西藏林芝地区不同品种葡萄果实表皮微生物群落结构组成分析

基于高通量测序技术对西藏林芝地区不同品种葡萄果实表皮细菌和真菌菌群的多样性及群落组成进行分析。结果表明,不同品种葡萄果实表皮微生物群落多样性差异明显,‘夏黑’葡萄果实表皮微生物丰富度和多样性指数最高,‘火焰无核’葡萄果实表皮最低。优势细菌属（相对丰度>1%）主要有伯克霍尔德氏菌属（Burkholderia-Caballeronia-Paraburkholderia）、慢生根瘤菌属（Bradyrhizobium）、SJA-15、劳尔氏菌属（Ralstonia）、异样根瘤菌属（Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium）、寡养单胞菌属（Stenotrophomonas）、甲基杆菌属（Methylobacterium）和阿菲波菌属（Afipia）,优势真菌属主要为球腔菌属（Mycosphaerella）、曲霉属（Aspergillus）、枝孢属（Cladosporium）和枝顶孢属（Acremonium）。结果显示,不同品种葡萄果实表皮微生物群落结构组成及相对丰度差异明显,说明葡萄品种影响葡萄果实表皮微生物群落结构组成。     

传统腊肉的理化、微生物与风味特性

利用长春当地传统方法制作腊肉,并进行基本的理化指标测定和微生物指标分析。结果表明：腊肉pH 值为5.09±0.33,质构特性在肥瘦肉两侧存在一定差异;风味物质种类在腌制风干后明显增多;腊肉菌落总数为 2.86×107 CFU/g,通过高通量测序方法得出,随风干时间（0～4 周）的延长,腊肉中所含菌属种类有所变化,且 各菌属的含量也有所增减;Shannon指数和Chao 1指数测定结果表明,不同腌制风干时间腊肉的菌群物种丰富度和 群落多样性无显著差异。     

绿豆芽生产过程中微生物的生长、分布及消毒剂处理效果评价

研究绿豆芽生产过程中微生物的生长情况及种类分布,采用2种不同类型的消毒剂进行处理及效果评价,并用16SrRNA方法对豆芽中主要微生物进行鉴定。结果表明,成品绿豆芽中的微生物总量为5.8×10~7 CFU/g,微生物的种类主要为克雷伯氏菌(Klebsiella)、不动杆菌(Acinetobacter)、肠杆菌(Enterobacter)和假单胞菌属(Pseudomonas)等。有效氯浓度1 000~3 000 mg/L的NaClO处理芽菜30 min,菌落总数分别降至1.2×10~7~1.0×10~5 CFU/g,而直接处理单独培养的模型对照组则平均降至2.3×10~4~1.0×10~3 CFU/mL。25 mg/L AgNO_3可使菌落总数降至3.8×10~5 CFU/g,消毒效果同样差于模型对照组。扫描电子显微镜观察分析芽菜表面微生物可形成明显可见的生物膜,显著强化了细菌对消毒剂的耐受力。     

基于高通量测序解析不同品种蓝莓表面菌群结构的多样性

为了解蓝莓果实表面真菌菌群多样性,明确不同品种蓝莓表面的优势菌群,采用高通量测序技术分析北陆、蓝金、莱克西、奥尼尔蓝莓中ITS1区基因序列,比较不同品种蓝莓表面微生物群落结构组成及多样性差异。结果发现:蓝莓主要的优势属为枝孢属、链格孢属;优势种为枝孢菌、链格孢、枝孢霉。由Simpson指数可知微生物群落丰度及群落多样性由高至低为莱克西、北陆、奥尼尔、蓝金。不同品种蓝莓表面的菌群结构与品种存在一定的聚类关系:北陆和莱克西群落结构相似度较高,而其它样品间微生物群落差异较大,各品种蓝莓表面的主要菌落相对丰度差异较大,枝孢菌、链格孢、枝孢霉为主要差异菌种。研究初步明确了蓝莓表面菌群结构的多样性,为保证蓝莓贮藏期间的品质安全提供一定的理论支持。     

蔬菜盐渍过程中微生物分布的研究

本实验对总好氧菌、乳酸菌、总芽孢菌、肠道菌、真菌等在雪里蕻盐渍过程中的分布状态,以及清洗、盐浓度、盐渍温度对微生物的影响进行了研究。研究发现细菌总数,肠道菌、真菌总数、乳酸菌、总芽孢菌在盐渍开始时分别为>106CFU/g、>105CFU/g、>104CFU/g、>103CFU/g、<102CFU/g,并且当初始盐浓度为8%,盐渍温度为15℃,盐渍时间为12h时,细菌总数、肠道菌群、真菌等分别减少了0.5、0.4、0.9logCFU/g,乳酸菌与总芽孢菌总数在12h的盐渍时间内变化不显著。     

气体二氧化氯对葡萄杀菌效果及品质的影响

以‘巨峰’葡萄为试材,从其表面分离优势菌并鉴定其为大肠杆菌与考克氏菌,利用NaClO₂与干冰间接反应开发一种气体二氧化氯(ClO₂)处理方法以灭活葡萄表面优势菌,通过测定菌落总数对数值减少量评估气体ClO₂对葡萄的杀菌效果,同时通过测定4℃贮藏30 d期间葡萄样品的色泽参数、果实硬度、质量损失率以及可溶性固形物含量评估气体ClO₂对葡萄品质的影响。结果表明,气体ClO₂处理对接种于葡萄表面的大肠杆菌与考克氏菌具有良好杀灭效果。当ClO₂累积浓度从131μL/(L·h)提升至599μL/(L·h),大肠杆菌菌落总数对数值减少量从1.6(lg(CFU/g))升至3.5(lg(CFU/g)),考克氏菌菌落总数对数值减少量从1.0(lg(CFU/g))升至4.5(lg(CFU/g))。与对照组(S0)相比,经ClO₂处理后贮藏30 d期间葡萄色泽并没有明显变化,与S0和低累积浓度处理组(S1)相比,高累积浓度处理组(S2)在4℃贮藏30 d后...     

一株拮抗酿酒葡萄灰霉病的枯草芽孢杆菌T3筛选、鉴定及抑菌分析

为筛选出对酿酒葡萄灰霉病具有良好抑制作用的拮抗菌,从酿酒葡萄根际土壤进行微生物分离,以灰葡萄孢(Botrytis cinerea)为靶标菌株,采用牛津杯法对拮抗菌进行筛选,通过形态学、生理生化特征以及16S rDNA进行鉴定分析,对拮抗菌进行生长特性及抑菌分析。结果表明:筛选出一株对灰葡萄孢(Botrytis cinerea)具有显著抑制作用的T3菌株,经鉴定为枯草芽孢杆菌(Bacillus subtilis),培养24~48 h T3菌株进入稳定期,最适的生长温度为37 ℃,最适pH为7.0。T3菌株对灰葡萄孢(Botrytis cinerea)MIC为3.6×105 CFU/mL,MBC为3.6×106 CFU/mL,3.6×109 CFU/mL T3菌株对灰葡萄孢(Botrytis cinerea)抑菌直径为31.6±0.2 mm(P<0.05),抑菌效价为158.0±1.0 mm/mL(P<0.05)。扫描电镜观察T3菌株造成灰葡萄孢(Botrytis cinerea)菌丝畸形、扭曲、顶端膨大、原生质收缩、孢子稀松,抑制菌丝生长及孢子萌发。本实验为酿酒葡萄灰霉病的生物防治提供新的微生物资源,具有进一步研究价值。     

冰鲜鸡预冷过程中微生物菌群多样性动态分析

为了对肉鸡屠宰预冷过程中预冷水、鸡胴体的微生物群落结构进行动态分析,研究二者优势菌群的消长规律。本文对第一批屠宰鸡通过预冷水0、2、4、6、8 h时预冷池中一阶、二阶预冷水以及鸡胴体进行菌落总数(TVC)检测,然后采用高通量测序的方法对二者的群落结构进行动态分析。菌落总数测定结果表明:在预冷过程中,一阶预冷水菌落总数由103 CFU/mL增长到105 CFU/mL,二阶预冷水菌落总数由102 CFU/mL增长到104 CFU/mL,在预冷前,鸡胴体的菌落总数为4.53 lg CFU/g,经预冷后在6~8 h内,鸡胴体表面菌落总数高于预冷前,说明预冷水已经失去了清洗减菌的作用,还可能会对鸡胴体造成交叉污染。高通量测序发现:在预冷过程中,一阶预冷水中气单胞菌属减少,假单胞菌属、链球菌属增加;二阶预冷水的不动杆菌属减少,假单胞菌属增加;与预冷前相比,预冷过程中鸡胴体的魏斯氏菌和漫球菌属减少,金黄杆菌属和假单胞菌属增加。本研究表明预冷后期预冷水失去减菌作用,对鸡胴体造成污染,预冷工艺主要对气单胞菌属、魏斯氏菌和漫球菌属有较好的减菌效果,对金黄杆菌和假单胞菌属减菌效果不佳。这为宰后胴体预冷工艺的优化提供参考依据,同时为冰鲜鸡产品的质量安全提供保障。     

影响新疆酿酒葡萄浆果体表菌群结构研究进展

酿酒葡萄体表微生物菌群对葡萄酒的品质起着重要作用,不同气候格局、栽培模式不仅影响着葡萄的品质,也影响着葡萄体表微生物菌群的多样性。该文综述了不同栽培气候、葡萄品种、杀菌剂对葡萄体表微生物菌群的影响,描述了新疆地区葡萄栽培的气候产区区划及栽培模式,阐明了对提高葡萄酒品质的意义,揭示新疆酿酒葡萄不同产区环境因子（无霜期、干燥度）和生物因子（葡萄品种、杀菌剂）与微生物菌群分布的相关性具有重要的意义。     

基于传统分离培养和宏基因组测序分析不同产地小龙虾菌群的多样性

采用传统培养和宏基因组测序法研究浦口、太湖东山两产地小龙虾整体、表面、尾肉3个部位的初始菌群,解析其携带污染菌的差异。结果表明:浦口、太湖东山小龙虾的整体初始菌落总数差异不显著,分别为7.78 lg(CFU/g)和7.76 lg(CFU/g),而在选择性培养基上菌落数有明显差异,经16S rRNA序列鉴定,浦口小龙虾整体样品的优势菌为柠檬酸杆菌和气单胞菌,蜂房哈夫尼菌是太湖东山小龙虾整体样品的优势菌。在科水平上,宏基因组测序发现肠杆菌科肠杆菌是两产地小龙虾整体样品的优势菌;在属水平上,浦口小龙虾整体样品中的主要菌属为微小杆菌属和气单胞菌属,而太湖东山小龙虾整体样品的主要菌属为黄杆菌属、气单胞菌属和希瓦氏菌属。两产地小龙虾不同部位初始菌群的分析结果反映小龙虾潜在食品安全风险,并且不同产地的小龙虾污染菌群不同,为今后小龙虾保鲜、运输研究提供参考依据。     

Grape berry bacterial microbiota: Impact of the ripening process and the farming system

Wine grapes are a primary source of microbial communities that play a prominent role in the quality of grapes prior to harvesting, as well as in the winemaking process. This study investigated the dynamics and diversity of the epiphytic bacteria on the grape berry surface during maturation. The quantitative and qualitative effects of conventional and organic farming systems on this microbial community were investigated, using both cultivation-dependent and independent approaches. Analyses of grape berry bacterial microbiota revealed changes in the size and structure of the population during the berry ripening process, with levels rising gradually and reaching their highest value when the berries were over ripe. As the season progressed to maturity, Gram-negative bacterial communities (mostly Pseudomonas spp.) declined whereas Gram-positive communities (mostly Micrococcus spp.) increased. The 16S rRNA gene sequences of cultured isolates were analysed and over 44 species were identified from 21 genera in the Proteobacteria, Actinobacteria, and Firmicutes phyla. Copper concentrations originating from phytosanitary treatments varied according to the vineyard and farming system. A negative correlation between copper concentrations and cell densities provided clear evidence that copper inhibited bacterial communities. The bacterial community structure was analysed by targeting the 16S rRNA genes, using PCR-DGGE on cultivable populations and T-RFLP on whole communities in cell suspension. The results suggest that the farming system has a clear impact on the bacterial community structure.     

Influence of the farming system and vine variety on yeast communities associated with grape berries

Wine production in most countries is based on the use of commercial strains leading to the colonisation of the wineries by these strains with the consequent reduction of autochthonous biodiversity. This implies that wine styles could therefore become standardised. The vineyard could be an important source of native yeasts of oenological interest. For this reason the objective of this study was to compare two agronomic conditions with the aim of preserving yeast biodiversity in the vineyard. A three year sampling plan was designed to evaluate the influence of different agronomic parameters on the biodiversity of fermentative grape yeasts. Thus two vineyards, one organic and one conventional, with three different grape varieties (Shiraz, Grenache and Barbera) were chosen. In total, 27 samples were collected from both vineyards. Of these, 1080 colonies were isolated and a total of 9 species were identified. The strains identified as Saccharomyces cerevisiae were genotyped by microsatellite analysis obtaining nine different electrophoretic patterns. Classical ecology indexes were used to obtain the richness (S), the biodiversity (H') and the dominance (D) of the species studied. The results indicated a clear influence on grape associated yeast diversity of the phytosanitary treatment used in the vineyard. This is the first time that classical ecology indexes have been used to study the ecology of the spontaneous fermentation of grape musts and the species Candida sorbosa and Pichia toletana have been described in vineyards of the Madrid winegrowing region.     

Understanding the microbial ecosystem on the grape berry surface through numeration and identification of yeast and bacteria

Microbial species present on the surface of grape berries at harvest play an important role in winemaking, thus counting and identifying them is of great importance. The use of conventional microbial techniques and molecular methods allowed a quantitative and qualitative inventory of the different microbial species present on the grape berries. These experiments were carried out in several areas of the Bordeaux region on the red grape varieties Merlot, Cabernet Sauvignon and Cabernet Franc. Populations and species clearly varied according to berry development stage. The most widespread yeast species at berry set, Aureobasidium pullulans was never detected at harvest. Fermentative yeasts were detected at harvest and not in the first stage of grape growth. Oenoccocus oeni was detected on immature as well as on mature berries. Gluconobacter oxydans was detected mainly at harvest. Detection of Pediococcus parvulus , was dependent on the vineyard. Veraison appeared to be a key stage for yeast colonisation and the increase in population involved a change in the proportion of each species. The number of A. pullulans fell significantly at veraison as it was superseded by fermentative yeasts. Microbial populations peaked at harvest when the berry surface available for adhesion was largest and no agrochemical treatments had been applied for some weeks. Soil, grape variety and grapegrowing practices may also influence this microbial ecosystem. Based on these and published data, we formulated hypotheses to describe this microbial ecosystem, thus enabling us to develop the concept of a microbial biofilm.     

Isolation of epiphytic yeasts with potential for biocontrol of Aspergillus carbonarius and A. niger on grape

Antagonistic yeasts were isolated from the epiphytic flora associated with grape berries cv. Negroamaro and identified at species level using molecular methods. A total of 144 yeast isolates were tested in a preliminary screening on agar to select isolates showing a killer activity against Aspergillus carbonarius and A. niger, the main species responsible for the accumulation of ochratoxin A in grape. Twenty-eight yeast isolates were selected for their inhibitory effects on the above fungal species and assayed by an in vitro nutritional competition test for their antagonistic capacity towards three selected ochratoxigenic strains. Six yeast isolates belonging to five species, namely 2 isolates of Issatchenkia orientalis and one each of Metschnikowia pulcherrima, Kluyveromyces thermotolerans, Issatchenkia terricola and Candida incommunis, were finally selected and screened on wounded grape berries for their ability to inhibit infection by ochratoxigenic moulds. With the exception of the K. thermotolerans isolate, when inoculated at 10(9) CFU/wound, the other five challenger yeasts reduced the A. carbonarius and A. niger colonization on grape berry (P<0.05). In particular, the best antagonistic activity was shown by the two I. orientalis isolates. Results suggest that antagonist yeasts with the potential to control A. carbonarius and A. niger on grape can be found among the microflora associated with the berries.     

Effects of geographical origin,varietal and farming system on the chemical composition and functional properties of purple grape juices: A review

BackgroundGrape juice is a beverage derived from Vitis sp genus, mainly V. labrusca, V. vinifera and V. rotundifolia species, in which sales have increased steeply because of the alleged beneficial health effects it exerts when consumed regularly. However, the isolated and interlinked impacts of geographical origin, varietal, and farming system on the juice's chemical composition and functional properties still are not fully comprehended.Scope and approachThis paper aims to assess how the producing region, variety, and farming system of grapes (conventional, organic, and biodynamic) affect the quantitative and qualitatively the chemical composition and the functionality (in vitro, in vivo) of juices.Key findings and conclusionsData have shown that the effects of botanical and geographical origins of purple grapes on the chemical composition (especially phenolic compounds) and functional properties of juices are remarkable. On the other hand, organic and biodynamic grape juices have very similar composition and functional properties in vitro, while organic and conventional are somewhat different. This evidence is in line with in vivo animal studies and human trials on healthy individuals have shown: differences in functional properties, especially antioxidant effects, between organic and conventional grape juices are negligible from the nutritional and biochemical perspectives.     

Copper content of grape and wine from Italian farms

The copper content of grape and wine from 16 wine-farms in Italy was studied during the harvest of 2003. The influence of the number of copper applications, the period between the last application and harvest, and the total amount of copper applied was examined. Of the total number of samples analysed, 13% of grape samples and 18% of wine samples exceeded the maximum residue level (MRL). The total amount of copper applied and the number of days between the last application and harvest explained 44% of the concentration of copper in grape. This low correlation may be due to other influencing factors, such as meteorology and the application method. In 2003, conditions were unusually dry and the recommended safety interval for copper application (20 days) was not sufficient to guarantee a residue level in grape below the MRL (20 mg kg^-1). In order to reduce the probability of copper residues being close to the MRL, a period of 40-50 days between the last application and harvest is suggested. Furthermore, the copper content of grape and wine was not dependent on the pest management strategy of the farm (conventional, integrated or organic). A more important factor influencing copper residue levels may be that copper applications are made in response to the prediction of a disease outbreak rather than being dependent on the pest management strategy in place. No difference in copper content was observed between red and white grape or wine.     

葡萄籽提取物主要质量指标研究

目的 为了确定保健食品原料葡萄籽提取物主要质量指标的参数,本实验搜集30批次葡萄籽提取物并测定了其主要质量指标。方法 采用紫外分光光度法测定了葡萄籽提取物的原花青素值、多酚含量,采用液相色谱法测定了儿茶素、表儿茶素含量,参照食品安全国家标准,测定了灰分、水分、微生物指标(菌落总数、大肠杆菌、沙门氏菌、酵母菌、霉菌)、重金属和溶剂残留等主要质量指标。结果 所有葡萄籽提取物样品中原花青素值范围在81.1%至108.2%之间;儿茶素及表儿茶素含量范围为1.12%至8.58%;多酚含量在70.1%至106.1%之间;灰分含量范围在0.2%至1.8%;水分含量范围在3.76%至5.23%。变异系数由大到小的顺序为：儿茶素、表儿茶素＞灰分＞多酚＞水分＞原花青素值。所有样品菌落总数均小于10 CFU/g,大肠杆菌、沙门氏菌均未检出,酵母菌、霉菌总数均小于10 CFU/g,甲醇、乙醇残留均小于2 mg/kg,重金属(以pb计)小于0.05 mg/kg。结论 与我国目前唯一相关标准对比,所有质量指标中,仅原花青素值的限量需要进一步调整。     

Simulating consequences of choosing a breeding goal for organic dairy production

In Denmark, Finland, and Sweden, the Nordic Total Merit index is used as the breeding selection tool for both organic and conventional dairy farmers based on common economic models for conventional dairy farming. Organic farming is based on the principles of organic agriculture (POA) defined by the International Federation of Organic Agriculture Movements. These principles are not set up with an economic point of view, and therefore it may be questionable to use a breeding goal (BG) for organic dairy production based on economic models. In addition to economics and the principles of organic agriculture, it is important to look at farmers' preferences for improving BG traits when setting up a BG for organic farming. The aim of this research was to set up, simulate, and compare long-term effects of different BG for organic and conventional dairy production systems based on economic models, farmers' preferences, and POA, with particular emphasis on disease resistance or on roughage consumption and feed efficiency. The BG based on economic models and on farmers' preferences were taken from previous studies. The other BG were desired gains indices, set up by means of a questionnaire about relatedness between the POA and BG traits. Each BG was simulated in the stochastic simulation program ADAM. The BG based on POA, with particular emphasis on disease resistance or on roughage consumption and feed efficiency, caused favorable genetic gain in all 12 traits included in this study compared with 6 traits for the other BG. The BG based on POA, with particular emphasis on disease resistance or on roughage consumption and feed efficiency, were very different from BG for organic and conventional production based on economic models and farmers' preferences in both simulated genetic change and correlations between BG. The BG that was created based on the principles of organic agriculture could be used as a specific index for organic dairy farming in Denmark, but this index was economically not very sustainable. Hence, an intermediate breeding goal could be developed by breeding companies to address both economics and the principles of organic agriculture.     

Co-existence of GM,conventional and organic crops in developing countries: Main debates and concerns

ABSTRACT

The co-existence approach of GM crops with conventional agriculture and organic farming as a feasible agricultural farming system has recently been placed in the center of hot debates at the EU-level and become a source of anxiety in developing countries. The main promises of this approach is to ensure “food security” and “food safety” on the one hand, and to avoid the adventitious presence of GM crops in conventional and organic farming on the other, as well as to present concerns in many debates on implementing the approach in developing countries. Here, we discuss the main debates on (“what,” “why,” “who,” “where,” “which,” and “how”) applying this approach in developing countries and review the main considerations and tradeoffs in this regard. The paper concludes that a peaceful co-existence between GM, conventional, and organic farming is not easy but is still possible. The goal should be to implement rules that are well-established proportionately, efficiently and cost-effectively, using crop-case, farming system-based and should be biodiversity-focused ending up with “codes of good agricultural practice” for co-existence.