagnesium bromide is proven to be beneficial for the in situ formation of smaller AgBr and AgCl particles step by step in comparison with traditional inorganic halide ions, and then promote the final growth of ultrafine Ag NWs.

Objective: The objective of the present experiment was to construct self-draining beds to keep surface bedding materials clean and dry for beef cattle comfort in a temperate climate.Methods: In Experiment 1, a self-draining bed was covered with sand at depths of 10 cm (S-10a), 15 cm (S-15), and 20 cm (S-20) respectively. In Experiment 2, self-draining beds of different sizes were covered with 10 cm of sand (S-10b) and wood shavings (WS) at depths of 15 cm and 20 cm (WS-15 and WS-20). Fifteen cattle were engaged to evaluate the comfort of self-draining beds covered with different bedding materials.Results: No cattle lay in the feed alley and cattle spent more time lying on S-10a than S-15 or S-20 in Experiment 1 (p0.05). In Experiment 2, no cattle selected the feed alley as the lying area. Cattle preferred WS-15 as the lying area and time spent lying on WS-20 was slightly higher than on S-10b (p

The yak (Bos grunniens) is a ruminant animal with strong regional adaptability. However, little is known about the adaptation of the rumen microbial community of yaks at different altitudes and the adaptation mechanism of the host and intestinal microorganisms to the habitat. We investigated the adaptability of the rumen microorganisms of yaks at high and low altitudes. We also compared and analyzed the abundance and diversity of core microorganisms and those that varied between different animals. The aim was to compare the rumen bacterial and fungal communities of grazing yak living at two elevations. Bacteroidetes, Firmicutes, Ascomycota, and Chytridiomycota were the dominant bacteria in the plateau and low-altitude regions. Significant differences between the dominant microorganisms in the rumen of yaks were evident in the two regions. The proportion of fiber-degrading bacteria was significantly different between yaks dwelling at high-altitude and low-altitude regions. The abundance of starch-degrading bacteria was not significantly different with altitude. Species clustering similarity analysis showed that the rumen microorganisms in the two areas were obviously isolated and clustered into branches. Functional prediction showed significant differences in rumen microbial methane metabolism, starch and sucrose metabolism, ion-coupled transporter and bacterial secretion system at different altitudes. Overall, the results of this study improved our understanding of the abundance and composition of microorganisms in the rumen of yak at different altitudes.

There are many kinds of microorganisms in the gastrointestinal tract of mammals, some of which are closely related to the host. Rumen microorganisms are essential for normal physiological activities of their host by decomposing plant crude lignin and providing essential nutrients. The composition and diversity of this microbial population are influenced by the host, environment, and diet. Despite its importance, little is known about the effects of factors such as altitude variation on rumen microbial population abundance and diversity in different ruminants. Here, we described the changes in overall rumen bacteria in four groups of cattle, including the Zhongdian yellow cattle and Zhongdian yaks, grazing at high altitudes (3600 m); the Jiangcheng yellow cattle and Jiangcheng buffalo were kept at an altitude of 1100 m. We found that there was a significant difference in rumen bacterial abundance of the Zhongdian yellow cattle and Zhongdian yaks at high altitude and there was obvious homogeneity in rumen bacterial abundance and diversity in the Jiangcheng yellow cattle and Jiangcheng buffalo at low altitude. Therefore, our research concluded that under the same dietary environment, there were differences in the abundance and diversity of certain bacteria in the rumen of different breeds of cattle, indicating that host genetic factors and intestinal microorganisms related to altitudinal variation had a greater influence on rumen bacterial abundance in the cattle.

Herein, silica carrier was used as underlying structure to prepare composite material loaded with rare earth element Er and Zn. Rare earth elements can improve antimicrobial effects of ZnO due to their specific electronic structure. Er–ZnO/SiO2 hybrid antibacterial material was prepared through sol-gel method and its structure and morphology were characterized by scanning electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, inductively coupled plasma emission spectroscopy and Brunauer-Emmett-Teller measurements. E. coli and S. aureus were selected as model bacteria to assess antibacterial activity of prepared hybrid material by plate coating method. Er–ZnO/SiO2 exhibited good antibacterial activity towards E. coli and S. aureus. Increase in Er3+ concentration from 0.12% to 1.10% led to increase in antibacterial performance followed by subsequent decrease. Improving effect of Er relied on the molar ratio of Er doped in ZnO/SiO2 hybrid material. The optimal sample was found to be 0.60%Er–ZnO/SiO2, with antibacterial rates of 93.71% and 70.46% against E. coli and S. aureus, respectively. Antibacterial mechanism was assessed by fluorescence detection of reactive oxygen species. In addition, flame atomic absorption spectrometry was used to measure the amount of released Zn2+. Results also showed that 0.60%Er–ZnO/SiO2 hybrid material generated more reactive oxygen species, released more Zn2+ ions, and had the largest surface area, which improved its antibacterial rate. Thus, Er enhanced antibacterial properties of ZnO/SiO2, providing these composite materials with great potential as antibacterial products.

Gayal (Bos frontalis) of the Yunnan region is well adapted to harsh environmental conditions. Its diet consists predominantly of bamboo, reeds, and woody plants, suggesting that the rumen of this species contains many fiber-degrading bacteria and cellulases. The aim of this study was to identify and modify specific cellulases found in the gayal rumen. In the present study, a directed evolution strategy of error-prone PCR was employed to improve the activity or optimal temperature of a cellulase gene (CMC-1) isolated from gayal rumen. The CMC-1 gene was heterologously expressed in Escherichia coli (E. coli) BL21, and the recombinant CMC-1 protein hydrolyzed carboxyl methyl cellulose (CMC) with an optimal activity at pH 5.0 and 50 °C. A library of mutated ruminal CMC-1 genes was constructed and a mutant EP-15 gene was identified. Sequencing analysis revealed that EP-15 and CMC-1 belonged to the glycosyl hydrolase family 5 (GHF5) and had the highest homology to a cellulase (Accession No. WP_083429257.1) from Prevotellaceae bacterium, HUN156. There were similar predicted GH5 domains in EP-15 and CMC-1. The EP-15 gene was heterologously expressed and exhibited cellulase activity in E. coli BL21 at pH 5.0, but the optimum temperature for its activity was reduced from that of CMC-1 (50 °C) to 45 °C, which was closer to the physiological temperature of the rumen (40 °C). The cellulase activity of EP-15 was about two times higher than CMC-1 at 45 °C or PH 5.0, and also was more stable in response to temperature and pH changes compared to CMC-1. This study successfully isolated and modified a ruminal cellulase gene from metagenomics library of Yunnan gayal. Our findings may obtain a useful cellulase in future applications and present the first evidence of modified cellulases in the gayal rumen.

With the continuous improvement in medical science in modern times, the spread of bacterial infection has become a matter of global concern. Therefore, the search for biological medical materials with antibacterial function has become a focus of intense research. In this work, pure SnO2 and Ag-doped SnO2 hollow nanofibers were fabricated by a combination of an electrospinning method and a calcination procedure, and the effects of the doped Ag on antibacterial activity and catalytic antibacterial activity were subsequently investigated. Through the process of high-temperature calcination, a high heating rate would lead to the formation of a hollow tubular structure in SnO2 fibers, and Ag2O would be reduced to Ag0 by a facile process with appropriate thermal treatment. Additionally, the existence of SnO2 as a tetragonal rutile structure was confirmed. On the basis of pure SnO2, doping with Ag greatly improved the antibacterial activity and catalytic antibacterial activity of hollow nanofibers. The formation mechanism and the antibacterial mechanism of pure SnO2 and Ag-doped hollow nanofibers are also discussed. This study has broad application prospects for biological medicine.

The gut microbial community of mammals, especially ruminants, plays an important role at different ages of the host. Ruminants have a unique compartment in their digestive tract; the rumen, comprising microorganisms that can effectively decompose plant fibers for the host to transform into milk and for growth and development, which is important for meat production. Colonization of rumen microorganisms is closely related to host developmental stage and affects host performance production. There is little information regarding initial colonization and subsequent changes of the rumen microbial population in wild grazing animals, from birth to adulthood. This study investigated the rumen bacterial and fungal populations of grazing yaks in five experimental groups, ranging from a few days after birth to adulthood using amplicon sequencing. Results indicated that rumen microbial communities of these yaks undergo a gradual change from 5 to 180 days after birth, with the bacterial and fungal diversity stabilizing at the age of 2 years. Additionally, Ruminococcus was detected in 5-day-old yak rumens, with a high percentage of Penicillium and other microbial species are important for normal rumen function detected in the adult rumen. The changes to the yak rumen microbial community after birth were reflected in the increased anaerobic fiber degradation group, and decreased aerobic and facultative anaerobic bacteria. Microbial diversity and abundance in the yak rumen increased with age. Rumen microbial composition of 6-month and 2-year-old yaks had obvious homogeneity. There were some differences in dominant rumen microorganisms among the different age groups. Further studies are required to confirm the functions of these differential and dominant microorganisms in each age group.

Tibetan pigs, indigenous to Tibetan plateau, are well adapted to hypoxia. So far, there have been not any definitively described genes and functional sites responsible for hypoxia adaptation for the Tibetan pig. The whole genome-wide association studies in human suggested that genetic variations in TMPRSS6 was associated with hemoglobin concentration (HGB) and red cell counts (RBC). Here we conducted resequencing of the nearly entire genomic region (40.1 kb) of the candidate gene TMPRSS6 in 40 domestic pigs and 40 wild boars along continuous altitudes and identified 708 SNPs, in addition to an indel (CGTG/----) in the intron 10. We conduct the CGTG indel in 838 domestic pigs, both the CGTG deletion frequency and the pairwise r2 linkage disequilibrium showed an increase with elevated altitudes, suggesting that TMPRSS6 has been under Darwinian positive selection. As the conserved core sequence of hypoxia-response elements (HREs), the deletion of CGTG in Tibetan pigs decreased the expression levels of TMPRSS6 mRNA and protein in the liver revealed by real-time quantitative PCR and western blot, respectively. We compared domestic pigs and Tibetan pigs living continuous altitudes, found that the blood-related traits with the increase of altitude, however, the HGB did not increase with the elevation in Tibetan pigs. Genotype association analysis results dissected a genetic effect on reducing HGB by 13.25 g/L in Gongbo'gyamda Tibetan pigs, decreasing mean corpuscular volume (MCV) by 4.79 fl in Diqing Tibetan pigs. In conclusion, the CGTG deletion of TMPRSS6 resulted in lower HGB and smaller MCV, which could reflect a blunting erythropoiesis and improving blood viscosity as well as erythrocyte deformability. It remains to be determined whether a blunting of erythropoiesis for TMPRSS6 or others genetic effects are the physiological adaptations among Tibetan pigs.

Silica microspheres with narrow particle size distribution and average diameter of 80–200 nm were prepared by hydrolysis and condensation of tetraethylorthosilicate (TEOS) in ethanol solution. Effects of TEOS, ammonia, water concentration, and temperature on particle size and morphology were investigated by laser particle size analysis and field emission scanning electron microscopy (FESEM), in conjunction with energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) spectroscopy. Monomer addition model was employed to analyze the stability of intermediate, as well as the direction in which the reaction of silica particles proceeds. The analysis was carried out by Materials Studio. Results demonstrated that particle size of Nano-silica increases with increasing concentrations of H2O, NH3·H2O, and TEOS in ethanol, whereas the size decreases with increasing temperature. In addition, when proper amount of H2O was added into NH3·H2O at suitable temperature, silicic acid can act as a nucleus for amorphous Nano-silica particles, forming microspheres with round and smooth surfaces. By contrast, when TEOS was used, resultant nanoparticles have poor surfaces. During nucleation process of Nano-silica, silicic acid can also acts as nucleating agent providing a platform for the growth of nanoparticles with symmetric structure. Findings further indicated that the reaction proceeds by first silicic acid participates in the reaction, and the dimer and trimer molecules then react with the surface of silicic acid molecules; the same products could be produced by different reactions.

The selection of raw milk with high levels of functional components that have health-promoting activities is very important for the exploitation and production of functional milk, but the differences in the functional components of whey from buffalo raised at different altitudes have not been thoroughly investigated. Here, we detected the effects of altitudes on the functional components in whey from dairy buffalo farms situated at low altitude (LA), medium altitude (MA), and high altitude (HA) sites with data-independent acquisition proteomic approaches. In 33 samples, 9331 peptides corresponding to 1008 high-confidence proteins were detected. HA-whey had a lower level of angiotensinogen than that of the LA- and MA-whey, and conversely contained higher levels of immune-enhancing components than for the latter two groups. Differential proteins were involved in vascular smooth muscle contraction, complement and coagulation cascades, and the secretion, production and regulation pathways in immune components. LA-whey showed higher levels of lymphocyte antigen and selenoprotein F than that of the HA-whey. Owing to the biological functions of their most abundant components, HA- and LA-whey are suitable for the processing of functional milk for lowering blood pressure, and the production of immune milk, respectively.

Background Comfortable beds play an important role in increasing the ruminant and milk production efficiency of dairy buffalo. In loose housing systems, cow lying comfort depends on both the bedding materials and bed base. Results Buffaloes spent more lying time on sand beds at depth of 15 and 20 cm (S-15 and S-20) than on beds of 10 cm (S-10a) beds or in the feed alley in Exp1 (P P P Conclusion A depth of sand or wood shavings at 15 cm can meet the lying comfort requirements of dairy buffaloes when bedding materials are used above drainage beds.

Twelve representative buffalo were selected from 22 suckling calves, 41 weaned calves, 57 reserve bulls and 20 adult bulls for slaughter. The study aims to assess the effect of age on dressing percentage, meat percentage and carcass meat yield and physico-chemical properties of longissimus dorsi and biceps femoris, and to evaluate the correlation between live weight and marbling, backfat thickness, rib eye area. The results showed that the slaughter performance and meat quality of Binlangjang male buffalo showed an obvious change with age. The dressing percentage decreased from 54.93% to 51.22% with the increase of age, while meat percentage and carcass meat yield increased gradually with age, which were 34.58–38.59%, 62.95–75.34%; Marbling, backfat thickness and rib eye area increased with age, and there was significant difference between the situation before 3 months and after 12 months of age (P 0.05). The levels of fat, protein, cholesterol and inosine acid were significantly different before 3 months of age from those after 12 months (P

A total of 6,219 positive clones were obtained by constructing a BAC library of uncultured ruminal fungi of gayal, and two clones (xynF1 and eglF2) with lignocellulolytic enzyme activity were selected. The sequencing results showed that xynF1 and eglF2 had 903-bp, and 1,995-bp, open reading frames likely to encode β-xylanase (XynF1) and β-glucosidase (EglF2), respectively. The amino acid sequence of XynF1 had 99% coverage and 95% homology to the endo-β-1,4-xylanase encoded by the cellulase gene of Orpinomyces sp. LT-3 (GenBank accession No. AEO51791.1). The amino acid sequence of EglF2 had 99% coverage and 93% homology to the β-glucosidase encoded by the cellulase gene of Piromyces sp. E2 (GenBank accession No. CAC34952.1). Analysis using the SMART software showed that XynF1 contains a glycoside hydrolase family 11 functional module and a carbohydrate-binding module, while EglF2 contains a glycoside hydrolase family 1 functional module. XynF1 showed the highest relative enzymatic activity, up to 95%, at 45°C and pH 4.2, while EglF2 showed the highest relative enzymatic activity, up to 95%, at 55°C and pH 6.2. In this study, we achieved efficient expression of the xynF1 and eglF2 genes in Pichia pastoris, which laid a foundation for the practical application of the lignocellulolytic enzymes.

Domesticated buffaloes have been integral to rice-paddy agro-ecosystems for millennia, yet relatively little is known about the buffalo genomics. Here, we sequenced and assembled reference genomes for both swamp and river buffaloes and we re-sequenced 230 individuals (132 swamp buffaloes and 98 river buffaloes) sampled from across Asia and Europe. Beyond the many actionable insights that our study revealed about the domestication, basic physiology and breeding of buffalo, we made the striking discovery that the divergent domestication traits between swamp and river buffaloes can be explained with recent selections of genes on social behavior, digestion metabolism, strengths and milk production.

With the continuous improvement in medical science in modern times, the spread of bacterial infection has become a matter of global concern. Therefore, the search for biological medical materials with antibacterial function has become a focus of intense research. In this work, pure SnO2 and Ag-doped SnO2 hollow nanofibers were fabricated by a combination of an electrospinning method and a calcination procedure, and the effects of the doped Ag on antibacterial activity were subsequently investigated. Through the process of high-temperature calcination, a high heating rate would lead to the formation of a hollow tubular structure in SnO2 fibers, and Ag2O would be reduced to Ag0 by a facile process with appropriate thermal treatment. Additionally, the existence of SnO2 as a tetragonal rutile structure was confirmed. On the basis of pure SnO2, doping with silver greatly improved the antibacterial activity of hollow nanofibers. The formation mechanism and the antibacterial mechanism of pure SnO2 and Ag-doped hollow nanofibers are also discussed. This study has broad application prospects for biological medicine.

Zinc oxide (ZnO) is an antibacterial agent with great potential in biological applications. However, the antibacterial activity of ZnO is relatively weak. In this study, a method of doping ZnO crystals with metals was employed to improve its antibacterial activity. V2O5-ZnO@SiO2 (V-ZS) composites, made of mesoporous SiO2 as a carrier, ZnO as an antibacterial agent, and V as a doping element, were fabricated by hydrothermal method at atmospheric pressure. The morphology, crystal structure, and chemical composition of V-ZS composites were analyzed by SEM, TEM, XRD, N2 physisorption, and XPS and the results were compared with those of undoped ZnO@SiO2 composites. UV–vis and PL spectroscopies were used to investigate the enhancement effect of V2O5 on the visible light absorption and separation of photogenerated carriers. The doping of ZnO with V was found to be favorable for the release of Zn2+ from ZnO particles and promoted the generation of reactive oxygen species. The enhancement effect of V2O5 depended on the percent of V2O5 in V2O5/ZnO nanostructures. The results of antibacterial activity revealed that even at a small amount of V2O5 (8%), action of the resulting 8%V-ZS composite increased against bacteria. It was also shown that V-ZS composites can mechanically damage the bacteria. The results obtained herein sustain the use of the developed composites in antibacterial products.

Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38 400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene , which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase (GenBank accession no. YP_004310852.1) from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coli BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45°C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.

Shade shelter is widely used to reduce heat load for farm animals, but little research focus on the cooling effects for Dehong buffalo calves. The aim of this experiment was to evaluate the impacts of shade cooling on the physiological and behavioural parameters for Dehong buffalo calves. Shade with 99% blockage of solar radiation was covered on steel frame with a 2.5m height above the loafing area floor (shaded). The loafing area of the control group was the same as the shaded group, apart from no shade shelter above the loafing area (non-shaded). Twenty-four Dehong buffalo calves were randomly and averagely allocated to the non-shaded and shaded groups. Six Dehong buffalo calves were loose housed in a barn, and they had access to loafing area. The results showed that the average daily air temperature of the loafing area under shade was lower 1.5°C than that of the loafing area without shade shelter. No difference in rectal temperature was found between the non-shaded and shaded calves ( P >0.05), but respiration rate of the non-shaded group in noon (12:00h, 31.6 breaths) and evening (18:00h, 28.5 breaths) was greater than for calves in shaded group (26.1 and 25.3 breaths, respectively) ( P P P

Chronic heat stress (HS), aggravated by global warming, reduces the production efficiency of the buffalo dairy industry. Here, we conducted a proteomic analysis to investigate the adaptation strategies used by buffalo in response to heat stress. Seventeen differentially abundant proteins with known functions were detected using label-free quantification (LFQ), and five of these differentially expressed proteins were validated with parallel reaction monitoring (PRM). These five proteins were associated with various aspects of heat stress, including decreased heat production, increased blood oxygen delivery, and enhanced natural disease resistance. Lipase (LPL), glutathione peroxidase 3 (GPX3), cathelicidin-2 (CATHL2, LL-37), ceruloplasmin (CP), and hemoglobin subunit alpha 1 (HBA1) were shown to play cooperative roles in the tolerance of chronic HS in dairy buffalo. We found that high levels of HBA1 increased blood oxygen transport capacity. Our results increase our understanding of the adaptation of dairy buffalo to chronic heat stress.

Chronic heat stress (CHS) reduces the production efficiency of the buffalo dairy industry. Relatively low-abundance proteins with particular functions in biological processes are changed by CHS. The present study aimed to quantify the differences in low-abundance proteins of crossbred dairy buffaloes under CHS and thermal-neutral (TN) conditions. With label-free quantification, 344 low-abundance proteins were identified in serum. Of these, 17 differentially expressed low-abundance proteins with known functions were detected, and six of the differentially expressed proteins related to heat stress were validated with parallel reaction monitoring. Lipase (LPL), glutathione peroxidase 3 (GPX3), cathelicidin-2 (CATHL2), ceruloplasmin (CP), and hemoglobin subunit alpha 1 (HBA1) cooperatively played roles in the thermal fitness of dairy buffalo by decreasing heat production and increasing blood oxygen delivery. Also, dairy buffaloes may adapt to CHS and hypoxia with high levels of RBCs, HBA1 and CP to increase blood oxygen delivery capacity.

Heat stress (HS) threatens the worldwide dairy industry by decreasing animal production performance and health. Holstein cows and dairy buffaloes are the most important dairy animals, but their differences in the metabolic mechanism of thermotolerance remain elusive. In this study, we used serum metabolomics to evaluate the differences in thermotolerance between Holstein cows and crossbred dairy buffaloes under chronic heat stress (HS) and thermal-neutral conditions. In response to HS, the body temperatures and respiratory rates were increased more for Holstein cows than for dairy buffaloes (38.78 vs 38.24 °C, p < 0.001; 43.6 vs 32.5 breaths/min, p < 0.001). HS greatly affected serum metabolites associated with amino acids, fatty acids, and bile acids. The enriched metabolic pathways of these serum metabolites are closely related to HS. We demonstrated that buffaloes adapt to HS by adopting a metabolism of branched-chain amino acids and ketogenic amino acids and gluconeogenesis, but Holstein cows decrease the effect of HS with citrulline and proline metabolism. Both physiological parameters and serum metabolic profiles indicate that dairy buffaloes are more thermotolerant than Holstein cows, providing the feasibility to vigorously develop the buffalo dairy industry in tropical and subtropical regions.

Tibetan pigs, indigenous to Tibetan plateau, are well adapted to hypoxia. So far, there have been not any definitively described genes and functional sites responsible for hypoxia adaptation for the Tibetan pig. Here we conducted resequencing of the nearly entire genomic region (40.1 kb) of the candidate geneTMPRSS6(Transmembrane protease, serine 6) associated with hemoglobin concentration (HGB) and red blood cell count (RBC) in 40 domestic pigs and 40 wild boars from five altitudes along the Tea-horse ancient road and identified 708 SNPs, in addition to an indel (CGTG/----) in the intron 10. Both the CGTG deletion frequency and the pairwise r2linkage disequilibrium showed an increase with elevated altitudes in 838 domestic pigs from five altitudes, suggesting thatTMPRSS6has been under Darwinian positive selection. As the conserved core sequence of hypoxia-response elements (HREs), the deletion of CGTG in Tibetan pigs decreased the expression levels ofTMPRSS6mRNA and protein in the liver revealed by real-time quantitative PCR and western blot, respectively. To explore whether reducedTMPRSS6expression level could improve blood viscosity, the relationship between CGTG indel and hematologic and hemorheologic parameters in 482 domestic pigs from continuous altitudes was detected and dissected a genetic effect on reducing HGB by 13.25g/L in Gongbo’gyamda Tibetan pigs and decreasing MCV by 4.79 fl in Diqing Tibetan pigs. In conclusion, the CGTG deletion ofTMPRSS6resulted in lower HGB and smaller MCV, thereby blunting erythropoiesis and improving blood viscosity as well as erythrocyte deformability.

Figure S1. Alignments of EDN3 protein with different species. Figure S2. Prediction secondary structure of EDN3 protein. t, refers to Beta turn; c, to Random coil; h, to Alpha helices; e, to Extended strand. (DOCX 13238 kb)

Table S1. EDN3 genome sequencing screening results. Table S2. The 16 SNPs obtained from online database. OAR refers to: Ovis aries; Chr: chromosome. (DOCX 18 kb)

A total of 6,219 positive clones were obtained by constructing a BAC library of uncultured ruminal fungi of gayal, and two clones (xynF1 and eglF2) with lignocellulolytic enzyme activity were selected. The sequencing results showed that xynF1 and eglF2 had 903-bp, and 1,995-bp, open reading frames likely to encode β-xylanase (XynF1) and β-glucosidase (EglF2), respectively. The amino acid sequence of XynF1 had 99% coverage and 95% homology to the endo-β-1,4-xylanase encoded by the cellulase gene of Orpinomyces sp. LT-3 (GenBank accession No. AEO51791.1). The amino acid sequence of EglF2 had 99% coverage and 93% homology to the β-glucosidase encoded by the cellulase gene of Piromyces sp. E2 (GenBank accession No. CAC34952.1). Analysis using the SMART software showed that XynF1 contains a glycoside hydrolase family 11 functional module and a carbohydrate-binding module, while EglF2 contains a glycoside hydrolase family 1 functional module. XynF1 showed the highest relative enzymatic activity, up to 95%, at 45°C and pH 4.2, while EglF2 showed the highest relative enzymatic activity, up to 95%, at 55°C and pH 6.2. In this study, we achieved efficient expression of the xynF1 and eglF2 genes in Pichia pastoris, which laid a foundation for the practical application of the lignocellulolytic enzymes.

The first animal with black traits to be studied was the Black-boned chicken. In China, we identified a flock of sheep with black traits. In the present study, the complete coding sequences of three genes, PSPH, SNRPA1 and TPM1, of Black-boned sheep (Ovis aries) were amplified using the reverse transcription polymerase chain reaction, according to the conserved sequence information for cattle and other mammals and known highly homologous sheep ESTs. The results showed that the Black-boned sheep PSPH gene encodes a protein of 225 amino acids, which has high homology with the PSPH proteins of seven species: cattle (99%), humans (94%), rat (93%), mouse (93%), African clawed frog (77%), Atlantic salmon (70%) and zebrafish (68%). The Black-boned sheep SNRPA1 gene encodes a protein of 255 amino acids, which has high identity with the SNRPA1 proteins of four species: cattle (99%), humans (99%), mouse (97%) and chicken (91%). The Black-boned sheep TPM1 gene encodes a protein of 284 amino acids, which has ...

The Chinese yakow is the offspring of yak (Bos grunniens) and Yellow cattle (Bos taurus). The melanocortin 1receptor gene (MC1R) plays a crucial role in determining coat colour of mammals. To investigate the relationship of polymorphism of the MC1R with coat colour in the Chinese yakow, the coding sequence (CDS) and the flanking region of MC1R were sequenced from 84 Chinese yakow samples and compared with the sequences of the MC1R from other bovid species. A fragment of 1134 base pair (bp) sequences including the full CDS (954bp) and parts of the 5'- and 3'-untranslated regions (162 and 18bp, respectively) of the Chineseyakow MC1R were obtained. A total of 13 single nucleotide polymorphisms (SNPs) including 4 SNPs (T-129C, A-127C, C-106T, G-1A) in the 5'-untranslated region and 9 SNPs (C201T, T206C, C340A, C375T, T663C, G714C, C870T, G871A and T890C) in the CDS were identified, revealing high genetic variability. Four novel SNPs including T206C, G714C, C870T and T890C, which have not been reported previously in bovid species, were retrieved. Within 9 coding SNPs, C201T, C375T, T663C and C870T were silent mutations, while T206C, C340A, G714C, G871A and T890C were mis-sense mutations, corresponding to amino acid changes p.L69P, p.Q114K, p.K238N, p.A291N and p.I297T, respectively. Amino acid sequences alignment showed a more than 96% similarity with other ruminates. However, three classical bovine MC1R loci the E(D), E(+) and e were not retrieved in the Chinese yakow, indicating other genes or factors could be involved in affecting coat colour in this species.

The gene coding for insulin-like growth factor-binding protein 3 (IGFBP3) is important for regulation of growth, development and metabolism in mammals. The present investigation was conducted to study nucleotide polymorphism of the IGFBP3 in gayal (Bos frontalis) and to compare the variations with those which occur in other ruminants. A fragment of 645 base pairs of the IGFBP3 covering a part of exon 2, the complete intron 2 and exon 3 and a part of intron 3 was amplified, sequenced (n=46) and digested (n=79) with HaeIII restriction enzyme from 125 collected gayal samples. Nine single nucleotide polymorphisms (SNPs) [C14T, A122C, C137T, G144C, C155T, G213A, C279A, G334A and G460A] were identified and located in intron 2, revealing high genetic variability. The alignment of nucleotide sequences was found to be very similar to those for other bovid species. Sequencing and HaeIII digestion showed that frequency of alleles C and A [consisting of fragments of sizes 56, 64, 228, 264, 282, 298 and 497 bp (CC genotype)] was 0.96 and 0.04 for the SNP C279A. Moreover, the genotype frequency of the SNP C279A in gayal was compared with that in other ruminants and it appears that this polymorphism may be associated with low fat content and rapid growth in this rare species.

The dominant rumen bacteria in Gayals, Yaks and Yunnan Yellow Cattle were investigated using PCR-DGGE approach. The analysis of DGGE profiles, identification of dominant bands and phylogenetic analysis 16S rDNA sequences in DGGE profiles were combined to reveal the dominant bacterial communities and compared the differences between those cattle species. DGGE profiles revealed that Gayals had the most abundant dominant bacteria and the lowest similarity of intraspecies between individuals than other two cattle species. A total of 45 sequences were examined and sequence similarity analysis revealed that Gayals had the most sequences appeared to uncultured bacteria, accounting for 85.0% of the total sequences, Yaks and Yunnan Yellow Cattle had 44.4 and 68.8% uncultured bacterial sequences, respectively. According to phylogenetic analysis, the rumen dominant bacteria of Gayals were mainly phylogenetically placed within phyla firmicutes and bacteroidetes, and the known bacteria were mainly belonged to the genera Lachnospiraceae bacterium, Ruminococcus flavefaciens and Clostridium celerecrescens. Moreover, the dominant bacteria of Yaks were also mainly belonged to phyla firmicutes and bacteroidetes, and the known dominant bacteria were including Ruminococcus flavefaciens, Butyrivibrio fibrisolvens, Pseudobutyrivibrio ruminis, Schwartzia succinivorans and Clostridiales bacterium, most of them are common rumen bacteria. In addition, the dominant bacteria in Yunnan Yellow Cattle were belonged to phyla firmicutes, bacteroidetes and Actinobacteria, and the known dominant bacteria containing Prevotella sp., Staphylococci lentus, Staphylococcus xylosus and Corynebacterium casei. Present study first detected Staphylococcus lentus and Staphylococcus xylosus in the rumen of cattle.

Melanocortin-1 receptor (MC1R) plays a major role in pigmentation in many species. To investigate if the MC1R gene is associated with coat color in water buffalo, the coding region of MC1R gene of 216 buffalo samples was sequenced, which included 49 black river buffalo (Murrah and Nili-Ravi), 136 swamp buffalo (Dehong, Diandongnan, Dechang, Guizhou, and Xilin) with white and gray body, and 31 hybrid offspring of river buffalo Nili-Ravi (or Murrah) and swamp buffalo. Among the three variation sites found, SNP684 was synonymous, while SNP310 and SNP384 were nonsynonymous, leading to p.S104G and p.I128M changes, respectively. Only Individuals carrying homozygote E(BR)/E(BR) were black. The genotype and phenotype analysis of the hybrid offspring of black river buffalo and gray swamp buffalo further revealed that the river buffalo type allele E(BR) or the allele carrying the amino acid p.104S was important for the full function of MC1R. The in silico functional analysis showed that the amino acid substitutions p.G104S and p.M128I had significant impact on the function of MC1R. Above results indicate that the allele E(BR) or the allele carrying the amino acid p.104S was associated with the black coat color in buffalo.

Complete coding sequences of three Black-boned sheep (Ovis aries) genes Rab2A, Rab3A and Rab7A were amplified using reverse transcription polymerase chain reaction (RT-PCR) based on the conserved sequence information of cattle or other mammals known to be highly homologous to sheep ESTs. The Black-boned sheep Rab2A gene encodes a protein of 226 amino acids which contains the conserved putative RabL2 domain and is highly homologous to the Rab2A proteins of seven other species—cattle (96%), human (83%), Sumatran orangutan (82%), rat (81%), mouse (80%), African clawed frog (72%) and zebrafish (71%). The Black-boned sheep Rab3A gene encodes a protein of 220 amino acids that contains the conserved putative Rab3 domain and is very similar to the Rab3A proteins of four species—cattle (99%), African clawed frog (99%), Western clawed frog (98%) and zebrafish (95%). And the Black-boned sheep Rab7A gene encodes a protein of 207 amino acids that contains the conserved putative Rab7 domain and has high homology with the Rab7A proteins of six other species—human (99%), dog (99%), Sumatran orangutan (99%), zebrafish (97%), rabbit (97%) and African clawed frog (96%). Analysis of the phylogenetic tree has demonstrated that the Black-boned sheep Rab2A, Rab3A and Rab7A proteins share a common ancestor and the tissue expression analysis has shown that the corresponding genes are expressed in a range of tissues including leg muscle, kidney, skin, longissimus dorsi muscle, spleen, heart and liver. Our experiment is the first to provide the primary foundation for a further insight into these three sheep genes.