The chemical characteristics of different sodium iron ethylenediaminetetraacetate sources and their relative bioavailabilities for broilers fed with a conventional corn‑soybean meal diet

Background Our previous studies demonstrated that divalent organic iron(Fe)proteinate sources with higher complexation or chelation strengths as expressed by the greater quotient of formation(Qf)values displayed higher Fe bioavailabilities for broilers.Sodium iron ethylenediaminetetraacetate(NaFeEDTA)is a trivalent organic Fe source with the strongest chelating ligand EDTA.However,the bioavailability of Fe when administered as NaFeEDTA in broilers and other agricultural animals remains untested.Herein,the chemical characteristics of 12 NaFeEDTA products were determined.Of these,one feed grade NaFeEDTA(Qf=2.07×10^(8)),one food grade NaFeEDTA(Qf=3.31×10^(8)),and one Fe proteinate with an extremely strong chelation strength(Fe-Prot ES,Qf value=8,590)were selected.Their bioavailabilities relative to Fe sulfate(FeSO_(4)·7H_(2)O)for broilers fed with a conventional corn-soybean meal diet were evaluated during d 1 to 21 by investigating the effects of the above Fe sources and added Fe levels on the growth performance,hematological indices,Fe contents,activities and gene expressions of Fe-containing enzymes in various tissues of broilers.Results NaFeEDTA sources varied greatly in their chemical characteristics.Plasma Fe concentration(PI),transferrin saturation(TS),liver Fe content,succinate dehydrogenase(SDH)activities in liver,heart,and kidney,catalase(CAT)activity in liver,and SDH mRNA expressions in liver and kidney increased linearly(P<0.05)with increasing levels of Fe supplementation.However,differences among Fe sources were detected(P<0.05)only for PI,liver Fe content,CAT activity in liver,SDH activities in heart and kidney,and SDH mRNA expressions in liver and kidney.Based on slope ratios from multiple linear regressions of the above indices on daily dietary analyzed Fe intake,the average bioavailabilities of Fe-Prot ES,feed grade NaFeEDTA,and food grade NaFeEDTA relative to the inorganic FeSO_(4)·7H_(2)O(100%)for broilers were 139%,155%,and 166%,respectively.Conclusions The bioavailabilities of organic Fe sources relative to FeSO_(4)·7H_(2)O were closely related to their Qf values,and NaFeEDTA sources with higher Qf values showed higher Fe bioavailabilities for broilers fed with a conventional corn-soybean meal diet.

Gut microbiome and serum metabolome analyses identify Bacteroides fragilis as regulators of serotonin content and PRL secretion in broody geese

Broody behavior is regulated by hypothalamic prolactin secretion,which seriously affects egg production in poulty production.Numerous studies have provided evidence that animal behavior is governed by dynamic bidirectional communication between specific gut bacteria and their host via the brain-gut-microbiome axis.However,little research focused on how the gut microbiota influence broody behavior in poultry.In this study,Zhedong white geese in laying and brooding phases were selected.Ten differentially abundant bacteria in cecum were detected between brooding and laying geese through metagenomic analyses and 16S rRNA sequencing(P<0.05),and Bacteroides fragilis was specifically identified as a key driver species in the brooding geese.Moverover,the serum metabolites were quantified,and the 313 differentially abundant metabolites were found between the two groups of different physiological geese.They were primarily enriched in the tryptophan metabolism pathways.Pearson correlation analyses revealed there was a significant positive correlation between B.fragilis abundance and the context of 11 tryptophan metabolism-related metabolites(such as serotonin,etc.)in broody geese,which hinted that those tryptophan metabolites might be produced or driven by B.fragilis.Finally,the serum hormone levels were also measured.We found there was a positive correlation between B.fragilis abundance and content of serotonin.Besides,prolactin secreted by the pituitary gland was greater in brooding geese than that in laying geese,which was also highly correlated with B.fragilis abundance.This result implied that B.fragilis could promote the secretion of prolactin by the pituitary gland.Together,the current study findings provided the information on gut microbiota influencing broody behavior,B.fragilis produced or driven more serum serotonin,and stimulated the pituitary gland to secret more prolactin,which potentially offered a new enlightenment for the intervention of broody behavior in poultry.

Mechanisms underlying the role of endoplasmic reticulum stress in the placental injury and fetal growth restriction in an ovine gestation model

Background Exposure to bisphenol A(BPA),an environmental pollutant known for its endocrine-disrupting properties,during gestation has been reported to increase the risk of fetal growth restriction(FGR)in an ovine model of pregnancy.We hypothesized that the FGR results from the BPA-induced insufficiency and barrier dysfunction of the placenta,oxidative stress,inflammatory responses,autophagy and endoplasmic reticulum stress(ERS).However,precise mechanisms underlying the BPA-induced placental dysfunction,and subsequently,FGR,as well as the potential involvement of placental ERS in these complications,remain to be investigated.Methods In vivo experiment,16 twin-pregnant(from d 40 to 130 of gestation)Hu ewes were randomly distributed into two groups(8 ewes each).One group served as a control and received corn oil once a day,whereas the other group received BPA(5 mg/kg/d as a subcutaneous injection).In vitro study,ovine trophoblast cells(OTCs)were exposed to 4 treatments,6 replicates each.The OTCs were treated with 400μmol/L BPA,400μmol/L BPA+0.5μg/m L tunicamycin(Tm;ERS activator),400μmol/L BPA+1μmol/L 4-phenyl butyric acid(4-PBA;ERS antagonist)and DMEM/F12 complete medium(control),for 24 h.Results In vivo experiments,pregnant Hu ewes receiving the BPA from 40 to 130 days of pregnancy experienced a decrease in placental efficiency,progesterone(P4)level and fetal weight,and an increase in placental estrogen(E2)level,together with barrier dysfunctions,OS,inflammatory responses,autophagy and ERS in type A cotyledons.In vitro experiment,the OTCs exposed to BPA for 24 h showed an increase in the E2 level and related protein and gene expressions of autophagy,ERS,pro-apoptosis and inflammatory response,and a decrease in the P4 level and the related protein and gene expressions of antioxidant,anti-apoptosis and barrier function.Moreover,treating the OTCs with Tm aggravated BPA-induced dysfunction of barrier and endocrine(the increased E2 level and decreased P4 level),OS,inflammatory responses,autophagy,and ERS.However,treating the OTCs with 4-PBA reversed the counteracted effects of Tm mentioned above.Conclusions In general,the results reveal that BPA exposure can cause ERS in the ovine placenta and OTCs,and ERS induction might aggravate BPA-induced dysfunction of the placental barrier and endocrine,OS,inflammatory responses,and autophagy.These data offer novel mechanistic insights into whether ERS is involved in BPA-mediated placental dysfunction and fetal development.

Finding biomarkers of experience in animals

At a time when there is a growing public interest in animal welfare,it is critical to have objective means to assess the way that an animal experiences a situation.Objectivity is critical to ensure appropriate animal welfare outcomes.Existing behavioural,physiological,and neurobiological indicators that are used to assess animal welfare can verify the absence of extremely negative outcomes.But welfare is more than an absence of negative outcomes and an appropriate indicator should reflect the full spectrum of experience of an animal,from negative to positive.In this review,we draw from the knowledge of human biomedical science to propose a list of candidate biological markers(biomarkers)that should reflect the experiential state of non-human animals.The proposed biomarkers can be classified on their main function as endocrine,oxidative stress,non-coding molecular,and thermobiological markers.We also discuss practical challenges that must be addressed before any of these biomarkers can become useful to assess the experience of an animal in real-life.

Marine antifouling behavior of the surfaces modified by dopamine and antibacterial peptide

Marine biofouling causes serious harms to surfaces of marine devices in transportation,aquaculture,and offshore construction.Traditional antifouling methods pollute the environment.A novel and green antifouling strategy was developed to prevent effectively the adhe sion of bacteria and microalgae.An antifouling surface was fabricated via coating Turgencin BMox2(TB)onto dopamine-modified 304stainless steel(304 SS).The surface physical and chemical properties before and after modification were characterized by Fourier transform infrared spectrometer(FTIR),X-ray photoelectron spectrometer(XPS),contact angle measurement(CA),3D optical profilometer,ellipsometer,and atomic force microscope(AFM).Antimicrobial peptide was coated onto the surface of 304 SS successfully,and the surface morphology and wettability of the modified sample were modified.Moreover,cytocompatibility of the peptide was evaluated by co-culture of peptide and cells,indicating promising cell biocompatibility at the modified sample surface.At last,antifouling performance and electrochemical corrosion were tested.Results show that the adhesion rates of Vibrio natriegens and Phaeodactylum tricornutum on the antifouling surface were reduced by 99.85% and 67.93%,respectively from those of untreated samples.Therefore,the modified samples retained superior corrosion resistance.The study provide a simply and green way against biofouling on ship hulls and marine equipment.

Composition and antioxidant activity of Paeonia lactiflora petal flavonoid extract and underlying mechanisms of the protective effect on H_(2)O_(2)-induced oxidative damage in BRL3A cells

Flavonoids have attracted considerable attention due to their health benefits. This study aimed to investigate the flavonoid profiles and antioxidant activity of Paeonia lactiflora petal flavonoid extract(PPF). The UHPLC-ESI-Q-Exactive HF MS/MS method was established for characterization, and 21 predominant flavonoid compounds were tentatively identified in PPF. Among them, isoscutellarein-7-(6’-acetylallosyl-(1->2)-glucoside) and scutellarin methylester were discovered in PPF for the first time. Pretreatment with PPF significantly reduced H2O2-induced cell damage, ROS accumulation, and malondialdehyde content and increased the activity of SOD, CAT, and GSH-Px in buffalo rat liver 3A(BRL3A) cells. Moreover, the expression of nuclear Factor E2-related factor(Nrf2) was upregulated by PPF, whose expression trend was consistent with that of heme oxygenase-1(HO-1), glutamate-cysteine ligase catalytic subunit(GCLC), and NAD(P)H quinone oxidoreductase-1(NQO1). These findings suggested that herbaceous peony flavonoids can be used as a natural bioactive agent to prevent oxidative stress.

Establishment and evaluation of the primary cultured tibial osteoblast model of broiler chicks

Osteoblasts are considered as a major factor contributing to bone development and mineralization,however,few studies have been done to establish and evaluate the primary cultured tibial osteoblast model of broiler chicks.Therefore,in the present study,two experiments were conducted to establish and evaluate the primary cultured tibial osteoblast model of broiler chicks.In experiment 1,osteoblasts were isolated from the tibia of one-day-old Arbor Acre male broiler chicks using the explant method and identified through the cell morphology,alkaline phosphatase(ALP)and alizarin red staining.Experiment 2 was carried out to evaluate the vitality and mineralization of primary cultured tibial osteoblasts of broilers on days 4,8,12,16,20,24,28 and 32 after incubation,respectively.The results from experiment 1 demonstrated that primary cultured tibial osteoblasts of broilers showed a spindle-shaped,triangular or polygonal morphology.More than 95%of the cells were stained blue-black after ALP staining,and mineralized nodules were formed after 4 days of continuous incubation.In experiment 2,lactate dehydrogenase(LDH)activity stayed at a relatively stabilized level although incubation time affected(P=0.0012)it during the whole culture period.Additionally,incubation time affected(P≤0.0001)the number and proportion of the area of mineralized nodules.They increased linearly and quadratically(P<0.04)with the increase of incubation time,and remained at a stabilized level from 24 to 32 days of incubation.The estimates of the optimal incubation time were 17 and 26 days based on the best fitted broken-line or quadratic models(P<0.0001)of the number and proportion of the area of mineralized nodules,respectively.These results indicate that the primary cultured tibial osteoblast model of broilers has been established successfully by the explant method,and it showed typical osteoblast morphology and characteristics of ALP activity and mineralization,and could maintain a relatively stabilized vitality from 4 to 32 days of incubation;and the optimal incubation time of primary tibial osteoblasts was 17 to 26 days.Therefore,it could be used to further study the underlying mechanisms of bone development and mineralization of broiler chicks.

Transcriptome-based analysis of key signaling pathways affecting the formation of primordial germ cell in chickens

Bone morphogenetic protein 4(BMP4)can induce the formation of chicken primordial germ cells(PGCs)in vitro;however,its regulatory mechanism in poultry remains unknown.This study aimed to use RNA-seq to analyze PGCs in chicken embryos and iPGCs induced by BMP4 in vitro,clarify the internal regulatory factors of PGCs,analyze the mechanism of the formation of PGCs,and lay a theoretical foundation for the further optimization of PGCs induction systems.Embryonic stem cells(ESCs),PGCs and iPGCs induced by BMP4 in vitro were collected.The transcriptional maps of the three cell types were studied using RNA-seq.The results showed 6,142 genes differentially expressed between PGCs and iPGCs,of which 2,728 were upregulated in iPGCs and 3,414 were downregulated in iPGCs.Compared to that in ESCs,BMP4 was significantly upregulated in PGCs and iPGCs.KEGG results showed that both the TGF-βand Wnt signaling pathways were activated during the formation of PGCs in vitro and in vivo,and the activation was more significant during iPGCs induced by BMP4.The expression of Nodal,an inhibitory factor of TGF-βsignaling,was significantly decreased in PGCs and iPGCs,but was not expressed in iPGCs,which further supports our conclusion.Additionally,the Lysosome and PI3K-AKT signaling pathways were significantly enriched in PGCs and iPGCs,respectively.Further,transmission electron microscopy(TEM)results showed that the number of autolysosomes was significantly higher after the addition of BMP4,which is consistent with the KEGG results.Furthermore,the number of PGCs was significantly reduced after ATG14 was interfered in vivo and in vitro.In conclusion,this study screened out the key signaling pathways during the formation of PGCs,aiming to provide help for enriching the mechanism network regulating PGCs formation in chicken and laying a theoretical foundation for further improving the efficiency of inducing PGCs in vitro.

Changes of Proline Content,Activity,and Active Isoforms of Antioxidative Enzymes in Two Alfalfa Cultivars Under Salt Stress

The plants of two elfalfa （Medicago sativa L.） cultivars differing in salt tolerance were subjected to three salt treatments, 70, 140, and 210 mM NaCl for 7 days. Root, shoot, and leaf growths were inhibited by increased salt treatments in both cultivars, and at 140 and 210 mM salt treatments, Zhongmu 1 had significantly higher root, shoot, and leaf dry weights per plant than Deft. The malondialdehyde （MDA） accumulation in Deft was considerably greater than in Zhongmu 1, indicating a higher degree of lipid peroxidation at 140 and 210 mM salt treatments. The changes in the activity and active isoforms of antioxidant enzymes such as superoxide dismutase （SOD, EC 1.15.1.1）, catalase （CAT, EC 1.11.1.6）, peroxidase （POD, EC 1.11.1.7）, and ascorbate peroxidase （APOX, EC 1.11,1.11）, accumulation of free proline, and rate of lipid peroxidation in leaves of two alfalfa cultivars were also investigated. After stress, the activity and active isoforms of antioxidative enzymes were altered and the extent of alteration varied between the cultivar Deft and Zhongmu 1. The proline accumulation in Deft was considerably greater than in Zhongmu 1 at 210 mM salt treatment. This indicated that proline accumulation may be the result, instead of the cause, of salt tolerance.

Effects of Limiting Amino Acids on Rumen Fermentation and Microbial Community In vitro

Three goats fitted with cannula were used to provide rumen liquor to investigate the effects of limiting amino acids on rumen fermentation and microbial community in vitro. The removal method was used in the current experiments. Treatments are total essential amino acid （TEAA）, His-removal, Lys-removal, Met-removal, and branch chain amino acid （BCAA）- removal. Results indicated that, pH-value ranged between 5.9 and 6.8, with the highest mean value for the group with BCAA-removal （6.54） in the culture. Concentration of NH3-N ranged between 10.99 to 30.51 mg 100 mL^-1, with the group of TEAA recording the highest average NH3-N concentration （17.85 mg 100 mL^-1）. Yields of microbial protein and limiting degree on microbial growth varied with treatments （P 〈 0.01）, and the lowest accrued in treatment with BCAA-removal （0.1389, 0.1772, and 0.3161 mg mL^-1 for bacteria, protozoa, and mixed microbes, respectively）, compared to the group with TEAA, microbial production of mixed microbes decreased by 44.52%. As for micro-flora, protozoa to bacteria ratio was the lowest for the group with Lys-removal （89.12%）, while the highest for the group with BCAA-removal （127.60%） （P 〈 0.01）. Furthermore, PCR-SSCP analysis revealed that, microbial profile subjected to substrates within bacteria and protozoa groups. It was therefore concluded that, dietary amino acid influenced both rumen fermentation and microbial characteristics.

Energy and protein requirements for maintenance of Hu sheep during pregnancy

This study aimed to determine the effect of stage and level of feed intake on energy metabolism, carbon-nitrogen （C-N） balance, and methane emission to determine energy and protein requirements for maintenance of maternal body including pregnancy tissues during pregnancy using the method of C-N balance. Twenty-one ewes carrying twin fetuses were randomly divided into three groups of seven ewes each in the digestion and respirometry trial at d 40, 100, and 130 of gestation, respectively. Three groups were fed a mixed diet either for ad lib#urn intake, 70 or 50% of the ad libfum intake during pregnancy. The results showed that the apparent digestibility of C and N were increased as feeding levels decreased at each stage of gestation. The daily net energy requirements for maintenance （NEro） were 295.80, 310.09, and 323.59 kJ kg-1 BW0.75 （metabolic body weight） with a partial efficiency of metabolisable energy utilization for maintenance of 0.664, 0.644, and 0.620 at d 40, 100, and 130 of gestation, respectively. The daily net protein requirements for maintenance were 1.99, 2.35, and 2.99 g kg-1 BW0.75 at d 40, 100, and 130 of gestation, respectively. These results for the nutritional requirements of the net energy and protein may help to formulate more balanced diets for Hu sheep during pregnancy.

Associations of Gonadotropin-Releasing Hormone Receptor (GnRHR) and Neuropeptide Y(NPY) Genes’Polymorphisms with Egg-Laying Traits in Wenchang Chicken

Single nucleotide polymorphisms （SNP） of chicken gonadotropin-releasing hormone receptor （GnRHR） and neuropeptide Y （NPY） were selected to identify the genotypes of Wenchang （Chinese indigenous breed） chicken with restricton fragment length polymorphisms. The associations of the SNPs with the total egg production （NE）, average days of continual laying （ADCL）, and number of double-yolked eggs （DYE） traits were analyzed. The frequency of restriction enzyme A/a alleles in the population was for GnRHR 0.69 （Bpu1102 Ⅰ A） and 0.31 （Bpu1102 Ⅰ a） and for NPY 0.46 （Dra Ⅰ B） and 0.54 （Dra Ⅰ b）. Trait data from a total of 120 hens, which were purebred introduced from Hainan Province, China from one generation were recorded. Two significant effects of genes＇ marker were found： for GnRHR and number of eggs （dominant; t= 2.67, df= 116） and NPY and number of eggs （additive; t= 1.97, df= 116）. The current research supports the effects of GnRHR and NPY genes on egg-laying traits of chickens.

Gas stunning with CO_2 affected meat color,lipid peroxidation, oxidative stress, and gene expression of mitogen-activated protein kinases, glutathione S-transferases,and Cu/Zn-superoxide dismutase in the skeletal muscles of broilers

Background: Meat color and lipid peroxidation are important traits related to meat quality. CO_2 concentration is a critical factor that can affect meat quality in the commercial use of gas stunning(GS). However, the effect and mechanism of CO_2 stunning on meat color and lipid peroxidation during long-term storage remain poorly studied. We aimed to study the effects of GS methods, especial y CO_2 concentration, on meat color and meat lipid peroxidation in broilers during long-term storage at 4 °C and to explore the potential mechanism of meat color change via lipid peroxidation and the inner lipid peroxide scavenging system.Methods: Eighteen broilers were sacrificed after exposure to one of the following gas mixtures for 90 s: 40% CO_2+21% O_2+ 39% N2(G40%), 79% CO_2+ 21% O_2(G79%), or no stunning(0% CO_2, control). Meat color, serum variables,enzyme activities, and the gene expression of mitogen-activated protein kinase(MAPK), nuclear factor-erythroid2-related factor 2(Nrf2), glutathione S-transferase(GST) and superoxide dismutase(SOD) were determined.(Continued on next page)Results: The concentrations of serum triiodothyronine(T3, P = 0.03) and the ratio of serum free triiodothyronine/free thyroxine(FT3/FT4, P < 0.01) were decreased, whereas levels of serum cortisol(P < 0.01) were increased in the 40%CO_2 group compared with the control group. Additionally, the thiobarbituric acid-reactive substances(TBARS)3 d(P < 0.01) and TBARS6 d(P = 0.01) in breast meat and the TBARS3 din thigh meat(P < 0.01) were increased in the40% CO_2 group compared with the control group. Serum T3 was negatively correlated with TBARS6 dboth in the breast and thigh meat(r =-0.63, P < 0.01 and r =-0.47, P = 0.05 respectively). T3/T4 was negatively correlated with TBARS6 din the breast meat and in the thigh meat(r =-0.57, P = 0.01; and r =-0.53, P = 0.03 respectively). Compared with the control group, Lightness(L*)1 d(P = 0.03) and L*9 d(P < 0.01) were increased, whereas total chromatic aberration(E*)1 d(P = 0.05) and E*3 d(P < 0.01) were decreased in the breast meat of both the G40% and G79% groups. The values of yel owness(b*)3 d(P = 0.01), b*6 d(P < 0.01) and E*6 d(P < 0.01) in the thigh meat were lower in both the G40% and G79% groups than in the control group. In the breast muscle, the m RNA levels of c-Jun N-terminal kinase 2(JNK2, P = 0.03),GSTT1(P = 0.04), and SOD1(P = 0.05) were decreased, and the m RNA levels of JNK1(P = 0.07), Nrf2(P = 0.09), and GSTA3(P = 0.06) were slightly lower in both the G40% and G79% groups compared with the control group. However, among these genes, only the m RNA level of JNK1 was decreased in the G40% group compared with the control group and the G79% group(P = 0.03) in the thigh muscle.Conclusions: Compared with the control group, meat color quality in the breast meat was decreased, and the expression of genes in the MAPK/Nrf2/ARE(antioxidant responsive element) antioxidant pathway in breast muscle was partly suppressed by GS of both 40% and 79% CO_2. However, oxidative stress and meat lipid peroxidation during storage were aggravated by GS with 40% CO_2 compared to GS with 79% CO_2 and no GS.

CNV analysis of Meishan pig by nextgeneration sequencing and effects of AHR gene CNV on pig reproductive traits

Background:Reproductive performance of livestock is an economically important aspect of global food production.The Chinese Meishan pig is a prolific breed,with an average of three to five more piglets per litter than European breeds;however,the genetic basis for this difference is not well understood.Results:In this study,we investigated copy number variations(CNVs)of 32 Meishan pigs and 29 Duroc pigs by nextgeneration sequencing.A genome-wide analysis of 61 pigs revealed 12,668 copy number variable regions(CNVRs)that were further divided into three categories based on copy number(CN)of the whole population,i.e.,gain(n=7,638),and loss(n=5,030)CNVRs.We then compared Meishan and Duroc pigs and identified 17.17Mb of 6,387 CNVRs that only existing in Meishan pigs CNVRs that overlapped the reproduction-related gene encoding the aryl hydrocarbon receptor(AHR)gene.We found that normal AHR CN was more frequent than CN loss in four different pig breeds.An association analysis showed that AHR CN had a positive effect on litter size(P<0.05)and that a higher CN was associated with higher total number born(P<0.05),number born alive(P<0.05),number of weaned piglets,and birth weight.Conclusions:The present study provides comprehensive CNVRs for Meishan and Duroc pigs through large-scale population resequencing.Our results provide a supplement for the high-resolution map of copy number variation in the porcine genome and valuable information for the investigation of genomic structural variation underlying traits of interest in pig.In addition,the association results provide evidence for AHR as a candidate gene associated with reproductive traits that can be used as a genetic marker in pig breeding programs.

Analysis of genetic diversity and population structure of Chinese yak breeds (Bos grunniens) using microsatellite markers

Nine Chinese yak breeds （Maiwa, Tianzhu White, Qinghai Plateau, Sibu, Zhongdian, Pali, Tibetan High Mountain, Jiulong, and Xinjiang） and Gayal were analyzed by means of 16 microsatellite markers to determine the level of genetic variation within populations, genetic relationship between populations, and population structure for each breed. A total of 206 microsatellite alleles were observed. Mean F-statistics （0.056） for 9 yak breeds indicated that 94.4% of the genetic variation was observed within yak breeds and 5.6% of the genetic variation existed amongst breeds. The Neighbor-Joining phylogenetic tree was constructed based on Nei＇s standard genetic distances and two clusters were obtained. The Gayal separated from the yaks far away and formed one cluster and 9 yak breeds were grouped together. The analysis of population structure for 9 yak breeds and the Gayal showed that they resulted in four clusters; one cluster includes yaks from Tibet Autonomous Region and Qinghai Province, one cluster combines Zhongdian, Maiwa, and Tianzhu White, and Jiulong and Xinjiang come into the third cluster. Pali was mainly in the first cluster （90%）, Jiulong was mainly in the second cluster （87.1%）, Zhongdian was primarily in the third cluster （83%）, and the other yak breeds were distributed in two to three clusters. The Gayal was positively left in the fourth cluster （99.3%）.

Effects of neutral salt and alkali on ion distributions in the roots,shoots,and leaves of two alfalfa cultivars with differing degrees of salt tolerance

The effects of neutral salt and alkali on the ion distribution were investigated in two alfalfa （Medicago sativa L.） cultivars, including Zhongmu 1, a high salt-tolerant cultivar, and Algonquin, a low salt-tolerant cultivar. The alkali stress expressed more serious growth inhibition than the neutral salt stress at the same Na＋ concentration. Compared with Algonquin, Zhongmu 1 did not exhibit a higher alkali tolerance under the Na2CO3-NaHCO3 treatment with the low Na＋ concentration （50 mmol L-l）. The alkali increased the accumulation of Na＋, Ca2＋, and Mg2＋ in the root and changed the Ca2＋ and Mg2＋ balance in the entire alfalfa plant. The salt and alkali stresses decreased the K＋ and Fe3＋ contents of the roots and leaves, the root Mn2＋ content, and the shoot Zn2＋ content, but they increased the Fe3＋ accumulation of the shoots, the shoot and leaf Cu2＋ contents, and the leaf Zn2＋ content in both alfalfa cultivars. Based on the results obtained under the conditions of this experiment, we found that the salt and alkali stresses reduced the plant growth in both alfalfa cultivars, while the alkali caused a stronger stress than the neutral salt in alfalfa. Thus, we conclude that under hydroponic conditions, the deleterious effects of the alkali on plants are due to the distribution change of some trophic ion balance in the roots, shoots, and leaves of the plants by causing of Na＋, CO3^2-, and/or HCO3- stresses.

Replacement of Forage Fiber Sources with Dried Distillers Grains with Solubles and Corn Germ Meal in Holstein Calf Diets

This study was carried out to determine the effect of replacement of forage fiber sources from alfalfa and Leymus chinensis with nonforage fiber sources（NFFS） from dried distillers grains with solubles and corn germ meal on calves growth, rumen development and blood parameters. 48 female and 12 male calves（（110.55±15.36） kg of body weight and 12 wk of age） were assigned randomly to four dietary treatments（15 calves/treatment） in a completely randomized design. Experimental diets were： 0% NFFS（control）, 9% NFFS（group 1）, 18% NFFS（group 2）, 27% NFFS（group 3）, and contained equivalent neutral detergent fiber and total digestible nutrients, respectively. The dry matter intake was similar among diets, averaging 3.33 kg d-1, and no differences were detected for body weight, withers height, body length and heart girth. In addition, the development of rumen, reticulum, omasum and abomasum also were similar among diets. Dry matter, crude protein, and neutral detergent fiber digestibilities increased with the increasing levels of NFFS in the experimental diets, but had no significant effect. Blood urea nitrogen, total protein and glubulin were not affected by the dietary treatment, but group 3 resulted in the highest（P〈0.05） concentrations of glucose and the lowest（P〈0.05） concentrations of triglycerides and albumin. In conclusion, dried distillers grains with solubles（DDGS） and corn germ meal（CGM） were available and alternative fiber sources for Holstein calf diets.

The protective roles of tea tree oil extracts in bovine mammary epithelial cells and polymorphonuclear leukocytes

Background: Tea tree oil(TTO) plays an important role in antibacterial activity and alleviating the inflammatory responses. Bovine mammary epithelium and polymorphonuclear leukocytes(PMNL) can actively respond to bovine mastitis infection. However, regulatory effects of TTO extracts on the innate immune response of bovine mammary epithelial cells(BMECs) and PMNL remain not reported. Therefore, aim of the study was to evaluate the effects of TTO extracts on the m RNA levels of the genes involved in the innate immune response of BMECs and PMNL.Results: Our results demonstrated that addition of 0.025% and 0.05% TTO increased the proliferation of BMECs, and significantly enhanced(P < 0.05) the viability of BMECs exposed to Staphylococcus aureus(S. aureus). An inhibitory effect was observed against the growth of S. aureus by TTO incubation. The 0.05% TTO reduced S. aureus biofilm formation, association and invasion of S. aureus to BMECs, and changed the morphological and structural features of S. aureus. The proinflammatory cytokines IL-1β, IL-6, and TNF-α were decreased(P < 0.001) by the incubation of TTO. Interestingly, the expression of IL-8 known for PMNL chemotactic function was elevated(P < 0.05) by 0.05%TTO treatment. Consistently, 0.05% TTO increased the migration of PMNL in S. aureus-exposed BMECs when compared with S. aureus treatment alone(P < 0.05). In addition, PMNL incubated with 0.05% TTO decreased the levels of NFKB inhibitor alpha(NFKBIA) and TNF-α.Conclusions: Our results indicate that use of TTO can relieve the BMECs pro-inflammatory response caused by S.aureus and promote the migration of PMNL to mount the innate immune responses, and it may be novel strategy for the treatment of bovine mastitis caused by S. aureus.

Effects of formic acid and corn flour supplementation of banana pseudostem silages on nutritional quality of silage,growth,digestion,rumen fermentation and cellulolytic bacterial community of Nubian black goats

The objective of this study was to evaluate the effects of adding formic acid and corn flour supplementation to banana pseudostem silages on the nutritional quality of these silages,growth,digestion,rumen fermentation and cellulolytic bacterial community of Nubian black goats fed these silages.Banana pseudostem silage was prepared either conventionally without any additives(CON)or mixed with 0.6% formic acid(F),10% corn flour(C),or both(F+C).Four experimental diets containing 40% of the corresponding silages were designed with roughage to concentrate ratio of 50:50(dry matter(DM)basis).A total of 48 Nubian black castrated goats(body weight(BW),(22.64±1.82)kg;4-mon-old)were randomized into one of the four treatment groups with 12 replicates of one castrated goat per replicate for each treatment in a completely randomized design.Each group was fed on one of the four experimental diets for 40 days.A factorial arrangement of treatments of 2(formic acid levels:0 and 0.6%)×2(corn flour:0 and 10%)was adopted.Formic acid supplementation increased(P<0.05)average daily gain,as well as lactic acid,propionate and butyric acid and water-soluble carbohydrate concentrations,but decreased(P<0.05)the feed conversion rate,pH value,acetate/propionate ratio,and butyric acid concentration relative to the CON group.Corn flour supplementation increased(P<0.05)the apparent digestibility of crude protein,neutral detergent fiber,and non-fibrous carbohydrate and Fibrobacter succinogenes,Ruminococcus albus,and Butyrivibrio fibrisolvens populations,but decreased(P<0.05)the Ruminococcus flavefaciens population relative to the CON group.There were no F×C treatment interactions(P>0.05)for any of the other indices except for the apparent digestibility of non-fibrous carbohydrate(NFC)(P<0.05).The results demonstrated that adding 0.6% formic acid and 10% corn flour supplementation to banana pseudostem silages improved the nutritional quality of these silages and enhanced the growth performance of Nubian black castrated goats by improving apparent nutrient digestibility,and regulating ruminal fermentation and bacteria populations.

Effects of selenium source and level on growth performance, antioxidative ability and meat quality of broilers

This experiment was conducted to investigate the effects of selenium(Se) source and level on growth performance, carcass traits, antioxidative ability and meat quality of broilers. A total of 320 one-d-old Arbor Acres commercial broilers were randomly assigned to 1 of 5 treatments with 8 replicates in a completely randomized design involving a 2×2 factorial arrangement of treatments plus one Se-unsupplemented basal diet control for 42 d. The two Se sources were sodium selenite and Se yeast, and the two supplemental Se levels were 0.20 and 0.40 mg Se kg^–1. The results showed that broilers fed the Se-supplemented diets had higher(P<0.05) average daily gain and average daily feed intake from 22 to 42 d of age, eviscerated yield and abdominal fat percentages, Se concentrations and glutathione peroxidase(GSH-Px) activities in breast and thigh muscles on d 42, and lower(P<0.05) feed/gain from 1 to 21 and 22 to 42 d of age, mortality from 22 to 42 d of age and malondialdehyde(MDA) concentration in thigh muscle on d 42 than those fed the control diet. Broilers fed the diets supplemented with Se yeast had higher(P<0.05) p H value and lower(P<0.05) shear force in thigh muscle than those fed the diets supplemented with sodium selenite. Additionally, broilers fed the diets supplemented with 0.40 mg Se kg^–1 had lower(P<0.05) shear force in thigh muscle and higher(P<0.05) GSH-Px activities in breast and thigh muscles than those fed the diets supplemented with 0.20 mg Se kg^–1. Furthermore, broilers fed the diet supplemented with Se yeast at 0.40 mg Se kg^–1 had higher(P<0.05) Se concentrations in breast and thigh muscles than those fed the diet supplemented with Se yeast at 0.20 mg Se kg^–1, but no differences(P<0.05) were observed in these indices of broilers fed the diets supplemented with sodium selenite between 0.20 and 0.40 mg Se kg^–1. The results from the present study indicated that supplemental Se could increase the growth performance, muscle Se concentration and antioxidative ability of broilers;and the Se from Se yeast was more effective than the Se from sodium selenite in improving meat quality of broilers.