Depletion study and estimation of the withdrawal period for enrofloxacin in pacu (Piaractus mesopotamicus)

Defining the pharmacokinetic parameters and depletion intervals for antimicrobials used in fish will help in the development of important guidelines for future regulations by Brazilian agencies on the use of these substances in fish farming. This paper presents a depletion study for enrofloxacin (ENR) and its main metabolite, ciprofloxacin (CIP), in pacu (Piaractus mesopotamicus) fillets. The depletion study was carried out under monitored environmental conditions, with the temperature controlled at 27 °C to mimic the fish farming conditions in Brazil. ENR was administered orally via medicated feed for 10 consecutive days at daily dosages of 10 mg/kg body weight (b.w.). The fish were slaughtered at 6, 12, and 24 h and 2, 3, 5, 8, 12, 17, and 24 days after the medication period. Considering a maximum residue limit of 100 μg/kg for the sum of the ENR and CIP residues in the fillet, the results obtained in the depletion study allowed the estimation of a half‐life for ENR of 2.75 days and a withdrawal period of 23 days. The results obtained in this study are important for the farming of pacu in tropical regions.     

Depletion study and withdrawal period calculation of florfenicol in the crucian carp (Carassius auratus) following multiple intramuscular injections

The previously adopted marker residue for florfenicol (FF) in China was only florfenicol amine (FFA); however, the marker residue has been changed to FF plus FFA since the end of 2017. The previous official withdrawal period determined based on the only concentration of FFA may no longer be suitable. Therefore, the present study aimed to determine the depletion profiles of FF and FFA and further calculate the withdrawal period in the crucian carp (Carassius auratus) based on the new marker residues. Florfenicol was intramuscularly administered at 10 mg/kg bodyweight daily for five consecutive days to crucian carps reared in freshwater at 10°C. After the last dose, plasma and tissue samples were randomly collected from 10 fish at different time points. The FF and FFA concentrations were simultaneously determined by high-performance liquid chromatography (HPLC) with a fluorescence detector and further subjected to noncompartmental analysis. The elimination half-life (h) of FF in different tissues decreased as follows: liver (39.1) > kidney (36.3) > skin plus muscle (34.6) > plasma (31.7), whereas that of FFA decreased as follows: kidney (41.4) > skin plus muscle (39.4) > liver (39.3) > plasma (35.7). Considering a maximum residue limit of 1 μg/g for the total concentration of FF and FFA in the skin plus muscle, a withdrawal period of 6 days was calculated based on the upper limit of the one-sided 95% confidence interval.     

农业部发布兽药停药期规定

为加强兽药使用管理 ,保证动物性产品质量安全 ,根据《兽药管理条例》规定 ,农业部组织制定了兽药国家标准和专业标准中部分品种的停药期规定 ,并确定了部分不需制定停药期规定的品种 ,于 2 0 0 3年 5月 2 2日以农业部公告第 2 78号予以公告。本公告自发布之日起执行。以前发布过的与本公告同品种兽药停药期不一致的 ,以本公告为准。兽药停药期规定兽药名称执行标准停药期1乙酰甲喹片兽药规范 92版牛、猪 3 5日2二氢吡啶部颁标准牛、肉鸡 7日 ,弃奶期 7日3二硝托胺预混剂兽药典 2 0 0 0版鸡 3日 ,产蛋期禁用4土霉素片兽药典 2 0 0 0版牛、羊…     

Critical review on the withdrawal period calculation for injection site residues

This review concerns a statistical method for calculating withdrawal period for injection site residues. A recently adopted Committee for Medicinal Products for Veterinary Use/European Agency for the Evaluation of Medicinal Product (CVMP/EMEA) guideline recommends to apply the same method for the calculation of withdrawal period for injection site residues as for other edible tissues. For reasons in this study developed below, this approach is deemed to be inappropriate for the injection site residues. The injection site residues often violate regression assumptions with regard to homoscedasticity (same variance in residue concentrations for different slaughter times) and linearity (of the mean depletion curve in log(e)-scale). The currently recommended method cannot adequately handle these aspects. An alternative pragmatic method taking into account the last slaughter time with all data below the reference threshold, combined with a safety span, is proposed for injection site residues. A nonparametric approach for calculating the withdrawal period is also presumed to be a sound alternative. The references commonly used are the Maximum Residue Limit (MRL) and the Acceptable Daily Intake (ADI). Unfortunately these references are not relevant to the acute risk exposure associated with injection site consumption. The use of alternative references, such as the Acceptable Single Dose Intake (ASDI) or the Acute Reference Dose (ARD) are thought to be more appropriate.     

Edwardsiella ictaluri as the causative agent of mortality in cultured Nile tilapia

Edwardsiella ictaluri was consistently isolated from the spleens, livers, and head kidneys of diseased Nile tilapia Oreochromis niloticus from a farm experiencing mortality events in several culture ponds. We describe the first published outbreak of E. ictaluri-induced edwardsiellosis in Nile tilapia. Pure cultures of the isolated bacteria were characterized both biochemically and molecularly. Biochemical analysis was performed using the API-20E and RapID One systems, and antimicrobial susceptibility was determined by the broth microdilution method. Molecular analysis involved sequencing of the 16S rRNA gene, species-specific real-time polymerase chain reaction (PCR), and PCR-mediated genomic fingerprinting (rep-PCR). Pairwise sequence analysis of the 16S rRNA gene identified the case isolates to be a 100% match to E. ictaluri cultured from channel catfish in the southeastern United States. However, rep-PCR analysis identified the case isolates to be genetically different from representative strains isolated from disease outbreaks in cultured channel catfish in Mississippi. Infectivity challenges (intraperitoneal injection and immersion) demonstrated that a representative E. ictaluri strain isolated from tilapia was pathogenic to naive tilapia, reproducing clinical signs and mortality, thereby establishing Koch's postulates.     

Hematology and plasma chemistry reference intervals for cultured tilapia (Oreochromis hybrid)

Abstract: Tilapia are a commonly aquacultured fish yet little is known about their normal physiology and response to disease. In this study we determined the results of complete hematologic (n = 40) and plasma biochemical profiles (n = 63) in production tilapia ( Oreochromis hybrids). The fish were raised in recirculating systems with a high stocking density (120 g/L), and were in the middle of a 15-month production cycle. Blood was analyzed using standard techniques, and reference intervals were determined using nonparametric methods. Non-production tilapia (n = 15) from low-density tanks (4 g/L) also were sampled; the clinical chemistry results were compared to reference intervals from the fish raised in high-density tanks. Differences were noted in plasma protein, calcium and phosphorus concentrations, such that reference intervals for high-density production tilapia were not applicable to fish raised under different environmental and management conditions.     

Cross-sectional study of hepatic CYP1A and CYP3A enzymes in hybrid striped bass, channel catfish and Nile tilapia following oxytetracycline treatment

Terramycin for Fish® (oxytetracycline, OTC) is one of three approved drugs for therapeutic treatment of fish in the United States. Nothing is known, however, of the effects of this therapeutic on drug metabolizing enzymes in fish post-treatment. The main purpose of the study was to examine whether the fish CYP1A and CYP3A enzymes would cross-react with antibodies to known mammalian cytochrome P-450 forms (CYP1A1 and CYP3A). Observational feeding studies of OTC effects were conducted in hybrid striped bass, channel catfish and Nile tilapia. Oxytetracycline was mixed into the feed to achieve a daily dose of 82.8 mg per kg body weight at a feeding rate of 1% body weight per day. Hepatic microsomes of each fish were prepared and Western blotting of CYP1A1 and CYP3A4 and enzyme assays of CYP1A2 and CYP3A4 were performed prior to OTC treatment and on post-treatment days 1, 6, 11 and 21. Both goat anti-rat CYP1A1 and rabbit anti-human CYP3A4 showed good cross-reactivity with all three species in this study. All three species exhibited distinct perturbations in one or more of the variables examined on day 1 post-treatment. Immediately following the 10-day medication period, relative liver weight (RLW) of hybrid striped bass was increased 44% and remained elevated through post-treatment day 21. Increased CYP3A4 enzyme activity and protein abundance were noted in channel catfish and Nile tilapia, respectively. This observational approach demonstrated species differences both in control activities and in the timing and extent of hepatic responses to OTC. The unique perturbations of hepatic CYP450 enzymes in different fish species to OTC treatment observed in this study may have relevance for the use of additional antibiotics or other therapeutics used in aquaculture.     

Evaluation of cimaterol for finishing swine including a drug withdrawal period

One hundred fifty crossbred pigs (55 kg) were allotted by weight, sex and litter to a randomized complete-block design with five dietary treatments, six blocks per treatment and five pigs per pen with sex equalized across treatments. Corn-soybean meal-based diets (.65% lysine) with 0, .25 and .5 mg/kg cimaterol were fed, on an ad libitum basis, to pigs slaughtered at an average pen weight of 104 kg/pig. Drug withdrawal prior to slaughter was 1, 3 and 5 d for pigs fed cimaterol at .25 mg/kg and 1 d for those fed cimaterol at .5 mg/kg of diet. Dietary cimaterol level influenced (quadratic, P less than .01) average daily gain during the first 42 d on test; however, daily feed intake and feed conversion ratio were not affected (P greater than .1). Pigs fed .25 mg/kg cimaterol with a 1-d drug withdrawal had 6.8, 7.7 and 13.5% less 10th rib fat depth and 11.1, 6.1 and 13.3% less P2 fat depth than those subjected to either a 3- or 5-d drug withdrawal or those fed the 0 mg/kg cimaterol diet (control), respectively. Overall, pigs fed cimaterol had 7.9% larger longissimus muscle area and 2.6% more kilograms of muscle than pigs fed the control diet. Cimaterol fed at .5 mg/kg resulted in higher (P less than .05) Warner-Bratzler shear force values and altered the proportion of saturation in some long-chain fatty acids, although the total saturated:unsaturated fat ratio was not affected. Pigs fed no cimaterol had less thaw loss (P less than .05) than did those fed other treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genetic parameter estimation for pre- and post-weaning traits in Brahman cattle in Brazil

Beef cattle producers in Brazil use body weight traits as breeding program selection criteria due to their great economic importance. The objectives of this study were to evaluate different animal models, estimate genetic parameters, and define the most fitting model for Brahman cattle body weight standardized at 120 (BW120), 210 (BW210), 365 (BW365), 450 (BW450), and 550 (BW550) days of age. To estimate genetic parameters, single-, two-, and multi-trait analyses were performed using the animal model. The likelihood ratio test was verified between all models. For BW120 and BW210, additive direct genetic, maternal genetic, maternal permanent environment, and residual effects were considered, while for BW365 and BW450, additive direct genetic, maternal genetic, and residual effects were considered. Finally, for BW550, additive direct genetic and residual effects were considered. Estimates of direct heritability for BW120 were similar in all analyses; however, for the other traits, multi-trait analysis resulted in higher estimates. The maternal heritability and proportion of maternal permanent environmental variance to total variance were minimal in multi-trait analyses. Genetic, environmental, and phenotypic correlations were of high magnitude between all traits. Multi-trait analyses would aid in the parameter estimation for body weight at older ages because they are usually affected by a lower number of animals with phenotypic information due to culling and mortality.     

Factors affecting the accuracy of the live animal swab test for detecting urine oxytetracycline and predicting oxytetracycline residues in calves

The live animal swab test (LAST) was compared with quantitative oxytetracycline (OTC) assay of urine samples and tissue specimens to determine the accuracy of the LAST in detecting OTC in bovine urine and predicting violative residues in tissues. When urine OTC concentration was greater than 4.3 micrograms/ml, the LAST result was 100% accurate. When urine OTC concentration was less than 4.3 micrograms/ml, the LAST result was 60% accurate; 20% of the LAST results were false-positive, and 20% were false-negative. Urine osmolarity was highest (P less than 0.05) in samples with false-positive results and lowest (P less than 0.05) in samples with false-negative results. A similar trend was observed for urine pH, but was not statistically significant. Urine samples with false-positive results apparently had osmolarity and pH conditions that inhibited growth of Bacillus subtilis when OTC was lacking. False-negative results probably were obtained because urine osmolarity and pH conditions were favorable for the growth of bacteria even in the presence of OTC or because OTC concentration was below the limit of detection by the LAST. The LAST was inconsistent in detecting urine OTC in small concentrations and correspondingly failed to accurately predict OTC residues in tissues.     

The present and future of withdrawal period calculations for milk in the European Union: focus on heterogeneous,nonmonotonic data

Harmonization of the method for calculating the withdrawal period for milk dates from the 1990s. European harmonization has led to guidance with three accepted methods for determining the withdrawal period for milk that are currently applicable. These three methods can be used by marketing authorization holders, but, in some cases, their diversity can lead to very different withdrawal periods. This is particularly the case when concentrations in milk are nonmonotonic and heterogeneous, meaning that concentrations strictly increase and then strictly decrease with significant interindividual variability in the time to reach the maximal concentration. Here, we first describe the concepts associated with the different methods used in the harmonized approach currently applicable for the determination of milk withdrawal periods, and then, we propose the application of a modern pharmacometric tool. Finally, with a nonmonotonic heterogeneous dataset, we illustrate the usefulness of this tool in comparison with the three currently applicable methods and discuss the limitations and advantages of each method.     

The present and future of withdrawal period calculations for milk in the European Union: dealing with data below the limit of quantification

The assessment of withdrawal periods for milk is affected by the occurrence of data below the lower analytical quantification limit (BLQ data) and the resulting uncertainty. The current regulatory approach for dealing with BLQ residues is simple and easy: BLQ data (and missing data) are arbitrarily reassigned a value of one‐half the LOQ before any calculation on the data with one of the three currently applicable methods. Here, we reconsider the determination of the withdrawal period of milk with data below the limit of quantification. Theoretical background on analytical limits and pharmacometric considerations will be established. Then, we analyze the uncertainty problems caused by the current approach and propose a calculation solution (maximum‐likelihood estimation handling left‐censored data) included in nonlinear mixed‐effects modeling. Finally, we illustrate this issue using a case example.     

Comparison of erythromycin and oxytetracycline for the treatment of ovine footrot

A microbiological study of 25 cases of ovine footrot was performed. Cultures belonging to Dichelobacter nodosus were isolated in 48% of the sampled animals. The sensitivity of the 99 strict anaerobic bacterial isolates to 5 antibiotics (penicillin G, amoxycillin, spiramycin, erythromycin and oxytetracycline) was studied. The percentage of resistant cultures was in all cases higher than 30%. The efficacy of erythromycin and oxytetracycline in the treatment of ovine footrot was studied. To conduct this test, an intramuscular injection was applied, of one antimicrobial or the other, at the beginning of the treatment. The tolerance of animals to the antimicrobials, the success rate of treatment and the severity of lameness were evaluated. The percentage of animals cured within 15 days was around 75%. In contrast, only 44% improvement was achieved in the lameness. No differences were found between the two antimicrobials in the above indices.     

A piscirickettsiosis-like syndrome in cultured Nile tilapia in Latin America with Francisella spp. as the pathogenic agent

In 2004, cultured Nile tilapia Oreochromis niloticus in several Latin America farms began to succumb to a disease similar to the piscirickettsiosis-like syndrome previously reported in tilapia in Taiwan and the United States. Mortality increased during 2005; reductions in tilapia biomass ranged from 5% to 80% in individual ponds and averaged 50% overall. All ages of fish have been involved. Clinical signs include lethargy, loss of appetite, petechia, exophthalmia, and abnormal swimming behavior. Gross lesions have included splenomegaly, renomegaly, and numerous white nodules observed in the spleen, kidney, testes, heart, ovaries, and occasionally the liver. A previously unreported black granulomatous lesion was reported in up to 30% of the fillets. Histologically, granulomatous infiltrates were observed in the kidney, spleen, liver, testes, ovary, and choroid gland, and rarely in the brain and heart. A small pleomorphic bacterium was observed in Giemsa-stained blood smears and spleen imprints. The bacterium did not grow on standard microbiological media and has not been isolated in cell culture. We obtained a near-complete 16S ribosomal DNA sequence with high similarity to Francisella spp. sequences previously identified in tilapias Oreochromis spp. (Taiwan), Atlantic cod Gadus morhua (Norway), and three-line grunts Parapristipoma trilineatum (Japan).     

Pharmacokinetic and tissue distribution study of oxytetracycline in rainbow trout following bolus intravenous administration

Oxytetracycline pharmacokinetics and tissue distribution were studied in rainbow trout following bolus i.v. administration at 5 mg/kg. The mean serum (log) drug concentration data were plotted against time (linear). The decay curve was described by a three-component exponential decay function and a three-compartment model. The t1/2 of rapid distribution was 0.9 h, the t1/2 of the slow distribution was 5.9 h and the t1/2 elimination was 81.5 h. Clearance was 25.4 ml/kg/h and Vd(area) 2988 ml/kg. Regression analysis of the serum levels for the three intervals, 0.5-2.0 h, 6.0-18.0 h, and 24-96 h, indicated that the rates of decay for each interval were 0.6151 h-1, 0.0564 h-1 and 0.0088 h-1 respectively. Rates of equilibration between tissues and serum were determined. Kidney equilibrated the fastest with t1/2 to equilibration of 1.1 h for H (anterior) kidney and 1.98 h for P (posterior) kidney. The highest drug levels were found in the liver and the lowest were in the brain.     

Bayesian inference for genetic parameter estimation on growth traits for Nelore cattle in Brazil,using the Gibbs sampler

This data set consisted of over 29 245 field records from 24 herds of registered Nelore cattle born between 1980 and 1993, with calves sires by 657 sires and 12 151 dams. The records were collected in south‐eastern and midwestern Brazil and animals were raised on pasture in a tropical climate. Three growth traits were included in these analyses: 205‐ (W205), 365‐ (W365) and 550‐day (W550) weight. The linear model included fixed effects for contemporary groups (herd‐year‐season‐sex) and age of dam at calving. The model also included random effects for direct genetic, maternal genetic and maternal permanent environmental (MPE) contributions to observations. The analyses were conducted using single‐trait and multiple‐trait animal models. Variance and covariance components were estimated by restricted maximum likelihood (REML) using a derivative‐free algorithm (DFREML) for multiple traits (MTDFREML). Bayesian inference was obtained by a multiple trait Gibbs sampling algorithm (GS) for (co)variance component inference in animal models (MTGSAM). Three different sets of prior distributions for the (co)variance components were used: flat, symmetric, and sharp. The shape parameters (ν) were 0, 5 and 9, respectively. The results suggested that the shape of the prior distributions did not affect the estimates of (co)variance components. From the REML analyses, for all traits, direct heritabilities obtained from single trait analyses were smaller than those obtained from bivariate analyses and by the GS method. Estimates of genetic correlations between direct and maternal effects obtained using REML were positive but very low, indicating that genetic selection programs should consider both components jointly. GS produced similar but slightly higher estimates of genetic parameters than REML, however, the greater robustness of GS makes it the method of choice for many applications.     

Efficacy of long-acting oxytetracycline for the prevention of tick-borne fever in calves

Twenty-six calves which had not previously grazed tick-infested pasture were divided into two equal groups. On May 26, 1988 (day 0) they were turned out into a field of rough grazing where cases of redwater fever had occurred the previous spring. Seven, 14, 21 and 28 days after the start of the trial the animals in one group each received an intramuscular injection of 20 mg/kg bodyweight of long-acting oxytetracycline. During the 60 days of the trial the animals received a severe tick-borne fever challenge, in some cases combined with a redwater fever challenge. An unforeseen complicating factor was the presence of animals persistently infected with bovine viral diarrhoea virus, present in almost equal numbers in both groups. At the end of the trial the treated group weighed on average 16 kg more than the control group, a difference which was attributed to the suppression of tick-borne fever by oxytetracycline.     

Long-acting antibiotic formulations in the treatment of calf pneumonia: a comparative study of tilmicosin and oxytetracycline

The treatment of an outbreak of acute pneumonia in 50 four- to eight-month-old Friesian and Friesian cross calves is described. At the first visit (day 0) 16 calves received 20 mg/kg bodyweight of oxytetracycline dihydrate intramuscularly and 15 received 10 mg/kg of the macrolide tilmicosin subcutaneously. The remaining 19 in-contact animals were not considered ill enough to be included in the trial and received 20 mg/kg of oxytetracycline dihydrate. The rectal temperature, demeanour, respiratory rate and respiratory effort of each calf was assessed on days 1, 2, 3, 9, 14, 21 and 28, and calves which had not responded were given repeat injections of the same antibiotic. All the calves recovered from the outbreak and of the 19 calves treated strategically, three required a second injection. Among the calves with clinical pneumonia, fewer treatments (P less than 0.01) were required by those treated with tilmicosin. The rectal temperatures of both groups decreased (P less than 0.05) after the first injection, but on day 3 the decrease was greater (P less than 0.05) in the group treated with tilmicosin. Respiratory rates varied widely but respiratory effort was less (P less than 0.05) on day 2 in the calves treated with tilmicosin. When long-acting antibiotic injections are used to treat enzootic pneumonia it is suggested that a second visit should be made on day 3 to assess the animals' response to treatment.