应用不同方法检测食品中大肠菌群结果的比较

评价检测食品中大肠菌群不同方法。比较国家标准、行业标准和显色培养基检测方法检测大肠菌群结果的差别。国家标准和行业标准检测结果基本一致,但有差异,应用显色培养基检测大肠菌群优于目前使用的国家标准和出口食品检验行业标准方法。检测食品中大肠菌群,显色培养基检测方法快速、灵敏、特异。     

水中粪大肠菌群测定方法-纸片快速法与多管发酵法的比较

2015年10月国家环境保护部发布了行业标准《水质总大肠菌群和粪大肠菌群的测定纸片快速法》(HJ755-2015)(以下简称纸片快速法)。之前环境监测部门一直在使用国家环境保护部于2007年3月发布的行业标准《水质粪大肠菌群的测定多管发酵法和滤膜法(试行)》(HJ/T347-2007)(以下简称多管发酵法)。纸片快速法操作简单快速,在标准中对采样瓶的处理、样品的保存、水样的稀释接种等做出了详细的规定,而多管发酵法中没有相应规定。本文对两个标准的异同点进行比较,提出了将这两种方法的优点应用于实际环境监测工作中。     

浅析影响食品中大肠菌群测定结果的几点因素

大肠菌群是评价食品卫生情况及受污染程度的最常用指标之一。大肠菌群的检测已是食品安全检测中必不可少的一个项目。本文主要对影响食品中大肠菌群测定结果的几点因素进行分析,以保证食品中大肠菌群检测结果的准确性和可靠性。     

宠物饲料中细菌污染状况调查

参照国家卫生标准规定方法检测了我国某地区宠物饲料的大肠菌群、沙门氏菌。结果是宠物饲料半成品的大肠菌群不合格率为49.06%,沙门氏菌阳性率为7.55%,成品中大肠菌群不合格率为29.27%,水 氏菌阳性率为0.81％。其中碎皮成形类饲料大肠菌群不合格率为54.76%,并检得沙门氏菌1株;皮压骨类饲料大肠菌群不合格率为26.67%;皮节骨类饲料大肠菌群不合格率为9.80%。     

用污染指示菌评价城市水体污染的研究

选择湖南省常德市江北城区水体为代表开展本项研究。结果表明,水体中污染指示菌,总大肠菌群（ＴＣ）和粪大肠菌群（ＦＣ）密度同ＢＯＤ＿５负荷以及综合污染指数Ｐ呈高度正相关。所以,污染指示菌指标,特别是粪大肠菌群指标足综合评价城市污水,尤其是生活污水污染的一个必不可少的重要参数。对该城区水体细菌污染的评价结果是,生活污水排放愈集中的地区,细菌污染愈严重。其中护城河水的ＴＣ密度超出我国地面水Ⅲ级标准４个数量级;ＦＣ密度超过ＷＨＯ娱乐用水标准３个数量级。护城河是细菌污染最严重水体。包括滨湖公园内湖在内的接纳生活污水多或较多的水体,细菌污好也相当严重。只是远离城区的柳叮湖受污染程度较轻。     

二级水灌溉对土壤及马铃薯块茎中大肠菌群数量的影响

采用多管发酵法检测二级水灌溉对马铃薯种植土壤中大肠菌群数量,结果表明:充分灌溉后的土壤中大肠菌群的数量均高于分根交替灌的土壤;地下滴灌的土壤中大肠菌群数量高于沟灌土壤的大肠菌群数量;二级水经加氯消毒后,土壤中大肠菌群数量明显下降。同样采用多管发酵法检测马铃薯块茎,结果发现:采用加氯消毒后的二级水灌溉的小区E和F中生长的马铃薯,其块茎组织中大肠菌群数量低于其他小区,采用地下滴灌灌溉的小区I和J所种植出的马铃薯块茎中大肠菌群的数量高于采用沟灌灌溉的小区K和L。结果表明:马铃薯块茎中大肠菌群的数量与其生境中大肠菌群的数量有密切关系。     

应用定量微生物风险评估海水浴场人体健康风险

【背景】定量微生物风险评估作为定量评估游泳人群暴露于病原微生物后健康风险的方法,在国外已得到广泛应用,但目前国内的应用处于起步阶段且缺乏所需的游泳人群暴露数据。【目的】收集游泳人群暴露数据,并在海水浴场中进行应用,评估粪大肠菌群作为风险评估指标的可行性。【方法】通过对6个典型海水浴场的水质状况、粪大肠菌群浓度与环境因子的相关性进行分析,并发放调查问卷收集国内游泳人群的暴露数据,进而应用定量微生物风险评估方法,得出各个海水浴场的胃肠道疾病患病风险。【结果】6个海水浴场中粪大肠菌群浓度均与水温、气温及总云量具有显著相关性(P<0.01)。位于南方的海水浴场粪便污染情况较北方严重,粪大肠菌群浓度第95百分位数远高于国内“差”类水质标准的阈值。儿童、成年男性、成年女性单次沐浴事件吞下海水的体积分别为35.1 mL (95%置信区间为32.4-37.8,α=0.578,β=0.016),45.0 mL (95%置信区间为31.1-59.3,α=0.532,β=0.012),35.7 mL (95%置信区间为29.7-41.8,α=0.753,β=0.032)。6个海水浴场患胃肠道疾病的风险...     

环凯食(饮)具大肠菌群检验纸片检测能力测试研究

研究以营养琼脂培养基的计数结果为基准,采用幅度卫生部食检所监制的两种不同厂家生产的食（饮）具大肠菌群检验纸片作为对照,对环凯食（饮）具大肠菌群检验纸片进行了检测能力测试研究,在测试中采用10cells/mL,50cells/mL和100cells/mL3个不同菌液浓度,环凯纸片的检测结果和营养琼脂平板计数结果基本一致。与其它两种纸片的检测结果无明显差异。完全可以用于食（饮）具大肠菌群的监督检验。     

微生物肥料中大肠菌群的测定及鉴别

为了正确区分微生物肥料产品中的大肠菌群,对近期检测的样品在伊红美兰(EMB)平板上得到的52株细菌进行了验证,并根据菌落的颜色和形态挑选出18个菌株,用Biolog微生物自动鉴定系统进行鉴定,以大肠杆菌科的4属7株标准菌株进行对照,鉴定结果和标准菌株一致。同时对微生物肥料中常用的13株革兰氏阳性(或可变)菌株和8株革兰氏阴性菌株也作了大肠菌群检测试验。其中,革兰氏阳性(或可变)菌株在乳糖胆盐发酵管和EMB平板上结果均为阴性,革兰氏阴性菌8株中有3株能利用乳糖,但都不产气,可以通过验证试验加以区分。明确了微生物肥料产品中常用菌种利用乳糖的情况以及在EMB平板上经常出现的不同菌落的分类地位,对大肠菌群的判定具有指导意义。     

PCR技术检测水体中大肠菌群的研究

大肠菌群被广泛用作饮用水的卫生学检测指标。传统的检测方法耗时长、专一性差、干扰因素多。因此 ,迫切需要开发快速、灵敏的新方法。近年来 ,应用PCR技术检测水体中大肠菌群的研究报道不断增多。利用PCR技术扩增编码lacZ基因 ( β 半乳糖苷酶基因 )和uidA基因 ( β D 半乳糖苷酶基因 )的DNA片段 ,可以分别检测总大肠菌群和E .coli。但目前利用PCR进行定量分析还缺乏精度 ,需要进行更深入的研究。     

Automated electrical impedance technique for rapid enumeration of fecal coliforms in effluents from sewage treatment plants.

Fecal coliforms growing in a selective lactose-based broth medium at 44.5 degrees C generate a change in the electrical impedance of the culture relative to a sterile control when populations reach 10(6) to 10(7) per ml. The ratio of these changes was measured automatically, and the data were processed by computer. A linear relation was found between the log10 of the number of fecal coliforms in an inoculum and the time required for an electrical impedance ratio signal to be detected. Pure culture inocula consisting of 100 fecal coliforms in log phase or stationary phase were detected in 6.5 and 7.7 h, respectively. Standard curves of log10 fecal coliforms in wastewater inocula versus detection time, based on samples collected at a sewage treatment plant over a 4-month period, were found to vary from one another with time. Nevertheless, detection times were rapid and ranged from 5.8 to 7.9 h for 200 fecal coliforms to 8.7 to 11.4 h for 1 fecal coliform. Variations in detection times for a given number of fecal coliforms were also found among sewage treatment plants. A strategy is proposed which takes these variations into account and allows for rapid, automated enumeration of fecal coliforms in wastewater by the electrical impedance ratio technique.     

Automated electrical impedance technique for rapid enumeration of fecal coliforms in effluents from sewage treatment plants.

Fecal coliforms growing in a selective lactose-based broth medium at 44.5 degrees C generate a change in the electrical impedance of the culture relative to a sterile control when populations reach 10(6) to 10(7) per ml. The ratio of these changes was measured automatically, and the data were processed by computer. A linear relation was found between the log10 of the number of fecal coliforms in an inoculum and the time required for an electrical impedance ratio signal to be detected. Pure culture inocula consisting of 100 fecal coliforms in log phase or stationary phase were detected in 6.5 and 7.7 h, respectively. Standard curves of log10 fecal coliforms in wastewater inocula versus detection time, based on samples collected at a sewage treatment plant over a 4-month period, were found to vary from one another with time. Nevertheless, detection times were rapid and ranged from 5.8 to 7.9 h for 200 fecal coliforms to 8.7 to 11.4 h for 1 fecal coliform. Variations in detection times for a given number of fecal coliforms were also found among sewage treatment plants. A strategy is proposed which takes these variations into account and allows for rapid, automated enumeration of fecal coliforms in wastewater by the electrical impedance ratio technique.     

Repair detection procedure for enumeration of fecal coliforms and enterococci from seafoods and marine environments.

The repair detection procedure of Speck et al. (Appl. Microbiol. 29:549-550, 1975) was adapted for the enumeration of coliforms, fecal coliforms, and enterococci in seafood and environmental samples. Samples were pour plated with Trypticase soy agar, followed by a 1- to 2-h incubation to effect repair; the plates were then overlaid with the selective medium and incubated. Violet red bile agar and an incubation temperature of 45 degrees C were used as the selective conditions for fecal coliforms, and KF streptococcal agar was used for the enumeration of enterococci. The method was more efficient than the standard most-probable-number method for fecal coliform enumeration and also allowed enumeration of the injured cells, which might have remained undetected when selective medium in the most-probable-number method was used. The repair detection method effectively recovered the injured portion of the population of enterococci capable of growing on KF streptococcal agar. The repair enumeration method was not suitable for coliforms in marine samples because associative marine bacteria mimicked coliforms in violet red bile agar plates incubated at 35 degrees C. The marine bacteria did not grow at 45 degrees C and therefore did not interfere with fecal coliform enumeration.     

Repair detection procedure for enumeration of fecal coliforms and enterococci from seafoods and marine environments.

The repair detection procedure of Speck et al. (Appl. Microbiol. 29:549-550, 1975) was adapted for the enumeration of coliforms, fecal coliforms, and enterococci in seafood and environmental samples. Samples were pour plated with Trypticase soy agar, followed by a 1- to 2-h incubation to effect repair; the plates were then overlaid with the selective medium and incubated. Violet red bile agar and an incubation temperature of 45 degrees C were used as the selective conditions for fecal coliforms, and KF streptococcal agar was used for the enumeration of enterococci. The method was more efficient than the standard most-probable-number method for fecal coliform enumeration and also allowed enumeration of the injured cells, which might have remained undetected when selective medium in the most-probable-number method was used. The repair detection method effectively recovered the injured portion of the population of enterococci capable of growing on KF streptococcal agar. The repair enumeration method was not suitable for coliforms in marine samples because associative marine bacteria mimicked coliforms in violet red bile agar plates incubated at 35 degrees C. The marine bacteria did not grow at 45 degrees C and therefore did not interfere with fecal coliform enumeration.     

Comparison of the hydrophobic-grid membrane filter procedure and standard methods for coliform analysis of water

The hydrophobic-grid membrane filter (HGMF) has been proposed as an alternate method to the standard membrane filter (MF) procedure for the detection and enumeration of coliforms from water. Eight samples of nonchlorinated wastewater effluents were analyzed by the HGMF, standard MF, and tube fermentation most-probable-number methods for fecal coliforms, and eight samples each of polluted surface and dosed drinking waters were analyzed by the same methods for total coliforms. The drinking waters were dosed with coliforms and other heterotrophs concentrated from nonchlorinated domestic wastewater and treated with chlorine to reduce the numbers of organisms and simulate stress caused by chlorination. Statistical analyses determined that recoveries of fecal coliforms were significantly higher by the filtration methods for the nonchlorinated domestic wastewaters but not for the other waters. The results also indicated that recoveries of fecal and total coliforms did not differ significantly when either MFs or HGMFs were used. Total coliform results obtained with HGMFs having greater than 100 positive grid cells were significantly more precise than estimates obtained by the standard MF method only for polluted surface waters.     

Comparison of the hydrophobic-grid membrane filter procedure and standard methods for coliform analysis of water.

The hydrophobic-grid membrane filter (HGMF) has been proposed as an alternate method to the standard membrane filter (MF) procedure for the detection and enumeration of coliforms from water. Eight samples of nonchlorinated wastewater effluents were analyzed by the HGMF, standard MF, and tube fermentation most-probable-number methods for fecal coliforms, and eight samples each of polluted surface and dosed drinking waters were analyzed by the same methods for total coliforms. The drinking waters were dosed with coliforms and other heterotrophs concentrated from nonchlorinated domestic wastewater and treated with chlorine to reduce the numbers of organisms and simulate stress caused by chlorination. Statistical analyses determined that recoveries of fecal coliforms were significantly higher by the filtration methods for the nonchlorinated domestic wastewaters but not for the other waters. The results also indicated that recoveries of fecal and total coliforms did not differ significantly when either MFs or HGMFs were used. Total coliform results obtained with HGMFs having greater than 100 positive grid cells were significantly more precise than estimates obtained by the standard MF method only for polluted surface waters.     

Comparison of four membrane filter methods for fecal coliform enumeration in tropical waters.

Four membrane filter methods for the enumeration of fecal coliforms were compared for accuracy, specificity, and recovery. Water samples were taken several times from 13 marine, 1 estuarine, and 4 freshwater sites around Puerto Rico, from pristine waters and waters receiving treated and untreated sewage and effluent from a tuna cannery and a rum distillery. Differences of 1 to 3 orders of magnitude in the levels of fecal coliforms were observed in some samples by different recovery techniques. Marine water samples gave poorer results, in terms of specificity, selectivity, and comparability, than freshwater samples for all four fecal coliform methods used. The method using Difco m-FC agar with a resuscitation step gave the best overall results; however, even this method gave higher false-positive error, higher undetected-target error, lower selectivity, and higher recovery of nontarget organisms than the method using MacConkey membrane broth, the worst method for temperate waters. All methods tested were unacceptable for the enumeration of fecal coliforms in tropical fresh and marine waters. Thus, considering the high densities of fecal coliforms observed at most sites in Puerto Rico by all these methods, it would seem that these density estimates are, in many cases, grossly overestimating the degree of recent fecal contamination. Since Escherichia coli appears to be a normal inhabitant of tropical waters, fecal contamination may be indicated when none is present. Using fecal coliforms as an indicator is grossly inadequate for the detection of recent human fecal contamination and associated pathogens in both marine and fresh tropical waters.     

Validity of fecal coliforms, total coliforms, and fecal streptococci as indicators of viruses in chlorinated primary sewage effluents.

Quantities of combined chlorine that usually destroyed more than 99.999% of the indigenous fecal coliforms, total coliforms, and fecal streptococci in primary sewage effluents destroyed only 85 to 99% of the indigenous viruses present. Viruses were recovered from five of eight chlorinated primary effluents from which fecal coliforms were not recovered by standard most-probable-number procedures. The limited volumes of such chlorinated effluents that can be tested for indicator bacteria with currently available multiple-tube and membrane filter techniques restrict the value of fecal coliforms, fecal streptococci, and even total coliforms as indicators of viruses in these effluents. Although fecal coliforms and fecal streptococci are useful indicators of viruses in effluents from which these bacteria are recovered, the absence of these bacteria and even total coliforms from disinfected effluents (in standard tests) does not assure that viruses are also absent.     

Validity of fecal coliforms, total coliforms, and fecal streptococci as indicators of viruses in chlorinated primary sewage effluents.

Quantities of combined chlorine that usually destroyed more than 99.999% of the indigenous fecal coliforms, total coliforms, and fecal streptococci in primary sewage effluents destroyed only 85 to 99% of the indigenous viruses present. Viruses were recovered from five of eight chlorinated primary effluents from which fecal coliforms were not recovered by standard most-probable-number procedures. The limited volumes of such chlorinated effluents that can be tested for indicator bacteria with currently available multiple-tube and membrane filter techniques restrict the value of fecal coliforms, fecal streptococci, and even total coliforms as indicators of viruses in these effluents. Although fecal coliforms and fecal streptococci are useful indicators of viruses in effluents from which these bacteria are recovered, the absence of these bacteria and even total coliforms from disinfected effluents (in standard tests) does not assure that viruses are also absent.     

Anaerobic incubation of membrane filter cultures for improved detection of fecal coliforms from recreational waters.

Anaerobic incubation of membrane filter cultures significantly enhanced detection of fecal coliforms in surface-water samples from recreational beaches. In contrast to standard aerobic incubation, anaerobic incubation suppressed overgrowth of masking, noncoliform bacteria but did not increase the frequency of fecal coliform recovery.