LC-MS/MS法分析生猪异位发酵床垫料中5大类兽用抗菌药物残留及蓄积趋势

建立了生猪异位发酵床垫料中四环素类、喹诺酮类、磺胺类、酰胺醇类和大环内酯类共54种抗菌药物残留量的LC-MS/MS检测方法.经超低温冷冻粉碎后的垫料样本,费休氏-库仑滴定法测定水分,Mcllvaine-Na2 EDTA缓冲液和乙腈提取,亲水亲脂平衡固相萃取小柱净化,LC-MS/MS法测定抗菌药物残留量,最终结果以折水后...     

抗菌肽对抗生素耐药菌株的抑菌活性研究

[目的]研究抗菌肽对抗生素耐药菌株的抑菌活性。[方法]利用抗性平板划线法从腹泻病牛血便中筛选分离出1株耐药菌,通过16S rDNA序列进行鉴定,采用琼脂孔穴扩散法通过梯度盐酸壮观霉素（spectinomycin,Spe⁺）、氨苄青霉素（ampicillin,Amp⁺）、硫酸卡那霉素（kanamycin,Kan⁺）和氯霉素（chloramphenicol,Cm⁺）试验确定该菌药敏特性,并利用1种抗菌肽制剂对该菌株进行药敏试验。[结果]经BLAST比对分析该菌16S rDNA序列,鉴定该耐药菌为科氏葡萄球菌（Staphylococcus cohnii）,此菌对Amp⁺敏感,但对试验中其他抗生素均有耐药性,各梯度抗菌肽对该耐药菌均具有明显的抑菌活性。[结论]抗菌肽能有效抑制耐药科氏葡萄球菌的生长,有望在畜牧生产中代替抗生素使用。     

Youssoufenes A2 and A3, Antibiotic Dimeric Cinnamoyl Lipids from the ΔdtlA Mutant of a Marine-Derived Streptomyces Strain

Two new dimeric cinnamoyl lipids (CL) featuring with an unusual dearomatic carbon-bridge, named youssoufenes A2 (1) and A3 (2), were isolated from the ΔdtlA mutant strain of marine-derived Streptomyces youssoufiensis OUC6819. Structures of the isolated compounds were elucidated based on extensive MS and NMR spectroscopic analyses, and their absolute configurations were determined by combination of the long-range NOE-based ¹H-¹H distance measurements and ECD calculations. Compounds 1 and 2 exhibited moderate growth inhibition against multi-drug-resistant Enterococcus faecalis CCARM 5172 with an MIC value of 22.2 μM.     

府谷县兽用抗菌药减量化实施经验总结

为深入贯彻落实习近平总书记关于食品药品安全“四个最严”要求,全面提升畜禽绿色健康养殖水平,府谷县以《全国兽用抗菌药使用减量化行动方案（2021—2025年）》为契机,与西北农林科技大学畜禽无抗养殖技术团队合作,签订“府谷县畜禽减抗替抗养殖技术示范推广”项目合作协议,在府谷县全面开展抗生素减量化示范推广,得到了府谷县委县政府和榆林市科技局的大力支持,从饲料端禁抗、养殖端减抗替抗和粪污端无抗三个方面遏制抗菌药的使用,经过2年的技术示范推广,取得了一定的经验和成果。     

群体感应调控因子ComA对枯草芽胞杆菌NCD-2抑菌物质产生和生物膜形成的影响

为明确群体感应系统在枯草芽胞杆菌NCD-2菌株抑菌活性和生物膜形成中的作用,通过同源重组技术对群体感应系统中的comA基因进行定位缺失突变,分别比较了NCD-2菌株和comA基因突变子在胞外酶合成、抑菌活性、脂肽抗生素(丰产素)产生以及生物膜形成上的差异,并进一步利用实时荧光定量RT-PCR技术比较了二者生物膜基质编码基因epsA和tasA的表达情况。结果表明,通过同源重组技术获得了comA基因突变子MA-4,同菌株NCD-2相比,其在胞外蛋白酶和纤维素酶的合成能力、对番茄灰霉菌Botrytis cinerea的抑菌活性、丰产素的合成量和生物膜的形成能力上均明显下降;生物膜基质编码基因tasA的表达量下降了70%,而epsA的表达量变化不明显。表明ComA是NCD-2菌株脂肽抗生素和生物膜形成中的重要调控因子。     

应用发光细菌法检测饲用抗生素单一及联合毒性的研究

以HgCl2为毒性参照,以等毒性配比法研究了饲用盐酸金霉素、吉他霉素、盐霉素和黄霉素对发光细菌的单一及联合毒性。结果表明:盐酸金霉素、吉他霉素、盐霉素、黄霉素对明亮发光杆菌的急性毒性EC50分别为9.21、146.13、4.24 mg/L和134.68 mg/L,发光细菌的相对发光强度(RLU)随着抗生素浓度的提高而降低,并分别在2~16、25~200、1~8 mg/L和25~200 mg/L范围内呈良好线性关系;多元饲用抗生素对发光细菌具有联合毒性的作用,吉他霉素与盐酸金霉素、盐霉素和黄霉素3种抗生素的二元组合对发光细菌的联合毒性表现为较强的协同作用,而盐酸金霉素、盐霉素和黄霉素两两分别组合对发光细菌的联合毒性表现为拮抗作用;三元及四元抗生素混合体系对发光细菌的联合毒性均表现为拮抗作用。     

一株猪源大肠埃希氏菌卡那霉素耐药基因的研究

提取一株猪源大肠埃希氏菌卡那霉素耐药菌株E.coliBJ96147的质粒DNA进行转化,转化子能稳定表达其全部抗生素抗性,表明E.coliBJ96147对卡那霉素（K）、庆大霉（GM）、链霉素（S）、氯霉素（C）、四环素（Tc)、复方新诺明等六种抗生素的抗性由耐药性R质粒介导;根据已发表的编码卡那霉素抗性的磷酸转移酶[APH（3′）-Ⅱ]钝化酶基因的核苷酸序列,设计合成了一对引物P1和P2,用PVR技术检测其钝化酶基因,结果该引物扩增出了预期的578bp大小的PCR产物,淫限制性内切酶HindⅢ进行酶切分析,发现产物被切成二条带,与预出现的389bp及189bp二条带相符,说明PCR可用于检测猪源大肠埃希氏菌卡霉那素磷酸转移酶[APH（3′）-Ⅱ]钝化酶基因。     

Isolation and Identification of Pathogenic Bacteria Causing Dairy Cow Mastitis in Liaoning and Drug Sensibility Test of Pathogenic E.coli

In order to understand the main bacterial pathogen species causing dairy cow mastitis in Liaoning, as well as the characteristics of drug sensitivity of the pathogenic E.coli,the milk samples from 75 dairy cows with clinical manifestations for mastitis in certain large-scale dairy farm in Liaoning were collected.The bacteria in milk were cultured and isolated with biochemical methods and in vitro drug sensitivity tests were processed with the isolated E.coli strains.The results showed that the main bacterial pathogen for dairy cow mastitis were E.coli（separation rate 58.7%),S.aureus（64.0%）and S.agalactiae（54.7%),and multiple infection including double and triple infection were identified.The drug sensitivity tests on the isolated E.coli indicated that the E.coli isolates were highly resistant to sulfonamides（resistance rate>85%）and chloramphenicol（resistance rate>30%),and they were relatively low resistant to ampicillin（9.5%),ciprofloxacin（9.5%),ceftiofur（7.1%）and ofloxacin（4.8%).The results was able to provide reliable theoretical basis for prevention and control of dairy cow mastitis in Liaoning area.     

First report of Shewanella sp. and Listonella sp. infection in freshwater cultured loach,Misgurnus anguillicaudatus

A novel disease outbreak occurred among cultured loach, Misgurnus anguillicaudatus in China. The diseased fish displayed lethargy, loss of appetite, black discolouration, anasarca and ventral skin hyperaemia. Clinical signs were observed in most of the fish examined, which consisted of discoloured liver, swollen spleen and petechial haemorrhage in the intestine and peritoneal membranes. The causative agent was found to be two species of bacteria by experimental infection, identified as Shewanella sp. and Listonella sp. The result showed that these were likely to be Shewanella putrefaciens and Listonella anguillarum respectively. Bacterial identification consisted of physiological and biochemical tests as well as 16S rRNA sequence analysis. In addition, antibiotic susceptibility testing was performed, revealing that Shewanella putrefaciens and Listonella anguillarum were susceptible to enrofloxacin, ciprofloxacin, levofloxacin, pipemidicacid and norfloxacin. This is the first report of S. putrefaciens and L. anguillarum from cultured loach.     

Occurrence of multiple antibiotic resistance and genotypic characterization in Edwardsiella tarda isolated from cage‐cultured hybrid red tilapia (Oreochromis sp.) in the Ping River,Northern Thailand

Edwardsiella tarda is a pathogen that causes edwardsiellosis in aquatic animals. The emergence of multiple antibiotic‐resistant strains makes antibiotic treatment difficult. This study aimed to investigate the antibiotic susceptibility patterns and the genotypic characterization of E. tarda isolated from cage‐cultured red tilapia in Thailand. A total of 30 isolates were identified as E. tarda using biochemical and molecular analysis. The disc diffusion method for testing antibiotic susceptibility showed all the isolates were resistant to colistin sulphate and oxolinic acid. High levels of resistance to amoxicillin, ampicillin, ceftazidime, oxytetracycline and sulphamethoxazole/trimethoprim were observed as well. The multiple antibiotic resistance index ranged from 0.25 to 0.92, indicating that these isolates had been exposed to high risk sources of contamination where antibiotics were commonly used. All the isolates carried the bla_TEM gene based on polymerase chain reaction (PCR). The tetA and sul3 genes were detected in 90% (27/30) and 26.7% (8/30) of the isolates respectively. Nine different genetic groups of isolates were obtained using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC‐PCR). A correlation between genetic types and multiple antibiotic‐resistant patterns was found. These results highlight the potential risks of multiple antibiotic‐resistant isolates for humans and the environment.