水生动物肌醇营养研究进展

肌醇是大多数水生动物保持健康、促进生长发育必需的营养素,它在细胞膜磷脂合成、神经递质传递和肝脏脂质转运等方面发挥重要作用。饲料中肌醇缺乏导致水生动物生长不良、体表充血、鳍条糜烂、体色发黑、脂肪肝等症状。本文综述了肌醇的理化性质,肌醇在水生动物营养中营养生理作用、需要量及影响因素等,同时还介绍了肌醇的来源和测定方法,并展望了肌醇在水生动物营养中进一步的研究和应用方向。     

基于卷积神经网络马铃薯叶片病害识别和病斑检测*

针对自然环境下马铃薯叶片病害识别率低和晚疫病斑定位难的问题,基于大田环境中采集的马铃薯叶片图像,首先对马铃薯叶片病害进行识别,对比AlexNet、VGG16、InceptionV3、ResNet50、MobileNet五种神经网络模型,结果表明InceptionV3模型的识别效果准确率最高,可达98.00%。其次对马铃薯叶片的晚疫病斑进行检测,提出一种改进型的CenterNet-SPP模型,该模型通过特征提取网络获取对象的中心点,再通过中心点回归获得中心点偏移量、目标大小等图像信息,训练后的模型在验证集下的mAP可达90.03%,以F₁为评价值分析对比其它目标检测模型,CenterNet-SPP模型的效果最好,准确率为94.93%,召回率为90.34%,F₁值为92.58%,平均检测一张图像耗时0.10 s。为自然环境下马铃薯叶片病害识别和检测提供较为全面的深度学习算法和模型研究基础。     

间接免疫荧光染色检测鸭源副黏病毒及在鸭体内的抗原定位

以抗新城疫病毒F蛋白的单克隆抗体为一抗,建立了间接免疫荧光染色法(IFA)检测石蜡切片中鸭副黏病毒(DPMV)的方法。以建立的IFA对DPMV人工感染鸭的各组织器官进行检测,结果显示:各组织器官均能检测到DPMV,但不同时间取样,阳性信号分布的器官不同。脾脏、胸腺、法氏囊、肠道、肝脏、肺脏DPMV的阳性检出率较高,表明这些器官为DPMV的主要靶器官。在所检测的阳性组织中,病毒抗原分布在细胞浆中。IFA检测石蜡切片中的DPMV具有直观、特异性强的优点,是对DPMV进行检测和抗原定位的良好方法。     

Practical use of response surface methodology for optimization of veterinary antibiotic removal using UV/H2O2 process

Three of the most commonly used veterinary antibiotics—enrofloxacin (ENR), sulfamethoxazole (SMX), and oxytetracycline (OTC)—were chosen as representative antibiotics for UV/H₂O₂ treatments. The objective was to determine the optimization of UV/H₂O₂ to remove antibiotics from aquaculture discharge water using response surface methodology. The degradation of the antibiotics was investigated under varying UV/H₂O₂ conditions in environments with different levels of pH, water matrices, humic acid, and constituent ions. The degradation results demonstrated that increasing the H₂O₂ dosage facilitated ENR degradation at a neutral pH while facilitating degradation of SMX and OTC at a slightly acidic pH. The optimum removal conditions for ENR, which was used in all influential effect experiments and the contact tank experiments, was obtained at 10 mM H₂O₂, a pretreated COD of 87.51 mg L⁻¹, and an initial pH of 6.15. Among the tested anions, only the presence of Cl^- showed slight positive effects on ENR degradation, due to the generation of secondary active radicals. During the reaction, the hydroxyl radical (OH) was present at a higher pH while singlet oxygen (¹O₂) was slightly present at a lower pH. The experimental results from H₂O₂ sequential addition indicated that freshly added H₂O₂ could quench the recently generated OH and therefore a high H₂O₂ concentration with frequent adding was not necessary. Our contact system reduced the ENR concentration in both the effluent reservoir and in the UV irradiation zone. The overall results supported the use of the UV/H₂O₂ system to treat remnant antibiotics in the discharge water.     

Soil Phosphorus Extracted by Bray 1 and Mehlich 3 Soil Tests as Affected by the Soil/Solution Ratio in Mollisols

Different relationships between soil-test methods results have been reported in several agricultural regions. Differences in the same soil-test procedure (e.g., soil/solution ratio) exist between soil-testing laboratories from different agricultural regions. Our objectives were to (1) determine the effect of soil/solution ratio on the amount of phosphorus removed by Bray 1 and Mehlich 3 methods, (2) compare the amounts of phosphorus removed by Bray 1 and Mehlich 3 in Mollisols from the Pampean region, and (3) determine whether soil/solution ratio affects the relationship between Bray 1 and Mehlich 3. Soil phosphorus availability was determined with two extractants (Bray 1 and Mehlich 3), using two soil/solution ratios (1:10 and 1:8, wt/v) in 72 soils (noncalcareous, loess-derived Molisolls) from the Pampean region. The amount of phosphorus removed was 20–24% greater when using 1:10 than 1:8 (wt/v) soil/solution ratio. This effect was significantly greater in Bray 1 than in Mehlich 3 (p = 0.04). When compared using the same soil/solution ratio, Mehlich 3 removed 4 to 8% more phosphorus than Bray 1. The soil/solution ratio used in the comparison affected the relationship between both extractants. The difference between extractants was slightly greater with a soil/solution ratio of 1:8 than of 1:10 (p = 0.03). Our results showed that even when using the same method, changes in the procedure (like soil/solution ratio) may cause different soil-test results and also differences in the relationship between two extracting solutions. Therefore, reported relationships between two methods are only valid for the soils and region where the relationship was developed and should not be extrapolated to other regions, even with similar soils.     

猪急性腹泻综合征冠状病毒SYBR Green荧光定量PCR检测方法的建立及应用

为建立快速、灵敏且特异的检测猪急性腹泻综合征冠状病毒（swine acute diarrhea syndrome coronavirus,SADS-CoV）检测方法,本试验扩增SADS-CoV N基因保守区域将其克隆至pMD18-T载体。所构建的重组质粒pMD18-T-SADS-qN作为阳性质粒标准品,以其为模板建立一种SYBR Green荧光定量PCR检测方法。结果显示,所建立方法在3.31×10¹~3.31×10⁷拷贝·μL^-1模板量时,呈良好的线性关系,相关系数（R²）为0.997,斜率为-3.318。该方法特异性检测SADS-CoV;而猪流行性腹泻病毒（PEDV）、猪传染性胃肠炎病毒（TGEV）、猪德尔塔冠状病毒（PDCoV）和猪繁殖与呼吸综合征病毒（PRRSV）检测结果均为阴性。所构建的标准品检测灵敏度下限可以达到3.31×10¹拷贝·μL^-1,组内和组间变异系数均小于1%,表明其具有良好的灵敏性和重复性。用该方法检测SADS-CoV感染IPI-2I和IPEC-J2细胞后不同时间点和不同接毒剂量的复制情况,结果显示,SADS-CoV感染细胞后2 h病毒含量较低,在12~36 h病毒含量迅速增长,36 h后增长速度减缓且病毒含量维持在较高水平。分别用0、0.1、1 MOI SADS-CoV感染细胞结果显示病毒的mRAN转录水平呈现剂量依赖性增加,当MOI为1时,IPI-2I和IPEC-J2细胞病毒含量分别为10^6.7、10^5.3拷贝·mL^-1。进一步利用所建立的方法对经口服攻毒SADS-CoV仔猪的临床样本进行检测,结果发现病毒在空肠回肠含量较高,表明病毒主要定殖于空肠和回肠。综上表明,本研究建立SYBR Green荧光定量PCR检测方法能灵敏特异地检测SADS-CoV,为SADS-CoV的诊断和病毒相关基础研究提供可靠的检测手段。     

中华蜜蜂囊状幼虫病毒检测方法研究进展

中蜂囊状幼虫病是一种由中蜂囊状幼虫病毒引起的严重威胁中蜂健康的传染性疾病。目前由于该病尚无有效的药物治疗方式,因此,预防与及时检测对于遏制该病传播显得尤为重要。本文主要从临床诊断、分子生物学和免疫学方面总结中蜂囊状幼虫病检测与鉴定的方法,包括形态诊断、RT-PCR技术、实时荧光定量PCR、巢式PCR、胶体金免疫快速诊断技术、ELISA、血清学技术和基因芯片等,为中蜂囊状幼虫病毒检测方法的改进和优化提供参考。     

山东省菏泽市某规模猪场不同猪群伪狂犬病野毒感染情况调查

为了解规模猪场不同猪群的伪狂犬病病毒(PRV)感染情况及场区病毒载量分布特点,在山东省菏泽市某规模猪场,利用实时荧光定量PCR方法,对该猪场不同孕龄母猪、不同阶段生长猪群,以及各生产阶段场区环境,采集猪鼻拭子和场区环境拭子进行PRV-gE核酸检测,并对检测结果进行统计分析。结果显示:在294份猪鼻拭子中,检出42份PRV-gE核酸阳性,总阳性率为14.28%;母猪群PRV-gE阳性率为17.83%(23/129),且妊娠前后各阶段阳性率呈先上升后下降趋势,中期较高(24.00%),但差异不明显(P>0.05);不同阶段生长猪群的平均PRV-gE阳性率为11.52%(19/165),随日龄增加,阳性率也呈先上升后下降趋势,80~90日龄育肥猪最高(28.13%),与其他生长阶段猪群差异明显(P<0.05);除饲料、水源、上猪台和出猪台外,其他大部分场区环境均检测到PRV-gE核酸阳性;育肥猪群病毒载量最高,为3.6×105拷贝数/mL,其猪舍环境的病毒载量也较高,与其他环境及生长阶段猪群差异均明显(P<0.05)。结果表明,不同猪群包括免疫猪群均可遭受PRV野毒感染,尤其是母猪妊娠中期和猪育肥阶段早期,且大部分场区环境均可被污染。结果提示,规模猪场要加强各种猪群尤其是育肥猪群的伪狂犬病免疫,通过监测来调整和优化免疫程序,同时要加强生物安全,防止病毒传入和扩散。     

响应面法优化沙棘再制奶酪加工工艺

本试验以新鲜沙棘果及奶酪为主要原料,探讨了沙棘再制奶酪的加工工艺。首先经护色、打浆、精磨、过滤、调酸、均质等工序,制备澄清均一的沙棘果汁。以样品感官品质和Vc含量为评价指标,通过单因素试验以及响应面试验,得出最佳配方及工艺为：35.00%天然奶酪、18.56%沙棘果汁（料水质量比1∶1）、1.54%复合乳化盐（m_柠檬酸钠∶m_{多聚磷酸钠}∶m_焦磷酸钠∶m_{六偏磷酸钠}=4∶2∶2∶1）、12.00%黄油、6.00%脱脂乳粉、6.00%白砂糖、0.50%复合稳定剂（m_卡拉胶∶m_黄原胶=4∶1）,其余20.40%为纯净水,乳化温度为84 ℃,乳化时间为11.4 min,搅拌速度为3 000 r/min。经此工艺制成的成品,感官评分为57.5分（满分60分）,Vc含量达16.61 mg/100 g。成品组织结构细腻光滑、色泽橘黄、口感润滑、营养丰富,具有沙棘果的风味,符合《GB25192—2010 再制奶酪》要求。     

发酵乳中沙门氏菌依赖解旋酶恒温基因扩增快速检测方法的建立

建立依赖解旋酶恒温基因扩增（helicase-dependent isothermal DNA amplification,HDA）快速检测发酵乳中沙门氏菌方法。以沙门氏菌invA基因序列为目的基因,设计特异性引物,优化反应体系中UvrD解旋酶及T4 gp32添加量,建立最优反应体系。通过HDA方法直接检测发酵乳中沙门氏菌,扩增其产物后进行电泳检测,验证方法特异性。结果表明：采用HDA快速检测法检测发酵乳中沙门氏菌特异性良好,优化后反应体系体积50 μL时,UvrD解旋酶添加量为0.10 μg,T4 gp32添加量为5.0 μg,得到与设计序列长度（304 bp）一致的扩增产物,检出限为2.6×102 CFU/g;该方法用于快速检测发酵乳中沙门氏菌能够满足检测需求,具有较高的灵敏度、易操作,可作为一种基础且快速的方法检测发酵乳中沙门氏菌。