The evaluation of endophyte toxin residues in sheep fat

Abstract

AIM: To monitor changes in concentrations of lolitrem B and epoxy-janthitrems in the fat of sheep grazing perennial ryegrass infected with wild-type- and AR37-endophyte, respectively, during the time of year when ryegrass staggers would be expected to be observed.

METHODS: Ten 5-month-old lambs with no previous exposure to endophytes were grazed on either wild-type (containing lolitrem B, n = 5) or AR37 (containing epoxy-janthitrems, n = 5) endophyte-infected perennial ryegrass (Lolium perenne L.) pastures between October 2008 and June 2009. Animals were regularly assessed for ryegrass staggers using the Keogh scale (0 = no signs, 5 = severe tremors). When a score of > 3.5 was observed animals were removed from the treatment pastures for 1 month. Fat biopsy samples were taken from each animal at approximately monthly intervals and analysed for endophyte metabolites using high performance liquid chromatography (HPLC) methods developed during this study. Regular herbage samples were also taken and concentrations of endophyte metabolites measured.

RESULTS: Efficient and reproducible methods to analyse both lolitrem B and epoxy-janthitrems in fat were developed. Concentrations of lolitrem B and epoxy-janthitrems in herbage and in sheep fat increased from late November to peak in mid-February. Ryegrass staggers was observed in both groups of sheep at this time. Following 1 month of grazing non-infected pasture mean concentrations in fat of lolitrem B decreased by 43% from 61.8 to 35.3 ppb, and of epoxy-janthitrems by 38% from 1032.0 to 639.5 ppb. Maximum concentrations in herbage of epoxy-janthitrems (35.7 ppm) were higher than of lolitrem B (3.4 ppm), but signs of staggers were less severe in sheep grazing pasture containing the former compared with the latter (median Keogh scores in late February were 2 and 3, respectively), consistent with epoxy-janthitrems being low potency toxins.

CONCLUSION: This study demonstrated that concentrations of epoxy-janthitrems and lolitrem B in sheep fat increased quickly during the initial phase of the study when concentrations in pasture increased, and decreased when animals were removed from pastures containing these compounds. These data will be used in the risk assessment of the endophyte metabolites.     

Selenium residues in sheep meat

Selenium did not exceed 108 ng/g in muscle after single subcutaneous dosesof 5 mg selenium into weaned lambs. Levels in liverand kidney exceeded 2000 ng/g only during the first day after treatment.     

万古霉素在动物性食品中残留的风险评价

近年来万古霉素滥用导致耐甲氧西林葡萄球菌感染的发生率逐年增高。由于其在饲养过程中使用后可对动物有一定的促生长作用,动物性食品存在生产者非法添加的可能性。为了规范生产者行为、防范风险,本文对万古霉素存在风险加以评价并提出建议。     

气相色谱法测定羊组织中溴氰菊酯残留

用气相色谱法对羊肌肉、肝脏、肾脏等组织中的溴氰菊酯残留进行检测.用石油醚、乙醚进行提取,硅胶柱净化,采用DB-5弹性石英毛细管柱(30 m×0.25 mm×0.25 μm)气相色谱法测定.溴氰菊酯标准曲线线性范围为25～1 000 ng/mL,相关系数为0.999 6,线性关系良好,在各组织中的回收率为84%～108%,适用于羊组织中溴氰菊酯残留的检测.     

A physiologically based pharmacokinetic model for oxytetracycline residues in sheep

A physiologically based pharmacokinetic model (PBPK) for oxytetracycline (OTC) residues in sheep was developed using previously published data from a combined serum pharmacokinetic and tissue residue study [Craigmill et al. (2000) J. Vet. Pharmacol. Ther.23, 345]. Physiological parameters for organ weights and tissue blood flows were obtained from the literature. The tissue/serum partition coefficients for OTC were estimated from the serum and tissue residue data obtained at slaughter. The model was developed to include all of the tissues for which residue data were available (serum, kidney, liver, fat, muscle and injection site), and all of the remaining tissues were combined into a slowly perfused compartment with low permeability. Total body clearance of OTC calculated in the previous study was used as the starting value for clearance in the PBPK model, with the kidney being the only eliminating organ. The model was built using ACSL (Advanced Continuous Simulation Language) Graphic Modeler, and the model was fit to the serum and tissue data using the ACSL Math/Optimizer software (AEgis Technologies Group, Inc., Huntsville, AL, USA). A sensitivity analysis was also performed to determine which parameters had the greatest effect on the goodness of fit. Numerous strategies were tested to model the injection site, and a model providing a biexponential absorption of the drug from the injection bolus gave the best fit to the experimental data. The model was validated using the clearance parameters calculated from the traditional pharmacokinetic model for each individual animal in the PBPK model. This simple PBPK model well predicted OTC residues in sheep tissues after intramuscular dosing with a long-acting preparation and may find use for other species and other veterinary drugs.     

Pharmacokinetics, safety and tissue residues of sustained-release sulfamethazine in sheep

Concentration-time profiles and the rates of absorption, extent of distribution and half-lives of sulfamethazine (SMZ), administered intravenously, orally as a water solution and as a sustained-release formulation (CalfSpan) were determined in 10 healthy sheep. The geometric mean half-life of elimination of i.v. SMZ was 10.8 h, compared to 14.3 h for the sustained-release preparation (CalfSpan) and 4.3 h for the oral water solution. Blood levels of SMZ were at or above 50 micrograms/ml for more than 48 h for CalfSpan, for 24 h after i.v. SMZ (100 mg/kg body wt), and for less than 24 h after p.o. SMZ (100 mg/kg body wt). The mean bioavailability of the oral SMZ solution was 58.3% (AUCp.o./AUCi.v.). The estimated bioavailability of the CalfSpan preparation was 52.5%. The safety of the sustained-release preparation was tested by dosing sheep with multiples (one, three and five times) of the recommended dose (one tablet, 8 g SMZ, per 20 kg body wt), once a day for 3 days. Clinical blood chemistries showed a significant increase in serum iron, and a decrease in serum phosphorus in animals treated at the 3x and 5x dose levels. Necropsies of the 5x dose animals did not show any gross signs that could be attributed to SMZ, and histological examination of tissues from the 5x animals revealed no organ pathology. Residues of SMZ in liver, fat, kidney and skeletal muscle were measured in 20 animals that received one bolus per 20 kg body wt. The results indicate that SMZ residues are cleared rapidly, and are at or below the tolerance level of 0.1 mg/kg within 8 days after dosing so that the 18-day withdrawal time used in cattle would provide an appropriate margin of safety if used in sheep.     

Milk depletion of dicloxacillin residues in cows and sheep following intramammary administration

The excretion rate of dicloxacillin from milk was studied after intramammary administration of a suspension of the drug active in vegetable oil. Eight cows and eight sheep, four of each group in low and four in high milk production, were dosed with 200 mg dicloxacillin/quarter in cows and 100 mg dicloxacillin/quarter in sheep, three times at 12 h intervals. The dicloxacillin concentrations in milk were quantified by high performance liquid chromatography (HPLC). In cows, time until dicloxacillin was undetectable was 48 h and no difference was observed between the groups. In sheep, dicloxacillin was undetectable 72 h and 84 h after the treatment in low and in high milk production groups, respectively. The implications of several factors affecting the possible milk withdrawal period were studied. The results indicated that the pharmaceutical vehicle and the coefficient of lipid solubility exerted major effects on depletion time.     

Pharmacological evaluation of sheep lung strip

Isolated sheep lung parenchymal strips responded to histamine > carbachol > PGF_2a > 5-HT with contractions, and to isoproterenol (Isop), and to large doses of epinephrine (E), norepinephrine (NE) and phenylephrine (PE) with relaxations. PGF_2a-contracted lung strip responded to PGE₁ and PGE₂ with relaxation. The strips which were partially contracted to histamine, PGF_2a, 5-HT and carbachol also responded to isop, E and NE with relaxations. Histamine responses were not modified by metiamide (an H₂-receptor antagonist). Mepyramine and atropine selectively antagonized contractions to histamine and carbachol, respectively. After β-blockade with propranolol, lung strips responded to NE > E > PE > isop with contractions, which were inhibited or reversed by phentolamine and dibenzyline. It is concluded that H₁ receptors are present in sheep peripheral airway smooth muscles, and that a- and β-adrenoceptors mediate contraction and relaxation, respectively, in sheep lung strips.     

Concentration of conjugated linoleic acid in grazing sheep and goat milk fat

The conjugated linoleic acid (CLA) content of grazing sheep and goat milk fat, throughout their lactation period, was examined. Six sheep and six goat representative farms were selected at random and milk samples were taken at monthly intervals for fatty acids profile determination. Sheep and goat nutrition was based on natural grazing and on supplementary feeding during the winter months. From April onwards, grazing native pastures was the only source of feed for sheep and goats. The University farm whose sheep are kept indoors all year round without any grazing, was also used as reference. Fifteen individual milk samples were also taken in April from a sheep and goat farm respectively, in order to see the variability of CLA inside the farm. The results showed that: a. the CLA content of grazing sheep and goat milk fat increased significantly in April–May (early growth stage of grass) and then declined while that of indoors kept sheep was more or less constant during the same period, b. the isomers cis-9, trans-11 and trans-10, cis-12 of CLA were found in grazing sheep milk fat, while in indoors kept sheep and goats' milk fat only the cis-9, trans-11 isomer was found, c. the CLA content of sheep milk fat was much higher than that of goats, d. a negative correlation between sheep milk fat and CLA content was found and e. there was considerable variation in milk fat CLA content between sheep and goat farms and inside the farms.     

Computed tomographic evaluation of abdominal fat in minipigs

Computed tomography (CT) exams were conducted to determine the distribution of abdominal fat identified based on the CT number measured in Hounsfield Units (HU) and to measure the volume of the abdominal visceral and subcutaneous fat in minipigs. The relationship between the CT-based fat volumes of several vertebral levels and the entire abdomen and anthropometric data including the sagittal abdominal diameter and waist circumference were evaluated. Moreover, the total fat volumes at the T11, T13, L3, and L5 levels were compared with the total fat volume of the entire abdomen to define the landmark of abdominal fat distribution. Using a single-detector CT, six 6-month-old male minipigs were scanned under general anesthesia. Three radiologists then assessed the HU value of visceral and subcutaneous abdominal fat by drawing the region of interest manually at the T11, T13, L1, L3, and L5 levels. The CT number and abdominal fat determined in this way by the three radiologists was found to be correlated (intra-class coefficient = 0.9). The overall HU ranges for the visceral and subcutaneous fat depots were -147.47 to -83.46 and -131.62 to -90.97, respectively. The total fat volume of the entire abdomen was highly correlated with the volume of abdominal fat at the T13 level (r = 0.97, p < 0.0001). These findings demonstrate that the volume of abdominal adipose tissue measured at the T13 level using CT is a strong and reliable predictor of total abdominal adipose volume.     

Effects of dose and duration of therapy on gentamicin tissue residues in sheep

The effects of different doses and dosage regimens on gentamicin pharmacokinetics and tissue residues were determined. Five groups of 12 sheep each were given gentamicin IM: group I, 2 mg of gentamicin sulfate/kg once; group II, 6 mg/kg once; group III, 18 mg/kg once; group IV, 6 mg/kg every 24 hours for 3 doses; and group V, 2 mg/kg every 8 hours for 9 doses. Serum concentrations were determined serially until sheep were killed and necropsied. Three sheep from each group were killed at 1, 4, 8, and 12 days after the last dose was administered. Renal cortex, renal medulla, liver, spleen, lung, skeletal muscle, and skeletal muscle at the injection site were assayed for gentamicin. An exponential equation was fitted to the serum concentrations, and various pharmacokinetic variables were determined. Serum clearance tended to increase as the single dose increased (P = 0.0588). Steady-state volume of distribution increased as the single dose was increased (P less than 0.05). Renal cortex contained the highest concentration of gentamicin which decreased in a biexponential manner. Concentrations in all tissues, except the injection site, were dependent upon the amount of the total dose, not the size of the injected dose (P less than 0.05). Concentrations at the injection site were up to 29 micrograms/g of tissue at 1 day after the last dose was given and were dependent upon the amount of total dose from multiple injections, not on the amount of each injected dose (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Regulation of murine macrophage phagocytosis of sheep erythrocytes by T-2 toxin

The effect of a single oral dose of 4 mg of T-2 toxin/kg of body weight on in vivo phagocytosis of sheep RBC by peritoneal macrophages was evaluated in nonsensitized mice and in mice sensitized with sheep RBC. T-2 toxin treatment had no effect on the viability or phagocytic activity of resident peritoneal macrophages in nonsensitized mice. However, a significant (P less than 0.005) increase in phagocytic activity occurred in cells from mice treated with toxin and subsequently sensitized with sheep RBC. In contrast, phagocytosis of sheep RBC was significantly (P less than 0.05) suppressed in cells from mice treated with toxin after sensitization. Toxin treatment induced necrosis of lymphocytes and significant decreases in thymus and spleen weights. Seemingly, T-2 toxin, administered at a dose that caused marked lymphoid depletion, suppressed or enhanced in vivo macrophage phagocytic activity in antigenically sensitized mice, and enhancement or suppression of phagocytosis was a function of the time of toxin treatment in relation to antigenic stimulation.     

禾草内生真菌与草坪草改良

论述禾草内生真菌具有增强禾草的抗病虫害和环境适应性方面的作用，同时提出了内生真菌在草坪草改良中需解决毒性、侵染方法、侵染率、种子保存和寄主范围等问题。     

细胞因子对小尾寒羊胎盘成熟的影响

为了研究细胞因子对小尾寒羊胎盘成熟的影响 ,本实验采用液相竞争法和平衡法 ,对小尾寒羊空怀期 (n=5 )和妊娠期 (n=13)第 85天、10 5天、12 5天、14 0天和 15 0天 (足月 )时的血清白细胞介素 - 1β(IL - 1β)、白细胞介素 - 6 (IL - 6 )和肿瘤坏死因子(TNF)的含量分别进行检测。结果表明 ,IL - 1β和 IL - 6含量在 12 5天时与对照组间差异极显著 (P<0 .0 1) ,TNF含量在各时期与对照组差异均显著 (P<0 .0 1或 P<0 .0 5 )。随着妊娠过程的进展 ,三种细胞因子含量逐渐增加 ,12 5天时达到最高值 ,分别为 0 .390± 0 .196 ng/ ml,5 79.8± 15 2 .8pg/ ml和 2 .348± 0 .396 ng/ ml。而后逐渐下降 ,到足月前略有回升 ,但仍低于 12 5天时的最高值。由此看出小尾寒羊在妊娠期间 ,细胞因子与妊娠之间有着密切的关系 ,前者对于维持妊娠起到重要作用。由于 14 0天至足月3种细胞因子基本稳定 ,这表明胎盘于 14 0天左右达到成熟 ,IL- 1β和 IL- 6与启动分娩有关 ,而 TNF可能参与小尾寒羊分娩的启动。     

阿勒泰羊杂交改良和田羊生产肥羔效果

阿勒泰羊原产区为新疆北部的福海、富蕴、青河和阿勒泰等县 ,是哈萨克羊中的一个分支 ,以体格大、肉脂生产性能高而著称。羔羊具有突出的早熟特性 ,早期生长发育快 ,抓膘能力强。近年来应用阿勒泰羊改良当地土种绵羊 ,杂交优势明显。鉴于和田地区羊体格小、生长慢、胴体产肉量低的情况 ,农十四师四十七团于上世纪 90年代初引进阿勒泰种公羊与当地土种绵羊和田羊在塔里木盆地南缘进行了杂交改良 ,取得了显著的效果。所以用阿勒泰羊改良当地土种羊是提高养羊经济效益、发展羔羊肉生产的方向。1　适应性纯种阿勒泰羊产于冬季严寒漫长、夏季凉爽…