Sulfamethazine residues in swine: comparison of on-farm monitoring methods

Three sulfamethazine-residue detection methods were used to evaluate samples collected from five swine farms over a 12-month period. All cooperating farms included sulfamethazine in swine diets at various stages of production, for growth promotion or disease control, and followed recommended drug withdrawal periods. Swine finishing ration, swine urine, and swine serum from market-weight animals were tested monthly for the presence of sulfamethazine. Thin-layer chromatograph (TLC) analysis of swine urine was the gold standard by which three other test method-sample combinations were compared. Samples were analyzed for sulfamethazine using TLC (feed), competitive enzyme immunoassay (serum), and agar-diffusion swab test (urine). The relative sensitivities and specificities of sulfamethazine-residue detection for the three combinations were: (1) TLC analysis (27%, 94%); (2) competitive enzyme immunoassay analysis (58%, 59%); (3) agar-diffusion swab test (78%, 12%). None of the three methods tested was individually adequate for on-farm monitoring of sulfonamide residues. Sulfamethazine residues in swine urine were found on 43.3% of the monthly farm visits and in 19.7% of all swine tested.     

Sulfamethazine blood/tissue correlation study in swine

Seventy market-weight hogs (90 to 113 kg) were used in a feeding study to determine the correlation of serum sulfamethazine concentrations with sulfamethazine concentrations in liver and muscle at time of slaughter. Test groups were fed medicated feeds prepared from commercial medicated premixes containing 110 g of sulfamethazine/metric ton for 30 days. Fifteen days before hogs were slaughtered, test groups were given maintenance feeds containing 1.1 to 13.9 g of sulfamethazine/metric ton and were fed these diets until slaughtered. Comparison of data from positive- and negative-control groups indicated that total withdrawal of sulfamethazine in the feed was not necessary for the liver to contain less than the allowed tolerance of 0.1 mg of sulfamethazine/kg of liver at slaughter. Feed concentrations of up to 2 g of sulfamethazine/metric ton could be tolerated in withdrawal feeds before liver sulfamethazine values exceeded 0.1 mg/kg of liver. Serum/tissue sulfamethazine ratios were erratic in hogs given 1.1 to 2.7 g of sulfamethazine/metric ton, but became less variable in hogs given greater than 5.7 g/metric ton. Feed concentrations greater than 8 g of sulfamethazine/metric ton produced values greater than 0.1 mg/kg of muscle and values of about 0.4 mg/kg of liver. When serum sulfamethazine concentrations alone were used as a predictor for tissue sulfamethazine values, 100% of the liver values exceeded 0.10 mg/kg of liver when sulfamethazine in serum was greater than 0.45 mg/L. However, 57.4% of samples having serum concentrations between 0.10 and 0.45 mg/L had associated sulfamethazine values greater than 0.1 mg/kg of liver. All hogs having serum sulfamethazine concentrations less than 0.1 mg/L had sulfamethazine concentrations less than 0.1 mg/kg of liver.     

Sulfamethazine residues in uncooked edible tissues of port following recommended oral administration and withdrawal.

Commercial feed rations containing sulfamethazine at the level of 110 ppm were fed for a period of 65 days to market pigs in a study simulating normal farm practices. The levels of sulfamethazine at the end of medication were in excess of 10 ppm in liver and kidney and up to 2.6 ppm in muscle tissues. Concentrations of sulfamethazine in tissues from pigs after withdrawal of medicated feed depleted to 0.1 ppm within nine days. The rate of depletion was similar for all tissues. It was observed that storage of tissue samples at freezer temperature (-20 degrees C) for 30 days further reduces sulfamethazine levels by 3 to 20% of their original value.     

Survey of chloramphenicol residues in diseased swine.

Tissue samples from 279 hogs suspected of having received antibiotic treatment were collected at federally-inspected abattoirs and submitted for chloramphenicol residue analysis during August and September 1984. Injection sites (when present), kidneys or muscle samples were tested by one of two gas chromatographic methods. Kidney samples were also tested at the abattoirs by the Swab Test On Premises. Thirty-one animals (11%) were found with detectable levels ranging from 1 part per billion to 5727 ppb. Highest levels were found at the injection sites, while levels in muscle tissue did not exceed 500 ppb. None of the kidneys from animals found to contain chloramphenicol residues produced a positive Swab Test On Premises result attributable to the presence of chloramphenicol. Twelve kidneys from animals free of chloramphenicol residues produced positive Swab Test On Premises results. Of these, five contained penicillin or streptomycin, but antibiotic residues were not detected in the remaining seven. In addition to the samples collected for this survey, samples from eight hogs representing a herd which had been treated for pneumonia were submitted by an abattoir in Manitoba in November 1984. Chloramphenicol levels in these animals ranged from 0.1 to 73 parts per million in the injection sites, and from 0.04 to 21 ppm in the muscle tissues. The survey data indicated that there were a significant number of animals reaching the abattoirs with detectable chloramphenicol residues, and that the Swab Test On Premises procedure was ineffective in detecting these animals.     

Elimination of sulfamethazine residues from swine.

Tissue concentrations of sulfamethazine in swine fed the drug at the rate of 500 g/ton of ration (550 g/1,000 kg) for a 30-day period depleted to 0.1 ppm or less within 4 to 10 days after withdrawal of medicated feed. Depletion from the tissues and plasma of treated pigs showed a linear relationship with time when the concentrations were plotted on a semilogarithmic graph. Six untreated pigs that were placed on bedding in pens formerly occupied by the treated group developed tissue residues at or above 0.1 ppm sulfamethazine; the mean plasma concentration of sulfamethazine reached 2.8 ppm by day 15.     

液相色谱法测定猪组织中阿维菌素类药物残留量

动物组织中残留的阿维菌素、依维菌素和多拉菌素经乙腈提取,碱性氧化铝固相萃取柱净化后,与荧光衍生化试剂反应,产物用高效液相色谱(荧光检测器)进行检测,实现了阿维菌素类药物的残留分析.本方法在猪组织中测定阿维菌素类药物的检测限为1μg/kg;在猪组织中添加浓度为1、2、5μg/kg时的回收率为55%～75%.     

高效液相色谱-质谱法测定猪组织中硝基咪唑类药物残留量

建立了一种高灵敏度和高选择性的检测猪组织中硝基咪唑类药物残留的高效液相色谱-大气压化学电离质谱(APCI)法,可同时进行猪组织中3种硝基咪唑类药物残留的确证分析.本方法检测猪肌肉、肝脏、肾脏和皮下组织中硝基咪唑类药物的检测限为0.5～1μg/kg;空白组织添加硝基咪唑类药物水平为0.5、1、2或4μg/kg时,测得回收率:肌肉为61.1%～85.4%,肝脏为62.3%～80.4%,肾脏为61.6%～82.0%,皮下组织为63.6%～84.1%.     

猪尿液中抗菌药物快速筛选拭子法的建立

为防止抗菌药物在动物性食品中的残留,以枯草芽孢杆菌为受试菌,建立一种检测活体动物尿液中抗菌药物残留的快速筛选拭子法,进行宰前活体检疫.添加试验测定猪尿液中5类抗菌药物最低检测限分别为:β-内酰胺类青霉素和氨苄青霉素均为0.05mg/L;氨基糖苷类庆大霉素0.05mg/L、新霉素0.4mg/L;四环素类金霉素0.1mg/L;大环内酯类红霉素0.05mg/L和氟喹诺酮类恩诺沙星0.2mg/L.各抗菌药物添加回收率范围均在64.0%～107.7%,变异系数均小于15%.假阴性结果显示,除青霉素(5%)和氨苄青霉素(4%)出现假阴性外,红霉素、庆大霉素、新霉素、恩诺沙星和金霉素均未出现假阴性.与国外同类试剂盒比较,结果显示两者对10种抗菌药物的检测限一致.     

高效液相色谱法测定猪组织中维吉尼亚霉素M1的残留

建立了猪组织中维吉尼亚霉素M1残留的高效液相色谱测定方法.样品用甲醇-0.2mol/L NH4H2PO4溶液提取,正己烷洗涤,经C18固相萃取柱净化后做HPLC分析.选用二极管矩阵检测器,波长245 nm;色谱柱为ODS-3 C18柱(4.6 mm×250mm,5 μm);流动相为乙腈-水(37.5:62.5,V/V).维吉尼亚霉素M1标准曲线线性范围在0.1～2.0 μg/mL.对不同组织的3种加药浓度进行回收率测定,回收率在72%～85%,变异系数在10%以内.最低检测限为10 ng/g.     

鸡血清中磺胺二甲嘧啶残留的酶免疫法测定

用直接竞争酶免疫测定法测定磺胺二甲嘧啶（ＳＭ２）在鸡血清中的残留,血清中的ＳＭ２与酶标ＳＭ２竞争酶联板上的抗ＳＭ２抗体的结合位点,最低检出限为５０μｇ／Ｌ。在１００μｇ／Ｌ处,回收率为９１％。对１１种磺胺类和非磺胺类药物进行测定,最大交叉反应只有０．５％。     

磺胺二甲嘧啶免疫抗原的合成研究

试验旨在研究磺胺二甲嘧啶（SM2）免疫抗原的合成。用重氮化方法将SM2分别与牛血清蛋白（BSA）和卵清蛋白（OVA）偶联,合成了免疫抗原SM2-BSA和包被抗原SM2-OVA。紫外扫描和聚丙烯胺凝胶电泳结果表明, SM2-BSA的紫外吸收光谱与BSA、SM2相比均发生了改变,证明偶联成功,并测得SM2-BSA的结合比为13∶1;用SM2-BSA免疫BALB/c小鼠,获得了高效价和特异的多克隆抗体。结果表明,半抗原SM2和载体蛋白偶联成功,并可获得高价、敏感的多克隆抗体。     

恩诺沙星与磺胺二甲嘧啶联合毒性探究

本研究旨在探究恩诺沙星与磺胺二甲嘧啶的联合毒性。选取SD大鼠为试验对象,将其分成6组,每组12只,分别为高剂量联合用药组（500 mg/kg体重）、中剂量联合用药组（250 mg/kg体重）、低剂量联合用药组（50 mg/kg体重）、恩诺沙星单药组（250 mg/kg体重）、磺胺二甲嘧啶单药组（250 mg/kg体重）及对照组（等量0.5%羧甲基纤维素钠溶液）,配制相应的药物进行灌胃,最后一次给药1 d后对大鼠进行称重、麻醉、心脏采血并剖检取肝脏组织,然后对大鼠增重率、血常规指标、血清生化指标及肝脏病理学变化几个方面进行分析,并利用SPSS 22.0软件评估2种药物的联合作用效果。结果显示,在雌性大鼠中,高、中剂量联合用药组使大鼠增重率显著下降（P<0.05）,雄性大鼠增重率无显著变化（P>0.05）;雄性大鼠给药组白细胞数量均显著增加（P<0.05）,高剂量联合用药组中性粒细胞和淋巴细胞占比分别表现为显著升高及下降（P<0.05）;高、中剂量联合用药组大鼠外周血中谷草转氨酶含量显著增加（P<0.05）,且高、中剂量联合用药组肝脏病理切片视野可见不同程度的炎性细胞浸润,并伴有不同程度的损伤。本研究结果表明,恩诺沙星和磺胺二甲嘧啶联用可使毒性增加,对大鼠的免疫系统状态有一定的影响,且能造成一定程度的肝脏损伤,剂量越高影响越大。本研究为恩诺沙星和磺胺二甲嘧啶联合用药机制的研究提供数据支持,并为二者的临床应用提供参考,提示新的食品安全评估应考虑药物联合暴露带来的影响。     

尿样中莱克多巴胺残留ELISA检测试剂盒的比对实验

酶联免疫吸附技术（ELIsA）由于其快速、简便、适合同时处理大量样品的特点，被国内外普遍应用于进行口一兴奋剂等各种兽药残留的筛选检测。国内目前销售的菜克多巴胺残留ELISA试剂盒主要有4种，据悉新的试剂盒还将推出。但国内外尚缺乏对不同莱克多巴胺ELISA试剂盒的检测性能的权威评估。由于不同试剂盒的检测效果可能存在差异，检测试剂不统一，可能导致检测结果差异较大，将不利于政府残留监控计划的实施和企业维护自身权益。本研究拟通过比对检测试验，对不同莱克多巴胺ELISA试剂盒进行评估和比对，为制定合理的莱克多巴胺残留检测技术方案和准确评判检测结果提供科学依据。     

Drug residues in food animals II. Plasma and tissue kinetics of oxytetracycline in young cross-bred swine

Tissue distribution and elimination kinetics of oxytetracycline in sixteen organs and body fluids were determined in young pigs following intravenous and oral administration. Seventeen non-fasted pigs, 8–10 weeks of age, weight range 16.4–34.5 kg were dosed intravenously at a dose rate of 11 mg/kg bodyweight. An additional seventeen weaning pigs, 12–14 weeks of age, weight range 27.2–36.3 kg were dosed orally at a dose rate of 48–65 mg/kg bodyweight. Oxytetracycline was rapidly distributed (half-life, 6.71 ± 1.13 min) in swine. The mean volume of distribution was 1.26 ± 0.18 l/kg and overall body clearance was 3.82 ± 0.59 ml/kg/min. The elimination half-life of oxytetracycline in pigs was 3.87 ± 0.62 h, which is shorter than has been observed in other domestic animal species. Oxytetracycline became rapidly and efficiently involved in enterohepatic cycling, with as much as 70% of a total intravenous dose being available for reabsorption from the gastrointestinal tract within 1 h after administration. This high degree of enterohepatic recycling prolonged the half-life, and the large amount of drug that entered the enteric tract contributed to the high volumes of distribution and high k ₁₂/ k ₂₁ ratios. The excellent tissue penetration of this drug further contributed to the high volume of distribution and high k ₁₂/ k ₂₁ ratios obtained. Relationships between plasma and tissue depletion for several major edible organs were found to be statistically significant. Blood plasma is proposed as a body fluid for monitoring oxytetracycline tissue residues.     

基于UPLC-MS/MS同时测定猪粪便中5种兽药残留量的研究

为了建立同时测定猪粪便中二甲氧苄啶、磺胺对甲氧嘧啶、恩诺沙星、金霉素、泰乐菌素5种兽药残留量的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法,样品经McIllvaineNa_2EDTA缓冲溶液提取,C18粉吸附杂质,HLB固相萃取小柱净化,浓缩富集后用0.1%甲酸甲醇溶液复溶,过滤上机,基质液外标法定量。结果显示,5种兽药在10~500μg/kg的质量浓度范围内线性良好,在添加浓度50~500μg/kg,平均回收率为53%~85%,RSD为1.87%~18.81%。该方法具有较好的准确度与精密度,能快速、准确测定猪粪便中5种兽药残留,为养殖环节实时监测兽药使用情况提供了技术支持。