Effects of Fermented Nano Chinese Herbal Medicines Replacing Antibiotics on Production Performance and Carcass Quality of Growing-Finishing Pigs

[Objectives]To study the effects of fermented nano Chinese herbal medicines replacing antibiotics on the production performance and carcass quality of growing-finishing pigs.[Methods]200 healthy and disease-free Duroc-Landrace-Yorkshire(DLY)growing-finishing pigs with similar birth time(45±2 d)and body weight of about 31.5 kg were randomly divided into group A(fed with non-antibiotic basal diet),group B(fed with antibiotic basal diet),group C(fed with non-antibiotic basal diet+0.1%fermented nano Chinese herbal medicine preparation),group D(fed with non-antibiotic basal diet+0.3%fermented nano Chinese herbal medicine preparation)and group E(fed with non-antibiotic basal diet+0.5%fermented nano Chinese herbal medicine preparation).The experiment lasted for 90 d.At the end of the experiment,the production performance and carcass indicators were measured.[Results]Compared with group A,the final weight,average daily gain(ADG)and average daily feed intake(ADFI)of groups B,C,D and E were extremely significantly increased(P<0.01),while the feed-to-gain ratio(F/G)was extremely significantly decreased(P<0.01).Compared with group B,the ADG of group E was significantly increased(P<0.05),while the F/G was significantly decreased(P<0.05).Compared with group A,the slaughter rate and carcass length of pigs in groups B,C,D and E were extremely significantly increased(P<0.01);compared with group B,the slaughter rate of groups D and E was extremely significantly increased(P<0.01).[Conclusions]The fermented nano Chinese herbal medicine preparation has better effects in promoting growth than that of antibiotic additives,and it can significantly improve the carcass quality of growing-finishing pigs.

Cloning and Sequence of Glycoprotein H Gene of Duck Plague Virus

The glycoprotein H （gH） gene homologue of duck plague virus （DPV） was cloned by degenerate polymerase chain reaction （PCR） and sequenced. It was located immediately downstream from the thymidine kinase gene （TK）. In addition, the 3＇-end of the gene homologue to herpesvirus UL21 was located downstream from the gH gene. DPV gH gene open reading frame （ORF） was 2 505 bp in length and its primary translation product was a polypeptide of 834 amino acids long. It possessed several characteristics of membrane glycoproteins, including an N-terminal hydrophobic signal sequence, an external domain containing eight putative N-linked glycosylation sites, a C-terminal transmembrane domain, and a charged cytoplasmic tail. Comparison with other herpesvirus revealed identities of 20.2, 25.1, 23.0, 23.0, 26.5 and 26.0% with the gH counterparts of the human herpesvirus virus 1 （HSV1）, equine herpesvirus 4 （EHV4）, bovine herpesvirus 1 （BHV1）, pseudorabies virus （PRV）, gallid herpesvirus 2 （GHV2） and gallid herpesvirus 3 （GHV3）, respectively.

Association between the Phenotypes and Genotypes of Antimicrobial Resistance in Haemophilus parasuis Isolates from Swine in Quang Binh and Thua Thien Hue Provinces, Vietnam

Haemophilus parasuis (H. parasuis) is one of the bacterial pathogens of great concern as it causes huge economic losses to the swine industry worldwide. One of the reasons why the control of H. parasuis has failed is the increase in antimicrobial resistance (AMR). The country of Vietnam has the second-largest pig production in Asia. However, there is still a lack of data about the AMR prevalence of H. parasuis in Vietnam.The purpose of this study is to investigate the prevalence of AMR and analyze the association between AMR and AMR genes (ARGs). The H. parasuis strains used in this research were isolated from swine in the Quang Binh and Thua Thien Hue Provinces, Central Vietnam, as reported in our previous study. All of the strains were tested for AMR against 25 antibacterial agents using the broth microdilution method and for the presence of ARGs using the polymerase chain reaction (PCR) method. The tested strains were shown to have a high frequency of resistance to trimethoprim/sulfamethoxazole (94.6%), followed by resistance to colistin, chloramphenicol, gentamicin, penicillin, lincomycin, and amoxicillin. The most prevalent ARGs in these strains were blaTEM-1 (94.6%), int (76.8%), gyrA (58.9%), and rmtD (50.0%). Cefuroxime, chloramphenicol, and tobramycin resistances were strongly correlated with the presence of the ARGs blaROB-1 (odds ratio (OR) = 26.3, 95% confidence interval (CI) 2.7–255.7, p = 0.002), catl (OR = 25.1, 95% CI 2.4–258.9, p = 0.004), and strB (OR = 23.5, 95% CI 2.6–212.6, p = 0.001), respectively.This study reveals for the first time the current situation of H. parasuis AMR in Central Vietnam, which is helpful for the clinical control of this disease, as well as for the development of policies and clinical practice guidelines to reduce AMR in swine production in Central Vietnam.

A Simple Procedure for Extraction of Surface Protein of Salmonella Serotypes and Escherichia coli Strains Isolated from Poultry and Pigs

Salmonella and E.coli possess different surface protein structures that can induce protective immune responses.Identification of these proteins capacitates development of diverse applications in prevention and diagnosis that contribute to effectively control disease-causing enterobacteria pathogens such as Salmonella and E.coli.A simple procedure for obtaining protein complexes of Salmonella serotypes and E.coli is performed in this study.A sonication process with heat treatment of whole bacteria induced the release of protein complexes.Concentration of the protein extract was quantified using protein quantification Kits-Rapid,and protein complex profile was obtained by SDS-PAGE(Sodium dodecyl sulfate polyacrylamide gel electrophoresis)and silver staining.The concentrations of protein ranged from 29.45 to 45.35μg/mL in the Salmonella protein extracts,and from 25.35 to 36.72μg/mL in the E.coli protein extracts.Six major groups of proteins from E.coli(YfiO,NipB,OmpF,YfgL,Talc,YaeT)and four major groups of proteins from Salmonella(Flagellin,OmpA,Porin,SEF21)were preliminarily determined by a simple procedure of extraction based on the molecular weight.

Determination of the Administration Routes,Doses and Appropriate Age to Vaccinate With Ornitin Triple Vaccine For Cross-breed Colored Broilers in Vietnam

The study was to determine the appropriate dose and administration route of Ornitin Triple vaccine in cross-breed coloured broilers in Vietnam by evaluating the antibody titer against Ornithobacterium rhinotracheale(ORT)and local reactions at injection sites on chickens after vaccination.The study was divided into 2 trials.Both trials were designed with 3 vaccine dose groups:0.0ml(control group),0.25ml and 0.5ml and 2 different administration routes:subcutaneous at neck(SC)and intramuscular at breast(IM)injection.The result showed that,no statistically significant difference was found between antibody titer of two administration routes as well as 2 vaccine doses until 13-week-old.Local reactions at the injection sites of IM route was less severe than SC at neck and in higher dose would produce a more severe swelling reaction.Daily weight gain was found to have a slight decrease in the vaccinated groups within 2 weeks after vaccination,however,no statistically significant difference was found in later stage(P>0.05).In conclusion,Ornitin Triple can be used to vaccinate by IM with the dose of 0.25ml for coloured broilers at early age(3-week-old),or 0.5ml for older birds and should be careful for some reactions at the injection sites.

Dietary encapsulated essential oils and organic acids mixture improves gut health in broiler chickens challenged with necrotic enteritis

Background:The poultry industry is in need of effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE)due to Clostridium perfringens.In the present study,we investigated the effects of dietary supplementation with a blend of encapsulated essential oils and organic acids(BLJ)on growth performance and gut health using a coinfection model of NE in broiler chickens.Methods:Two hundred and eighty-eight one-day-old male Arbor Acres broiler chicks were randomly assigned using a 2×2 factorial design into two groups fed either 0 or 500 mg/kg dietary BLJ and co-challenged(or not challenged for the control)with Eimeria spp./C.perfringens.Results:Infected birds fed the BLJ-supplemented diet exhibited an improved feed conversion ratio throughout the trial(P<0.01),a higher villus height and villus height/crypt depth ratio,and reduced intestinal C.perfringens counts,liver C.perfringens carriage,gut lesion scores and serum fluorescein isothiocyanate dextran(FITC-D)concentrations at 7 d postinfection compared with those of birds without BLJ supplementation(P<0.05).NE-infected birds fed BLJ exhibited significantly upregulated claudin-1 and IGF-2 mRNA levels(P<0.05),increased A20 mRNA expression and significantly downregulated TRAF-6,TNFSF15 and TOLLIP mRNA levels in the jejunum at 7 d post-infection compared with those in birds without BLJ supplementation(P<0.05).Compared with the uninfected and untreated birds,the uninfected birds fed BLJ displayed increased relative abundances of Lactobacillus and Coprococcus but reduced Rikenellaceae levels.Compared with the unsupplemented NE-challenged birds,infected birds fed BLJ showed an increased relative abundance of Unclassified_Lachnospiraceae and a significantly decreased relative abundance of Erysipelotrichaceae.Conclusion:BLJ supplementation improved growth performance and gut health in NE-infected broiler chickens by strengthening the intestinal barrier function,positively modulating the gut microbiota community and differentially regulating intestinal immune responses.Our results also suggested that adding BLJ effectively controlled NE infections after experimental Eimeria and Clostridium perfringens coinfection.

The ultra-sonication of minerals in swine feed

A sample preparation method based on ultrasound assisted-extraction （UAE） of Ca, Mg and P from swine feed has been described. The experiment was performed to cover the variables influencing the sonication process and, the method validation using standard reference material. Final solutions obtained upon sonication were analyzed by flame atomic absorption spectrometry （for Ca and Mg） and by UV-vis spectrophotometry （for P）. The best conditions for metal extraction were as follows： sample mass： 100 mg in 20 mL 0.10 mol/L HCI, a particle size： 〈60 lam, sonication time： 5 cycles of 10 s and ultrasound power： 102 W. The UAE method was applied in digestibility assays in different piglet feeds and their results showed that it is highly comparable （P 〉 0.05） to the other methods used for such purposes, as block digestion, and offered a Ca, Mg and P method of quantification limit of 10.6, 12.4 and 14 mg/kg, respectively. The major advantages of the UAE method compared to other methods are the high treatment rate, low reagent usage in the extracts and, it does not generate toxic residues that might negatively affect human health and the environment, accompanied by good precision and accuracy.

PRRS in Vietnam and Its Diagnosis

Genotyping 42 serum samples taken from the pigs in the oubreaks from 2009 to 2013 with RT-PCR and nested RT-PCR, showed more than 80% of samples were positive with Chinese PRRSV clade, and the others were European and North American classical PRRSV genotypes. Ten serum samples from unnapprent PRRS herds were examined for antibodies against PRRSV with ELISA and also for PRRSV with RT-PCR. It was clearly that the titer of antibodies against PRRSV by ELISA test could not be used for interpreting PRRSV infection. In despite of PRRS vaccination or non-vaccination, a risk of PRRSV infection and re-infection exist, utilizing RT-PCR in combination with serology will give the producer and veterinarian PRRSV more exact situation in the herds.

Investigation of Two Precipitation Methods for Extracting Immunoglobulin Y(IgY)from Egg Yolks

Two groups of hens(control and immunization group)were arranged in an experimental design with an immunization schedule of 3 injections of BSA antigen.IgY antibodies were extracted from egg yolks by two precipitation processes(chloroform and polyethylene glycol precipitates)and quantified using a standard curve of protein concentration.The purification of IgY was confirmed by SDS-PAGE.Total protein extracted from egg yoks were less contaminated with yellow pigments(lutein and zeaxanthin)nd by using chloroform precipitate.The 2 week post-immunization,IgY-1 concentration increased respectively to 3903±726μg.ml(chloroform-1 extraction process)and 2937±294μg.ml(PEG extraction process)(P<rd 0.01).After 3 immunization,IgY level obtaining from in immunization group extracted by chloroform process(6633±1166μg.ml-1)increased 2.7 times higher than that in control group(2482±414μg.ml-1).Whereas IgY concentrations obtained from PEG extraction process were not significantly different between the experimental group and control group.Chloroform and PEG precipitation methods had the same protein profile on the SDS-PAGE.IgY antibody was identified by the presence of bands corresponding with IgY heavy chain(67-70 kDa)and IgY light chain(25 kDa)for both precipitation processes.

Dietary coated essential oil and organic acid mixture supplementation improves health of broilers infected with avian pathogenic Escherichia coli

Colibacillosis caused by avian pathogenic Escherichia coli(APEC)is a very prevalent disease in poultry farms in China.The exploration of effective non-antibiotic substances is of great significance for the control of APEC infections.This experiment evaluated the efficacy of coated essential oil and organic acid(EOA)supplementation to prevent E.coli O78 infection in broiler chickens.A total of 288 one-day-old male broiler chicks were randomly distributed into 4 groups with 6 replicates per group.Chickens were fed a diet either supplemented with EOA(500 mg/kg feed)or not,and either uninfected or infected with E.coli O78 intratracheally.Results showed that E.coli O78 infection reduced body weight gain,increased mortality and the ratio of feed to gain along with cecal and liver E.coli load,damaged gut mucosa,induced local and systemic inflammation,and altered cecal microbial composition,diversity and function(P<0.05).Supplemental EOA improved feed conversion efficiency,lowered gross lesion scores and cecal E.coli population,enhanced intestinal goblet cells and serum IgG concentration,and tended to decrease serum IL-12 production(P<0.05).Essential oil and organic acid addition downregulated IFN-γmRNA,tended to decrease mucin-2 mRNA levels while upregulating IL-10 mRNA,and tended to increase ZO-1 gene expression in the jejuna of infected birds at 7 d after E.coli O78 challenge(P<0.05).The 16S rRNA gene sequencing indicated that both EOA addition and E.coli O78 challenge altered the diversity and composition of the cecal microbiota community.Furthermore,infected birds fed EOA showed decreased Bacteroidetes and genus Lactobacillus abundance compared with the infected control.LEfSe analysis showed that Firmicutes,Ruminococcaceae,Clostridiales,Clostridia,Lactobacillus,Lactobacilaceae,and cc-115 were enriched in the non-infected but EOA-treated group(P<0.05).Collectively,dietary EOA supplementation could mildly alleviate E.coli-induced gut injury and inflammation.

用去透明带仓鼠卵穿透试验检测牛冷冻精子的受精能力

研究了用去透明带仓鼠卵穿透试验检测牛冷冻精子的受精能力。对 36头西门塔尔、 2 3头夏洛来和 12头利木赞牛冷冻精子的实验结果表明 :三个品种的牛冻精均能穿入去透明带仓鼠卵 ,并形成发育良好的雄原核。穿透率在 70 %～ 80 %左右 ,有原核卵百分率在 30 %左右。

Nutritional evaluation of integral cassava root silages for growing pigs

The experiment aimed at determining the nutritional value of integral cassava root silages with yogurt as inoculant or wastewater(manipueira) by liquid addition ensiling process. Eighteen crossbred piglets(Large White x Landrace), castrated males with an initial average weight of 50 kg were allocated in metabolism cages throughout 11-day trial duration(6 days for animal's adaptation to the cages and to the experimental diets, and 5 days of urine and faeces collection). The experimental design was a randomized block arrangement with 3 treatments and 6 replicates: basal diet(100%); mixture composed of75% basal diet and 25% integral cassava root silage with yogurt as inoculant; a mixture composed of 75%of basal diet and 25% integral cassava root silage with wastewater. The silages with wastewater and yogurt presented the following values of apparent digestibility of dry matter 89.96% and 90.01%,apparent digestibility of crude protein of 60.67% and 66.43%, apparent digestibility of gross energy of90.43% and 91.48%, gross energy metabolizability coefficients of 87.88% and 88.93%, digestible energy values of 3,705 and 3,783 kcal/kg on a dry matter(DM) basis, and metabolizable energy values of 3,600 and 3,676 kcal/kg DM, respectively. The results have demonstrated that integral cassava root silages with wastewater or yogurt have a high nutritional value and can be used as an alternative energy source in growing pig's diets.

Nano-selenium alleviates cadmium-induced cerebellar injury by activating metal regulatory transcription factor 1 mediated metal response

This study aims to investigate the role of metal regulatory transcription factor 1(MTF1)-mediated metal response in cadmium(Cd)-induced cerebellar injury,and to evaluate the antagonistic effects of nanoselenium(Nano-Se)against Cd toxicity.A total of 80 chicks(1 d old,male,Hy-Line Variety White)were randomly allocated to 4 treatment groups for 3 months:the control group(fed with a basic diet,n=20),the Nano-Se group(basic diet with 1 mg/kg nano-Se 1 mg/kg Nano-Se in basic diet,n=20),the NanoSe+Cd group(basic diet with 1 mg/kg Nano-Se and 140 mg/kg Cd Cl_(2),n=20)and the Cd group(basic diet with 140 mg/kg Cd Cl_(2),n=20).The results of the experiment showed that the Purkinje cells were significantly decreased with their degradation and indistinct nucleoli after Cd exposure.Moreover,exposure to Cd caused a significant accumulation of Cd and cupper.However,the contents of Se,iron,and zinc were decreased,thereby disturbing the metal homeostasis in the cerebellum.The Cd exposure also resulted in high levels of malondialdehyde(MDA)and down regulation of selenoprotein transcriptome.Furthermore,the expressions of MTF1,metallothionein 1(MT1),MT2,zinc transporter 3(ZNT3),ZNT5,ZNT10,zrt,irt-like protein 8(ZIP8),ZIP10,transferrin(TF),ferroportin 1(FPN1),ATPase copper transporting beta(ATP7B),and copper uptake protein 1(CTR1)were inhibited by Cd exposure.However,all these changes were significantly alleviated by the supplementation of Nano-Se.This study proved that Cd could disorder metal homeostasis and induce oxidative stress,whereas Nano-Se could relieve all these negative effects caused by Cd via activating the MTF1-mediated metal response in the cerebellum of chicken.