救必应水提物与抗菌药联用对产ESBLs细菌的抑制效果

【目的】探讨救必应水提取物与抗菌药联合诱导细菌传代的体外抑菌活性,为救必应的综合开发利用提供参考依据。【方法】采用96孔反应板二倍微量稀释法体外检测救必应水提取物与抗菌药的最小抑菌浓度（MIC）,并以1/2 MIC的救必应水提取物与抗菌药联合诱导产超广谱β-内酰胺酶（Extended spectrum beta-lactamases,ESBLs）细菌传代,然后用微量棋盘稀释法测定救必应水提取物与抗菌药联合作用后的部分抑菌浓度指数（FICI）。【结果】救必应水提取物的MIC为1.0 g/mL,以0.5g/mL的救必应水提取物与阿莫西林、左氧氟沙星、克林沙星、磺胺间甲氧嘧啶、加替沙星、头孢噻呋钠6种抗菌药联合诱导产ESBLs细菌传代后MIC显著降低,联用后的FICI分别为0.375、0.750、0.750、0.515、0.750和0.750。【结论】救必应水提取物可明显增强抗菌药对产ESBLs细菌的抗菌活性,且呈协同或相加作用。     

牧区奶干渣组合式发酵剂响应面法优化

以脱脂牦牛乳为试验材料,发酵3.5 h滴定酸度为试验指标,在不同菌种最佳组合方式筛选的研究基础上,利用响应面Box-Behnken中心组合试验研究了4株筛自牧区、发酵性能优良的菌种:保加利亚乳杆菌(MGD1-3)、嗜热链球菌(MGB39-5)、嗜热链球菌(G81-1)、植物乳杆菌(BM5152)不同接种比例对牧区奶干渣发酵效果的影响。结果表明,优化得到最优组合式发酵剂为:MGD1-3体积分数3.06%、MGB39-5体积分数0.64%、G81-1体积分数0.55%、BM5152体积分数2.04%,滴定酸度为75.87°T。为了便于实际操作,确定最终体积接种比为:MGD1-3∶MGB39-5∶G81-1∶BM5152=30∶6∶5∶20,接种量体积分数为6.0%,该条件下进行3次平行试验,得到滴定酸度平均值为74.21°T;将接种优化后组合式发酵剂制成的奶干渣(试验组)与牧区采集奶干渣(对照组)进行感官与理化指标比较,结果表明该组合式发酵剂发酵性能优良。     

烤烟K326种子可培养内生细菌的分离与鉴定

采用NA、KB、R2A和TSA 4种培养基分离烟草品种K326种子中的可培养内生细菌,并选择其中的50株外观形态特征有显著差异的菌株进行了16S r DNA测序与分析。结果表明,烟草K326种子中携带有大量可培养内生细菌,培养获得菌落总数约为8.1×105cfu/g;主要分为假单胞菌属、寡养单胞菌属、芽孢杆菌属、贪铜菌属和杆菌属等5个属,其中优势内生菌为嗜麦芽寡养单胞菌、假单胞菌和芽孢杆菌,分别占菌种总数的34%、32%和26%。     

大型海藻的细菌降解试验

研究了褐藻酸降解菌的筛选及利用其降解海带和裙带菜的方法及适宜条件。结果表明,褐藻酸降解菌更易降解嫩海藻,其降解能力与菌液或酶液的pH、盐度有很大的关系,先培养细菌离心后降解海藻、培养细菌后不离心降解海藻、培养细菌与降解海藻同时进行这三种方式对海藻都有降解能力,但效率依次降低。     

光合细菌对三角帆蚌养殖水体水质的影响

将红螺菌科的红假单胞菌应用于三角帆蚌(Hyriopsis cum ingii)养殖水体,测定水化学环境因子和微生态群变化情况。结果表明,光合细菌可稳定养殖水体pH,去除氨氮、亚硝基氮、总氮,降低COD,改变水体氮磷比;光合细菌能有效控制异养细菌、弧菌、气单胞菌数量,对真菌的增殖也有一定抑制作用,避免养殖水体水质恶化。     

Evaluation of the nutritive value of Leucaena leucocephala leaf meal, inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings

Nine isonitrogenous (35% crude protein approximately) and isocaloric (18.37 kJ g^?1) experimental diets (RLL20–BCFL40) were formulated with either raw or treated (inoculated with fish intestinal bacteria) Leucaena leucocephala leaf meal at 20%, 30% and 40% levels replacing other ingredients partially from a fish meal based reference diet (RD). Two specific strains of fish intestinal bacteria, Bacillus subtilis (isolated from Cyprinus carpio) and B. circulans (isolated from Oreochromis mossambicus) having extracellular cellulolytic and amylolytic activities, were used to inoculate Leucaena leaf meal for 15 days at 37°C. The crude fibre, cellulose and hemicellulose contents and the antinutritional factors, tannin, phytic acid and mimosine in the leaf meal decreased due to inoculation. However, free amino acids and fatty acids increased in the treated leaf meal. The response of rohu, Labeo rohita, fingerlings fed the experimental diets for 80 days was compared with fish fed a RD. Both the inclusion level and type of Leucaena leaf meal in diets significantly affected the growth performance of rohu. Fish fed diets containing inoculated Leucaena leaf meal performed better in comparison with those with the RD. On the basis of growth response, feed conversion ratio, protein efficiency ratio and apparent net protein utilization, diet formulated with 30%Leucaena leaf meal inoculated with B. circulans resulted in the best performance of rohu fingerlings followed by diet with 40%B. subtilis inoculated Leucaena leaf meal. The apparent protein digestibility (APD) was better in fish fed diets containing B. circulans inoculated leaf meal. An increasing level of raw Leucaena leaf meal was associated with a decrease in the carcass protein content of rohu fingerlings. The activity of α‐amylase increased with the increasing level of treated leaf meal in diets. Cellulase activity increased with increasing level of inclusion of raw leaf meal, and was comparatively lower in fish fed diets with treated leaf meal. Activities of protease and lipase were higher in fish fed the RD. The results showed that it is possible to incorporate Leucaena leaf meal inoculated with enzyme‐producing fish intestinal bacteria in carp diets up to 40% level of inclusion.     

Benthic microbial indicators of fish farm impact in a coastal area of the Tyrrhenian Sea

We studied the impact of organic loads due to the biodeposition of a fish farm in a non-impacted coastal area of the Tyrrhenian Sea (Western Mediterranean). Sediment chemistry and benthic microbial community were investigated from July 1997 to February 1998 on monthly basis at two stations: one was located under the fish farm, while the second was about 1 km away, and served as a reference site. The presence of the cage induced rapid changes in the benthic conditions: the sediments were rapidly (after 6 weeks) reduced. A significant accumulation of biopolymeric carbon was observed beneath the cage both 2 weeks after the initial cage deployment, on non-impacted sediments, and 5–7 months after, and appeared to be related to the fish farm production cycle. The density of microbial communities beneath the fish farm increased only during the first month of farming activity (July) and partially 7 months after cage disposal (i.e. in January), when there was a significant sediment organic enrichment. Additional evidence of rapid impact of the fish farm on the benthic bacterial communities is provided by the increase in the numbers of autofluorescent microbial cells. The photosynthetic eukaryotic cells displayed a highly reduced contribution to total autofluorescent microbial density, indicating that their decrease is related with biopolymeric carbon accumulation, and probably was due also to “shadow effect” induced by suspended material, coming from the fish farm, on the environment below. We propose here, to apply the ratio of culturable heterotrophic bacteria to microbial direct counts (CFU/MDC) to detect fish farm impact. In cage sediments, organic enrichment and the consequent modification of the characteristics of the benthic environment, determined an increase in aerobic heterotrophic bacteria and vibrio density indicating that they are efficient colonizers of organic-rich sediments. Densities of Escherichia coli and Enterococci were not significantly higher than in the reference site, and are likely of terrestrial origin. Thus they cannot be used as specific indicators of fish farm impact.     

分泌抗金黄色葡萄球菌细菌素乳酸菌的分离鉴定

为分离和筛选产抗金黄色葡萄球菌乳酸菌素的优势乳酸菌,利用乳酸菌分离培养基MRS从收集的各种腌制菜汁中分离培养乳酸菌,通过细菌培养特性、革兰氏染色特点、生理生化特性初步鉴定,同时根据Genbank中乳酸菌的16SrDNA序列设计特异性引物,采用PCR方法进一步鉴定,并以金黄色葡萄球菌为指示菌对乳酸菌的发酵上清液进行抑菌特性研究。结果表明,从腌渍菜汁中分离获得90株产酸菌,通过形态学、生理生化特性和PCR鉴定,结果73株产酸菌为乳酸杆菌;分泌产物抑菌试验表明,有10株菌具有抑制金黄色葡萄球菌活性,经酸排除和过氧化氢排除试验,仍然有5株乳酸菌的分泌产物具有抑制金黄色葡萄球菌活性。可见,从腌渍菜汁分离到的乳酸菌具有抑制金黄色葡萄球菌活性的特性,主要是通过分泌乳酸菌素来发挥作用。     

荧光定量PCR法比较研究健康和子宫内膜炎奶牛阴道菌群结构

为揭示奶牛阴道菌群结构与子宫内膜炎发生关系,本实验选择健康奶牛和患有子宫内膜炎奶牛各5头,采集其阴道粘液样品,提取样品总DNA,利用SYBR Green Ⅰ荧光定量PCR方法对这两类奶牛阴道菌群结构的差异进行了检测.检测结果显示,健康奶牛阴道内乳杆菌属(p＜0.05)、芽孢杆菌属(p＜0.05)和魏斯氏菌属(p＜0.01)的细菌数量显著或极显著高于子宫内膜炎奶牛,而子宫内膜炎奶牛阴道内大肠杆菌数量显著高于健康奶牛(p＜0.05).研究结果表明,阴道菌群结构失衡可能是奶牛子宫内膜炎发生的原因.