乳酸菌培养液中活菌数与吸光度的关系研究

通过平板计数法测定脱脂乳为培养基的发酵液中乳酸菌的活菌数,并测其对应的吸光度,研究两者之间的相关关系。结果表明:在发酵过程中以脱脂乳为培养基的发酵液中乳酸菌的活菌数与吸光度之间呈线性相关关系。因此,通过测定发酵液的吸光度可以评定发酵乳制品中乳酸菌的活菌数。     

The dynamics of bacterial community in a polyculture aquaculture system of Penaeus chinensis,Rhopilema esculenta and Sinonovacula constricta

Polyculture with Chinese shrimp Penaeus chinensis, jellyfish Rhopilema esculenta and razor clam Sinonovacula constricta (S.J.R) is a common aquaculture system in Dandong, China. Investigation on the composition and function of microbial communities and their relationship with environmental factors in aquaculture systems have crucial implications. Therefore, the sequencing of 16S rRNA gene was applied to profile the bacterial community composition of water and sediment in S.J.R polyculture system in the present study. Proteobacteria was the dominant phylum in S.J.R polyculture ponds following with Actinobacteria and Bacteroidetes in water; in contrast, more diverse phyla were detected in sediments. Water temperature, pH and dissolved oxygen were determined the bacterial community composition in the S.J.R polyculture systems. Specially, Planktomarina, PS1 clade, Candidatus Aquiluna and Fibrobacteres, which can resist the phytoplankton blooms or degrade the cellulose, were more abundant in water at the early stage of cultivation in the S.J.R polyculture system. Moreover, the functions of bacterial communities in water were mainly associated with cell communication mediated, while the functions related to basal metabolism were enriched in sediments. The findings of this study allowed us to better understand the bacterial community in the S.J.R polyculture system and accumulated data for the improvement of cultural management.     

耐胃肠道环境及肠道病原菌拮抗的猪源乳酸菌的分离与筛选

从健康猪粪便筛选分离到对肠道致病菌有较强抑制特性的3株乳酸菌,分别鉴定为屎肠球菌(Enterococcus faecium)、粪肠球菌(Enterococcus faecalis)和干酪乳杆菌(Lactobacillus casei)。体外益生特性研究表明:3株乳酸菌能耐受胃肠道环境,能黏附猪小肠表面粘液,对猪源产肠毒素型大肠杆菌(Escherichia coli O139,E. coli O139)、E. coli C83905、E. coli C83524、E. coli C83529、伤寒沙门氏菌(Salmonella typhimurium O4Hi,ST O4 Hi)、金黄色葡萄球菌(Staphylococcus aureus,SA)均有较强的抑制特性。对小白鼠肠道大肠杆菌的数量进行研究,结果证明对照组大肠杆菌数量没有明显降低,而添加乳酸菌组大肠杆菌数量明显降低。对小白鼠腹腔注射E. coliC83905,乳酸菌组死亡率明显小于对照组。结果证明3株乳酸菌在猪饲料添加剂方面有较好的应用前景。     

罗非鱼"突眼"病细菌性病原的分离与鉴定

从患"突眼"症状的罗非鱼体内分离到多株细菌,经人工感染证实其中TL60829NA菌株为病原菌,经生理、生化和API细菌鉴定系统以及分子生物学试验,鉴定其为痢疾志贺氏菌(Shigella dysenteriae)。其主要生理生化特征为革兰氏阴性短杆菌,不具动力,氧化酶、过氧化氢酶、V-P反应、吲哚试验、苯丙氨酸、甘露醇、KCN均为阴性;对氨苄西林、羧苄西林、红霉素、多西环素、氟哌酸、卡那霉素敏感,对呋喃唑酮、复方新诺明、磺胺和庆大霉素不敏感。     

Design and validation of a RT‐qPCR procedure for diagnosis and quantification of most types of infectious pancreatic necrosis virus using a single pair of degenerated primers

Infectious pancreatic necrosis virus (IPNV) is an important virus which affects the salmonid aquaculture industry worldwide; therefore, it is important to develop rapid and reliable methods of diagnosis to detect the disease at early stages. Nowadays, RT‐qPCR is replacing other methods because it provides additional information on the viral load, which is important to have a better understanding of the virus replication level and of the stage of the infection and its risk level. The main problem stems from the high diversity of this virus, which can compromise the reliability of the diagnosis. In this study, we have designed an RT‐qPCR procedure for diagnosis and quantification of IPNV based on a single pair of primers targeted to segment B. The procedure has been validated, in vitro and in vivo, testing two different types of standards against seven reference strains and 23 field isolates from different types. The procedure is reliable for the detection of any type, with a detection limit of 31 TCID₅₀ mL^?1, 50 pfu mL^?1 or 66 RNA copies mL^?1, depending on the standard. All the standard curves showed high reliability (R² > 0.95). The results support the high reliability of this new procedure for the diagnosis and quantification of IPNV.     

红壤溶磷菌的筛选及溶磷机制

采用以磷酸铝为磷源的蒙金娜(PVK)液体培养基研究了从红壤土中筛选出的4种溶磷菌的溶磷效果,选出其中的优势菌株B1,并对其溶磷机理做出初步探讨。结果表明,所筛选出的4株溶磷菌在液体培养条件下均有显著的溶磷效果,其中菌株B1在培养4 d后有效溶磷量最大,达到292.8 mg L-1。各处理培养液pH在培养期间均有显著下降,pH从7.0下降至3.2~4.7。高效液相色谱测定发现,各菌株培养液中有机酸的种类与含量随培养时间变化而不同,其中菌株B1主要分泌草酸和苹果酸,培养1 d后有机酸总量可达到5 mmol L-1;通过添加有机酸对磷酸铝活化的试验表明,分泌有机酸溶磷仅是菌株B1溶磷机制之一,可能还存在其他溶磷机制。菌株B1生长的适宜pH范围为5~9,最适培养温度为30℃,100 ml三角瓶的最适装液量为30~40 ml。经鉴定,菌株B1与苏云金芽孢杆菌有99.9%的相似性。     

Fungal endophyte Phomopsis liquidambari B3 enriches the diversity of nodular culturable endophytic bacteria associated with continuous cropping of peanut

Previously, the application of the fungal endophyte Phomopsis liquidambari B3 exhibited notable effect in alleviating soil obstacles caused by continuous cropping of peanut. It was supposed that P. liquidambari B3 could enhance the efficiency of nodulation and nitrogen fixation by enriching the diversity of nodular endophytic bacteria under continous monocropping system. To verify this hypothesis, a pot experiment was conducted under natural conditions. Four treatments were set as follow: discontinuous cropping soil (NS), continuous cropping soil (CK), continuous cropping soil inoculate with actived P. liquidambari B3 (CSB3), and continuous cropping soil inoculate with sterilized P. liquidambari B3 (CSIB3). As a result, a total of 120 isolates were obtained. Our study clearly declared that the addition of fungal endophyte effectively enriched the genetic diversity and the community composition in CSB3. In contrast, a low genetic diversity level and simplex community structure were exhibited in CK and CSIB3. Meanwhile, the positive percentage rate and corresponding community composition of the plant growth-promoting isolates in NS and CSB3 were larger and more abundant than the other. This is the first time to describe the effects of fungal endophytes on the diversity of the nodular culturable endophytic bacteria associated with continuous cropping of peanut.     

酶解牦牛血发酵液制备抗氧化肽工艺优化

为进一步提高牦牛血枯草芽胞杆菌发酵液多肽含量,对牦牛血发酵液进行酶解,以酶解液的·OH清除率、多肽含量为主要指标,筛选酶解发酵液的最适蛋白酶,通过单因素和响应面试验优化酶解工艺,并对比菌酶联合制备产物与发酵液体外抗氧化活性。结果表明,碱性蛋白酶酶解发酵液最佳条件为:酶解时间3 h、p H值9.5、酶解温度60℃和酶底比190 U·g-1,此条件下制备得到酶解液多肽含量为5.52mg·mL~(-1),较发酵液提高2.39倍。菌酶联合制备产物对·O2-及脂质过氧化的IC50分别为6.22、4.87mg·mL~(-1),发酵法制备产物对·O2-及脂质过氧化的IC50分别为8.42、11.71 mg·mL~(-1),且菌酶联合制备产物的还原力优于发酵制备产物。因此,菌酶联合制备牦牛血抗氧化肽法优于传统单一发酵法。本研究结果对提高牦牛血抗氧化肽得率,增加牦牛产品附加值具有重要的意义。     

Tomato fruit quality as influenced by the interactions between agricultural techniques and harvesting period

Tomato (Solanum lycopersicum Mill) is an important crop in terms of its economic and nutritional value. Many factors, including cultivar, climate, geography, geochemistry, and agricultural practice, can affect its nutrient concentrations. An HJ‐biplot study was performed to examine the effects of cultivar (Dorothy, Boludo, Dominique, Thomas, and Dunkan), agricultural practices, climatic factors, and their interactions. Significant differences were analyzed using a one‐way ANOVA. All samples were collected and assayed at the same degree of ripeness. In the conventional and organic tomato samples, those harvested from December to April had the highest concentrations of fructose, glucose, citric acid, malic acid, ascorbic acid, protein, Na, and Mg, while those harvested in October had the highest concentrations of lycopene and hydroxycinnamic acid. There were high concentrations of Ca, P, Zn, and Cu in the no‐soil tomatoes. Conventional and organic cultivation practices showed similar results with respect to the collection period, both presenting high organic compound concentrations, while high mineral concentrations seemed to correspond to the no‐soil practice. No clear pattern was observed among the different cultivars, perhaps due to all the samples having been collected at the same degree of ripeness.     

根表功能细菌生物膜及其在土壤有机污染控制与修复中的潜在应用价值

土壤中的有机污染物可从根系进入植物体内,并可进一步通过食物链富集,从而威胁人群健康。植物根际微生物种类繁多、数量巨大,其中很多根际细菌可通过成膜作用在植物根表形成细菌生物膜,协助植物抵抗外界的不良环境或促进植物生长。有机污染物在被植物根系吸收的过程中,多需经过根表细菌生物膜这一特殊界面。综述了根际细菌在植物根表的成膜作用,以及根表功能细菌生物膜对污染物根际环境过程的影响及作用机理,分析了利用根表功能细菌生物膜调控植物吸收有机污染物的可行性,试图为防治土壤有机污染、降低作物污染风险、保障农产品安全等提供理论依据。