P10B抗菌肽和硫酸小檗碱对肉鸡源大肠杆菌的体外联合抗菌作用研究

为了解P10B抗菌肽与硫酸小檗碱联合抗菌作用,进行P10B抗菌肽与硫酸小檗碱联用的体外抑菌试验。试验菌株为20株肉鸡源大肠杆菌菌株。采用微量肉汤稀释法测定P10B与硫酸小檗碱对大肠杆菌的最低抑菌浓度;棋盘稀释法测定两个药的分级抑菌浓度指数,判断联合抗菌效果。结果表明,在联合药敏试验中,两药呈协同作用的占10%,呈相加作用的占85%,呈无关作用的占5%。平均分级抑菌浓度指数为0.825,两种药联和用药效应主要表现为累加作用。     

Amoxicillin—current use in swine medicine

Amoxicillin has become a major antimicrobial substance in pig medicine for the treatment and control of severe, systemic infections such as Streptococcus suis. The minimum inhibitory concentration 90% (MIC 90) is 0.06 μg amoxicillin/ml, and the proposed epidemiological cut‐off value (ECOFF) is 0.5 μg/ml, giving only 0.7% of isolates above the ECOFF or of reduced susceptibility. Clinical breakpoints have not been set for amoxicillin against porcine pathogens yet, hence the use of ECOFFs. It has also been successfully used for bacterial respiratory infections caused by Actinobacillus pleuropneumoniae and Pasteurella multocida. The ECOFF for amoxicillin against A. pleuropneumoniae is also 0.5 μg/ml demonstrating only a reduced susceptibility in 11.3% of isolates. Similarly, P. multocida had an ECOFF of 1.0 μg/ml and a reduced susceptibility in only 2.6% of isolates. This reduced susceptibility disappears when combined with the beta‐lactamase inhibitor, clavulanic acid, demonstrating that it is primarily associated with beta‐lactamase production. In contrast, amoxicillin is active against Escherichia coli and Salmonella species but using ECOFFs of 8.0 and 4.0 μg/ml, respectively, reduced susceptibility can be seen in 70.9% and 67.7% of isolates. These high levels of reduced susceptibility are primarily due to beta‐lactamase production also, and most of this resistance can be overcome by the combination of amoxicillin with clavulanic acid. Currently, amoxicillin alone is considered an extremely valuable antimicrobial in both human and animal medicine and remains in the critically important category of antibiotics alongside the fluoroquinolones and macrolides by the World Health Organization as well as the third‐ and fourth‐generation cephalosporins, but these cephalosporins show marked resistance to basic beta‐lactamase production and are only destroyed by the extended‐spectrum beta‐lactamases. Amoxicillin alone and in combination with clavulanic acid are currently classed together in Category 2 in the European Union. By reviewing the pharmacodynamic data and comparing this with pharmacokinetic data from healthy and infected animals and clinical trial data, it can be seen that the product has a good efficacy against S. suis and A. pleuropneumoniae, in spite of usage over many years. However, it may be much less efficacious on its own against E. coli, due to reduced susceptibility and resistance associated with beta‐lactamase production, which is largely overcome by the use of clavulanic acid. It is felt that this differentiation may be useful in future classification of amoxicillin alone, in comparison with its combined use with clavulanic acid and thereby preserve the use of the more critically important antibiotics in veterinary medicine and reducing the risk of their resistance being transmitted to human.     

Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs

Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis.     

Effect on Rendement Napole genotype on metabolic markers in Ossabaw pigs fed different levels of fat

We investigated effects of Rendement Napole (RN ) genotype on metabolic markers in Ossabaw pigs fed diets with different levels of dietary fat. Thirty‐two pigs, belonging to either the wild‐type (WT , rn⁺/rn⁺) or carrier (CAR , RN ^?/rn⁺) genotypes (n  = 16/genotype), were divided into two dietary groups, (high fat [HF ] or low fat [LF ]) diets, for 12 weeks (n  = 8 pigs/genotype/diet) after which pigs were killed for gene expression analysis by RT ‐PCR . Feeding HF diet caused increased daily gain (ADG , p  <  .05) and final body weight (BW ) (p  <  .05) in comparison with the LF diet (p  <  .05). Feed efficiency (gain:feed) was higher (p  <  .05) in pigs on the HF and was higher (p  <  .05) in CAR pigs compared to WT . There was genotype × diet interaction (p  =  .05) on final BW such that CAR animals on LF diet had the same final BW as animals of both genotypes on HF diet. Carrier pigs on LF diet had higher (p  <  .05) average daily gain and gain:feed than WT pigs. There was a trend (p  <  .08) for a higher feed consumption in pigs on the LF diet. Backfat thickness was higher (p  <  .01) in pigs on the HF diet. Serum triglyceride was higher (0.62 vs. 0.33 mg/dl, p  <  .01) in pigs on HF diet. Serum insulin was higher (p  <  .05) in CAR versus WT pigs (0.40 vs. 0.015 μg/ml). Pigs on the HF diet had a higher (p  <  .05) serum insulin compared to those on the LF diet (0.032 vs. 0.023 μg/ml). Carnitine palmitoyl transferase 1‐alpha was higher (p  <  .05) in the longissimus dorsi and semitendinosus muscles of pigs on HF diet. Acyl‐CoA oxidase I was elevated (p  <  .05) in the liver of pigs on HF diet. Fatty acid synthase was lower in the longissimus dorsi muscle, liver and mesenteric fat (p  <  .05) of carrier pigs. The RN gene regulates specific metabolic markers in the Ossabaw pigs.     

Effect of feed intake level and dietary protein content on the body temperature of pigs housed under thermo neutral conditions

Feed intake and diet composition appear to affect the body temperature of pigs. Two trials were conducted to analyse the effect of feed intake level and dietary protein content on the intestinal temperature (IT) of pigs housed under thermo neutral conditions. Ten pigs (64.1 ± 1.3 kg initial body weight) fitted with an ileal cannula were used. A thermometer set to register the IT at 5‐min intervals was implanted into the ileum through the cannula. In both trials, the ambient temperature ranged from 19.1 to 21.6°C and the pigs were fed at 07:00 and 19:00 hr (same amount each time). In trial 1, the pigs were fed daily 1.2 or 1.8 kg of a wheat–soybean meal diet. The IT followed a similar pattern along a 24‐hr period regardless the feed intake level. The IT rapidly increased up to 0.61 and 0.74°C after the morning meal and up to 0.53 and 0.47°C after the evening meal in pigs fed 1.2 and 1.8 kg/d respectively. The postprandial IT was higher in pigs fed 1.8 kg after each meal (p < .05). In trial 2, pigs were fed daily 1.8 kg of a low (11%) or a high (22%) crude protein diet. The IT followed a similar pattern along the 24‐hr period regardless the dietary protein level. The postprandial IT did not differ between pigs fed the low protein or the high protein (p > .10). The IT rapidly increased up to 0.66 and 0.62°C after the morning meal in pigs fed the high‐ and low‐protein diet (p < .05), but there was no change after the evening meal (p > .10). In conclusion, the feed intake level affected the IT of pigs housed under TN conditions, but the dietary protein content had no effect.     

β‐hydroxy‐β‐methyl butyrate promotes leucine metabolism and improves muscle fibre composition in growing pigs

The aim of this study was to investigate the effects of excess leucine (Leu) vs. its metabolites α‐ketoisocaproate (KIC) and β‐hydroxy‐β‐methyl butyrate (HMB) on Leu metabolism, muscle fibre composition and muscle growth in growing pigs. Thirty‐two pigs with a similar initial weight (9.55 ± 0.19 kg) were fed 1 of 4 diets for 45 days: basal diet, basal diet + 1.25% L‐Leu, basal diet + 1.25% KIC‐Ca, basal diet + 0.62% HMB‐Ca. Results indicated that relative to the basal diet and HMB groups, Leu and KIC groups exhibited increased Leu concentrations and decreased concentrations of isoleucine, valine and EAAs in selected muscle (p < 0.05) and had lower mRNA levels of MyHC I and higher expression of MyHC IIx/IIb (p < 0.05), and there was no significant difference between the basal and HMB‐supplemented groups. Moreover, the mRNA expression levels of AMPKα and UCP3 were higher but the myostatin mRNA levels were lower in the soleus muscle of the HMB group than those from other groups (p < 0.05). These findings demonstrated that doubling dietary Leu content exerted growth‐depressing effects in growing pigs; dietary KIC supplementation induced muscular branched‐chain amino acid imbalance and promoted muscle toward a more glycolytic phenotype; while dietary HMB supplementation promoted the generation of more oxidative muscle types and increased muscle growth specially in oxidative skeletal muscle, and these effects of HMB might be associated with the AMPKα‐Sirt1‐PGC‐1α axis and mitochondrial biogenesis.     

Lipopolysaccharide and cytokines modulate leukotriene (LT)B4 and LTC4 production by porcine endometrial endothelial cells

Uterine inflammatory response is mediated by inflammatory mediators including eicosanoids and cytokines produced by immune and endometrial cells. Interactions between lipopolysaccharide (LPS) and cytokines, and leukotrienes (LTs) in endothelium, important for the host defence during the inflammation, are unknown. We studied the effect of LPS, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐4 and IL‐10 on 5‐lipooxygenase (5‐LO), LTA₄ hydrolase (LTAH) and LTC₄ synthase (LTCS) mRNA and protein expression, LTB₄ and LTC₄ release from porcine endometrial endothelial cells, and cell viability. For 24 hr, cells were exposed to LPS (10 or 100 ng/ml of medium) and cytokines (each 1 or 10 ng/ml). 5‐LO mRNA/protein expression augmented after incubation with larger doses of LPS, TNF‐α, IL‐4 and IL‐10 and smaller dose of IL‐1β. Larger dose of TNF‐α, smaller doses of LPS and IL‐1β and both doses of IL‐10 increased LTAH mRNA/protein expression. LTAH protein content was up‐regulated by larger dose of LPS, but it was reduced in response to both doses of IL‐4. LTCS mRNA expression was elevated by larger doses of LPS, IL‐4 and IL‐10 or both doses of TNF‐α and IL‐1β. LTCS protein level increased after treatment with both doses of IL‐1β, IL‐4 and IL‐10, smaller dose of LPS and larger dose of TNF‐α. Both doses of LPS and larger doses of TNF‐α and IL‐10 increased LTB₄ release. LPS, IL‐1β and IL‐10 at smaller doses, or TNF‐α and IL‐4 at larger doses stimulated LTC₄ release. Smaller doses of TNF‐α and IL‐1β or both doses of IL‐4 enhanced the cell viability. This work provides new insight on the participation of LPS, TNF‐α, IL‐1β, IL‐4 and IL‐10 in LTB₄ and LTC₄ production/release from porcine endometrial endothelial cells, and the effect of above factors on these cells viability. The used cellular model gives the possibility to further establish the interactions between inflammatory mediators.     

基于“多基因聚合-早世代组配”策略的水稻分子改良研究

传统水稻育种技术最主要瓶颈是选择效率低和周期长。为了提高水稻优异材料选育和杂交稻组配效率,创新水稻分子育种策略,利用分子标记辅助选择多基因聚合和早世代杂交组配,展开水稻恢复系分子改良和杂交新组合的调查评价。供体亲本H318与恢复系亲本华占进行杂交,通过选择和设计关键有利基因的分子标记,利用毛细管电泳基因分型技术,将Wxb、fgr、Xa23、Pi2、Pi46以及Pita 6个稻米品质、香味、抗白叶枯病和抗稻瘟病相关功能基因进行聚合利用。通过多世代的田间生物学性状调查,米质、抗性等表型鉴定,获得14份以恢复系华占为遗传背景,目标基因纯合的优质、双抗和香型稳定的水稻株系。依据材料稳定遗传特性,将改良后代株系与生产应用的不育系进行测配和组合的调查评价,筛选获得潜在优良杂交稻。在育种进程中采用"多基因聚合-早世代组配"策略,实现多个有利基因快速聚合,定向改良稻米品质和抗病性等关键性状,并且促进杂交水稻组合的高效选育。     

外源氮添加对湿地土壤N_2O排放量的影响

为搞清湿地土壤驱动N2O排放的关键氮源类型,有效减少湿地N2O的排放,本文通过室内控制温湿度,用气相色谱法分析不同外源氮素对湿地N2O排放的影响。结果表明:外加氮源组总是高于对照组N2O排放量(4.4 mg·m-3)。在设定的剂量范围内,单独添加尿素或尿素与硝酸铵1∶1配合时N2O排放量呈现先增后减的单峰分布趋势,峰值分别为10.6 mg·m-3和229.0 mg·m-3;单独添加硝酸铵时N2O排放量(32.6~111.0 mg·m-3)随着氮素添加量增加呈现持续上升趋势。单独添加尿素或硝酸铵、尿素与硝酸铵1∶1配合均促进N2O的排放,但硝酸铵尿素混合添加对N2O排放量的贡献单独添加硝酸铵单独添加尿素。这为预测内蒙古高原区农牧交错带湿地氮素输入可能带来的温室效应和有效减排提供科学依据。