细菌生物被膜与抗生素耐药机制研究进展

细菌引发的感染已成为感染性疾病的主要原因之一,具有耐药性和难治性的特点,而且还可以对抗机体的免疫清除作用,对人类和动物的健康造成很大的危害,也给人们生活的其他方面带来了越来越严重的影响,引起了基础和临床研究的极大关注。但是细菌生物被膜对抗生素的耐药机制目前还不十分清楚,现对近年来细菌生物被膜对抗生素耐药形成的几种可能机制进行综述。     

Identification of the tet(B) resistance gene in Streptococcus suis

The tetracycline resistance gene, tet(B), has been described previously in Gram negative bacteria. In this study tet(B) was detected in plasmid extracts from 17/111 (15%) Streptococcus suis isolates from diseased pigs, representing the first report of this resistance gene in Gram positive bacteria.     

Measurement of C‐peptide concentrations and responses to somatostatin,glucose infusion,and insulin resistance in horses

Reasons for performing study: Hyperinsulinaemia is detected in horses with insulin resistance (IR) and has previously been attributed to increased pancreatic insulin secretion. Connecting peptide (C‐peptide) can be measured to assess pancreatic function because it is secreted in equimolar amounts with insulin and does not undergo hepatic clearance. Hypothesis: A human double antibody radioimmunoassay (RIA) detects C‐peptide in equine serum and concentrations would reflect responses to different stimuli and conditions. Methods: A validation procedure was performed to assess the RIA. Six mature mares were selected and somatostatin administered i.v. as a primed continuous rate infusion, followed by 50 nmol human C‐peptide i.v. Insulin and C‐peptide concentrations were measured in horses (n = 6) undergoing an insulin‐modified frequently sampled i.v. glucose tolerance test, and in horses with insulin resistance (n = 10) or normal insulin sensitivity (n = 20). Results: A human RIA was validated for use with equine sera. Endogenous C‐peptide secretion was suppressed by somatostatin and median (range) clearance rate was 0.83 (0.15–1.61) ml/min/kg bwt. Mean ± s.d. C‐peptide‐to‐insulin ratio significantly (P = 0.004) decreased during the glucose tolerance test from 3.60 ± 1.95 prior to infusion to 1.03 ± 0.18 during the first 20 min following dextrose administration. Median C‐peptide and insulin concentrations were 1.5‐ and 9.5‐fold higher, respectively in horses with IR, compared with healthy horses. Conclusions: Endogenous C‐peptide secretion decreases in response to somatostatin and increases after dextrose infusion. Results suggest that relative insulin clearance decreases as pancreatic secretion increases in response to dextrose infusion. Hyperinsulinaemia in insulin resistant horses may be associated with both increased insulin secretion and decreased insulin clearance. Potential relevance: Both C‐peptide and insulin concentrations should be measured to assess pancreatic secretion and insulin clearance in horses.     

Variation in resistance to the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in Pietrain and Miniature pigs

Porcine reproductive and respiratory syndrome (PRRS) is one of the economically most important diseases of swine. Viraemia and the prolonged persistence of the virus are among the most critical factors. Virus replication and severity of disease vary with virus isolates, and there is rising evidence for a genetic component of the host susceptibility. Dissecting the genetic basis of resistance/susceptibility to PRRS virus (PRRSV) might lead to improved knowledge on the molecular mechanisms of PRRS and the establishment of genetic markers for future disease control. The aim of this study was to establish a porcine model with emphasized genetic differences in PRRSV susceptibility. Seven ‘Wiesenauer Miniature’ pigs (MI), a local German breed and eight commercial Pietrain (PI) pigs were challenged with 10⁵ TCID₅₀ of an attenuated PRRSV strain (Ingelvac^® PRRSV MLV). Clinical status, viraemia and seroconversion of the pigs were compared. No clinical signs were observed during the experiment. Viraemia peaked on day 6 p.i., with 100% of viraemic pigs in PI and on day 12 p.i with 87% of viraemic MI. Viraemia lasted for up to 35 days in MI and for at least 72 days in PI. This surprising result was confirmed by a second study with another four MI. MI and PI showed maximum virus titres of 10^2.5 TCID₅₀/ml of serum and 10^4.5 TCID₅₀/ml, respectively, indicating a virus replication in MI of approximately 3.3% that of PI over the complete period. MI were more efficient in antibody production. With such pronounced breed differences, the model is of high relevance for the genetic dissection of PRRS pathogenesis and susceptibility.     

鸭大肠杆菌超广谱β-内酰胺酶的检测及药物敏感性分析

分析鸭大肠杆菌的耐药性与超广谱β-内酰胺酶的关系,为临床相关感染的治疗提供理论依据。采用CLSI推荐的初步筛选试验和表型确证试验方法,检测了临床分离的12株鸭大肠杆菌产超广谱β-内酰胺酶的情况,用微量稀释法测定了环丙沙星等9种抗菌药对其的抗菌活性。临床分离的12株鸭大肠杆菌中,有4株产超广谱β-内酰胺酶,检出率为33%。产超广谱β-内酰胺酶的菌株对多种药物耐药,对氟喹诺酮类药物之间产生交叉耐药,对氟喹诺酮类、氨基糖苷类、头孢菌素类产生多重耐药性,产酶菌的耐药性明显大于非产酶菌。产超广谱β-内酰胺酶是鸭大肠杆菌对抗菌药物产生耐药性的主要机制之一。     

蜡样芽孢杆菌芽孢萌发及其对常规灭菌方法抵抗力的研究简

芽孢杆菌属细菌在芽孢状态下,对外界环境及理化因素具有高度抵抗力。适宜的温度、湿度及营养条件能促进芽孢萌发,影响其对常规灭菌方法的抵抗力。本试验选取蜡样芽孢杆菌作为研究对象,在适宜培养条件下使其形成芽孢,通过研究萌发后芽孢对温度、H₂O₂及含氯消毒剂等条件的抵抗力,证实了适当的条件（温度37 ℃、湿度40%、pH 7.4、30 min）能有效促进蜡样芽孢杆菌芽孢萌发,致使芽孢对高温（80 ℃、2 h）、H₂O₂（1.5 mol/L、2 h）及含氯消毒剂（有效氯含量约800 mg/L、2 h）等常规灭菌方法抵抗力明显降低,并能被有效杀灭和灭活,相对有效杀灭率分别达98.4%、98.9%及99.1%,有效杀灭率分别为88.3%、88.7%及88.9%,这对进一步研究芽孢杆菌属细菌的杀菌消毒条件具有重要参考意义。     

辽宁地区猪源大肠埃希菌整合子携带情况调查与分析

查明辽宁地区整合子在猪源大肠埃希菌中的分布及整合子携带耐药基因盒的种类,可为该病的综合防控提供科学依据。本研究利用整合酶基因PCR扩增法检测整合子,并对整合子可变区进行扩增测序。结果表明,71.43%(40/56)的菌株为Ⅰ类整合子阳性,1.79%(1/56)的菌株同时为Ⅰ类和Ⅱ类整合子阳性,未检测到Ⅲ类整合子;87.8%(36/41)的菌株表现为Ⅰ类整合子可变区扩增阳性,扩增产物大小在116bp~2 307bp之间,100%(1/1)菌株表现为Ⅱ类整合子可变区扩增阳性,大小为2 106bp;整合子可变区含有编码对氨基糖苷类抗生素耐药的基因(aadA1、aadA2、aadA5、aadA22、aadB、aacA4和sat2),编码对磺胺类抗生素耐药的基因(dfrA1、dfrA12、dfrA17),编码对氯霉素抗生素耐药的基因(cmlA1、cmlA6)。因此,整合子在大肠埃希菌中广泛存在,辽宁地区大肠埃希菌中整合子主要携带编码对氨基糖苷类、磺胺类和氯霉素耐药基因盒。     

夏、冬两季猪源葡萄球菌耐药情况比较

本研究旨在比较夏、冬两季某规模化养猪场猪源葡萄球菌的耐药情况。分别在夏、冬两季对猪场同一猪采集其直肠肛拭子样和鼻腔鼻拭子样,采用微量肉汤稀释法对分离出的葡萄球菌进行临床9种常用抗菌药物的药敏试验。结果显示,夏季鼻腔分离的葡萄球菌对阿米卡星、氟苯尼考、氧氟沙星、苯唑西林和克林霉素5种抗菌药物的耐药率均高于冬季分离的鼻腔菌。夏季直肠分离的葡萄球菌对阿莫西林/克拉维酸、苯唑西林、庆大霉素、氧氟沙星、四环素、克林霉素和利福平7种抗菌药物的耐药率低于冬季分离的直肠菌。且夏、冬两季鼻腔分离的葡萄球菌多药耐药种数在0~9耐均有分布,夏、冬两季直肠分离的葡萄球菌多药耐药种数在1~9均耐有分布。冬季鼻腔分离的葡萄球菌6耐的多药百分率显著高于夏季分离菌的多药百分率（P＜0.05）,冬季直肠分离的葡萄球菌8耐的多药百分率显著高于夏季分离菌的多药百分率（P＜0.05）。结果表明,夏、冬两季猪不同部位分离的葡萄球菌对大多数被检抗菌药物的耐药率情况不同,其耐药率与养殖场季节性用药呈现相关性。     

Anthelmintic resistance and management of nematode parasites on beef cattle-rearing farms in the North Island of New Zealand

AIM: To provide information on current farmers’ opinions and farming practices thought to be related to anthelmintic resistance, and to test for associations between the presence of anthelmintic resistance and management practices on beef cattle- rearing farms in the North Island of New Zealand.

METHODS: A study using an interview-based questionnaire about management of internal parasites was conducted on 62 beef cattle-rearing farms in the North Island of New Zealand, using case-control analyses to test for associations between management practices and the presence or absence of resistance to ivermectin or albendazole. Resistance was inferred from faecal nematode egg count (FEC) reduction (FECR) tests (FECRTs) when there was <90% reduction in FEC 7-10 days after treatment of calves <12 months of age.

RESULTS: Of the 59 farmers who completed the questionnaire, most (n=40) ranked parasites highly, and at about the same level as quality and quantity of feed, as important production-limiting factors for their enterprises. In contrast, anthelmintic resistance was not perceived to be a problem on 13 farms, and its importance was rated low on 24, moderate on 15, and high on only six farms. Despite all farms having planned parasite control programmes, there was heavy reliance on clinical signs of parasitism to determine frequency of treatments. About one in three farmers with beef breeder cows routinely treated their calves at marking, one in five treated mixed-age cows, and almost half treated rising 2-year-old cows before calving. One in four farmers used anthelmintics on calves on 8–12 occasions in their first year of life. Co-grazing with other species was rare, but follow-on grazing within 3 months after older cattle or sheep was common. On most farms, grazing cattle was restricted to part of the farm, a finding with implications for parasite control and persistence of larvae in refugia. Macrocyclic lactone (ML) anthelmintics or their combinations with other action families were currently, and for the past 5 years, used more frequently than benzimidazoles and levamisole, and benzimidazole-levami- sole combinations. The prevalence of resistance to ivermectin was high (82%) and no plausible model of associations could be constructed from the data. The prevalence of resistance to albendazole was 60%, and the risk of resistance increased as the number of rising 1-year-old cattle present mid-winter increased, and decreased as the number of breeding cows >2 years old present mid-winter increased.

CONCLUSION: It is clear that in practice anthelmintic resistance is a secondary consideration to obtaining productivity advantages from the use of anthelmintics in beef cattle. Farmers’ opinions were divided on many issues and the overall impres- sion was of confused and diverse thinking regarding the principles of the use of anthelmintics. The overall outlook regarding anthelmintic resistance in cattle is bleak unless the need for integrated and long-term research activities is acted upon soon.     

Influence of ante‐ and peri‐mortem factors on biochemical and physical characteristics of turkey breast muscle

Summary

The course of post‐mortem breakdown of glycogen and ATP in turkey pectoralis major muscle was markedly influenced by several ante‐ and peri‐mortem variables. Application of a proper stunning procedure was highly effective in preventing peri‐ and post‐mortem muscle stress reactions.

The physiological level of glycogen and ATP was not significantly affected by road transportation covering 260 km. Birds which rested for 24 hrs following transportation had lower glycogen and ATP levels at the moment of slaughter than non‐rested birds.

According to the changes in the rate and extent of post‐mortem biochemical reactions, several meat characteristics such as water‐holding capacity, colour, and tenderness were significantly changed.

Furthermore, the results also indicate that turkey breast muscle is susceptible to a PSE‐like condition as described in pork.