Stability of Domoic Acid in 50% Methanol Extracts and Raw Fecal Material from Bowhead Whales (Balaena mysticetus)

Domoic acid (DA), the toxin causing amnesic shellfish poisoning (ASP), is produced globally by some diatoms in the genus Pseudo-nitzschia. DA has been detected in several marine mammal species in the Alaskan Arctic, raising health concerns for marine mammals and subsistence communities dependent upon them. Gastrointestinal matrices are routinely used to detect Harmful Algal Bloom (HAB) toxin presence in marine mammals, yet DA stability has only been studied extensively in shellfish-related matrices. To address this knowledge gap, we quantified DA in bowhead whale fecal samples at multiple time points for two groups: (1) 50% methanol extracts from feces, and (2) raw feces stored in several conditions. DA concentrations decreased to 70 ± 7.1% of time zero (T₀) in the 50% methanol extracts after 2 weeks, but remained steady until the final time point at 5 weeks (66 ± 5.7% T₀). In contrast, DA concentrations were stable or increased in raw fecal material after 8 weeks of freezer storage (−20 °C), at room temperature (RT) in the dark, or refrigerated at 1 °C. DA concentrations in raw feces stored in an incubator (37 °C) or at RT in the light decreased to 77 ± 2.8% and 90 ± 15.0% T₀ at 8 weeks, respectively. Evaporation during storage of raw fecal material is a likely cause of the increased DA concentrations observed over time with the highest increase to 126 ± 7.6% T₀ after 3.2 years of frozen storage. These results provide valuable information for developing appropriate sample storage procedures for marine mammal fecal samples.     

猪肉源金黄色葡萄球菌毒力基因检测与耐药性分析

【目的】了解陕西关中地区猪肉中金黄色葡萄球菌(Staphylococcus aureus)的污染状况、耐药性及其毒素基因的分布。【方法】采集陕西关中6个地区的猪肉165份,按国标GB/T 4789.10-2010的方法,对其中的金黄色葡萄球菌进行分离,采用PCR方法对该菌进行确证并对其相关基因(如nuc、mecA、PVL、SEs和ETs)进行检测,最后采用琼脂稀释法检测金黄色葡萄球菌对11种抗菌药物的耐药性。另外,在BP平板中分别添加头孢西丁(4μg/mL)和苯唑西林(4μg/mL),分离耐甲氧西林金黄色葡萄球菌(MRSA)。【结果】165份样品的金黄色葡萄球菌污染率为33.33%(55/165);从中分离出103株金黄色葡萄球菌,但未检测出MRSA,这些菌对甲氧苄啶的耐药性最强,耐药率为100%;其次对红霉素和四环素的耐药率较高,分别为57.28%和34.95%;对苯唑西林、庆大霉素、氯霉素、环丙沙星的耐药率分别为2.91%,10.68%,2.91%和3.88%;所有菌株对头孢西丁、头孢哌酮、万古霉素、阿米卡星均敏感,同时得到21种耐药谱,多重耐药率达20.39%。猪肉金黄色葡萄球菌中杀白细胞素基因(Panton-valentine leu-kocidin,PVL)的检出率为34.95%,肠毒素基因(SEs)中sej的检出率最高,为98.06%,然后依次为sea(50.49%)、see(34.95%)、sed(31.07%)、sec(13.59%)、seh(8.74%)、sei(8.74%)、seg(6.80%)和seb(1.94%);同时得到71种毒素基因型,以sea+sej(11.65%)最为流行,分布地区不尽相同,其次为PVL+sea+see+sej(9.71%),耐红霉素的金黄色葡萄球菌含的毒素基因类型比较复杂,sej基因检出率高达98.68%。在BP平板中分别添加头孢西丁和苯唑西林,均未检测出MRSA。【结论】猪肉存在金黄色葡萄球菌的污染,其污染菌株存在多重耐药性并携带较多毒素基因,提示应加强猪肉金黄色葡萄球菌的监测。在BP平板中分别添加头孢西丁(4μg/mL)和苯唑西林(4μg/mL)筛选MRSA的方法不一定可靠,其可信度有待证明。     

饲料中T-2毒素直接竞争ELISA检测方法的建立

采用直接竞争ELISA方法建立了一种快速检测饲料样本中T-2毒素残留的方法,在0～81μg/L范围内,T-2毒素的Logit(B/B0)与T-2毒素质量浓度的对数呈显著的线性关系,并对该检测体系的检测限、准确度、精密度等性能进行了测定。结果显示,该方法的IC50(半数抑制质量浓度)为2.8μg/L,对饲料样本的检测限为10μg/kg,样本添加回收率为77.6%～98.2%,变异系数为5.1%～12.0%。该方法操作简便、准确,检测时间短(仅需15min),适用于饲料样本中T-2毒素的快速检测。     

猪流感病毒多表位融合蛋白对小鼠的免疫原性分析

作者拟利用表位多肽的抗原性及黏膜佐剂免疫增强作用,设计可通过黏膜途径免疫接种的猪流感病毒通用型疫苗.体外合成H1N1、H3N2亚型猪流感病毒表位抗原基因,C末端串联大肠杆菌热敏性肠毒素LTb基因,构建pET30(a)-ep-LTb表达载体,利用SDS-PAGE、Western blot分析重组融合蛋白表达及生物学特性,小鼠免疫试验分析融合蛋白免疫原性.SDS-PAGE检测表达蛋白相对分子质量约38 ku,主要以包涵体形式存在.重组蛋白可与抗His-tag抗体和CTB抗体发生特异性反应.ELISA及HI试验检测,经黏膜途径免疫的小鼠产生了针对重组表位模拟抗原蛋白及H1N1、H3N2亚型猪流感病毒的血清抗体及局部黏膜分泌型IgA抗体.利用猪流感病毒表位抗原与LTb基因串连获得的融合蛋白具有良好的免疫原性和反应原性,且在黏膜接种局部产生理想的分泌型IgA抗体,能有效阻断病原由黏膜局部感染,为研制猪流感病毒通用型疫苗奠定了基础.     

Emerging therapeutic approaches for canine sarcomas: Pushing the boundaries beyond the conventional

Sarcomas represent a group of genomically chaotic, highly heterogenous tumours of mesenchymal origin with variable mutational load. Conventional therapy with surgery and radiation therapy is effective for managing small, low‐grade sarcomas and remains the standard therapeutic approach. For advanced, high‐grade, recurrent, or metastatic sarcomas, systemic chemotherapy provides minimal benefit, therefore, there is a drive to develop novel approaches. The discovery of “Coley's toxins” in the 19th century, and their use to stimulate the immune system supported the application of unconventional therapies for the treatment of sarcomas. While promising, this initial work was abandoned and treatment paradigm and disease course of sarcomas was largely unchanged for several decades. Exciting new therapies are currently changing treatment algorithms for advanced carcinomas and melanomas, and similar approaches are being applied to advance the field of sarcoma research. Recent discoveries in subtype‐specific cancer biology and the identification of distinct molecular targets have led to the development of promising targeted strategies with remarkable potential to change the landscape of sarcoma therapy in dogs. The purpose of this review article is to describe the current standard of care and limitations as well as emerging approaches for sarcoma therapy that span many of the most active paradigms in oncologic research, including immunotherapies, checkpoint inhibitors, and drugs capable of cellular metabolic reprogramming.     

产志贺毒素多重耐药大肠杆菌噬菌体的生物学特性及全基因组分析

为探明大肠杆菌噬菌体的生物学特性和全基因组特征,本研究以在广西某猪场污水中分离得到的1株产志贺毒素且多重耐药的大肠杆菌GXEC010为宿主菌,同时在污水中分离到1株具有裂解作用的噬菌体,并通过透射电镜观察、最佳感染复数测定、一步生长曲线绘制、热敏感性与酸碱度稳定性评估、杀菌试验及全基因组测序等方法分析该噬菌体的生物学特性与基因组特征。结果表明,分离纯化得到能高效裂解大肠杆菌GXEC010的噬菌体,命名为vB_EcoM_BP10,其噬菌斑呈清晰透明,边缘光滑整齐;该噬菌体的最佳感染复数为1;一步生长曲线结果显示,感染宿主菌潜伏期为5 min,爆发期为65 min,爆发量为51;可耐受温度范围为30~60 ℃,在pH为5~8的环境中能维持稳定性;噬菌体杀菌试验结果显示,感染复数（MOI）为1时噬菌体vB_EcoM_BP10具有良好的杀菌效果。全基因组测序结果表明,噬菌体vB_EcoM_BP10基因组总片段长度为52 288 bp,GC含量为44.16%,含有71个阅读框（ORF）。GO功能注释表明,噬菌体vB_EcoM_BP10可参与小分子代谢、细胞氮化合物代谢等生物学过程中,具备核苷酸转移酶活性、离子结合、DNA结合等分子功能。与Escherichia phage ST32、 Enterobacteria phage phiEcoM-GJ1的亲缘关系较近。综上所述,噬菌体vB_EcoM_BP10具有清晰的宿主受体特异性,良好的热稳定性和酸碱稳定性,本研究结果能为有效防控产志贺毒素多重耐药大肠杆菌奠定一定的理论基础。     

赭曲霉毒素的毒理学试验

用含2.569mg/kg赭曲霉毒素A (Ochratoxin A.以下称OA)的饲料,对40日龄左右的星波罗肉用鸡进行了饲喂试验试验鸡表现为精神沉郁,食欲不振、肠炎、贫血、极度消瘦等症状,于喂后13～18天,先后衰竭死亡,剖检发现肾、肝和肠道等都有明显的病理变化。经用高效液相色谱仪(HPLC)测定,肾脏,肝脏内OA的残留量分别为0.179mg/kg和0.1302mg kg。用从饲料中提纯的OA对孵化5日龄的鸡胚进行了气室内接种。根据接毒后记录的鸡胚死亡数和死亡率。计算出半数致死量为0.128μg。     

霍乱肠毒素B亚单位在转基因番茄果实中特异性表达及其免疫原性的研究

 利用番茄果实特异性启动子-E8启动子,将霍乱肠毒素B亚单位基因(ctb)用根癌农杆菌浸润法转入番茄植株,并使其在果实中特异表达,然后对试验小鼠进行口服免疫,研究转基因番茄果实的免疫原性。结果表明,ctb基因在14株番茄植物基因组中被证实有整合,并在其中2株的番茄果实中有特异性表达,最高表达量分别为455和385 ng·g-1FW,口服免疫后的小鼠血液和肠道粘膜中均检测到抗CTB抗体。表明获得了在番茄果实中特异、高效表达霍乱肠毒素B亚单位基因(ctb)的植物口服疫苗候选植株。     

苏云金芽孢杆菌Cry8Ca2蛋白的纯化与活性的研究

cry8 Ca2基因是本实验室自行分离克隆的一种新型苏云金芽孢杆菌(Bacillus thuringiensis,Bt)杀虫晶体蛋白基因,其表达130 kDa蛋白对铜绿异丽金龟(Anomala corpulenta)和黄褐异丽金龟(A.exoleta)幼虫具有较高的活性。本研究对Cry8Ca2蛋白原毒素及胰凝乳蛋白酶消化片断的纯化条件及活性进行研究,最适的消化条件为质量比1∶20,37℃,消化1 h。通过阴离子交换层析得到纯的活性毒素蛋白。用原毒素与纯化的毒素对铜绿丽金龟幼虫进行生测,Cry8Ca2原毒素蛋白的LC₅₀为0.47μg/g土,纯化的毒素的LC50为0.08μg/g土,毒素的杀虫活性可以达到原毒素的6倍。     

玉米弯孢叶斑病菌毒素对寄主防御酶系活性的影响及诱导抗性效应

 本文系统地研究了玉米弯孢叶斑病菌毒素对不同玉米抗性品种苯丙氨酸解氨酶(PAL)、过氧化物酶(PO)和超氧化物歧化酶(SOD)活性的影响及在一定条件下对寄主抗性的诱导效应。研究结果发现,抗病品种的PAL和PO酶活性比感病品种易被毒素激活,但抗病品种被毒素处理后的SOD活性明显低于感病品种,而且峰值出现得晚,这可能与抗病品种SOD活性的下降有利于积累活性氧、进一步激发寄主的抗性有关。此外,研究发现毒素在一定浓度下可作为抗性的诱导因子,为深入研究抗性机制和防病手段提供了新途径。