细菌对氟苯尼考的耐药机制研究

氟苯尼考是一种治疗动物细菌性疾病的抗菌药物,在养殖业中的广泛使用已产生严重的耐药问题。文章对近十几年来国内外报道的关于细菌对氟苯尼考耐药机制研究进行归纳总结,论述了细菌对氟苯尼考产生耐药性及耐药性传播的机制,以期为临床上合理使用氟苯尼考提供参考。     

氟苯尼考酯（钠）对肉仔鸡致病性大肠杆菌的治疗对比试验

近几年，商品肉鸡养殖规模迅速发展，但由于饲养条件较差等原因，肉仔鸡大肠杆菌病发病率明显上升，鉴于大肠杆菌的耐药性日趋严重，目前很多抗菌药物都不能有效控制鸡群的大肠杆菌病。而氟苯尼考仍然是目前临床上治疗禽大肠杆菌的有效药物，但氟苯尼考不溶于水，现在临床上用的水溶性氟苯尼考有效成分含量低，效果不理想，且治疗成本高。     

猪源大肠杆菌O157∶H7体外氟苯尼考耐药诱导及对抗菌药物敏感性变化研究

为初步研究猪源大肠杆菌O157∶H7(E.coli O157∶H7)对氟苯尼考耐药性的产生和消除机制,本研究采用亚抑菌浓度体外耐药诱导的方法将两株猪源大肠杆菌O157∶H7诱导成氟苯尼考高度耐药菌株,采用无氟苯尼考压力下连续传代培养的方法将获得的氟苯尼考耐药菌株的氟苯尼考耐药性消除,检测耐药诱导菌和耐药消除菌对抗菌药物的敏感性,并检测菌株质粒携带的耐药基因。结果显示,经氟苯尼考耐药诱导,猪源大肠杆菌O157∶H7对氟苯尼考、阿莫西林、头孢唑啉、头孢拉定和头孢噻吩由敏感变为耐药,对头孢噻肟的敏感性由敏感变为中介,对氧氟沙星、环丙沙星和阿奇霉素由中介变为耐药;而经耐药消除后,菌株恢复对上述药物的敏感性;在菌株的质粒中检测到氟苯尼考耐药基因、喹诺酮类耐药基因和β-内酰胺酶基因,与耐药表型相符。结果表明,在氟苯尼考压力的长期存在下,猪源大肠杆菌O157∶H7对氟苯尼考产生耐药,且对青霉素类、头孢类和喹诺酮类药物产生交叉耐药,在去除氟苯尼考压力下连续培养,可消除菌株的部分耐药性。     

猪源大肠杆菌O157:H7体外氟苯尼考耐药诱导及对抗菌药物敏感性变化研究

为初步研究猪源大肠杆菌O157:H7 (E.coli O157:H7)对氟苯尼考耐药性的产生和消除机制,本研究采用亚抑菌浓度体外耐药诱导的方法将两株猪源大肠杆菌O157:H7诱导成氟苯尼考高度耐药菌株,采用无氟苯尼考压力下连续传代培养的方法将获得的氟苯尼考耐药菌株的氟苯尼考耐药性消除,检测耐药诱导菌和耐药消除菌对抗菌药物的敏感性,并检测菌株质粒携带的耐药基因。结果显示,经氟苯尼考耐药诱导,猪源大肠杆菌O157:H7对氟苯尼考、阿莫西林、头孢唑啉、头孢拉定和头孢噻吩由敏感变为耐药,对头孢噻肟的敏感性由敏感变为中介,对氧氟沙星、环丙沙星和阿奇霉素由中介变为耐药;而经耐药消除后,菌株恢复对上述药物的敏感性;在菌株的质粒中检测到氟苯尼考耐药基因、喹诺酮类耐药基因和β-内酰胺酶基因,与耐药表型相符。结果表明,在氟苯尼考压力的长期存在下,猪源大肠杆菌O157:H7对氟苯尼考产生耐药,且对青霉素类、头孢类和喹诺酮类药物产生交叉耐药,在去除氟苯尼考压力下连续培养,可消除菌株的部分耐药性。     

大肠杆菌O78对氟苯尼考产生耐药性的观察

氟苯尼考是美国先灵葆雅公司研制的氯霉素类兽用广谱抗生素。目前,氟苯尼考已在亚洲、欧洲、美洲的30多个国家上市,其剂型主要有注射剂、预混剂,适用于鱼、猪、牛及其他家畜。氟苯尼考于1999年4月份在我国批准上市,上市后因其抗菌谱广、安全、高效而取代残留毒性大而被禁止使用的氯霉素。随着广泛被应用,已有多种细菌对氟苯尼考产生耐药性。国外因为应用比较早,耐药性比较严重,曾有报道,从肉牛体内分离164株大肠杆菌,氟苯尼考对其最小抑菌浓度均大于256μg/mL。国内应用氟苯尼考比较晚,但也有耐药性的报道。试验采用不同的药物浓度对细菌进行诱导,并比较其诱导结果,现报道如下。     

黄颡鱼源嗜水气单胞菌对氟苯尼考的耐药性及其消失速率研究

在离体条件下通过测定最小抑菌浓度MIC值,研究4株黄颡鱼源嗜水气单胞菌对氟苯尼考的耐药产生速率与耐药性消失速率。结果表明:以37℃条件下培养48h为1代,HSY02菌株对氟苯尼考的最小抑菌浓度为3.906 3μg/m L,HSY01、HSY03、HSY04菌株对氟苯尼考的最小抑菌浓度为1.953 1μg/m L。在含有氟苯尼考的药物营养肉汤培养基中连续传代8代后,氟苯尼考对3株嗜水气单胞菌HSY01、HSY03、HSY04的最小抑菌浓度(MIC)由1.953 1μg/m L上升至62.50μg/m L,耐药性增长了32倍。HSY02菌株的最小抑菌浓度(MIC)由3.906 3μg/m L上升至62.50μg/m L,耐药性增长了16倍;且耐药性获得后保持稳定,其消失速率为0。     

氟苯尼考研究进展及其在猪场使用建议

氟苯尼考作为一种动物专用抗菌药因其优良的药效学特征,近15年被国内畜禽养殖业广泛使用,特别是其对猪呼吸道疾病的防治起到了重要作用。近年来不合理的用药方式导致细菌对其耐药性显著增强,这种情况严重影响了氟苯尼考的使用效果。本文从氟苯尼考的药理学特征、药效学特征以及现有制剂种类进行归纳,提出了氟苯尼考在猪场使用的几个建议。     

兔巴氏杆菌耐药性分析

对临床分离的36株兔巴氏杆菌,按照Kirby-Bauer药敏纸片法进行了22种抗菌药物的耐药性实验.结果:兔巴氏杆菌耐药性比较严重,而且多数为多重耐药,对临床常用的药物(如喹诺酮类、庆大霉素、卡那霉素、磺胺类药物等)多数产生了不同程度的耐药性,同时发现对青霉素类、氟苯尼考以及利福平敏感性较高.     

宠物犬源大肠杆菌氟苯尼考耐药基因floR的检测及药物敏感性分析

floR是氟苯尼考特异性耐药基因之一,当前宠物犬源大肠杆菌氟苯尼考耐药基因floR的研究报道很少,故本试验利用PCR方法对宠物犬源大肠杆菌氟苯尼考耐药基因floR进行检测,并采用微量肉汤稀释法测定宠物犬源大肠杆菌对10种抗菌药物的最小抑菌浓度(MIC)。结果显示,20株宠物犬源大肠杆菌中floR耐药基因的阳性菌株数为9株,阳性检出率为45%;对氟苯尼考的耐药率为65%,并呈现3~7重的多重耐药性。结果表明,随着氟苯尼考在兽医临床的广泛应用,其耐药率越来越高,需不断加强对氟苯尼考等抗菌药物的耐药性监控。     

氟苯尼考胁迫对土壤解磷菌耐药性的影响

建立土壤生态模型,考察氟苯尼考胁迫对土壤解磷菌耐药性的影响。在土壤中添加不同浓度氟苯尼考(Ⅰ组0 mg/kg,Ⅱ组0.1 mg/kg,Ⅲ组1.0 mg/kg,Ⅳ组10.0 mg/kg,Ⅴ组100.0 mg/kg),于加药前和加药后第7天、第21天、第49天采集土壤样品,分离解磷菌。分别采用微量肉汤二倍稀释法和K-B纸片扩散法测定解磷菌对氟苯尼考和17种抗菌药的敏感性。结果表明,随着氟苯尼考添加浓度的增大,耐氟苯尼考解磷菌的数量逐渐增多;加药后第49天,从Ⅳ组分离的解磷菌对氟苯尼考的耐药率最高为45%;17种抗菌药中,对氟苯尼考敏感的解磷菌和耐氟苯尼考解磷菌对其中10种抗菌药的敏感性差异极显著(P<0.01)。说明氟苯尼考一定程度上增强了解磷菌的耐药性。     

Pyrantel resistance in two herds of donkey in the UK

Resistance to currently available anthelmintics is a serious phenomenon which is prevalent globally. Cyathostomins are one of the major parasites, and are of primary concern in donkeys. There have been reports of emerging resistance to pyrantel, but the status of pyrantel resistance in donkey populations in the UK is largely unknown. This report investigates pyrantel resistance in two geographically isolated donkey herds in the South West of England. The first herd had suspected pyrantel resistance, with already established resistance to other anthelmintics. In the second herd the efficacy of pyrantel was not suspected at the time the study took place. Faecal Egg Count Reduction Test (FECRT) was carried out, revealing large scale resistance. Eighty one percent of the first herd and 73% of the second herd had a FEC of less than 95% after treatment, and anthelmintic resistance was confirmed using the World Association for the Advancement of Veterinary Parasitology guidelines. These findings indicate that anthelmintic resistance to pyrantel exists in both tested donkey populations and illustrate the continuing development of resistance through different classes of chemotherapeutics.     

Level of benzimidazole resistance in a strain of Ostertagia circumcincta studied over several infections in lambs

A benzimidazole-resistant strain of Ostertagia circumcinta (HFRO) was used experimentally to infect lambs. The level of resistance, measured by an egg hatch assay, was studied throughout each infection and also after treatment of the lambs with fenbendazole. The HFRO strain was highly resistant to benzimidazoles. There was day-to-day variation in the level of resistance throughout a single infection with a high level of resistance in the early part of the infection, around Day 27 post-inoculation of infective larvae, falling to a lower level later in the infection. Egg hatch assays on the 3 days immediately post-treatment with fenbendazole showed the resistance level was high then resistance fell to the pre-treament level after 7 days. Selection for benzimidazole resistance using fenbendazole treatment at the normal dose rate of 5 mg kg⁻¹ over five passages of the HFRO strain in lambs failed to increase the resistance level. Storage of larvae over a 5-months period at 4°C, prior to infection of lambs, did not produce any alteration in the resistance Level. The possible reasons for the variations in resistance found with the HFRO strain are discussed along with the implications for sheep parasite control and further development of benzimidazole resistance.     

广东地区猪链球菌的耐药性特点及四环素耐药基因携带情况

猪链球菌(Streptococcus suis,SS)是危害世界养猪业的重要细菌性病原菌之一,本研究旨在了解猪链球菌临床分离株的耐药性特点和四环素耐药基因的携带情况,为临床上该病的防控提供科学依据。本试验采用K-B法和CLSI推荐的肉汤微量稀释法检测猪链球菌广东分离株对18种抗菌药物的耐药性。结果显示,被检菌株对四环素类、磺胺类和氨基糖苷类抗菌药物表现较强的耐药性,尤其对四环素类药物的耐药率高达98.2%;对头孢菌素类药物比较敏感;菌株都耐受6种或6种以上的抗菌药物,其中以6和14重耐药菌株的数量最多,分别占20%和17%。本试验设计了4对引物检测四环素耐药基因携带情况,结果发现56株菌中以携带tetM基因为最多(85.71%)。结果表明tetM很可能是介导广东SS分离株对四环素产生极高耐药率的主要原因之一。     

Associations of antimicrobial use with antimicrobial resistance in Campylobacter coli from grow-finish pigs in Japan

To determine associations between antimicrobial use and antimicrobial resistance in Campylobacter coli, 155 isolates were obtained from the feces of apparently healthy grow-finish pigs in Japan. In addition, data on the use of antibiotics collected through the national antimicrobial resistance monitoring system in Japan were used for the analysis. Logistic regression was used to identify risk factors to antimicrobial resistance in C. coli in pigs for the following antimicrobials: ampicillin, dihydrostreptomycin, erythromycin, oxytetracycline, chloramphenicol, and enrofloxacin. The data suggested the involvement of several different mechanisms of resistance selection. The statistical relationships were suggestive of co-selection; use of macrolides was associated with enrofloxacin resistance (OR=2.94; CI(95%): 0.997, 8.68) and use of tetracyclines was associated with chloramphenicol resistance (OR=2.37; CI(95%): 1.08, 5.19). The statistical relationships were suggestive of cross-resistance: use of macrolides was associated with erythromycin resistance (OR=9.36; CI(95%): 2.96, 29.62) and the use of phenicols was associated with chloramphenicol resistance (OR=11.83; CI(95%): 1.41, 99.44). These data showed that the use of antimicrobials in pigs selects for resistance in C. coli within and between classes of antimicrobials.     

Management and environmental factors related to benzimidazole resistance in sheep nematodes in Northeast Spain

A survey to determine the level of parasite resistance to benzimidazoles (BZ) under field conditions was performed on 107 commercial sheep farms located in the Aragon region of northeast Spain. Resistance was measured using the discriminant dose, a simplified form of the in vitro egg hatch assay (EHA). Taking into account the spatial structure of the data, a multivariate approach was applied to management and environmental variables as well as to their relationships with BZ resistance levels compiled from each flock. Results estimated that 11% of flocks had resistant parasite populations, although we suspected the presence of BZ-resistant parasite strains in 98% of the sample. Resistance levels were more similar among the nearest flocks, suggesting a contagious spatial distribution of resistance (i.e., resistance at neighbouring farms was not independent from one another). Management variables such as frequency of deworming, grazing in private pastures and underdosing were positively related to resistance levels, whereas only the use of BZ was negatively related to resistance levels, likely because BZ were replaced by other anthelmintics in flocks where reduced BZ efficacy was suspected. In addition to climatic conditions and seasonality, land use was an environmental variable associated with observed BZ resistance levels. Generally, resistance was highest in cooler and wetter areas but was lower in flocks sampled during January-March compared to flocks sampled in April-June or October-December. Variation partitioning procedures showed that the variation of resistance explained by the effect of environmental variables was higher than management variables. The effects of both variable groups, however, highly overlapped with the spatial structure of resistant levels, which suggests that a considerable amount of the effects attributable to both variable groups may be actually due to the spatial distribution of resistance. The resistance variation explained by the spatial component suggested that other uncontrolled factors acting at short spatial scale (e.g., common management and environmental variables; the importation of resistant strains and their posterior spread across neighbouring flocks; the selection history of the worms carried out by historical management events previous to this survey; and genetic, physiological or both types of parasite population variation) could yield this contagious spatial structure of BZ resistance. Although further research is needed, both seasonal variation and the dependence of resistance levels among neighbouring flocks should be taken into account in the design of future research or observational resistance programmes to minimise spatial and temporal pseudo-replication. Thus, research would avoid biased estimations of resistance prevalence or of its relationship with putative factors.     

乌鲁木齐地区不同健康状态和饲养环境中宠物源沙门菌耐药特征分析

为探究不同健康状态和不同饲养环境下沙门菌对临床常用抗菌药物的耐药情况及耐药基因携带情况,采用琼脂稀释法对健康与患病、家养与流浪宠物粪样中分离出的99株沙门菌进行临床常用12种抗菌药物的耐药性检测;用PCR方法对获得的耐药菌株进行相关耐药基因的检测。结果:分离出的宠物源沙门菌对所有被检的抗菌药物耐药率均未超过10.0%,对头孢噻呋、安普霉素、阿米卡星、卡那霉素和硫酸庆大霉素100%敏感;患病动物源沙门菌对被检抗菌药物的耐药率高于健康动物源沙门菌,家养动物源沙门菌对被检抗菌药物的耐药率高于流浪动物源沙门菌(P<0.05)。耐药基因检测结果显示:floR的检出率为3.0%;ant(3″)-Ia、tetA、tetB检出率均为2.0%;blaTEM、oqxA、aac(6′)-Ib-cr基因检出率均为1.0%,未检出其他被检基因。结果表明,新疆乌鲁木齐市不同健康状态和不同饲养环境下宠物源沙门菌对被检抗菌药物的耐药率及耐药基因携带率低,仅有2株沙门菌存在ant(3″)-Ia+tetA+tetB 3种耐药基因共存情况。根据试验结果发现检出的耐药表型与耐药基因之间存在相关性,可根据药敏试验结果合理选用抗菌药物进行临床细菌病的治疗。     

动物源细菌耐药性评估在兽用抗菌药使用减量化行动中的应用

动物源细菌耐药性已引起广泛关注,全国正在逐步推进兽用抗菌药物使用减量化行动。为更好地推动减量化行动的实施,浙江省在耐药性的评估及应用方面进行探索。研究分析了耐药性评估在减量化行动中的作用,建立了耐药系数评估模型与应用方法,研判了目前耐药性监测可能存在的问题,以期更好地推进兽用抗菌药物使用减量化行动。     

Anthelmintic resistance in cattle nematode in the western Pampeana Region of Argentina

During the past three years there have been several reports of cattle anthelmintic resistance (AR) in Argentina. Twenty-five cattle herds in the west fattening area of the Pampeana Region were surveyed to assess the prevalence of AR and the species of nematode involved. The survey involved fattening and cow-calf systems with a mean of 6760 bovines. Resistance was determined in vivo by a faecal egg count reduction test (FECRT). Efficacies from the World Association for the Advancement of Veterinary Parasitology (WAAVP) formula were compared with those obtained from the Abbott formula. Sixty 6-12-month-old calves were selected from each property and divided into four groups, namely, IVM (ivermectin, 0.2mg/kg), BZD (fenbendazole, 5mg/kg or albendazole 7.5mg/kg), LVM (levamisole, 7.5mg/kg) and Control (untreated group). Nematode larvae were identified by four pooled faecal cultures for each group. Cooperia and Ostertagia were the predominant nematode larvae in pre-treatment cultures. The mean FECRT efficacies for IVM, BZD and LVM were 81.0+/-21%, 93.3+/-8% and 97.6+/-4%, respectively. AR was detected in 16 (64%) herds. IVM resistance was present in 15 herds and BZD resistance in eight herds (32%). Resistance to both ivermectin and fenbendazole was present in seven herds. LVM resistance was not detected. Cooperia was the predominant post-treatment nematode in all the herds with IVM resistance. In one herd with IVM resistance, Ostertagia counts were slightly increased over pretreatment counts. Ostertagia was the prevalent post-treatment genera in herds with BZD resistance. A high level of resistance to ivermectin was recorded in this fattening region of Argentina. Also several herds with resistance to benzimidazole were observed. These results show the importance of AR in Argentinian cattle production. High correlation between WAAVP and Abbott formulae was observed. However, under some uncertain diagnosis, both formulae could be applied as complementary to help result interpretations.     

湖州地区野桑蚕对有机磷农药抗性调查

2013年秋调查了野桑蚕对有机磷农药的耐药性,结果表明湖州地区野桑蚕对有机磷农药已经产生了抗性,不同地区抗性存在差异。抗性最强地区为南浔区和孚镇,其他依次为南浔区善琏镇＞菱湖镇＞千金镇＞吴兴区八里店镇。但湖州地区野桑蚕整体对有机磷农药抗性还处在较低水平。     

Molecular types and antibiotic resistance of Staphylococcus aureus isolates from bovine mastitis in a single herd in China

The molecular diversity, antibiotic resistance patterns and presence of resistance genes were determined in Staphylococcus aureus isolates from cases of bovine mastitis in a dairy cattle herd in China. Multiple locus variable number tandem repeat analysis was used for molecular typing. Resistance was determined through minimum inhibitory concentrations and resistance genes were detected by PCR. There was low molecular diversity; one predominant strain (type I) accounted for the majority of cases of S. aureus mastitis in the herd and this strain had a high frequency of resistance to penicillin and tetracycline. The most prevalent resistance genes were blaZ, ermC and tetM.