Exploring the genetic control of sweat gland characteristics in beef cattle for enhanced heat tolerance

Background Thermal stress in subtropical regions is a major limiting factor in beef cattle production systems with around$369 million being lost annually due to reduced performance.Heat stress causes numerous physiological and behavioral disturbances including reduced feed intake and decreased production levels.Cattle utilize various physiological mechanisms such as sweating to regulate internal heat.Variation in these traits can help identify genetic variants that control sweat gland properties and subsequently allow for genetic selection of cattle with greater thermotolerance.Methods This study used 2,401 Brangus cattle from two commercial ranches in Florida.Precise phenotypes that contribute to an animal's ability to manage heat stress were calculated from skin biopsies and included sweat gland area,sweat gland depth,and sweat gland length.All animals were genotyped with the Bovine GGP F250K,and BLUPF90 software was used to estimate genetic parameters and for Genome Wide Association Study.Results Sweat gland phenotypes heritability ranged from 0.17 to 0.42 indicating a moderate amount of the phenotypic variation is due to genetics,allowing producers the ability to select for favorable sweat gland properties.A weighted single-step GWAS using sliding 10 kb windows identified multiple quantitative trait loci(QTLs)explaining a significant amount of genetic variation.QTLs located on BTA7 and BTA12 explained over 1.0%of genetic variance and overlap the ADGRV1 and CCDC168 genes,respectively.The variants identified in this study are implicated in processes related to immune function and cellular proliferation which could be relevant to heat management.Breed of Origin Alleles(BOA)were predicted using local ancestry in admixed populations(LAMP-LD),allowing for identification of markers'origin from either Brahman or Angus ancestry.A BOA GWAS was performed to identify regions inherited from particular ancestral breeds that might have a significant impact on sweat gland phenotypes.Conclusions The results of the BOA GWAS indicate that both Brahman and Angus alleles contribute positively to sweat gland traits,as evidenced by favorable marker effects observed from both genetic backgrounds.Understanding and utilizing genetic traits that confer better heat tolerance is a proactive approach to managing the impacts of climate change on livestock farming.

Identification and Antibacterial Activity Study of a Terrestrial Strain of Brevibacterium aureus 431

This study isolated and purified strain 431 from an animal probiotic product.Through staining and microscopic examination,colony morphology analysis,biochemical reaction tests,and 16S rDNA sequence alignment,the strain was identified and named Brevibacterium aureus 431.The study focused on the production of biosurfactants by strain 431,and antibacterial activity tests were conducted on the strain and its secondary metabolites.The results showed that strain 431 exhibited no resistance to 10 commonly used drugs,and its concentrated secondary metabolites were highly sensitive to the indicator bacterium Escherichia coli.Oral administration of strain 431 to BALB/c mice resulted in normal mental state,diet,and bowel movements,with no signs of illness or death,indicating that strain 431 is highly safe and non-pathogenic to mice.The study suggests that Brevibacterium aureus 431 has significant research value as a new source of actinomycetes and that its secondary metabolites have potential application value in the development of antibacterial drugs.

Exploration of Ideological and Political Education in Veterinary Microbiology Experimental Technology

The veterinary microbiology experimental course plays a key role in education and practice,integrating“knowledge,skills,and qualities”into the talent cultivation program.The syllabus has been revised to emphasize the course’s ideological and political education objectives.The extraction of ideological and political elements focuses on developing a curriculum that includes ideological and political teaching materials and cases,ensuring coherence across classroom teaching,social practice,and online courses.By organically integrating ideological and political elements within stories,along with professional knowledge,operational skills,and social practice,the course aims to enhance the appeal and effectiveness of ideological and political education in the experimental course.

Robust identification of regulatory variants(eQTLs)using a differential expression framework developed for RNA‑sequencing

Background A gap currently exists between genetic variants and the underlying cell and tissue biology of a trait,and expression quantitative trait loci(eQTL)studies provide important information to help close that gap.However,two concerns that arise with eQTL analyses using RNA-sequencing data are normalization of data across samples and the data not following a normal distribution.Multiple pipelines have been suggested to address this.For instance,the most recent analysis of the human and farm Genotype-Tissue Expression(GTEx)project proposes using trimmed means of M-values(TMM)to normalize the data followed by an inverse normal transformation.Results In this study,we reasoned that eQTL analysis could be carried out using the same framework used for dif-ferential gene expression(DGE),which uses a negative binomial model,a statistical test feasible for count data.Using the GTEx framework,we identified 35 significant eQTLs(P<5×10^(–8))following the ANOVA model and 39 significant eQTLs(P<5×10^(–8))following the additive model.Using a differential gene expression framework,we identified 930 and six significant eQTLs(P<5×10^(–8))following an analytical framework equivalent to the ANOVA and additive model,respectively.When we compared the two approaches,there was no overlap of significant eQTLs between the two frameworks.Because we defined specific contrasts,we identified trans eQTLs that more closely resembled what we expect from genetic variants showing complete dominance between alleles.Yet,these were not identified by the GTEx framework.Conclusions Our results show that transforming RNA-sequencing data to fit a normal distribution prior to eQTL analysis is not required when the DGE framework is employed.Our proposed approach detected biologically relevant variants that otherwise would not have been identified due to data transformation to fit a normal distribution.

Diagnosis and Prevention of Glasserella parasuis

This paper introduced the characteristics of Glasserella parasuis from the aspects of pathogen,epidemiology,clinical symptoms and anatomical symptoms,and put forward its clinical and laboratory diagnosis methods.Moreover,the disease was differentiated from similar diseases such as mycoplasmal pneumonia of swine,swine flu infectious pleuropneumonia of swine,swine streptococcosis and porcine reproductive and respiratory syndrome.Finally,the prevention and treatment measures of the disease are proposed.

Molecular epidemiology of piliated pneumococcal isolates at a major tertiary hospital in the Klang Valley,Malaysia

Objective:To characterise a collection of pili-carrying and none pili-carrying pneumococcal isolates of clinical origin for serotypes,antibiotic resistance and genotype.Methods:In total,42 clinical isolates were collected between October 2017 and December 2019.Those isolates were analysed for antimicrobial susceptibility,serotype distribution,detection of pneumococcal virulence and pilus genes.Multilocus sequence typing was performed only for piliated isolates,followed by phylogenetic analysis.Results:The common isolation sites among the pneumococcal isolates were tracheal aspirate(28.6%),blood(26.2%),and sputum(23.8%).Fifty percent isolates were resistant to erythromycin,tetracycline(50.0%)and trimethoprim-sulfamethoxazole(43.0%).The most frequent were serotypes 19F(28.6%),6A/B(23.8%)and 19A(14.3%).Piliated isolates were detected in a small proportion(33.3%);64.3%were multidrug-resistant.ST320 was the prevalent sequence type among the piliated isolates and genetically related to the Pneumococcal Molecular Epidemiology Network clones Taiwan19F-14(CC271).In the phylogenetic analysis,some piliated isolates showed a close association having similar ST320,carrying serotype 19A and both pilus genes indicating their clonal spread.Conclusions:Pneumococcal lineages of piliated isolates have been globally disseminated and pili could have played a role in the spread of antibiotic resistant clones.

Bioengineering Downstream Technology Online Open Course Drives SPOC Hybrid Teaching Reform Based on Ideological and Political Education

In the context of emerging engineering disciplines,a hybrid teaching reform for the Bioengineering Downstream Technology course,based on ideological and political education and online open courses,is being carried out.This reform focuses on aspects such as“building a professional teacher team for ideological and political education,scientifically designing the ideological and political teaching system,innovating classroom teaching methods,and improving both formative and summative evaluation systems.”The“Craftsmanship in Education and Cultivating Soul and Roots”small private online course hybrid teaching reform for the Bioengineering Downstream Technology online open course provides a replicable model for the comprehensive implementation of ideological and political education in engineering courses and offers a reference for advancing ideological and political education and hybrid teaching reform in new engineering disciplines.

Effects of dietary supplementation with Clostridium butyricum on the growth performance and humoral immune response in Miichthys miiuy

The effects of dietary supplementation with Clostridium butyricum on growth performance and humoral immune response in Miichthys miiuy were evaluated. One hundred and fifty Miichthys miiuy weighing approximately 200-260 g were divided into five groups and reared in 15 tanks with closed circuiting culture system. The animals were fed 5 diets： basal diet only （control） or supplemented of the basal diet with C. butyricum at doses of 10^3 （CB1）, 10^5 （CB2）, 10^7 （CB3） or 10^9 （CB4） CFU/g. Compared with the control, the serum phenoloxidase activity was significantly increased by the supplementation （P〈0.05）, acid phosphatases activity was increased significantly （P〈0.05） at the doses of 10^9 CFU/g. Serum lysozyme activity peaked at dose of 10^7 CFU/g and in the skin mucus at dose of 10^9 CFU/g. Immunoglobulin M level in the serum and skin mucus was increased except at dose of 10^3 CFU/g （P〈0.05）. The growth at the dose of 10^9 CFU/g was higher than that of the control （P〈0.05）. It is concluded that supplementation of C. butyricum can mediate the humoral immune responses and improve the growth performance in Miichthys miiuy.

Effects of Dietary β-Glucan and Glycyrrhizin on Non-Specific Immunity and Disease Resistance of the Sea Cucumber (Apostichopus japonicus Selenka) Challenged with Vibrio splendidus

Sea cucumbers, Apostichopus japonicus Selenka, were fed diets containing non-immunostimulant (basal diet), 0.2% β-glucan and 0.02% glycyrrhizin in a recirculatory water system for 45 days, and subsequently challenged with Vibrio splendidus by injection at 1.0×108 cfu/sea cucumber for 15 days.Phagocytic capacity (PC), intracellular superoxide anion production (ISAP), ly-sozyme (LSZ) activity and superoxide dismutase (SOD) activity in the coelomic fluid were analyzed on the 0th, 5th, 10th and 15th days after injection.Results showed that after the 45-day feeding period, PC, ISAP, LSZ activity and SOD activity in sea cucumbers fed with dietary β-glucan or glycyrrhizin were significantly higher than in those fed with the basal diet.On the 5th day after infection, all the immune parameters examined in the sea cucumbers injected with V.splendidus decreased in value significantly.On the 15th day, PC, ISAP and LSZ activity returned to levels similar to those on the 0th day.For the sea cucumbers injected with saline, there were no significant differences in all the immune parameters examined and in the cumulative morbidity during the 15-day challenging trial.After injecting with V.splendidus, the cumulative morbidity of sea cucumbers fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin when challenged with V.splendidus challenged sea cucumber fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin.There was no significant difference in cumulative morbidity between the dietary β-glucan and glycyrrhizin treatments over time.

Immunogenicity of formaldehyde and binary ethylenimine inactivated infectious bursal disease virus in broiler chicks

Infectious bursal disease virus (IBDV) was inactivated by two different chemicals—formaldehyde and binary ethylenimine (BEI). Formaldehyde was used at 0.1% and 0.2%, while BEI was used at concentrations of 0.001 and 0.002 mol/L. These four vaccines were tested for their efficiency in generating humoral immune response in different groups of broiler chicks. Both BEI-inactivated vaccines gave relatively higher antibody titers and were almost twice as efficient as formaldehyde-inactivated ones.

Pyrolysis of different sewage sludge feedstocks for biochar products:Characterization and application

Four sewage sludge(SS)feedstocks with distinct properties were converted into biochars by pyrolysis at 300−700℃,in order to clarify the effects of the composition difference of SS feedstocks.The yields of biochars present a positive correlation with the contents of ash in SS.Notedly,the contents of organic matter(OM)in SS largely determine the quality of biochars.SS feedstocks with high content of OM are more likely to form stable biochars with higher aromaticity/carbonization degree,and the formed biochars possess higher calorific values.The contents of residual OM in biochars derived from SS feedstocks with low content of OM likely fail to meet the needs of soil improvement(10 wt.%).Most of heavy metals(HMs)existing in raw SS are remained in biochars after pyrolysis.The biochar produced from SS feedstocks with high content of HMs usually contains higher contents of HMs.Surprisingly,the leachability of HMs in biochars is all weakened to some extent compared to raw SS.In addition,the biochars show higher thermal stability and pH values,and P/K nutrients are enriched in biochars.The biochars prepared from four SS feedstocks exhibit different adsorption ability of methylene blue,especially at low dosage of biochar.

Gene cloning and induced expression pattern of IRF4 and IRF10 in the Asian swamp eel(Monopterus albus)

The Asian swamp eel （Monopterus albus） is one of the most economically important freshwater fish in East Asia, but data on the immune genes of M. albus are scarce compared to other commercially important fish. A better understanding of the eel＇s immune responses may help in developing strategies for disease management, potentially improving yields and mitigating losses. In mammals, interferon regulatory factors （IRFs） play a vital role in both the innate and adaptive immune system; though among teleosts IRF4 and IRFIO have seldom been studied. In this study, we characterized IRF4 and IRFIO from M. albus （malRF4 and malRFlO） and found that malRF4 cDNA consists of 1 716 nucleotides encoding a 451 amino acid （aa） protein, while malRFlO consists of 1 744 nucleotides including an open reading frame （ORF） of 1 236 nt encoding 411 aa. The malRFlO gene was constitutively expressed at high levels in a variety of tissues, while malRF4 showed a very limited expression pattern. Expression of malRF4 and malRFlO in head kidney, and spleen tissues was significantly up-regulated from 12 h to 48 h post-stimulation with polyinosinic： polycytidylic acid （poly I：C）, lipopolysaccharide （LPS） and a common pathogenic bacteria Aeromonas hydrophila. These results suggest that IRF4 and IRF10 play roles in immune responses to both viral and bacterial infections in M. albus.

Effects of the dietary supplementation with fructooligosaccharides on the excretion of nitrogen and phosphorus in Miichthys miiuy fries

Effects of dietary supplementation with fructooligosaccharides on the excretion of nitrogen and phosphorus in Miichthys miiuy fries were investigated. Nine hundred Miichthys miiuy fries were divided into 3 groups, each with triplicates. The basal diet and the basal diet supplemented with carnitine groups were considered as the negative and positive controls respectively. Results showed that the nitrogen concentration in excreted feces decreased significantly in fries fed the diet supplementation with 1000×10?6 fructooligosaccharides and 200×10?6 carnitine (P<0.05). The ammonic-nitrogen concentration decreased significantly in the carnitine group only (P<0.05), indicating the decreasing tendency caused by the supplementation with fructooligosaccha-rides. Supplementation with both did not have significant effects on the concentration of phosphorus in feces of Miichthys miiuy fries.

Effects of α-tocopheryl acetate supplementation in preslaughter diet on antioxidant enzyme activities and fillet quality of commercial-size Sparus macrocephalus

This study examined the effects of dietary α-tocopheryl acetate supplementation on antioxidant enzyme activities and fillet quality in commercial-size Sparus macrocephalus. Three hundred fish [main initial weight （350±12） g] were divided into three groups （E250, E500 and El000） and reared in 9 cages. The fish were fed for 8 weeks with three diets containing different levels of dietary α-tocopheryl acetate （289, 553, 1069 mg/kg）. Over the experimental period, fish were fed to satiation and reached a final mean weight of （465±28） g without significant body weight difference and proximate composition difference. Fillet α-tocopherol was significantly （P〈0.05） different between groups, reaching levels of 14.2, 22.1, 30.9 pg/mg fillet for groups E250, E500 and El000, respectively. Total serum superoxide dismutase （SOD） activity increased significantly （P〈0.05） in fish fed the diets high in α-tocopheryl acetate, but serum glutathione peroxidase （GPX） activity was unaffected. In storage on ice, fillets of fish fed the diets high in α-tocopheryl acetate exhibited significantly lower （P〈0.05） levels of oxidation. These results suggested that increased dietary α-tocopheryl acetate could increase its flesh deposition, increase the activity of SOD and prevent lipid peroxidation ofSparus macrocephalus fillets in retail storage on ice.

Extraction and Purification Process Optimization and Antioxidant Activity in vitro of Flavonoids in Amaranthus caudatus L.

[Objectives]To explore the optimal extraction and purification process of the flavonoids in Amaranthus caudatus L.and to study the antioxidant activity in vitro of the flavonoids in A.caudatus.[Methods]Taking A.caudatus as the raw material,flavonoids were extracted by alcohol extraction method,and AB-8 macroporous adsorption resin was selected for purification.The hydroxyl radical scavenging ability,DPPH radical scavenging ability,and O^2-radical scavenging ability were used as evaluation indicators,to explore the antioxidant activity in vitro of the flavonoids in A.caudatus.[Results]The optimal extraction process conditions of flavonoids in A.caudatus are:liquid-to-material ratio 40:1,extraction temperature 60℃,ethanol concentration 60%,ultrasonic power 320 W,extraction time 50 min.Under these conditions,the extraction yield of flavonoids in A.caudatus is(1.35±0.01)%.The optimal purification process conditions of flavonoids in A.caudatus are 2.5 g AB-8 macroporous adsorption resin,sample volume 5 mL,mass concentration of adsorption solution 1.60 mg/mL,pH value of adsorption solution 3.0,sample flow rate 3 BV/h,ethanol concentration in desorption process is 70%and the desorption flow rate is 3 BV/h.Under these conditions,the recovery rate reaches 88.35%±0.68%.[Conclusions]A.caudatus has a high content of flavonoids and has excellent free radical scavenging ability in vitro.This study is intended to provide important technical support for the research of flavonoid activity of A.caudatus and the development of functional products.

Calcium potentiates the effect of estradiol on PGF2α production in the bovine endometrium

Background： Estradiol （E2） is required for luteolysis in cows and its injection stimulates prostaglandin F2α （PGF2α） release. The main goal of our study was to investigate the ability of endometrial explants and cells treated with E2 and the calcium ionophore （CI） A23187 to synthesize PGF2α. Results： Treatment with E2 in vivo resulted in a 48.4% increase of PGF2α production by endometrial explants treated in vitro with A23187. Production of PGF20 was better stimulated with A23187 at concentrations of 10^-6 and 10^-5 mol/L compared with other concentrations used. The concentration of PGF2α for untreated bovine endometrial cell cultures was 33.1 pg/mL, while for cultures treated with E2, A23187, or a combination of E2 and A23187, the PGF2α concentration was 32.5, 92.4 and 145.6 pg/mL, respectively. Conclusions： Treatment with A23187 tended to stimulate PGF20 production. In the presence of E2, A23187 significantly stimulated PGF20 synthesis. It appears that A23187 potentiates the effects of E2 with respect to synthesis of endometrial PGF2α in cattle.

Comparative study of Anaplasma parasites in tick carrying buffaloes and cattle

A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Com- parative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.

Integration of association and computational methods reveals functional variants of LEPR gene for abdominal fat content in chickens

Leptin receptor(LEPR)plays a vital role in obesity in humans and animals.The objective of this study is to assess LEPR functional variants for chicken adipose deposition by integration of association and in-silico analysis using a unique chicken population,the Northeast Agricultural University broiler lines divergently selected for abdominal fat content(NEAUHLF).Five online bioinformatics tools were used to predict the functionality of the single nucleotide polymorphisms(SNPs)in coding region.Further,the possible structure–function relationship of high confidence SNPs was determined by bioinformatics analyses,including the conservation and stability analysis based on amino acid residues,prediction of protein ligand-binding sites,and the superposition of protein tertiary structure.Meanwhile,we analyzed the association between abdominal fat traits and 20 polymorphisms of chicken LEPR gene.The integrated results showed that rs731962924(N867I)and rs13684622(C1002R)could lead to striking changes in the structure and function of proteins,of which rs13684622(C1002R)was significantly associated with abdominal fat weight(AFW,P=0.0413)and abdominal fat percentage(AFP,P=0.0260)in chickens.Therefore,we are of the opinion that rs13684622(C1002R)may be an essential functional SNP affecting chicken abdominal fat deposition,and potentially applied to improvement of broiler abdominal fat in molecular marker-assisted selection(MAS)program.Additionally,the coupling of association with computer electronic predictive analysis provides a new avenue to identify important molecular markers for breeders.

Development and Application of a PCR Approach for Detection of Bovis, Sheep, Pig, and Chicken Derived Materials in Feedstuff

A PCR method for detection of bovis, sheep, pig, and chicken derived materials in feedstuff was established, and the existing method was improved according to the research on general primer and species-specific primers. First, general primer designed according to 16S rRNA gene sequence of hovis, sheep, pig, chicken, fish, and horse mtDNA was used for primary detection of animal derived materials in feedstuff. Species-specific primers designed according to conserved sequence of mtDNA of bovis, sheep, pig, and chicken were used for amplification of a 271,274, 149, and 266 bp fragment, respectively. Further confirmation of the detection result was then carried out. PCR method for detection of animal derived materials in unknown feedstuff was developed by using general primer, relevant PCR system, and PCR condition. Also a PCR method for detection of each species （bovines, sheep, pig, and chicken） was designed by using our speciesspecific primers. High sensitivity and specificity of our method were confirmed with a minimum detection level of 0.1%. Method for detection of animal derived materials in this research is not only cheap and easy for operation but also precise and reliable results can be obtained. It could be one of the effective methods for the detection of animal derived materials in feedstuff.

Effect of Incorporating Free or Encapsulated Ascorbic Acid in Chicken Frankfurters on Physicochemical and Sensory Stability

Sodium erythorbate, antioxidant and curing accelerator, used in meat products, have no vitamin functionality. Ascorbic acid （AA）, in contrast, is a powerful antioxidant, but very unstable. The application of encapsulated AA in frankfurters could allow the incorporation of an antioxidant with vitamin functionality and improved stability. The aim of this study was to evaluate the application ofAA microcapsules in frankfurters and the effects of their application on the product. Different analyses were conducted. The values obtained for mass loss, moisture content, water activity, pH and instrumental color were not significantly different among the treatments. The frankfurters containing the microcapsutes exhibited the lowest hardness value （approximately 5,000 g）, resulting in better scores in the sensory analysis. It was possible to apply the AA microcapsules without compromising the oxidative stability and physicochemical characteristics of the product, providing a promising method for protecting AA and producing fortified frankfurters.