Effects of the composition of bacteriological growth media on a chemiluminometric assay of β-galactosidase in Escherichia coli

The effects of the composition of bacteriological growth media on the light output in a chemiluminometric assay of β-galactosidase in Escherichia coli using 1,2-dioxetane substrates has been studied. In this assay a basic conflict exists between conditions that promote optimal bacterial growth and those conducive to maximal chemiluminescence. Common medium ingredients such as yeast or beef extract, protein hydrolysates and lactose suppress light emission and/or lead to high backgrounds. Quenching of light emission is probably partly due to light absorption by medium ingredients such as oxgall, and partly to interference with the reaction triggering the chemiluminescent process. Elevated backgrounds are caused by the presence of high concentrations of protein hydrolysates, which interact with the alkali in the accelerator solution. Only two purposely developed media, i.e. ILM and Colicult™ are shown to reconcile the requirements of growth support with that of optimal luminescent properties. © 1997 John Wiley & Sons, Ltd.     

化妆品中耐热大肠菌群和金黄色葡萄球菌检测能力验证分析

为增强实验室竞争力,考察实验室检测水平,本实验室参加了ACAS-pT743(2019)化妆品耐热大肠菌群及金黄色葡萄球菌的检测能力验证.本次能力验证中实验室共接收了四个样品,按照《化妆品安全技术规范》2015版对样品展开检测,并采用关键生理生化指标对可疑菌落加以确定以及16 S rDNA序列测定方法对结果进行复确认.结...     

旅游干扰的水质模拟对繁殖前期中国大鲵夜间活动节律的影响

动物的日活动节律是它们对各种环境条件昼夜变化的综合适应。研究旅游干扰对繁殖前期中国大鲵(Andrias davidianus)的夜间活动节律影响,有助于探明旅游干扰是否以及如何影响大鲵繁殖行为的机理。前期研究发现旅游干扰程度与水中溶解氧(Dissolved Oxygen,DO)含量显著负相关。在大鲵繁殖前期,通过控制大鲵生态繁育池水中的进水量(主要影响DO含量)来模拟重度与轻度旅游干扰下大鲵水环境,通过监测大鲵夜间出洞与进洞时间、水温(Water Temperature,WT)、DO和大肠菌群的动态变化,比较不同旅游干扰强度下大鲵夜间活动节律的差异,以明确旅游干扰导致的水质变化对大鲵夜间活动节律的影响。结果表明,雌/雄鲵出洞时间在模拟重度旅游干扰水质下显著晚于轻度水质中,雌鲵分别为19:10±0:22与18:53±0:24,雄鲵分别在18:29±0:20与18:13±0:18。重度干扰导致水质变化分别推迟了雌/雄大鲵出洞时间约17 min与16 min。但是,雌雄大鲵进洞时间在模拟重度与轻度旅游干扰水质下,差异不显著,雌鲵分别为24:10±0:13与24:18±0:17,雄鲵分别为6:44±0:23与6:40±0:29。此外,该研究还发现,雌/雄鲵出洞时间与三种因子(WT、DO与大肠菌群)显著相关,然而,雌/雄鲵进洞时间与以上3因子不相关,可能主要由其生物钟决定。重度旅游干扰导致的水质变化对大鲵繁殖前期活动节律影响小,仅对出洞时间有一定影响。     

Nutrient and Microbial Dynamics in Biosolids Amended Soils Following Rainfall Simulation

Municipal waste treatment plants are mandated by U.S. EPA to treat domestic wastewater prior to releasing it to receiving streams. The dewatering and high temperature drying processes at the plant are considered effective in reducing microbial contaminants in the waste. The resulting solid material (biosolid) is rich in nutrients that may serve as a value-added product for plant growth. In this study, we examined the nutrient value of biosolids, their potential biological and chemical risks that could result from surface application to two Mid-Atlantic soils: Bojac (coarse-loamy, mixed, thermic Typic Hapludult) and Cullen (clayey, mixed, thermic Typic Hapludult). Soils were placed on tilt beds and packed to their respective bulk density. Biosolids were added at a rate of 2.24 Mg/ha equivalent and mixed with the top 5 cm of the soil bed. Simulated rain was applied at a rate of 65 mm h^?1 for 45 minutes. Surface runoff and percolation water were collected and analyzed for elemental content, Escherichia coli (E. coli) and total coliform bacteria. Among the nutrient elements of concern (P, Zn, Mn, and Cu) in biosolids, none were found to be higher than the specified EPA limits. The concentration of P was highest in runoff and percolation water from beds packed with Bojac and biosolids. The combined effects of high clay (35%), Al (1.14%), and Fe (5.11%) in Cullen increased its P-adsorbing capacity. Low levels of E. coli and other coliform bacteria were present in samples from biosolids-treated beds packed with Cullen. Microbial counts in runoff and percolation samples varied with soil type; in some instances they were ten-fold higher in Bojac than in Cullen. The results obtained in this study suggest that surface runoff from land applications of biosolids might contribute to microbial contamination of receiving waters near agricultural fields.     

Evaluation of new chromogenic substrates for the detection of coliforms

Aims: To evaluate a new range of chromogenic substrates for the detection of β‐galactosidase activity in coliforms and to compare their performance in agar media and broths. Methods and Results: Sixteen novel galactoside substrates were prepared and incorporated into agar and broth. Their performance was compared using Escherichia coli (five strains), Salmonella (two strains), Enterobacter (two strains), Klebsiella, Pseudomonas, Listeria, Serratia, Shigella, Citrobacter, Proteus and Staphylococcus as well as pathological urine samples. The six substrates out of the initial 16 that showed the greatest sensitivity were VQE‐gal, VQM‐gal, VLPr‐gal, VLE‐gal, VLM‐gal and VBzTM‐gal, whose released chromophores were red, brown or purple. VQE‐gal and VLPr‐gal were studied in greater detail and were incorporated into agar medium. Coliform colonies appeared red and brown respectively, following incubation at 37°C for 24 h; however, positive results were obtained within a working day. The VQE‐gal medium was compared with some commercially available media. Conclusions: The range of substrates described can be used in broths as well as in agars. The VQE agar allows the detection of coliforms within a working day. VQE‐gal medium proved to be more sensitive when compared to other available chromogenic media and allows the unambiguous detection of coliforms.     

Identification of sources of Escherichia coli in South Carolina estuaries using antibiotic resistance analysis

Fecal pollution from nonhuman (pets, livestock or wildlife) and human sources is often one of the major factors associated with urbanization that contribute to the degradation of water quality. Methods to differentiate animal from human sources of fecal coliform contamination could assist resource managers in developing strategies to protect shellfish harvesting areas and recreational waters. In this study, surface water samples were collected from both a developed and an undeveloped watershed in coastal South Carolina. Influent and effluent samples from several wastewater treatment plants (WWTPs) in the same area were also collected. Most Probable Numbers (MPNs) of fecal coliforms were determined for all samples. Escherichia coli isolates were analyzed for antibiotic resistance (AR) to 10 antibiotics. Then, AR indices (no. of resistant/total no. of antibiotics tested), were calculated for each isolate and site. Results indicated that MPNs from the WWTP samples were significantly higher than those from the developed watershed which were significantly higher than those from the undeveloped watershed (p<0.0001). The AR analyses suggested that there was a trend toward increased antibiotic resistance in samples for the urbanized Broad Creek (BC) watershed. In the Okatee River (OR), E. coli isolates from three sites (20%) showed resistance to a single antibiotic (penicillin) but in BC, isolates from seven sites (47%) were resistant to multiple antibiotics, and the predominant resistance pattern was chlortetracycline-oxytetracycline-tetracycline. Raw sewage isolates from most WWTPs contained E. coli that exhibited resistance to multiple antibiotics. Cluster analysis indicated that all resistant OR sites had antibiotic resistant isolates that matched AR patterns found in isolates from WWTPs. Similarly, six of the seven sites in BC had AR patterns that matched with resistance patterns in WWTPs. These results suggest that AR testing may be a useful tool for differentiating E. coli from human and wildlife sources. Further testing of bacterial isolates from known animal sources is necessary to better assess the utility of this approach.     

Significance and suitability of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis> vs. fecal coliforms in assessing microbiological water quality

We examined the significance and suitability of Aeromonas hydrophila versus fecal coliforms in assessing microbiological water quality. For this, we used the membrane filtration method to simultaneously estimate the abundance level of A. hydrophila and fecal coliforms in waters from the Mfoundi river watershed at Yaoundé, and compared how fluctuations in A. hydrophila abundance matched those observed with fecal coliforms index as an indicator of water quality in the system under study. Our results revealed that waters were not safe according to the standards for water quality established by the Word Health Organization (WHO). They also indicated the prevalence of A. hydrophila as compared to fecal coliforms, and suggested that water from the Mfoundi River and its tributaries could be classified as hypereutrophic based on the density of Aeromonas. Moreover, the spatial distribution of fecal coliforms and A. hydrophila exhibited similar trends within the different water bodies investigated, suggesting that A. hydrophila can be used as indicator of water quality in highly polluted waters. We concluded that waters from the Mfoundi River watershed at Yaoundé represent a great potential risk of infection for users, and foresee that the next challenge will be to determine, among other factors, the physico-chemical factors influencing the observed spatial distribution.     

Evaluation of deep subsurface sampling procedures using serendipitous microbial contaminants as tracer organisms

Surface microbiological investigations are critically dependent on the procedures used to collect samples for study. It can be difficult to distinguish between indigenous organisms and those encountered as contaminants during the drilling process. We found that coliform bacteria contaminated drilling mud slurries. These bacteria proved useful as tracer organisms in evaluating the degree of microbial contamination accidentally encountered while drilling for subterranean samples. While these organisms were found in high numbers in both the circulating muds and in the mud reservoir, few subsurface samples harbored conforms. Subsurface slurries did not inhibit the growth of a known coliform inoculum. These results indicate that the methods used to collect and field‐process cores from Atlantic coastal plain sediments were sufficient to prevent a large degree of bacterial contamination in most samples. The microflora in drilling fluids did not quantitatively or qualitatively account for the number and diversity of bacteria in subsurface samples. We conclude that a large and viable bacterial community is present in deep regions of the terrestrial subsurface.