黄曲霉毒素B1、玉米赤霉烯酮和呕吐毒素的累加细胞毒性研究

由于饲料中多种霉菌毒素并存的几率比较高，本研究以仔猪肠上皮细胞（IPEC-J2）为模型，研究黄曲霉毒素B1（AFB1）、玉米赤霉烯酮（ZEA）和呕吐毒素（DON）的叠加细胞毒性。细胞毒性试验选用AFB1、ZEA和DON三种毒素作为响应面Box-Behnke设计的三个因素，以AFB1：10、20、30 μg/L，ZEA：150、300、450 μg/L，DON：500、1000、1500 μg/L作为Box-Behnke设计的三个编码水平。利用响应面设计构建得到17组复合霉菌毒素组合，以其对IPEC-J2细胞活力的影响作为参考指标，得到对细胞损伤程度最高和最低的霉菌毒素添加比例。结果表明：经方程预测后，得到细胞活力最低（霉菌毒素毒性最高）的AFB1、ZEA和DON组合为30、150 μg/L和1500 μg/L，经测定细胞活力为32.32%|得到细胞活力最高（霉菌毒素毒性最低）的AFB1、ZEA和DON组合为10、150 μg/L和600 μg/L，经测定细胞活力为53.01%。该结果为多种霉菌毒素叠加毒性的研究提供了依据。 [关键词] IPEC-J2细胞|黄曲霉毒素B1|玉米赤霉烯酮|呕吐毒素|细胞毒性     

高性能阻隔膜对新制蒸汽压片玉米储存过程防霉效果的研究

本试验利用普通隔膜和高性能阻隔膜材料储存新制蒸汽压片玉米,检测不同保存时间点蒸汽压片玉米的霉变情况,探讨利用高性能阻隔膜储存新制蒸汽压片玉米的可行性。试验以不密封的普通隔膜储存新制蒸汽压片玉米为未密封普通隔膜组,以密封的普通隔膜储存新制蒸汽压片玉米为密封普通隔膜组,以密封的高性能阻隔膜储存新制蒸汽压片玉米为高阻隔膜组。将各组新制蒸汽压片玉米分别在储存0、7、14、21、28、42、56 d后取出,进行感官评价并测定其化学成分含量、脂肪酸值及黄曲霉毒素B1(AFB1)、玉米赤霉烯酮(ZEA)和呕吐毒素(DON)含量。结果表明:1)随着储存时间的延长,未密封普通隔膜组和密封普通隔膜组相继出现霉变现象,高阻隔膜组储存至56 d时尚未出现霉变现象。2)随着储存天数的增加,不同膜处理的蒸汽压片玉米的粗脂肪、可溶性淀粉含量和淀粉糊化度呈降低趋势,粗灰分、粗蛋白质、中性洗涤纤维、酸性洗涤纤维含量在储存期间均没有显著变化(P0.05)。3)储存时间对蒸汽压片玉米的脂肪酸值存在极显著的影响(P0.01),高阻隔膜组蒸汽压片玉米的脂肪酸值增长最为缓慢,储存至56 d时其脂肪酸值为10.54 mg/kg。4)在储存期间,蒸汽压片玉米的AFB1、ZEA含量随储存时间延长呈显著上升趋势(P0.05),各组呕吐毒素含量没有显著差异(P0.05);储存至56 d时,未密封普通隔膜组、密封普通隔膜组、高阻隔膜组的AFB1、ZEA和DON含量分别为1.03、0.73、0.01μg/L,181.13、170.49、148.19μg/L和1.23、1.23、1.20μg/m L。由此可见,高性能阻隔膜能够有效抑制霉菌生长,延缓脂肪酸败,使新制蒸汽压片玉米安全储存至56 d。     

我国西南地区肉禽配合饲料中霉菌毒素的污染分布规律

本研究旨在了解我国西南地区肉禽配合饲料中霉菌毒素的污染分布规律。从西南地区(四川、重庆、贵州、广西和云南)不同规模饲料厂随机抽取100份肉禽(肉小鸭前期、肉仔鸡前期和肉仔鸡后期)配合饲料样品,用酶联免疫吸附试验(ELISA)试剂盒初筛和高效液相色谱法复查测定样品中黄曲霉毒素B_1(AFB_1)、玉米赤霉烯酮(ZON)、呕吐毒素(DON)和伏马毒素(FB)的含量。结果表明,肉禽配合饲料中AFB_1、DON、ZON和FB的检出率分别为89%、96%、85%和94.74%,AFB_1和ZON的超标率分别为18%和5%,而DON和FB无超标样品;不同地区的肉禽配合饲料中AFB_1、DON和ZON的平均含量之间存在差异,其中,肉禽配合饲料中AFB_1含量最高的是重庆(11.76μg/kg),DON含量最高的是四川(1.23 mg/kg),ZON含量最高的是云南(0.26 mg/kg);不同品种的肉禽配合饲料中AFB_1含量由高到底依次是肉小鸭前期、肉仔鸡后期和肉仔鸡前期配合饲料,DON和FB含量最高的是肉仔鸡前期配合饲料,而ZON含量最高的是肉仔鸡后期配合饲料,不同品种肉禽配合饲料中4种霉菌毒素含量均差异不显著(P0.05);从不同规模饲料厂来看,大规模饲料厂(年产量10万t)的饲料产品的污染水平比小规模饲料厂(年产量2万~5万t)低,但是不同规模饲料厂肉禽配合饲料中4种霉菌毒素的含量均差异不显著(P0.05)。由此可见,我国西南地区肉禽配合饲料普遍受到AFB_1、DON、ZON和FB的污染,其中AFB_1的超标率较高,4种霉菌毒素在不同地区、不同品种和不同饲料厂规模的饲料产品中的含量之间存在差异。     

Screening of Aflatoxins and Zearalenone in feedstuffs and complete feeds for pigs in Southern Vietnam

A survey ofaflatoxins and zearalenone (ZEA) in 120 samples, including cassava chip (12), maize (12), rice bran (24), broken rice (24), and complete feeds for growing pigs (24) and sows (24) was carried out in southern Vietnam. Aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) were determined by an HPLC method using an immunoaffinity column for clean-up. Zearalenone was analysed by Enzyme-Linked Immunosorbent Assay (ELISA) AgraQuant. High incidences of aflatoxins and ZEA in feedstuffs and pig feeds (83.3%-100%) were detected. Aflatoxins and ZEA were detected in all commodities analyzed. Aflatoxin B1 was found in 100, 92, 92, 83, 100 and 96%, respectively, of samples of cassava chip, maize, rice bran, broken rice, and complete feeds for growing pigs and sows, with mean concentrations of 0.86, 77.5, 1.3, 1.6, 4.7 and 7.5 microg/kg, respectively. AFB2 and AFG1 were found in low concentrations and co-occurred with AFB1 in all samples where they were found. ZEA was detected in 8%, 33%, 36%, 21%, 67% and 67%, respectively, of the samples of cassava chip, maize, rice bran, broken rice, and complete feeds for growing pigs and sows, with average concentrations of 10.0, 163.5, 83.1, 31.4, 86.0 and 101.4 microg/kg, respectively. The high incidence of mycotoxins found in feedstuffs highlights the need for periodic monitoring and prevention ofmycotoxins, not only in feeds, but also in food for human consumption.     

The administration of diets contaminated with low to intermediate doses of deoxynivalenol and supplemented with antioxidants and binding agents slightly affects the growth,antioxidant status,and vaccine response in weanling pigs

This study aimed to evaluate the impact of grading levels of deoxynivalenol (DON) in the diet of weaned pigs, as well as the effects of a supplementation with antioxidants (AOX), hydrated sodium calcium aluminosilicates (HSCAS), and their combination on the growth, AOX status, and immune and vaccine responses against the porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2). At weaning, 336 piglets were allocated to six dietary treatments according to a randomized complete block design. Treatments were as follows: basal diet (CTRL); basal diet containing DON at 1.2 mg/kg (DON1.2); basal diet containing DON at 2.4 mg/kg (DON2.4); DON2.4 diet + a mix of AOX which included vitamins A and E at 20,000 IU and 200 IU/kg feed respectively, selenized yeast at 0.3 mg/kg, and a grape seed extracts at 100 mg/kg feed (DON2.4 + AOX); DON2.4 diet + the mix of AOX and the modified HSCAS mentioned above (DON2.4 + AOX + HSCAS); DON2.4 + AOX + HSCAS. Pigs were vaccinated against PRRSV and PCV2 at 7 d; on 0, 14, and 35 d, growth performance was recorded, and blood samples were collected in order to evaluate the oxidative status, inflammatory blood markers, lymphocyte blastogenic response, and vaccine antibody response. Increasing intake of DON resulted in a quadratic effect at 35 d in the lymphocyte proliferative response to concanavalin A and PCV2 as well as in the anti-PRRSV antibody response, whereas the catalase activity decreased in DON2.4 pigs compared with the CTRL and DON1.2 groups (P ≤ 0.05). Compared with the DON2.4 diet, the AOX supplementation slightly reduced gain to feed ratio (P = 0.026) and increased the ferric reducing ability of plasma as well as α-tocopherol concentration (P < 0.05), whereas the association of AOX + HSCAS increased the anti-PRRSV IgG (P < 0.05). Furthermore, the HSCAS supplement reduced haptoglobin levels in serum at 14 d compared with the DON2.4 group; however, its concentration decreased in all the experimental treatments from 14 to 35 d and particularly in the DON2.4 + AOX pigs, whereas a different trend was evidenced in the DON2.4 + HSCAS group, where over the same period haptoglobin concentration increased (P < 0.05). Overall, our results show that the addition of AOX and HSCAS in the diet may alleviate the negative effects due to DON contamination on the AOX status and immune response of vaccinated weanling pigs.     

Effect of chronic lead intoxication on the distribution and elimination of amoxicillin in goats

A study of amoxicillin pharmacokinetics was conducted in healthy goats and goats with chronic lead intoxication. The intoxicated goats had increased serum concentrations of liver enzymes (alanine aminotransferase and γ-glutamyl transferase), blood urea nitrogen, and reactivated δ-aminolevulinic acid dehydratase compared to the controls. Following intravenous amoxicillin (10 mg/kg bw) in control and lead-intoxicated goats, elimination half-lives were 4.14 and 1.26 h, respectively. The volumes of distribution based on the terminal phase were 1.19 and 0.38 L/kg, respectively, and those at steady-state were 0.54 and 0.18 L/kg, respectively. After intramuscular (IM) amoxicillin (10 mg/kg bw) in lead-intoxicated goats and control animals, the absorption, distribution, and elimination of the drug were more rapid in lead-intoxicated goats than the controls. Peak serum concentrations of 21.89 and 12.19 µg/mL were achieved at 1 h and 2 h, respectively, in lead-intoxicated and control goats. Amoxicillin bioavailability in the lead-intoxicated goats decreased 20% compared to the controls. After amoxicillin, more of the drug was excreted in the urine from lead-intoxicated goats than the controls. Our results suggested that lead intoxication in goats increases the rate of amoxicillin absorption after IM administration and distribution and elimination. Thus, lead intoxication may impair the therapeutic effectiveness of amoxicillin.     

Interaction between the three frequently co‐occurring Fusarium mycotoxins in rats

To test the complex, acute biochemical effects of combined, naturally co‐occurring fusariotoxins, a 5‐day rat study was performed. Mycotoxin treatment was invented by intraperitoneal injection: FB₁ (F): 9 µg/animal/day (approx. 30 µg/kg bw/day), DON (D): 16.5 µg/animal/day (approx. 55 µg/kg bw/day) and ZEN (Z): 12.75 µg/animal/day (approx. 42.5 µg/kg bw/day). The binary groups (FB₁ and DON [FD], FB₁ and ZEN [FZ] and DON and ZEN [DZ]) as well as the ternary (FB₁, DON and ZEN [FDZ]) group were dosed at the same combined level as the individual mycotoxins. Body weight, feed intake and mortality were not affected by any of the treatments. FB₁ and DON in combination (FD) increased the plasma aspartate aminotransferase activity synergistically (compared to the individual FB₁ and DON). In the liver, both the total glutathione (GSH) and the glutathione peroxidase (GPx) activity were increased (p < 0.05) by the binary FB₁ and ZEN (FZ) and the DON and ZEN (DZ) groups as well as the ternary FB₁, DON and ZEA group (FDZ) compared to the control. The GSH level of the ternary group was significantly increased compared to the FB₁ group, whereas the GPx activity of the ternary group was significantly increased compared to all three the individual mycotoxin groups. The Bliss independence method revealed synergism between DON and ZEN (DZ), as well as FB₁ and DON (FD) on liver GPx activity. None of the toxins alone or in combination exerted strong genotoxicity on lymphocytes, neither on the gross histopathological characteristics. However, even at these low levels acute exposure of more than one of these mycotoxins (FB₁, DON and ZEN) affected metabolic and detoxification changes.     

白藜芦醇对玉米赤霉烯酮致小鼠肝脏氧化损伤及炎症的保护作用

为探究白藜芦醇（RSV）对玉米赤霉烯酮（ZEA）中毒小鼠肝脏损伤是否具有保护作用,试验将40只清洁级雄性BALB/c小鼠随机分为5组：对照组（生理盐水）、ZEA组（40 mg/kg）、不同浓度的RSV （50、100或200 mg/kg）与ZEA （40 mg/kg）共处理组,每组8只,各组均灌胃给药,试验期12 d。试验结束后采集小鼠肝脏样品,计算肝脏系数,采用苏木素-伊红（HE）染色和免疫组织化学染色法观察各组肝脏病理变化;检测肝脏NF-κB蛋白表达;比色法检测肝脏组织中丙二醛（MDA）含量、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性及血清中谷丙转氨酶（ALT）、谷草转氨酶（AST）和乳酸脱氢酶（LDH）活性,ELISA法检测血清中白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和IL-1β含量。结果显示,与对照组相比,ZEA组小鼠肝组织发生明显病理学变化;与ZEA组相比,不同浓度RSV与ZEA共处理组小鼠肝脏组织病理学变化都有所减轻。免疫组织化学结果显示,与对照组相比,ZEA组肝脏NF-κB蛋白表达增多;与ZEA组相比,不同浓度的RSV与ZEA共处理组肝脏NF-κB蛋白表达均有下降。与对照组相比,ZEA组肝脏系数和肝脏组织MDA含量极显著升高（P<0.01）,肝脏组织SOD和CAT活性显著或极显著降低（P<0.05;P<0.01）,血清中AST、ALT和LDH活性,IL-6、TNF-α和IL-1β含量均极显著升高（P<0.01）。与ZEA组相比,不同浓度RSV与ZEA共处理组肝脏组织CAT活性显著或极显著升高（P<0.05;P<0.01）,血清中AST、ALT和LDH活性及IL-6和IL-1β含量显著或极显著降低（P<0.05;P<0.01）;50 mg/kg RSV与ZEA共处理组肝脏组织SOD活性和血清中TNF-α含量分别显著升高（P<0.05）和极显著降低（P<0.01）;100和200 mg/kg RSV与ZEA共处理组肝脏系数和肝脏组织MDA含量显著或极显著降低（P<0.05;P<0.01）。结果表明,ZEA对小鼠肝脏有严重的氧化及炎症损伤作用,RSV对ZEA中毒的肝损伤具有一定保护作用,尤其以100 mg/kg RSV保护效果最佳。     

Studies on the application of enzyme immunoassays for the Fusarium mycotoxins deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone.

Polyclonal antisera against zearalenone (ZEA) were produced in rabbits after immunization with ZEA-oxime coupled to human serum albumin. Using these antibodies and a ZEA-oxime-horseradish peroxidase conjugate in a competitive direct enzyme immunoassay (EIA), the detection limit for ZEA was 70 pg/ml. The relative cross-reactivities of the assay with ZEA, alpha-zearalenol, beta-zearalenol, zearalanone, alpha-zearalanol, and beta-zearalanol, respectively, were 100%, 37.3%, 7.2%, 59.2%, 5.3%, and 3.9%, respectively. This EIA and two EIAs for deoxynivalenol (DON) and 3-acetyldeoxynivalenol(3-AcDON) (Usleber et al., 1991) were used to analyze wheat samples. The limits of determination for DON, 3-AcDON, and ZEA in wheat were 200 ppb, 50 ppb, and 20 ppb, respectively. The analysis of reference materials (wheat flour) containing DON by EIA showed good agreement with the nominal values. The EIA for ZEA was in addition used to analyze biological fluids, obtained during a feeding trial. Two lactating cows were administered 25 mg and 100 mg ZEA per day, respectively, over a period of 6 days. Serum, milk, urine, and feces were assayed in the ZEA-EIA with and without sample treatment with beta-glucuronidase prior to the analysis. Maximum toxin levels (ZEA-equivalents) found in milk were 0.4 and 1.2 ppb (glucuronides). The toxin concentration in milk decreased rapidly after the last toxin administration. In the urine, maximum levels of toxin-glucuronide conjugates were 23 ppb and 24 ppb, respectively. The serum toxin levels corresponded to those found in milk. In the feces, mean values were 150 ppb and 500 ppb, respectively, no conjugated toxins were found in feces.     

The effects of naturally deoxynivalenol-contaminated oats on the clinical condition,blood parameters,performance and carcass composition of growing pigs

A feeding trial with naturally deoxynivalenol (DON)-contaminated oats included in feed mixtures at graded levels was conducted in growing pigs. The DON concentrations were 0, 0.7, 1.7, and 3.5 mg/kg of complete feed mixture givenad libitum to different groups. The data recorded were feed consumption, body weight gain, slaughter weight, biochemical and haematological data including serum immunoglobulin A, clinical condition and post-mortem pathology including histopathology.Significantly decreasing body weight gain throughout the experimental period, decreased slaughter weight and reduced feed utilization efficiency were observed for the group fed a diet containing 3.5 mg/kg of DON. At the same DON concentration, there were increased liver weights and decreased concentrations of serum protein and albumin, and a temporary fall in packed blood cell volume, serum calcium and serum phosphorus. For the groups fed diets containing 1.7 and 3.5 mg/kg of DON, a statistically significant, dose-related decrease in daily feed consumption was observed. No other effects on haematological, biochemical or immunological parameters were recorded. The carcass quality was not affected in any group.It was concluded that significant effects in growing pigs may be observed at a dietary DON concentration of 1.7 mg/kg, originating from naturally contaminated oats included in a diet that was otherwise adequate and contained only minor traces of other mycotoxins.Abbreviations ALAT alanine aminotransferase - ASAT aspartate aminotransferase - CFU colony-forming units - DON deoxynivalenol - 3-ac-DON 3-acetyl-deoxynivalenol - F.U. feed unit - FUS-X fusarenon-X - HPLC high-pressure liquid chromatography - IgA immunoglobulin A - NFSA nutrient-free sporulation agar - NIV nivalenol - PDA potato dextrose agar     

玉米赤霉烯酮对断奶小母猪生产性能、血清抗氧化功能和免疫功能的影响

为了研究不同水平玉米赤霉烯酮(zearalenone,ZEA)污染饲粮对断奶小母猪生产性能、血清抗氧化功能、血清抗体水平及外周血淋巴细胞增殖率的影响。将40头健康三元杂交(杜×长×大)断奶小母猪按日龄[(35±1)日龄]和平均体重[(14.01±0.86)kg]分为4组,对照组饲喂基础饲粮(ZEA水平的测定值为0 mg/kg),试验组在基础饲粮中分别添加0.5、1.0及1.5 mg/kg ZEA[ZEA水平的测定值分别为(0.52±0.07)mg/kg、(1.04±0.03)mg/kg和(1.51±0.13)mg/kg]。预试期10 d,正试期35 d。结果表明:饲粮ZEA对断奶小母猪平均日采食量、平均日增重和料重比没有显著影响(P0.05)。与对照组相比,ZEA显著降低了血清谷胱甘肽过氧化物酶(GSH-Px,0.5、1.0和1.5 mg/kg ZEA)活性、猪瘟(1.0和1.5 mg/kg ZEA)和伪狂犬病病毒抗体水平(1.5 mg/kg ZEA)以及外周血淋巴细胞增殖率(1.0和1.5 mg/kg ZEA)(P0.05),而显著升高了血清丙二醛(MDA,0.5、1.0和1.5 mg/kg ZEA)含量(P0.05)。随着饲粮中ZEA水平的升高,断奶小母猪的料重比呈一次线性降低趋势(P=0.075),血清GSH-Px、超氧化物歧化酶(SOD)活性,血清病毒(猪瘟、伪狂犬病和高致病性猪蓝耳病病毒)抗体水平和外周血淋巴细胞增殖率均呈一次线性降低(P0.05),而血清MDA含量则呈一次线性升高(P0.05)。由此可见,饲粮中0.5 mg/kg的ZEA足以诱导小母猪的氧化应激反应,1.0 mg/kg的ZEA能够显著降低断奶小母猪的特异性体液免疫和细胞免疫功能。     

Isotype patterns of immunoglobulins: hallmarks for clinical status and tissue parasite density in Brazilian dogs naturally infected by Leishmania (Leishmania) chagasi

The role of anti-leishmanial immune response underlying the susceptibility/resistance during canine visceral leishmaniasis (CVL) has been recognized throughout ex vivo and in vitro investigations. Recently, we demonstrated that immunoglobulin levels (Igs), as well as the parasite load are relevant hallmarks of distinct clinical status of CVL. To further characterize and upgrade the background on this issue, herein, we have evaluated, in Leishmania (Leishmania) chagasi naturally infected dogs, the relationship between tissue parasitism (skin, bone marrow, spleen, liver and lymph node), the CVL clinical status (asymptomatic (AD), with no suggestive signs of the disease; oligosymptomatic (OD), with maximum three clinical signs-opaque bristles; localized alopecia and moderate loss of weight; symptomatic (SD), serologically positive with severe clinical signs of visceral leishmaniasis), and the humoral immunological profile of anti-Leishmania immunoglobulins (IgG, IgG1, IgG2, IgM, IgA and IgE). Our major statistically significant findings revealed distinct patterns of tissue parasite density within L. chagasi-infected dogs despite their clinical status, pointing out the spleen and skin as the most relevant sites of high parasitism during ongoing CVL. Parasite density of bone marrow and spleen were the most reliable parasitological markers to decode the clinical status of CVL. Moreover, the parasite density of bone marrow better correlates with most anti-Leishmania Igs reactivity. Additionally, a prognostic hallmark for canine visceral leishmaniasis was found, highlighting strong correlation between IgG1 and asymptomatic disease, but with IgA, IgE and IgG2 displaying better association with symptomatic disease. The new aspects of this study highlighted pioneer findings that correlated the degree of tissue parasite density (low (LP), medium (MP) and high (HP) parasitism) with distinct patterns of anti-Leishmania Igs reactivity. In this scope, our data re-enforce the anti-Leishmania IgG but with IgA reactivity as the better marker for overall tissue parasitism. The association between clinical status, Ig profile and the tissue parasitism support a novel investigation on the impact of humoral immune response and susceptibility/resistance mechanism during ongoing CVL.     

黄曲霉毒素B1、复方中药和硒对肉鸡肝组织匀浆中自由基代谢的影响

为观察黄曲霉毒素B1(AFB1)对肉鸡肝组织匀浆自由基代谢的变化,并研究复方中药与硒对AFB1的拮抗效应,将10日龄120只试验肉鸡随机分成4组,每组30只,分别为Ⅰ组(对照组)、Ⅱ组(AFB1组:AFB10.2mg/kgBW)、Ⅲ组(AFB10.2mg/kgBW+亚硒酸钠2mgSe/kgDM)和Ⅳ组(AFB10.2mg/kgBW+复方中药20g/kgDM)。分别处理后于17、24、31、38日龄宰杀5只肉鸡,提取肝组织,检测肝组织中的总抗氧化能力(T-AOC)、总超氧化物歧化酶(T-SOD)活性和过氧化氢酶(CAT)及丙二醛(MDA)含量的变化。结果显示,口服AFB1(0.2mg/kgBW)后T-SOD、T-AOC和CAT活性降低(P<0.05),MDA含量明显升高(P<0.05),而复方中药与硒治疗组T-SOD、T-AOC和CAT活性升高(P>0.05或P<0.05),MDA含量明显下降(P>0.05或P<0.05)。证实,由自由基增多而引起的脂质过氧化损伤在肉鸡AFB1慢性中毒发病机理中起着重要的作用,复方中药与硒均可有效降低脂质过氧化造成的损伤。     

脱氧雪腐镰刀菌烯醇与玉米赤霉烯酮联合暴露对体外培养鸡脾脏淋巴细胞内环境稳态的影响

本试验旨在研究脱氧雪腐镰刀菌烯醇(DON)与玉米赤霉烯酮与(ZEA)联合暴露对体外培养鸡脾脏淋巴细胞内环境稳态的影响。分别以0.012 50μg/mL DON+0.006 25μg/mL ZEA、0.050μg/mL DON+0.025μg/mL ZEA、0.2μg/mL DON+0.1μg/mL ZEA、0.8μg/mL DON+0.4μg/mL ZEA对体外培养鸡脾脏淋巴细胞进行联合暴露培养,48 h后测定细胞膜ATP酶(Ca~(2+)-ATP酶、Na~+/K~+-ATP酶)活性以及细胞内pH、Ca~(2+)水平和钙调蛋白(CaM)的mRNA表达水平。同时设不添加毒素的空白对照组。结果表明:添加毒素的各试验组间,细胞内Ca~(2+)水平、CaM mRNA表达水平随毒素浓度的升高而增加,且添加毒素的各试验组均显著或极显著高于空白对照组(P0.05或P0.01)。细胞内pH以及细胞膜Ca~(2+)-ATP酶与Na~+/K~+-ATP酶活性均随毒素浓度的升高而降低,且添加毒素的各试验组均显著或极显著低于空白对照组(P0.05或P0.01)。由此得出,DON、ZEA联合暴露导致体外培养鸡脾脏淋巴细胞内酸化、离子平衡失调等一系列细胞内环境稳态失衡,且呈剂量依赖性。     

Effects of purified zearalenone on growth performance, organ size, serum metabolites, and oxidative stress in postweaning gilts

Zearalenone (ZEA), an estrogenic mycotoxin, is produced mainly by Fusarium fungi. Previous studies indicated that acute ZEA exposure induced oxidative stress and damage in multiple organs. Therefore, the present study was designed to investigate the adverse effects of dietary ZEA (1.1 to 3.2 mg/kg of diet) on oxidative stress and organ damage in postweaning gilts. A total of 20 gilts (Landrace × Yorkshire × Duroc) weaned at d 21 with an average BW of 10.36 ± 1.21 kg was used in the study. Gilts were housed in a temperature-controlled room, divided into 4 treatments, and fed a basal diet only (control) or basal diet supplemented with purified ZEA at a dietary concentration of 1 (ZEA1), 2 (ZEA2), or 3 (ZEA3) mg/kg of diet for 18 d ad libitum. The actual ZEA contents (analyzed) were 0, 1.1 ± 0.02, 2.0 ± 0.01, and 3.2 ± 0.02 mg/kg for control, ZEA1, ZEA2, and ZEA3, respectively. Gilts fed different amounts of dietary ZEA grew similarly with no difference (P > 0.05) in feed intake. Vulva size increased linearly over the 18 d of feeding in gilts fed diets containing 1.1 mg of ZEA/kg or greater (P < 0.001). Relative weight of genital organs, liver, and kidney increased linearly (P < 0.05) in a ZEA-dose-dependent manner. Serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, γ-glutamate transferase, urea, and creatinine (P < 0.05), and malondialdehyde concentrations in both serum and liver (P < 0.001) were also increased linearly in a ZEA-dose-dependent manner. However, spleen relative weight (P = 0.002) and activities of total superoxide dismutase and glutathione peroxidase (in both serum and liver (P < 0.05) were decreased linearly as dietary ZEA increased. Results showed that besides genital organs, the liver, kidney, and spleen may also be target tissues in young gilts fed diets containing 1.1 to 3.2 mg of ZEA/kg for 18 d. Increased key liver enzymes in the serum suggest progressive liver damage caused by feeding ZEA, and an increase in oxidative stress in gilts is another potential impact of ZEA toxicity in pigs.     

Further survey of the occurrence of Fusarium toxins in wheat grown in southwest Germany.

A total of 53, 54, 57, 52 and 60 wheat samples for feed use were collected randomly after the 1989, 1990, 1991, 1992 and 1993 crops, respectively, from farms in an area of southwest Germany. Deoxynivalenol (DON), 3- and 15-acetyldeoxynivalenol (3-, 15-ADON), nivalenol (NIV), HT-2 toxin (HT-2), T-2 toxin (T-2), diacetoxyscirpenol (DAS), and fusarenon-X (FUS-X) were determined by gas chromatography, combined with mass selective detection (GC-MS), zearalenone (ZEA), alpha- and beta-zearalenol (alpha-beta-ZOL) were determined by HPLC. DON was the major toxin, with incidences at 77 to 93% and mean contents at 167 to 735 micrograms/kg. In contrast, incidences of ZEA, 3-ADON, NIV, HT-2, and T-2 were at 13 to 37%, 10 to 44%, 15 to 67%, 0 to 11%, and 0 to 12%, respectively, with mean contents in positive samples between 2 and 73 micrograms/kg, except for 948 micrograms/kg 3-ADON in samples from 1993. 15-ADON and FUS-X were assayed in samples from 1991, 1992 and 1993. 15-ADON was found in 0 to 11% of samples at mean levels < or = 17 mu/kg, DAS, alpha- and beta-ZOL, and FUS-X were not detected in any sample. Over the years, incidences and levels of toxins remained constant, decreased or increased, with most differences between years being slight and insignificant. The risk for livestock due to DON, HT-2 and ZEA was estimated based on maximum tolerated levels recommended for these toxins in some countries.     

Effects of ZEA,DON and Their Combined Effects on the Secretion of Cytokines in CTLL-2 Cells

To analyze the effects of ZEA or/and DON on the immunologic function,CTLL-2 cell was used as experimental materials. After exposing the CTLL-2 cell to different concentrations of ZEA (0,5,10,20 μg/mL),DON (0,0.5,1,2 μg/mL) and ZEA+DON (blank group,5 μg/mL ZEA,0.5 μg/mL DON,5 μg/mL ZEA+0.5 μg/mL DON) for 48 h,the concentration of GZMB,PFP,IFN-γ and TNF-α in CTLL-2 cell and supernatant fluid were detected by ELISA method. The result showed that DON and ZEA could reduce the concentration of PFP,GZMB and IFN-γ in the CTLL-2 cells and the culture supernatant fluids,and increase the concentration of TNF-α. The intoxicated group had significant or extremely significant difference compared to control group (P <0.05;P <0.01),which showed the dose effect. The group exposed to the combined of ZEA and DON showed an additive effect. The result indicated that ZEA,DON and the two combined could reduce the killing activity of immunological cell by affecting the secretion of immunological cytokines, and indirectly affect the negative feedback regulation of humoral immunity and cellular immunity,which led to the decline of immune function in animals.     

Effects of different glucocorticoid treatments on serum acute phase proteins in dogs

The serum concentrations of haptoglobin, caeruloplasmin, C-reactive protein and serum amyloid A were measured in three groups of seven healthy dogs. Group 1 received a single dose of 1.1 mg/kg methylprednisolone acetate, administered subcutaneously; group 2 received 1 mg/kg per day of prednisone administered orally for three weeks; and group 3 received 2.2 mg/kg per day of prednisone administered orally for seven days. Before the administration of the glucocorticoids the serum concentrations of all the acute phase proteins were within the authors' laboratory reference ranges. After the administration of the drugs there were significant increases in the concentration of haptoglobin in all three groups, the increases being larger in groups 2 and 3. In contrast, the concentrations of C-reactive protein, caeruloplasmin and serum amyloid A were not affected.     

ZEA、DON及其联合染毒对CTLL-2细胞因子分泌的影响

为研究镰刀菌毒素玉米赤霉烯酮（ZEA）、脱氧雪腐镰刀菌烯醇（DON）及其联合作用对动物免疫功能的影响,试验以CTLL-2细胞（细胞毒性T淋巴细胞株）为材料,用不同浓度的ZEA （0、5、10、20 μg/mL）、DON （0、0.5、1、2 μg/mL）及联合（空白组、5 μg/mL ZEA、0.5 μg/mL DON、5 μg/mL ZEA＋0.5 μg/mL DON）处理CTLL-2细胞48 h,采用ELISA法检测了细胞内及培养上清液中颗粒酶B （GZMB）、穿孔素（PFP）、干扰素γ（IFN-γ）和肿瘤坏死因子α（TNF-α）等细胞因子的含量。结果显示,ZEA、DON能够降低CTLL-2细胞胞内及培养上清液中PFP、GZMB、IFN-γ的浓度,增加TNF-α浓度,染毒组与对照组相比均有显著或极显著差异（P＜0.05;P＜0.01）,且均呈剂量效应关系;ZEA、DON联合染毒表现为加性效应。结果表明,ZEA、DON及其联合作用可通过影响免疫细胞因子的分泌,降低免疫细胞杀伤活力,间接影响机体体液免疫和细胞免疫的负调节,从而导致动物机体免疫机能下降。     

脱氧雪腐镰刀菌烯醇与T-2毒素对热应激蛋鸡肠道屏障功能及内质网应激和炎症反应的影响

本试验旨在研究脱氧雪腐镰刀菌烯醇（DON）与T-2毒素对热应激蛋鸡肠道屏障功能及内质网应激和炎症反应的影响。选择36周龄体重相近的京红1号蛋鸡96只，随机分成4组，每组8个重复，每个重复3只鸡。对照组（CON组）饲喂基础饲粮，DON组、T-2组、DON+T-2组分别在基础饲粮中添加3.00 mg/kg DON、 0.50 mg/kg T-2毒素、 1.50 mg/kg DON+0.25 mg/kg T-2毒素。预试期3 d，正试期28 d。试验期间白天平均温湿指数为84.87。结果表明：1）与CON组相比，T-2组十二指肠、空肠隐窝深度显著升高（P<0.05）,DON+T-2组空肠隐窝深度显著升高（P<0.05）,DON、T-2、DON+T-2组十二指肠绒毛高度/隐窝深度（V/C）值显著降低（P<0.05）,T-2、DON+T-2组空肠V/C值显著降低（P<0.05）,DON组回肠隐窝深度显著升高（P<0.05）。2）与CON组相比，DON、T-2、DON+T-2组血清脂多糖（LPS）含量显著升高（P<0.05）,DON+T-2组血清二胺氧化酶（DA...