A study of the pharmacokinetics and tissue residues of an oral trimethoprim/sulphadiazine formulation in healthy pigs

Twenty-six healthy female pigs weighing 19.5-33 kg were used in three separate experiments. The animals were fed individually twice a day. Trimethoprim/sulphadiazine (TMP/SDZ) formulation was added to feed in the amount of 6 mg/kg bw (TMP) and 30 mg/kg bw (SDZ). TMP and SDZ concentrations in blood plasma, muscles, liver and kidneys were measured. Pharmacokinetic parameters show that the absorption of TMP from the alimentary tract in pigs is faster than the absorption of SDZ, and the elimination of TMP is slower than that of SDZ. The absorption half-lives were 0.96 (TMP) and 2.24 h (SDZ), whereas elimination half-lives were 5.49 (TMP) and 4.19 h (SDZ). The observed TMP:SDZ ratios in blood plasma after multiple dose administration ranged from 1:11.4 to 1:23.2. One day after administration of the last dose of TMP/SDZ the plasma concentration ratio was 1:15.5, but in muscles, liver and kidneys it was much lower: 1:0.79, 1:0.14 and 1:1.53 respectively. The absolute TMP and SDZ tissue concentrations 1 day after the last multiple dose administration were very low (maximum TMP: 0.29 μg/g in liver; maximum SDZ: 0.23 μg/g in kidneys). Neither drug was detected in any tissue 8 days after the last administration of TMP/SDZ. Based on our results, it was concluded that there is no support for the TMP:SDZ pharmaceutical ratio 1:5 in oral formulations of these compounds for pigs. The administration of oral TMP/SDZ formulations once a day may result in the absolute tissue concentrations of these drugs being too low for antibacterial activity. The withdrawal period for such an oral TMP/SDZ formulation for pigs (according to accepted guidelines in Europe for MRL of TMP < 0.05 mg/kg of tissue) should not be less than 5 days.     

Determination of sulphachloropyrazine-diaveridine residues by high performance liquid chromatography in broiler edible tissues

Diaveridine (DVD) is used in combination with sulphachloropyrazine (SPZ) as an effective antibacterial agent and antiprotozoal agent, respectively, in humans and animals. To gain a better understanding of the metabolism of SPZ and DVD in the food-producing animals, a high performance liquid chromatography (HPLC) method to determine and quantify sulphachloropyrazine (SPZ) and diaveridine (DVD) suspension residues from broilers is reported. Thirty healthy chickens were orally administered with sulphachloropyrazine-diaveridine (SPZ-DVD) suspension in water of 300 mg/l (SPZ) per day for seven successive days. Six chickens per day were slaughtered at 0, 1, 3, 5 and 7 days after the last administration. This procedure permitted SPZ and DVD to be separated from muscle tissue, liver, kidneys and skin with fat after extraction with acetonitrile and acetone under slightly acidic conditions. From the detected residuals in different tissues, we found that SPZ was quickly eliminated in liver and muscle, and slowly eliminated in kidney and skin with fat. DVD was quickly eliminated in liver and slowly eliminated in kidney. The withdrawal period of SPZ was 3.26, 3.72, 4.39 and 5.43 days in muscle, liver, kidney and skin with fat, respectively. The withdrawal period of DVD was 4.77, 4.94, 6.74 and 4.58 days in muscle, liver, kidney and skin with fat, respectively. Therefore, the suggested withdrawal period for SPZ-DVD suspension should be 7 days after dosing for seven successive days.     

Plasma concentrations and tissue residues of bicozamycin given orally to pigs

Bicozamycin was dissolved in water and administered to pigs by stomach tube at 40 mg/kg once daily for 7 consecutive days. The plasma concentration was determined on days 1 and 7 of the dosing period. The mean (± SD) peak plasma concentrations were 2.06±0.36 µg/ml at 3.08±0.80 h on day 1 and 2.36±1.32 µg/ml at 2.80±0.74 h on day 7, the elimination half-lives being 3.80±0.92 h and 2.43±1.41 h, respectively. The mean areas under the plasma concentration-time curves were 15.88±2.18 (µg h)/ml on day 1 and 12.31±6.98 (µg h)/ml on day 7. These pharmacokinetic parameters did not differ between days 1 and 7, suggesting that there was no accumulation in the plasma after consecutive oral dosing. The residues in kidney, liver and muscle were examined in pigs slaughtered on days 1, 3 and 5 after the last dosing. One day after withdrawal, residues were found in the kidneys of all three pigs examined, at a mean concentration of 0.26 µg/g, and in muscle from one pig, but not in liver from any of the pigs. Bicozamycin was not detected in any of the samples taken 3 or 5 days after withdrawal.Abbreviations AUC area under the plasma concentration-time curve - C _max peak concentration - T _max time of the peak - t _1/2 elimination half-life     

猪肾脏等组织中盐酸莱克多巴胺的HPLC检测方法及其残留消除

本研究建立了猪肝脏、肾脏、肌肉和脂肪中盐酸莱克多巴胺的高效液相色谱检测方法。方法的检测限为1ng/g,定量限为2 ng/g,肝脏的平均回收率在73.4%～83.2%,变异系数在2.1%～6.6%;肾脏的平均回收率在73.1%～95.5%,变异系数在3.8%～4.0%;肌肉的平均回收率在80.7%～82.5%,变异系数在4.9%～9.1%;脂肪的平均回收率在73.3%～78.8%,变异系数在2.9%～6.7%。对60头健康猪以18 mg/kg的剂量混饲给药28d,停药饲喂14 d。在给药7、142、8 d和停药1、2、3、7、9、14 d分别屠宰6头猪,取各组织进行残留量测定。结果显示：肾脏中残留量最高,肝脏其次,残留量在停药期间较给药期间显著降低。其中肾脏和肝脏中的残留量分别在停药14 d、停药9 d后降至定量限以下;肌肉和脂肪中的残留量显著低于肾脏和肝脏,给药28 d时,残留基本低于定量限;停药1 d时均低至检测限以下。     

Determination of sulfadiazine,trimethoprim, and N4‐acetyl‐sulfadiazine in fish muscle plus skin by Liquid Chromatography–Mass Spectrometry. Withdrawal‐time calculation after in‐feed administration in gilthead sea bream (Sparus aurata L.) fed two different diets

This study presents a depletion study for sulfadiazine and trimethoprim in muscle plus skin of gilthead sea bream (Sparus aurata L.). N⁴‐acetyl‐sulfadiazine, the main metabolite of sulfadiazine (SDZ), was also examined. The fish were held in seawater at a temperature of 24–26 °C. SDZ and trimethoprim (TMP) were administered orally with medicated feed for five consecutive days at daily doses of 25 mg SDZ and 5 mg TMP per kg of fish body weight per day. Two different diets, fish oil‐ and plant oil‐based diets, were investigated. Ten fish were sampled at each of the days 1, 3, 5, 6, 8, 9, 10, and 12 after the start of veterinary medicine administration. However for the calculation of the withdrawal periods, sampling day 1 was set as 24 h after the last dose of the treatment. Fish samples were analyzed for SDZ, TMP, and acetyl‐sulfadiazine (AcSDZ) residues by liquid chromatography–mass spectrometry. SDZ and TMP concentrations declined rapidly from muscle plus skin. Considering a maximum residue limit of 100 μg/kg for the total of sulfonamides and 50 μg/kg for TMP residues in fish muscle plus skin, the withdrawal periods of the premix trimethoprim‐sulfadiazine 50% were calculated as 5 and 6 days, at 24–26 °C, in fish oil (FO) and plant oil (PO) groups, respectively. The investigation of this work is important to protect consumers by controlling the undesirable residues in fish.     

Evaluation of some coagulation parameters in hepatic coccidiosis experimentally induced with Eimeria stiedai in rabbits

The objective of this study was to evaluate some coagulation parameters in hepatic coccidiosis experimentally induced with Eimeria stiedai in rabbits. Fourteen healthy New Zealand rabbits were equally divided into two groups. One group received no treatment, the other group was orally inoculated with 40 000 sporulated oocysts of E. stiedai in a 1 ml inoculum using a catheter. At day 24 after inoculation, blood samples were collected into sodium citrate-containing tubes to evaluate some coagulation parameters. Although statistically not significant, infected rabbits had prolonged prothrombin time and activated partial thromboplastin time compared with rabbits in the control group. A significant reduction (P < 0.05) was observed in the level of fibrinogen of infected rabbits compared with that of the controls. A slight decrease in thrombocyte counts of infected rabbits was not statistically significant.     

灰黄霉素在家兔组织中的分布及残留消除规律研究

研究灰黄霉素在家兔组织中的分布及消除规律,为灰黄霉素用于治疗家兔真菌病的安全性评价提供依据。选择42只新西兰白兔,于饲料中添加灰黄霉素（800g／1000kg）混饲,连续饲喂14d,分别于停药后1、3、5、7、9、14、21d,各处死6只,取其肝、肾、肌肉、皮肤及脑组织。以盐酸普萘洛尔为内标,二氯甲烷提取后用高效液相色谱-串联质谱仪（HPLC—MS／MS）进行各种组织中灰黄霉素浓度分析。结果显示,连续用药14d后,灰黄霉素在家兔组织中的分布情况为：肝组织浓度最高（134．61μg／kg）,肾组织次之（54．09μg／kg）,脑组织浓度最低（未检出）;停药后,随着时间的延长,灰黄霉素在肌肉、肝、肾和皮肤组织中的浓度逐渐下降,且在肝脏中的消除速度最快,停药21d后,灰黄霉素在肾组织中浓度为3．39μg／kg,在肝组织中浓度为12．36μg／kg,其他组织低于定量限或未检出。建议家兔生产中慎用灰黄霉素。     

液相色谱-串联质谱法研究地克珠利在家兔组织中残留及消除规律

[目的]建立一种检测家兔组织中地克珠利含量的液相色谱-串联质谱法（LC—MS／MS）,研究地克珠利在家兔组织中的残留及消除规律。[方法]选择48只家兔（雌雄各半）,在饲料中加入10g／1000kg地克珠利混饲,连续饲喂30d。分别于休药后的第0、1、2、3、4、5、6天,每个时间点随机处死6只家兔（雌雄各半）,取肌肉、肝脏、肾脏组织,DMF,乙腈沉淀法提取并用LC—MS,MS法检测地克珠利残留量。[结果]地克珠利在肾脏中的残留量最高（休药0d为912．6μg／kg）,肝脏次之（休药0d为156．μg／kg）,肌肉最低（休药0d为43．1μg／kg）,在肾脏中的消除速度最快。[结论]该方法适于检测家免组织中的地克珠利含量。休药0d即可满足欧盟要求     

恩诺沙星可溶性粉在白羽乌骨鸡组织中残留消除规律研究

为研究恩诺沙星可溶性粉在乌骨鸡体内的残留消除规律。50只11日龄健康白羽乌骨鸡,随机分为2组,给药组40只,对照组10只,给药组混饮恩诺沙星水溶液,浓度为75 mg/L,2次/d,连续用药5 d,对照组不给任何抗菌药物,与给药组同环境饲养。在最后一次给药后9、20、40、70、106 d采集肌肉、肝脏、肾脏、皮脂等样本,经LC-MS/MS法测定组织中的恩诺沙星及环丙沙星残留量,并利用WT1.4软件计算休药期。恩诺沙星可溶性粉在乌骨鸡肌肉、肝脏、肾脏及皮脂中的休药期分别是102.48、0、41.93、116.38 d。为保证消费者健康和食品安全,推荐恩诺沙星可溶性粉在乌骨鸡体内的休药期应大于117 d。     

Fatal toxoplasmosis in brown hares (Lepus europaeus): possible reasons of their high susceptibility to the infection

Brown hares (Lepus europaeus) trapped in the countryside and domestic rabbits were experimentally infected with Toxoplasma gondii (K7 strain) oocysts. Hares (n=12) were divided into groups of 4 and infected with 10, 10(3) and 10(5) oocysts. Rabbits (n=12) were infected in the same way. The experimentally infected animals were monitored for 33 days after infection (p.i.). Most of the infected hares demonstrated behavioural changes, and all of them died between 8 and 19 days p.i. Three of the rabbits demonstrated only clinical changes related to the concurrent pasteurellosis. The typical pathological finding in the hares were haemorrhagic enteritis, enlargement and hyperaemia of mesenteric lymph nodes, splenomegaly and multiple miliary necrotic lesions in the parenchyma of the liver and other organs. Pathological changes in the rabbits were less pronounced than in the hares. In rabbit brains, tissue cysts of the T. gondii were found. The incidence of T. gondii antibodies both in the hares and the rabbits was first ascertained on day 7 p.i. On day 12 p.i., antibodies were already found in all the animals infected. Antibody titres in indirect fluorescence antibody test (IFAT) using the anti-rabbit conjugate were markedly higher in rabbits than in hares. In all hares, T. gondii was isolated post mortem from the liver, brain, spleen, kidney, lung, heart and skeletal muscles. Although T. gondii was also isolated in all rabbits, it was not always isolated in all their organs. In all hares, parasitemia was demonstrated on days 7 and 12 p.i. The percentage of rabbits with detected parasitemia was lower. In hares, a decrease in the numbers of leukocytes during the infection was observed. No such decrease was observed in the rabbits. The lymphocyte activity after the stimulation with non-specific mitogens showed significant differences between the hares and the rabbits even before the infection. After the infection, the hares infected with 10(3) and 10(5) doses and in rabbits infected with a 10(5) dose showed a decrease of lymphocyte activity. Rabbits infected with a 10(3) dose showed an increase of the lymphocyte activity. While in hares toxoplasmosis was an acute and fatal disease, the infection in rabbits had subclinical manifestations only and easily passed to a latent stage. The different courses of toxoplasmosis in the hare and the rabbit may be due to the differences in the natural sensitivity of the two species to the T. gondii infection or a negative impact of stress to the immune status of hares.     

氟苯尼考注射液在鸡体内残留消除研究

［摘 要］为研究氟苯尼考注射液在鸡体内的残留消除规律。本实验采用35只约3kg左右的健康白羽鸡,随机分为2组,给药组30只,对照组5只。给药组用药量为20mg/kg/次,每隔48小时用药1次,连用2次,对照组不给任何抗菌药物,与给药组同环境饲养。在最后一次给药6h、24h(1天)、72h(3天)、120h(5天)、168h(7天)时采集肉、肝、肾、皮脂样本,经LC-MS/MS法测定组织中的氟苯尼考及其标示物氟苯尼考胺残留量,并利用WT1.4软件计算休药期。结果显示：氟苯尼考注射液在鸡肌肉、肝脏、肾脏及皮脂中的休药期分别是3.41d、2.06d、2.25d、1.47d。为保证兽药使用安全、消费者健康和食品安全,推荐氟苯尼考注射液在鸡体内的休药期为4d。     

Preventive treatment of rabbit coccidiosis with alpha-difluoromethylornithine

The effect of alpha-difluoromethylornithine (DFMO) administered in drinking water (0.2 and 0.5%) against liver and intestinal coccidioses in experimentally infected rabbits was studied. Zootechnical and clinical parameters were used to assess the efficacy of the compound. In both coccidioses the lower dose did not show any important effect. The higher dosage (0.5%) however, reduced the lesions induced by coccidia to an important extent. The animals infected and treated with 0.5% DFMO showed similar weight gain, food intake and relative liver weight figures to the non-infected controls. Serum transaminase levels in the rabbits infected with Eimeria stiedai and treated with 0.5% DFMO were close to those of the non-infected animals. Infected rabbits treated with this dosage also showed reductions (over 80%) in faecal oocyst output.     

Residues of Sulfadimidine/Sulfanilamide and Sulfamethoxypyridazine in Sheep Tissue

Following treatment of sheep with different sulfonamides, residues in kidney, liver, and muscle have been determined by microbiological and chemical methods. By the microbiological method residues could be detected in kidney until the third day after the combined treatment with sulfadimidine/sulfanilamide. Using the chemical method, residues of about 3.80 p.p.m. could be found that day in kidney, while the concentrations in liver and muscle were about 1.90 and 1.20 p.p.m., respectively. On the eighth day after the last treatment traces of the medicine could be found in kidney, liver and muscle by using the chemical method.Residues of sulfamethoxypyridazine could be detected micro-biologically in kidney the second day after the last administration of the drug. The concentration at that time in kidney, liver and muscle determined by chemical analyses was about 4.66, 2.45 and 1.23 p.p.m., respectively. Traces of sulfamethoxypyridazine in kidney, liver and muscle could also be detected on the eighth day after the last medication.Considering altered metabolic rates of sick animals and variations in excretion rates between individuals of the same species, as well as variation in size of the doses applied, a 10 day withdrawal period for sulfonamides is proposed.     

氧氟沙星在乌骨鸡体内残留消除规律研究

采用高效液相色谱法分析氧氟沙星在乌骨鸡体内各组织中的残留消除规律。荧光检测器的激发波长为280nm,发射波长为450Dill。色谱柱为ttypersilODS—C18,柱温40~C,以磷酸（以三乙胺调pH至2．5）-乙腈（82：18）为流动相,流速1．0mL／min,进样量为20肛L。氧氟沙星在乌骨鸡肌肉、肝脏、。肾脏中的平均回收率分别为86．3％、83．2％、82．5％。检测限为10ng／g。当组织中氧氟沙星浓度在10～2000ug／kg范围时,线性关系良好。利用该方法对氧氟沙星在乌骨鸡体内的残留消除规律进行了研究,结果表明,氧氟沙星在鸟骨鸡体内的残留消除速度极为缓慢。40天休药期后,2日龄使用组可在肌肉、肝脏和肾脏中分别检出氧氟沙星146．8“g／kg、20．6“g／kg和437．4μg／kg;28日龄使用组分别检测出193．5ug／kg、45．0μg／kg和425．5μg／kg。2日龄使用组休药78天时（即停药71天）,肌肉和肝脏中还能分别检出氧氟沙星11．2μg／kg和12．0μg／kg。实验表明乌骨鸡使用氧氟沙星仅适用于10日龄以下给药并休药70天以上,或不给予该类药物。     

Disposition kinetics of doxycycline in chickens naturally infected with Mycoplasma gallisepticum

1. The pharmacokinetic properties of doxycycline were determined in healthy chickens and chickens naturally infected with Mycoplasma gallisepticum after a single intravenous (i.v.) and oral administration of the drug at 20 mg/kg body weight. Tissue residues of the tested drug after an oral dose of 20 mg/kg given twice daily for 5 consecutive days were also estimated in diseased chickens. 2. The plasma concentrations of doxycycline following single i.v. and oral administration were higher in healthy chickens than in diseased ones. Following i.v. injection, the elimination half-life (t1/2beta), distribution half-life and mean residence time (MRT) were longer in healthy chickens than in diseased birds. The values of total body clearance (ClB) and volume of distribution (Vdss) were larger in healthy chickens than in diseased birds. 3. After single oral administration, the absorption half-life (tl/2ab) and the elimination half-life were longer in normal birds than in diseased ones. The maximum plasma concentration of the drug was higher in normal chickens than in diseased ones. 4. Following repeated oral administration, the concentration of doxycycline in all tissues except muscle was higher than the corresponding concentrations in plasma. Concentrations of doxycycline in different tissues were in the following order: kidney > liver > lung > muscle. The drug was detected in liver and kidney in substantial concentrations on d 5 post administration of the last dose whereas, on d 7, its concentration in all tissues was below the lower limit of the sensitivity of the assay method used. Because of the low sensitivity of the microbiological assay method used in this study, a safe withdrawal time for doxycycline in diseased birds could not be estimated for the meanwhile.     

盐酸克伦特罗在羊主要脏器中残留量消除规律的研究

本试验对盐酸克伦特罗在休药期肉羊眼睛、心脏、肾脏、肺脏、脾脏等组织中的残留规律进行了研究。选择24头体重为(30±5)kg健康肉羊进行试验,在饲料中添加50 μg/kg盐酸克伦特罗,连续饲喂35 d后休药,通过液相色谱—质谱联用/质谱检测休药期肉羊组织中克伦特罗含量,研究其残留量消除规律。试验结果表明,肉羊眼睛中有高浓度的盐酸克伦特罗残留且其在肝脏中消除较慢;停药第14天眼睛中盐酸克伦特罗的浓度仍为42.42～63.48 μg/kg,脾脏中盐酸克伦特罗消除速度是最快的;停药3 d时,检测不到盐酸克伦特罗的残留量(低于检出限0.07 μg/kg),故眼睛可用作检测盐酸克伦特罗在肉羊生产上非法使用的靶标。     

Decreasing profile of residual sulphaquinoxaline in eggs

1. Sulphaquinoxaline (SQ) was added to the diet of laying hens at 200 mg/kg for 7 successive days. Contents (mg/kg) of SQ in albumen and egg yolk of eggs laid after drug withdrawal were determined by high pressure liquid chromatography (HPLC). The contents in the whole egg were calculated taking into consideration the respective weights of albumen and egg yolk. 2. A time-lag in the initiation of decrease of SQ from eggs after the withdrawal of dietary SQ was observed. 3. The decreasing pattern from whole egg could be well described by the following equation with a time-lag of 1.0 d, y = 2.07e-0.5620(t-1.0), where y is the SQ content in whole egg, t is time (d) after the withdrawal of dietary SQ and a constant of 2.07 is the SQ content in whole egg laid at the withdrawal. 4. Biological half-life of SQ in the whole egg was estimated to be 1.23 d. 5. From the above equation, SQ residue of whole egg laid at 9th d after withdrawal will be below the detection limit of 0.01 mg/kg.     

Tissue residues of ractopamine and urinary excretion of ractopamine and metabolites in animals treated for 7 days with dietary ractopamine

Ractopamine HCl is a beta-adrenergic leanness-enhancing agent recently approved for use in swine. Depletion of ractopamine in tissues, and elimination of ractopamine and its metabolites in urine, is of interest for the detection of off-label use. The objectives of this study were to measure the residues of ractopamine in livers and kidneys of cattle (n = 6), sheep (n = 6), and ducks (n = 9) after treatment with dietary ractopamine for seven (sheep, ducks) or eight (cattle) consecutive days and to measure the depletion of ractopamine from urine of cattle and sheep. Two cattle and sheep and three ducks were each slaughtered with withdrawal periods of 0, 3, and 7 d. Urine samples were collected daily from cattle and sheep. Tissue ractopamine concentrations were determined using the regulatory method (FDA approved) for ractopamine in swine tissues. Ractopamine residues in urine samples were measured before and after hydrolysis of conjugates. Analysis was performed with HPLC using fluorescence detection after liquid- (hydrolyzed samples) and(or) solid-phase extraction. No residues were detected in duck tissues. Liver residues in sheep averaged 24.0 and 2.6 ppb after 0- and 3-d withdrawal periods, respectively. Sheep liver residues after a 7-d withdrawal period were less than the limit of quantification (2.5 ppb). Sheep kidney residues were 65.1 and undetectable at 0- and at 3- and 7-d, withdrawal periods, respectively. Cattle liver residues were 9.3, 2.5, and undetectable after 0-, 3-, and 7-d withdrawal periods, respectively; kidney residues were 97.5, 3.4, and undetectable at the same respective withdrawal periods. Concentrations of parent ractopamine in sheep urine were 9.8+/-3.3 ppb on withdrawal d 0 and were below the LOQ (5 ppb) beyond the 2-d withdrawal period. After the hydrolysis of conjugates, ractopamine concentrations were 5,272+/-1,361 ppb on withdrawal d 0 and 178+/-78 ppb on withdrawal d 7. Ractopamine concentrations in cattle urine ranged from 164+/-61.7 ng/mL (withdrawal d 0) to below the LOQ (50 ppb) on withdrawal d 4. After the hydrolysis of conjugates in cattle urine, ractopamine concentrations were 4,129+/-2,351 ppb (withdrawal d 0) to below the LOQ (withdrawal d 6). These data indicate that after the hydrolysis of conjugates, ractopamine should be detectable in urine of sheep as long as 7 d after the last exposure to ractopamine and as long as 5 d after withdrawal in cattle.     

Relationship between liver disorders and protection against Eimeria stiedai infection in rabbits immunized with soluble antigens from the bile of infected rabbits

Soluble antigens exist in the bile of rabbits infected with Eimeria stiedai (E. stiedai) in the acute phase, and rabbits immunized with the antigens show resistance against the infection. In this study, the liver function of rabbits immunized either with the soluble antigens or PBS were examined following the parasite challenge. Rabbits immunized with PBS shed a number of oocysts and showed an increase in r-glutamyltransferase (GGT) activity and a decrease in blood Indocyanine green (ICG) clearance. However, rabbits immunized with the soluble antigens shed a lower number of oocysts and showed a transient increase of alanine-aminotransferase (ALT) activity on Day 8 post-challenge (p.c.). The blood Indocyanine green clearance of the rabbits showed no change throughout the experiment. By histopathological observation of the liver, a number of merozoites were found in the biliary ducts on Day 8 post-challenge in the non-immunized rabbits. In contrast, a number of lymphocytes and neutrophilic leukocytes assembled around the biliary ducts of the immunized rabbits, but few parasites were found there on Day 8 post-challenge. These results suggest that the soluble antigens stimulate local immune reactions, for example around the biliary ducts, resulting in elimination of the parasite's development.     

Persistence of chloramphenicol residues in calf tissues.

Twenty-one 5 to 18 day old calves were administered 11 mg chloramphenicol in propylene glycol per kg body weight intramuscularly twice daily for three days. Groups of calves were euthanized with a barbiturate overdose at 5, 21, 42 and 70 days after the last dose was administered. Serum, kidney, analyzed for the drug using a quantitative gas chromatographic method with a detection limit of five parts per billion. After five days of withdrawal, chloramphenicol was detected in all the injection sites and in 6 out of 16 of the other samples. After 21 days of withdrawal, chloramphenicol was detected in all the injection sites and in one each of the serum, liver and muscle samples. After 42 days of withdrawal, chloramphenicol was detected in the injection sites only, and after 70 days of withdrawal it was not detected in any of the samples.