MPN法测定医疗保健产品生物负载的探讨

本文从医疗保健产品微生物检测的实际需要出发,详细介绍了生物负载MPN法测定的应用进展、使用前提、方法实施及验证。最后总结了MPN法的结果计算公式,并讨论了结果计算中的一些特殊情况,可为生物负载检测提供参考。     

食品中单核细胞增生李斯特氏菌两种定量检测方法的比较

研究比较不同杂菌污染条件下平板计数(Plate counting)法和稀释培养计数(Most probable number counting)法检测食品中单核细胞增生李斯特氏菌(Listeria monocytogenes,LM)含量的准确性,并探讨冷藏和冷冻食品,MPN保存过程中LM的数量增长情况。采用国标法中的平板计数法和MPN计数法对不同程度人工污染杂菌的牛奶、凉拌菜和盐水鸭分别进行LM含量的检测,并用MPN计数法对冷藏(2~8℃)和冷冻(-20℃)条件下保存的牛奶、凉拌菜和盐水鸭进行LM含量的检测。结果表明,杂菌染菌浓度较低时(LM含量与杂菌含量比为10:1), LM检出限较高(≥ 100 CFU/g (mL)),平板计数法检测LM含量的准确性较高,而杂菌染菌的初始浓度较高时(LM含量与杂菌含量比为1:10),LM检出限较低(< 100 CFU/g (mL)),MPN计数法检测LM含量的准确性较高;牛奶、凉拌菜和盐水鸭在经过不同冷藏和冷冻保存时间后LM数量对比差异有统计学意义(p<0.05),且食品中LM数量随着冷藏和冷冻保存时间的增加而增多。     

Survival of enterohaemorrhagic Escherichia coli O157:H7 strain in Turkish soudjouck during fermentation, drying and storage periods

Soudjouck (a kind of Turkish sausage) batter was inoculated with Escherichia coli O157:H7 at a level of 10⁵ colony-forming unit (CFUg) and kept overnight at 4°C. After stuffing the soudjouck batter into natural casing, fermentation was carried out at 24±2°C and 90–95% relative humidity (RH) for 3 days with subsequent drying at 22±2°C and 80–85% RH for 5 days. Then, half of soudjouck samples were vacuum-packed in polyethylene bags and the rest were kept open. All samples were stored at 4°C (55% RH) for 3 months. E. coli O157:H7 and lactic acid bacteria counts, moisture contents and pH values of the samples were determined during fermentation, drying and storage periods. Results showed that count of E. coli O157:H7 decreased by 3 log unit during fermentation and drying periods. It was observed that this pathogen survived longer in vacuum-packaged samples (more than 2 months) than non-vacuum samples (more than 1 month).     

Low contamination of Campylobacter spp. on chicken carcasses in Minas Gerais state,Brazil: Molecular characterization and antimicrobial resistance

Here, we evaluated Campylobacter contamination on chicken carcasses and phenotypic and genotypic profiles of antimicrobial resistance of the isolated strains. A total of 95 of samples were collected from 19 slaughterhouses from Minas Gerais - Brazil, and analyzed by MPN-PCR method. Campylobacter was found in 16.8% of samples with microbial load ranging from 60 to 184 MPN/carcass. All isolates were resistant to at least 5 (31.2%) of the antimicrobials screened using the disk diffusion method. Thr-86-Ile gyrA mutation, bla_OxA-61 and tet(O) genes were found in 95%, 100% and 40% resistant isolates to ciprofloxacin, ampicillin and tetracycline, respectively. Almost all isolates (90%) showed the three genes required to synthesize the CmeABC efflux system. The use of efflux pump inhibitor (PAβN) resulted in a significant reduction in the MICs of antimicrobials (2–128 fold), indicating the importance of efflux systems in conferring antimicrobial resistance. Campylobacter were detected at low concentrations in Brazilian chicken carcasses. However, high-levels of antimicrobial resistance were observed and associated with several mechanisms. This study provides a baseline survey on contamination of Campylobacter in Brazilian chicken carcasses and its antimicrobial resistance, giving support for actions directed at reducing this pathogen in the food chain.     

某市给水管网铁细菌生长特征调查

给水管网中存在的微生物腐蚀现象会造成管网水水质恶化等一系列问题，因此有必要对给水管网中铁细菌的生长特征进行调查研究。选取某市给水管网，分别采用MPN法和平板计数法对管网水及水厂处理单元出水中铁细菌的数量进行测定。试验结果表明：管网水中的铁细菌含量为0～10^7 CFU／mL，变化范围较大，各处理单元出水中均有铁细菌存在；铁细菌数量与管线长度和温度没有相关性，和余氯值呈明显的负相关；铁细菌数量和管网水中总铁以及溶解性铁含量无明显相关性，但和颗粒性铁含量有一定的相关性。     

多管法检测大肠菌群MPN值─—HBG程序的探讨

应用多管法检测大肠菌群细菌数是一种适用各种水样的行之有效的检验方法。但此方法操作繁琐，数据处理较难。本文应用概率理论，推导出多种情况下大肠菌群最可能数（ＭＰＮ值）计算公式。并编制了方便可靠的ＨＢＧ程序，从而进一步提高了此法的实用价值。     

盐碱地小麦根际联合固氮菌数量分布研究

结合气相色谱仪和乙炔还原(ARA),采用最可能计数法(MPN)和荧光抗体(FA)染色法对盐碱地小麦根际联合固氮菌及优良固氮菌株Pseudomonassp.ChW1和Zoogloeasp.ChW6数量分布进行了测定。结果表明:小麦根际各部位均存在联合固氮菌类群,但数量相对较小、差异较大(102～106个g-1干土或根),分布趋势呈RP>RS>NRS>HP;菌株Pseudomonassp.ChW1是一种相对广谱的联合固氮菌株,在小麦根际不同部位均有分布,数量除在根系表面(RP)较多(105个g-1干根)外,其它部位较少(102～103个g-1干土或根);Zoogloeasp.ChW6菌株是一种分布谱相对较窄的联合固氮菌株,只分布于根表土壤(RS)和根系表面(RP),数量102～104个g-1干土或根。自然状况下,小麦根际固氮菌数量较少,有必要通过使用联合固氮菌接种剂提高小麦根际固氮菌种群数量,增强种群竞争力,增加根际微生物固氮总量。     

Populations of antibiotic-resistant coliform bacteria change rapidly in a wastewater effluent dominated stream

Incomplete elimination of bacteria and pharmaceutical drugs during wastewater treatment results in the entry of antibiotics and antibiotic-resistant bacteria into receiving streams with effluent inputs. In Mud Creek in Fayetteville, AR, ofloxacin, trimethoprim, and sulfamethoxazole have been detected in water and sediment, and tetracycline has been detected in sediment downstream of treated effluent input. These antibiotics have been measured repeatedly, but at low concentrations (< 1 μg/L) in the stream. To determine if effluent input results in detectable and stable changes in antibiotic resistances downstream of effluent input, antibiotic resistance in Escherichia coli and total coliform bacteria in Mud Creek stream water and sediment were determined using a culture-based method. Isolated E. coli colonies were characterized for multiple antibiotic resistance (MAR) patterns on solid media and to evaluate E. coli isolate richness by amplification of a partial uidA gene followed by denaturant gradient gel electrophoresis (DGGE). Despite temporal variability, proportions of antibiotic-resistant E. coli were generally high in effluent and 640 m downstream. The MAR pattern ampicillin-trimethoprim-sulfamethoxazole was associated with a DGGE profile that was detected in effluent and downstream E. coli isolates, but not upstream. Percent resistance among coliform bacteria to trimethoprim and sulfamethoxazole was higher 640 m downstream compared to upstream sediment and water (with one exception). Resistance to ofloxacin was too low to analyze statistically and tetracycline resistance was fairly constant across sites. Resistances changed from 640 m to 2000 m downstream, although dissolved nutrient concentrations within that stream stretch resembled effluent. Antibiotic resistant bacteria are entering the stream, but resistances change within a short distance of effluent inputs, more quickly than indicated based on chemical water properties. Results illustrate the difficulty in tracking the input and fate of antibiotic resistance and in relating the presence of low antibiotic concentrations to selection or persistence of antibiotic resistances.