牛奶中土霉素残留检测方法改进及临床消除规律探究

试验对牛奶中的土霉素残留检测方法进行了改进,并采用改进后的方法对牛奶中土霉素的临床消除情况进行了探究。样品前处理过程中,选取0.1mol/LNa₂EDTA-Mcllvaine缓冲溶液提取牛奶中的土霉素,正己烷除脂及HLB固相萃取柱净化浓缩。釆用Symmetry Shield RPC18(250mm×4.6mm,5μm)反相色谱柱,以乙腈和0.048%的磷酸溶液(pH2.5)为流动相,采用梯度洗脱的方式对样品进行分析。方法改进后,土霉素的出峰时间为6.5min,峰形尖锐,与样品中的杂质峰分离良好。土霉素标准溶液在5~2000μg/kg范围内线性关系良好,标准曲线R²=0.9999;牛奶中土霉素在10、20、100μg/kg3组浓度的加标回收率分别为87.9%、90.5%和87.8%,批内变异系数范围为2.2%~5.8%,批间变异系数范围为4.0%~5.1%。土霉素在牛奶中的检出限为5μg/kg,定量限为10μg/kg,适用于研究土霉素在牛奶中的残留消除规律。选取了29头产后患有子宫内膜炎的奶牛,分成4个不同剂量组(2、3、4和5g),通过子宫内灌注土霉素的方式进行治疗,然后采用优化后的方法检测牛奶中残留的土霉素。结果发现,4g及以下剂量组给药72h后,牛奶中的土霉素可以消除至残留限量以下;而当给药剂量达到5g时,72h后牛奶中的土霉素未消除至残留限量以下,因此不建议采用5g及以上剂量灌注病牛子宫。     

长效土霉素注射液的急性毒性及肌肉刺激性研究

为考察新型长效土霉素注射液的安全性,分别对小白鼠和家兔进行急性毒性和局部刺激性试验。随机将60只小白鼠分为6组,每组10只,其中雌雄各半,进行急性毒性试验考察;随机将9只家兔分为3组,每组3只,进行局部刺激性试验。结果表明,该新型长效土霉素注射液小鼠肌肉注射LD 50为600.32 mg/kg,LD 50的95%的可信区间为(600.32±2.44)mg/kg;随着时间延长和给药剂量增加,刺激性增强,注射长效土霉素7 d后,家兔股四头肌出现红肿、发紫、坏死、光泽消失现象,15 d后肌肉组织的坏死范围扩大。该产品对小白鼠表现为低毒,对家兔的股四头肌有局部刺激性。     

DOMs耦合生物质炭对土霉素的吸附行为研究

以650 ℃下制备甘蔗渣生物质炭（BC650）为研究对象,在模拟自然条件下,在BC650上负载环境中常见的溶解性有机质（DOMs）即小分子没食子酸（GA）和大分子黄腐酸（HA）,并对其性质进行表征;再以典型抗生素土霉素为目标污染物,探究外源DOMs耦合生物质炭对抗生素的吸附性能的影响。结果表明,DOMs的负载提高了生物质炭的比表面积,生物质炭的表面化学性质发生改变;预载DOMs后,生物质炭对土霉素的吸附能力明显降低,且GA负载的抑制作用较HA更为显著,就平衡吸附量而言,HA处理后其下降了10%,而GA的处理使其降低了30%,DOMs的预载对生物质炭吸附抗生素的影响可能是由DOMs处理改变了生物质炭的表面化学性质而致;DOMs负载前后生物质炭对土霉素表现出非线性吸附,氢键作用为主要吸附机制。所得结果可为生物质炭在抗生素污染控制中的实际应用提供重要的理论依据。     

Differential susceptibility to tetracycline,oxytetracycline and doxycycline of the calf pathogens Mannheimia haemolytica and Pasteurella multocida in three growth media

For clinical isolates of bovine Mannheimia haemolytica and Pasteurella multocida, this study reports minimum inhibitory concentration (MIC) differences for tetracycline, oxytetracycline and doxycycline between cation‐adjusted Mueller‐Hinton broth (CAMHB), foetal bovine serum (FBS) and Roswell Park Memorial Institute (RPMI) medium. MICs were determined according to CLSI standards and additionally using five overlapping sets of twofold dilutions. Matrix effect: (a) free drug MICs and minimum bactericidal concentrations (MBC) for all drugs were significantly higher in FBS than in CAMHB for both pathogens (p < 0.001); (b) MICs and MBCs were higher for CAMHB and FBS compared to RPMI for P. multocida only. Net growth rate for P. multocida in CAMHB was significantly slower than in FBS and higher than in RPMI, correlating to MIC and MBC ranking. Drug effect: doxycycline MICs and MBCs were significantly lower (p < 0.001) in both CAMHB and FBS than tetracycline and oxytetracycline for both pathogens. Only for M. haemolytica were oxytetracycline MIC and MBC significantly lower than tetracycline, precluding the use of tetracycline to predict oxytetracycline susceptibility in this species. Determining potencies of tetracyclines in a physiological medium, such as FBS, is proposed, when the objective is correlation with pharmacokinetic data for dosage determination.     

乌骨鸡肉及组织中土霉素、四环素残留量研究

利用高效液相色谱法(HPLC)测定乌骨鸡胸肉、腿肉、心、肝、肾、肌胃组织中的土霉素、四环素残留量.结果表明,土霉素、四环素标准品浓度在8.2～262.5 μg·mL-1范围内线性关系良好,土霉素线性方程为^y=19 392.82x-22.90(r=0.999 48),四环素线性方程为^y=14 089.84x-33.34(r=0.998 88).乌骨鸡胸肉的土霉素加样回收率平均为89.84%,RSD=1.03%;四环素加样回收率平均为90.65%,RSD=1.05%.     

Pharmacokinetic–pharmacodynamic integration and modelling of oxytetracycline for the calf pathogens Mannheimia haemolytica and Pasteurella multocida

A calf tissue cage model was used to study the pharmacokinetics (PK) and pharmacodynamics (PD) of oxytetracycline in serum, inflamed (exudate) and noninflamed (transudate) tissue cage fluids. After intramuscular administration, the PK was characterized by a long mean residence time of 28.3 hr. Based on minimum inhibitory concentrations (MICs) for six isolates each of Mannheimia haemolytica and Pasteurella multocida, measured in serum, integration of in vivo PK and in vitro PD data established area under serum concentration–time curve (AUC_0–∞)/MIC ratios of 30.0 and 24.3 hr for M. haemolytica and P. multocida, respectively. Corresponding AUC_0–∞/MIC ratios based on MICs in broth were 656 and 745 hr, respectively. PK‐PD modelling of in vitro bacterial time–kill curves for oxytetracycline in serum established mean AUC_0–24 hr/MIC ratios for 3log₁₀ decrease in bacterial count of 27.5 hr (M. haemolytica) and 60.9 hr (P. multocida). Monte Carlo simulations predicted target attainment rate (TAR) dosages. Based on the potency of oxytetracycline in serum, the predicted 50% TAR single doses required to achieve a bacteriostatic action covering 48‐hr periods were 197 mg/kg (M. haemolytica) and 314 mg/kg (P. multocida), respectively, against susceptible populations. Dosages based on the potency of oxytetracycline in broth were 25‐ and 27‐fold lower (7.8 and 11.5 mg/kg) for M. haemolytica and P. multocida, respectively.     

五种土霉素注射液在猪体内的药物代谢动力学比较研究

对五种土霉素注射液产品(A、B、C、D和得米先20%)进行了猪体内的药代动力学比较研究。五组猪分别按20 mg/kg的剂量耳后肌肉注射,HPLC测定血药浓度,3P97软件拟合血药浓度-时间数据,计算药代动力学参数。五种长效土霉素注射液均符合二室开放模型,峰浓度值分别为1.511、.772、.132、.022、.72μg/mL,AUC分别为48.456、0.22、52.41、66.045、9.8(μg.h)/mL。比较而言,得米先20%药动学最好,其次为产品C,产品B最差。     

Kinetic analysis of oxytetracycline residues in Chinese mitten crab,Eriocheir sinensis,muscle following intramuscular administration

Crab culture is a very important economic industry in China. An epidemic of tremor disease of Chinese mitten crabs, Eriocheir sinensis, has become a serious problem in recent years. A spiroplasm has been proved to be the causative agent of this disease. Oxytetracycline (OTC) is used widely in aquaculture and was confirmed to be very effective against this pathogen. In this study, the distribution and depletion patterns of OTC in crab muscle were evaluated following single intramuscular doses of 2, 8 and 40 mg kg^?1 body weight. OTC was detected with a validated HPLC method. Concentration–time profiles were well described by a three‐compartment model with first‐order absorption after a single dose of 8 and 40 mg kg^?1. For comparison, a non‐compartment model was employed. A withdrawal time of 48.29 and 55.92 days was suggested prior to consumption after receiving 8 and 40 mg kg^?1. A recommended therapeutic dose of OTC in theory was calculated to be 36.37 mg kg^?1. OTC was distributed well throughout the body. The elimination of OTC in muscle was slower compared with fish and other crustaceans. A dose of 40 mg kg^?1 is suggested for practical use.     

土霉素在土壤中的垂直迁移及其影响因素研究

采用土柱淋溶法,研究了土霉素在土壤中的垂直迁移特性,探讨了不同土壤类型、淋溶体积、淋溶液pH值、施药量等因素对土霉素在土壤中淋溶迁移的影响。结果表明,在红壤、黑钙土、赤红壤3种不同类型的土样中,土霉素的迁移深度为：赤红壤〉红壤〉黑钙土;土霉素主要富集在土壤表层,其含量随土壤深度增加而明显降低;同一深度的土壤中土霉素含量随着淋溶液体积的增加而增大,随pH值增大而减小;土霉素在土层中的含量分布、淋溶深度与其施药量均呈正相关;当淋溶液中含有一定量的土霉素溶液时,土霉素在土壤中的迁移深度明显增加。     

土霉素及镉污染对土壤呼吸及酶活性的影响

随着饲料工业以及畜禽养殖业的规模化发展,抗生素和重金属在土壤环境中同时存在的几率不断增大。为了分析抗生素和重金属对土壤微生物生态系统的影响,以土霉素(OTC)与镉(Cd)为污染物,采用室内培养法,研究了土霉素(OTC)与镉(Cd)单一处理及复合污染对土壤呼吸和酶活性的影响。结果表明,10mg/kg重金属镉单独污染对土壤微生物呼吸表现为先抑制后激活作用,且显著抑制了土壤蔗糖酶、脲酶、磷酸酶活性,对3种酶活性平均抑制率从大到小依次为:蔗糖酶磷酸酶脲酶;1mg/kg土霉素显著激活土壤微生物呼吸,50和200mg/kg土霉素对土壤微生物呼吸的影响呈现出先抑制后激活的规律。各处理浓度下的土霉素对蔗糖酶和脲酶活性均主要表现为抑制作用,对磷酸酶活性的影响呈现出一定的波动性;当土霉素的浓度为1和200mg/kg时,其与10mg/kg镉的复合污染对土壤微生物呼吸及3种酶活性的影响主要为拮抗作用,但当土霉素的浓度为50mg/kg时,与10mg/kg镉的复合污染对土壤微生物呼吸及3种酶活性的影响则主要为协同作用。微生物呼吸对土霉素与镉胁迫更为敏感,最高抑制率和激活率分别可达98.98%和300.82%,土壤酶活性受土霉素和镉污染的影响要弱于它们对土壤微生物呼吸的影响。