Pharmacokinetics and dosage regimen of cefotaxime in cross-bred calves following single intramuscular administration

The pharmacokinetics, penetration into erythrocytes and plasma protein binding of cefotaxime were investigated in cross-bred calves. Following a single intramuscular dose of cefotaxime (10 mg/kg), the absorption half-life and elimination half-life were 0.13±0.03 h and 2.97±0.72 h, respectively. The apparent volume of distribution and total body clearance were 3.28±0.72 L/kg and 0.78±0.08 L/kg per h, respectively. The extent of penetration into erythrocytes was 24–40% of the total blood concentration. Cefotaxime was bound to plasma proteins of calves to the extent of 25.5–33.6%. A satisfactory intramuscular dosage regimen for cefotaxime in calves would be 11 mg/kg followed by 10 mg/kg at 7 h intervals.Abbreviations ATCC American type cell culture - MIC minimum inhibitory concentration - PCV packed cell volume     

Antibiotic resistant Escherichia coli in southeastern Australian pig herds and implications for surveillance

To investigate public health implications of antibiotics to control post‐weaning scours, we surveyed 22 commercial pig herds in southeastern Australia. Fifty faecal samples per herd were collected from pre‐ and post‐weaned piglets. Presumptive Escherichia coli isolates were confirmed by MALDI‐TOF MS. Isolates (n = 325) were screened for susceptibility to 19 veterinary antibiotics using MIC broth microdilution. All 325 E. coli isolates underwent further testing against 27 antibiotics used in human medicine and were screened for ETEC adhesin and enterotoxin genes (F4 (K88), F5 (K99), F6 (987P), F18, F41, STa, STb, Stx2e and LT) by multiplex PCR. Isolates identified as phenotypically resistant to third‐generation cephalosporin (3GC) and aminoglycoside antibiotics were screened by multiplex PCR/reverse line blot to detect common β‐lactam and aminoglycosides resistance genes, confirmed by sequencing. Twenty (6.1%) of the E. coli isolates were resistant to 3GC antibiotics and 24 (7.4%) to the aminoglycoside antibiotic gentamicin. Genetic analysis revealed six different extended spectrum β‐lactamase (ESBL) genes (blaCTX‐M‐1, ‐14, ‐15, ‐27, blaSHV‐12 and blaCMY‐2‐like genes), four of which have not been previously reported in Australian pigs. Critically, the prevalence of 3GC resistance was higher in non‐pathogenic (non‐ETEC) isolates and those from clinically normal (non‐diarrhoeal) samples. This highlights the importance of non‐ETECE. coli as reservoirs of antimicrobial resistance genes in piglet pens. Antimicrobial resistance surveillance in pig production focused on diagnostic specimens from clinically‐affected animals might be potentially misleading. We recommend that surveillance for emerging antimicrobial resistance such as to 3GC antibiotics should include clinically healthy pigs.     

Phylogenetic groups and cephalosporin resistance genes of Escherichia coli from diseased food-producing animals in Japan

A total of 318 Escherichia coli isolates obtained from different food-producing animals affected with colibacillosis between 2001 and 2006 were subjected to phylogenetic analysis: 72 bovine isolates, 89 poultry isolates and 157 porcine isolates. Overall, the phylogenetic group A was predominant in isolates from cattle (36/72, 50%) and pigs (101/157, 64.3%) whereas groups A (44/89, 49.4%) and D (40/89, 44.9%) were predominant in isolates from poultry. In addition, group B2 was not found among diseased food-producing animals except for a poultry isolate. Thus, the phylogenetic group distribution of E. coli from diseased animals was different by animal species. Among the 318 isolates, cefazolin resistance (minimum inhibitory concentrations: ≥32 μg/ml) was found in six bovine isolates, 29 poultry isolates and three porcine isolates. Of them, 11 isolates (nine from poultry and two from cattle) produced extended spectrum β-lactamase (ESBL). The two bovine isolates produced bla_CTX-M-2, while the nine poultry isolates produced bla_CTX-M-25 (4), bla_SHV-2 (3), bla_CTX-M-15 (1) and bla_CTX-M-2 (1). Thus, our results showed that several types of ESBL were identified and three types of β-lactamase (SHV-2, CTX-M-25 and CTX-M-15) were observed for the first time in E. coli from diseased animals in Japan.     

植物组织培养中抗污染培养基新配方的探索

组织培养中培养基的污染问题一直是该领域的技术盲区,本实验旨在研发更高效抑制杂菌污染的植物组织培养系统。通过在MS培养基中添加不同浓度及组合的头孢霉素、利福平、青霉素等抗生素以及化学药物进行抗菌筛选,确定了以头孢霉素、百菌清、代森锰锌为主的抗菌培养基配方。之后评估甘薯(Ipomoea batatas)、烟草(Nicotiana tabacum)与拟南芥(Arabidopsis thaliana)三种组培材料在该培养基上的生长与污染情况,进而探索出一种高效培养基抗污染新方法,即药液包衣。利用含有25 mg/L头孢霉素+30 mg/L的代森锰锌与百菌清1:1混合液的培养基,并在外植体插入培养基前利用100 mg/L多菌灵药液或100 mg/L代森锰锌与百菌清1:1混合液进行浸润包衣,可显著降低组培材料被污染的风险,且对除愈伤组织以外的外植体的生长发育影响较弱。该方法的使用在对外植体伤害较小的情况下,对于已污染组培材料的挽救以及大田种质材料的实验室保存有重要借鉴意义。     

PRIME HLB固相萃取/超高效液相色谱-串联质谱法测定乳制品中头孢菌素类药物残留

建立牛乳、酸乳和乳粉3 种基质乳制品中采用超高效液相色谱-串联质谱（ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS）同时检测4 种头孢菌素类药物残留的方法。以5%甲酸乙腈溶液进行提取,经Oasis PRIME HLB固相萃取柱净化,氮吹后复溶,采用电喷雾离子源正离子模式及多反应监测模式进行检测。结果表明：牛乳和酸乳中头孢氨苄、头孢匹林、头孢洛宁和头孢喹肟的定量限为4 μg/kg,乳粉中为32 μg/kg     

头孢菌素类降解菌Achromobacter sp.YF-1的筛选与功能鉴定

为合理、无污染地利用头孢菌素类废弃药渣,以长期堆放的头孢菌素类药渣土壤为样品源,从中获得浸出液,通过第三代头孢药品头孢克肟驯化、筛选出具有头孢菌素类抗生素降解能力的菌株;利用革兰氏染色法、电镜超薄切片技术双染色法对其进行形态学观察,16S rDNA进行分子生物学序列分析;采用单因素控制变量法对菌株的生长特性及最佳降解条件进行了初步探讨。结果表明:该菌为杆状细菌,无芽孢;经16S rDNA序列比对,进一步确定该菌株属于Achromobacter属,并命名为Achromobacter sp. YF-1;该降解菌在温度37℃、pH 6～7、转速120 r/min培养条件下培养7 d,头孢菌素类抗生素降解率达92.71%;该菌株对头孢菌素类抗生素的降解能力稳定,可用于处理药渣中残留的头孢菌素类抗生素。     

头孢菌素C对蔬菜种子萌发的毒理效应

为探究头孢菌素菌渣无害化处置与肥料化利用过程中残留的头孢菌素C对蔬菜生长的毒理效应，通过恒温保湿培养法，采用生菜（Lactuca sativa Linn.var.ramosa Hort.）、油麦菜（Lactuca sativa var.longifoliaf.Lam）、白菜（Brassica pekinensis（Lour.）Rupr.）及油菜（Brassica campestris L.） 4种蔬菜种子为研究对象，以种子发芽率、幼苗的鲜质量、胚轴长、胚根长为毒性敏感指标，研究了7种浓度头孢菌素C溶液胁迫下4种蔬菜毒性的剂量-效应关系，比较了不同蔬菜在头孢菌素C毒害下的生态毒性差异和相对敏感的指标。结果表明：头孢菌素C溶液对4种蔬菜种子毒性敏感指标的影响程度依次为胚根 > 鲜质量 > 胚轴 > 发芽率；头孢菌素C溶液浓度与4种蔬菜胚根生长率呈显著的线性关系（P<0.01）；头孢菌素C对生菜、油麦菜、白菜以及油菜的最大无响应浓度（NOEC）分别为868.10、727.25、796.27 mg·L^-1和629.277 mg·L^-1，4种蔬菜对头孢菌素C的毒性敏感度依次为油菜 > 油麦菜 > 白菜 > 生菜，其中油菜IC₅₀值为1 538.36 mg·L^-1。研究表明，与油麦菜、白菜和生菜相比，油菜更适合作为后续实验的生态毒性指示植物。     

鸡肉中头孢类抗生素的MISPE-HPCE检测

以7-氨基头孢烷酸为模板分子,4-乙烯基吡啶为功能单体,乙二醇二甲基丙烯酸酯为交联剂,制备了7-氨基头孢烷酸分子印迹聚合物。以该分子印迹聚合物为固相萃取材料,以高效毛细管电泳为检测手段,进行头孢氨苄、头孢拉定、头孢哌酮、头孢唑啉等4种头孢类药物的色谱分析。结果表明,该方法能有效萃取和检测鸡肉中的药物。在试验条件下,4种头孢类药物的回收率为78.00%～83.04%,RSD为2.18%～3.79%。