Isolation,Identification and Biological Characteristics of Staphylococcus chromogenes from Tibetan Pigs

In order to understand the infection of Staphylococcus disease in healthy Tibetan pigs in Linzhi city,Tibet.232 samples were collected from the slaughterhouse in Bahe town,Gongbujiangda county,Linzhi city,Tibet,the teaching practice ranch at the Tibet Agriculture and Animal Husbandry University in Bayi district,Linzhi city,and the slaughterhouse in Bayi district,Linzhi city.These samples were isolated,cultured and identified,and one isolated strain was systematically studied for biological characteristics.Epidemiological survey results showed that 73 of 232 samples were positive for Staphylococcus,the positive rate was 31.47%,16 positive samples were detected in the slaughterhouse of Bahe town,Gongbujiangda county,Linzhi,Tibet,with a detection rate of 23.89%(16/67),43 positive samples were detected in the pastures of the teaching practice ranch at the Tibet Agriculture and Animal Husbandry University in Bayi district,Linzhi city,with a detection rate of 34.40% (43/125),14 positive samples were detected in the slanghterhouse in Bayi district,Linzhi city,with a detection rate of 35.00% (14/40).Through PCR identification,gene sequencing and biochemical identification,5 strains of Staphylococcus chromogenes were isolated,and one of them (Sa LZ1) was studied systematically.The biological characteristics of the isolated strains showed that the Sa LZ1 strain formed round white convex colonies on the agar medium,with regular edges,smooth surfaces,and opaque white colonies.The isolates were Gram-positive cocci bacteria.PCR sequencing results showed that the isolate had the highest homology with Staphylococcus chromogenes (GenBank accession No.:MG576253.1),positive for serum inulin,manicol,glucose,saccharose,lactose,negative for arabinose,sodium hippurate,raffinose,xylose,urea,etc,consistent with the biochemical characteristics of Staphylococcus chromogenes.The logarithmic growth period of isolated bacteria was 5 to 10 h.The pathogenicity test of mice showed that the mortality rate of mice reached 100% when the concentration of Sa LZ1 strain reached 3.5×10⁹ CFU/mL.And the results of pathological section showed that Sa LZ1 strain could cause the visceral tissue of mice to develop pathological changes.The above results provided a theoretical basis for the prevention and control of Staphylococcus suis in Linzhi,Tibet.     

Construction of Full-length cDNA of Recombinant Rabies Virus SRV9 Strain Expressing EgM123 Gene of Echinococcus granulosus

The aim of the experiment was to construct the recombinant rabies virus SRV₉ vaccine strain with EgM123 gene by reverse genetics technology and provide the technical means for effective prevention and control of rabies and hydatidosis in China's agricultural and pastoral areas.In this study,the structural protein N,P and L genes of rabies virus SRV₉ were synthesized using gene synthesis technology,which was based on the complete genome sequence of rabies virus SRV₉ and the fusion fragment of the N-P-M fusion fragment and the rabies G gene,through the carrier of enzyme insertion connection methods,the recombinant rabies virus L gene,N-P-M gene fusion fragment and G+EgM123+eGFP gene fusion fragment were successively recombined on the expression vector pcDNA3.1(-) to construct the full-length cDNA of recombinant rabies virus SRV₉ with EgM123 gene.The synthesized genes were constructed on pcDNA3.1(-) expression vector,and the results of transformation,plasmid digestion and gene sequencing showed that the length of N,P,L,N+P+M and G+EgM123+eGFP gene fragments were 1 365,1 107,6 471,3 160 and 3 256 bp,respectively.The full-length cDNA fragment of EgM123 gene recombinant rabies virus full-length cDNA was 12 465 bp,and the sequencing results of each gene fragment were 100%.In this experiment,the full-length cDNA fragment of recombinant EgM123 rabies and eukaryotic expression vectors of the N,P and L genes of rabies virus were successfully constructed,which could save EgM123 gene recombinant rabies by reverse genetics,it also provided the reference for the development of rabies and hydatid disease combined gene recombinant oral live vaccine.     

Effect of Supplementing Alfalfa Powder to Diet on Growth Performance and Meat Quality of Goose

The experiment was aimed to investigate the effects of dietary alfalfa powder on the growth performance,body size,slaughter performance,organ index and meat quality of Siji geese.A total of 256 15-day-old geese (half male and half female) with similar body weight were randomly divided into four groups with 4 replicates of 16 geese each.The geese in control group were fed with corn-soybean meal-based diet (control diet),and geese in the experimental groups were fed the basal diet replaced with 10%,20% and 30% alfalfa powder,respectively.The results showed that,compared with the control group,dietary alfalfa powder supplementation had no significant effect on the average daily intake,average daily gain,feed conversion ratio,body oblique length,keel length,shin circumference,semi diving length,chest width and shin length (P>0.05),and also had no effect on the carcass ratio,half-eviscerated carcass ratio and pectoral muscle ratio (P>0.05).However,dietary 20% and 30% alfalfa powder supplementation significantly increased the chest depth (P<0.05),dietary 30% alfalfa powder significantly decreased the full-eviscerated carcass ratio (P<0.05),and dietary 10% alfalfa powder significantly increased the leg muscle ratio (P<0.05) of geese.In addition,dietary alfalfa powder significantly decreased the abdominal fat ratio (P<0.05),raised the growth index of liver,jejunum,ileum,and cecum (P<0.05),and increased the water loss rate of pectoral muscle and leg muscle (P<0.05) compared with the control group.Dietary 10% and 30% alfalfa powder significantly increased duodenal growth index (P<0.05) of geese.In conclusion,dietary alfalfa powder supplementation might improve the slaughter performance (increase chest depth and leg muscle ratio,reduce abdominal fat rate) and organ growth index (increase the growth index of liver,duodenum,jejunum,ileum and cecum).However,dietary alfalfa powder increased water loss rates in pectoral muscle and leg muscle,and 30% alfalfa powder reduced the full-eviscerated carcass ratio of geese.The appropriate supplementation level of alfalfa powder was recommended at 10% in the present study.     

Evaluation of Thermal Environment of Cowshed and Correlation Analysis Between Thermal Parameter and Physiological Parameter of Dairy Cows in Central Plain of Hebei

The purpose of this study was to evaluate thermal environment of cowshed in four seasons in central plain of Hebei,and analyze the correlation between the temperature parameters and physiological indexes of dairy cows.Three dairy cow houses with different building structures were selected,the thermal parameters (ambient temperature (Ta) and relative humidity (RH)) and physiological parameters (respiratory rate,rectal temperature and body surface temperature) were detected.The results showed that the Ta,RH and index of temperature and humidity (THI) changed significantly in four seasons (P<0.05),the average daily temperature in summer was the highest (28.59 ℃),and the average daily temperature in winter was the lowest (1.55 ℃).In summer,dairy cows were suffering from mild heat stress for 15.5 h every day,and from moderate heat stress for 6.0 h.In winter,dairy cows were under mild cold stress for an average of 12.0 h per day.The Ta and THI of three cowsheds in each season had no significant differece (P>0.05),except for that in winter.Compared with the cowshed with low wall or roller blind,the average Ta in the cowshed with only roof was 0.80-1.27 ℃ higher in summer and 1.36-1.84 ℃ lower in winter.In addition,the physiological parameters of dairy cows were extremely significantly higher in summer than those in other seasons (P<0.01).The respiration frequency or rectal temperature of cows among cowsheds in summer were significantly different (P<0.05),and the seasonal difference in body surface temperature was significant (P<0.05).Correlation analysis of thermal parameters and cow physiological parameters showed that the respiratory frequency,rectal temperature and body surface temperature were positively correlated with THI and Ta (P<0.05).However,there was no significant correlation between physiological parameters and RH (P>0.05).The results provided scientific basis for environment evaluation of cows,and physiological status of dairy cows were inferred according to environmental thermal parameters,which provided data for occurrence and early warning of stress.     

Isolation and Identification of a Variant Porcine Epidemic Diarrhea Virus and the Immune Efficacy of Its Inactivated Vaccine

This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID₅₀,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×10^8.0 TCID₅₀/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×10^8.0TCID₅₀/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China.     

草坪草抗旱生理与节水技术研究进展

针对当前绿地草坪建设中水资源不足问题，分别论述了干旱胁迫对草坪草生长发育的影响及草坪草抵抗干旱的生理机制。并从节水灌溉、节水抗旱型品种选育、抗旱保水剂和草坪管理措施等方面综述了近年来在草坪业上的相关节水技术。     

紫花苜蓿品种比较试验

通过对26个紫花苜蓿（Medicago sativa）品种的生育期、株高、产量、越冬率等指标的试验比较，结果表明：公农一号、保丰、陇东、德福等几个品种的表现优于其他参试品种，适宜在辽西北风沙地地区推广种植。