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At least 271 records · Page 15Linked to original sources
USGS1995

Screening tests for assessing the anaerobic biodegradation of pollutant chemicals in subsurface environments

Screening methods were developed to assess the susceptibility of ground water contaminants to anaerobic biodegradation. One method was an extrapolation of a procedure previously used to measure biodegradation activity in dilute sewage sludge. Aquifer solids and ground water with no additional nutritive media were incubated anaerobically in 160-ml serum bottles containing 250 mg·l −1 carbon of the substrate of interest. This method relied on the detection of gas pressure or methane production in substrateamended microcosms relative to background controls. Other screening procedures involved the consumption of stoichiometrically required amounts of sulfate or nitrate from the same type of incubations. Close agreement was obtained between the measured and calculated amounts of substrate bioconversion based on the measured biogas pressure in methanogenic microcosms. Storage of the microcosms for up to 6 months did not adversely influence the onset or rate of benzoic acid mineralization. The lower detection limits of the methanogenic assay were found to be a function of the size of the microcosm headspace, the mean oxidation state of the substrate carbon, and the method used to correct for background temperature fluctuations. Using these simple screening procedures, biodegradation information of regulatory interest could be generated, including, (i) the length of the adaptation period, (ii) the rate of substrate decay and (iii) the completeness of the bioconversion.

Journal of Microbiological Methods↗
USGS1996

Simple method for the measurement of the hydrogenotrophic methanogenic activity of anaerobic sludges

The specific hydrogenotrophic activity of anaerobic sludges is usually assayed by gas chromatographic analysis for methane in the headspace of sealed test vials. Gas is sampled with a pressure lock syringe which allows quantification independent of the pressure prevailing in the vials. An alternative method was developed using pressure transducer monitoring of the decrease in headspace gas pressure as the H2/CO2 substrate is converted to CH4. Application of a simple formula related the decrease at each sample point to millilitres of CH4 produced and gave values for the specific hydrogenotrophic activity of granular anaerobic sludge which were in good agreement with the values obtained by the more labor-intensive gas chromatographic method. The simplicity of the method facilitates multiple replicate analyses and allows more accurate determination of initial rates than is achievable by the gas chromatographic method which is prone to analytical error at the very low concentrations of CH4 present in the headspace during the early stages of the assay. Mass transfer of H2 from headspace to liquid was found to be rate-limiting and to result in significant under-estimation of the specific hydrogenotrophic activity of the granular sludge. A test protocol, which used a vial volatile suspended solids concentration between 1.7 and 8 g l-1; a 1:5 ratio between liquid and headspace; incubation of the vials horizontally with vigorous shaking (180 rev./min) and an initial H2/CO2 (80/20) gas pressure of 100-150 kPa was found to give reproducible and maximal values for the specific hydrogenotrophic activity of the test sludge.

Journal of Microbiological Methods↗
USGS1993

Effect of organic loading on nitrification and denitrification in a marine sediment microcosm

The effects of organic additions on nitrification and denitrification were examined in sediment microcosms. The organic material, heat killed yeast, had a C/N ratio of 7.5 and was added to sieved, homogenized sediments. Four treatments were compared: no addition (control, 30 g dry weight (dw) m-2 mixed throughout the 10 cm sediment column (30 M), 100 g dw m-2 mixed throughout sediments (100M), and 100 g dw m-2 mixed into top 1 cm (100S). After the microcosms had been established for 7-11 days, depth of O2 penetration, sediment-water fluxes and nitrification rates were measured. Nitrification rates were measured using three different techniques: N-serve and acetylene inhibition in intact cores, and nitrification potentials in slurries. Increased organic additions decreased O2 penetration from 2.7 to 0.2 mm while increasing both O2 consumption, from 30 to 70 mmol O2 m-2 d-1, and NO3- flux into sediments. Nitrification rates in intact cores were similar for the two methods. Highest rates occurred in the 30 M treatment, while the lowest rate was measured in the 100S treatment. Total denitrification rates (estimated from nitrification and nitrate fluxes) increased with increased organic addition, because of the high concentrations of NO3- (40 ??M) in the overlying water. The ratio of nitrification: denitrification was used as an indication of the importance of nitrification as the NO3- supply for denitrification. This ratio decreased from 1.55 to 0.05 with increased organic addition.

FEMS Microbiology Ecology↗
USGS1994

Demethylation and cleavage of dimethylsulfoniopropionate in marine intertidal sediments

Demethylation and cleavage of dimethylsulfoniopropionate (DMSP) was measured in three different types of,intertidal marine sediments: a cyanobacterial mat, a diatom-covered tidal flat and a carbonate sediment. Consumption rates of added DMSP were highest in cyanobacterial mat slurries (59 ?? mol DMSP l-1 slurry h-1) and lower in slurries from a diatom mat and a carbonate tidal sediment (24 and 9 ??mol DMSP l-1 h-1, respectively). Dimethyl sulfide (DMS) and 3-mercaptopropionate (MPA) were produced simultaneously during DMSP consumption, indicating that cleavage and demethylation occurred at the same time. Viable counts of DMSP-utilizing bacteria revealed a population of 2 x 107 cells cm-3 sediment (90% of these cleaved DMSP to DMS, 10% demethylated DMSP to MPA) in the cyanobacterial mat, 7 x 105 cells cm-3 in the diatom mat (23% cleavers, 77% demethylators), and 9 x 104 cells cm-3 (20% cleavers and 80% demethylators) in the carbonate sediment. In slurries of the diatom mat, the rate of MPA production from added 3-methiolpropionate (MMPA) was 50% of the rate of MPA formation from DMSP. The presence of a large population of demethylating bacteria and the production of MPA from DMSP suggest that the demethylation pathway, in addition to cleavage, contributes significantly to DMSP consumption in coastal sediments.

FEMS Microbiology Ecology↗
USGS1995

A comparison of two nitrification inhibitors used to measure nitrification rates in estuarine sediments

Nitrification rates were measured using intact sediment cores from South San Francisco Bay and two different nitrification inhibitors: acetylene and methyl fluoride. Sediment oxygen consumption and ammonium and nitrate fluxes were also measured in these cores. Four experiments were conducted in the spring, and one in the fall of 1993. There was no significant difference in nitrification rates measured using the two inhibitors, which suggests that methyl fluoride can be used as an effective inhibitor of nitrification. Nitrification was positively correlated with sediment oxygen consumption and numbers of macrofauna. This suggests that bioturbation by macrofauna is an important control of nitrification rates. Irrigation by the tube-dwelling polychaete, Asychis elongata, which dominates the benthic biomass at this location, appears particularly important. Ammonium fluxes out of the sediment were greatest about one week after the spring bloom, while nitrification peaked about one month later.

FEMS Microbiology Ecology↗
USGS1980

Physio-chemical and morphological characteristics of avian encephalomyelitis virus

Avian encephalomyelitis virus (AEV) was purified from infected chick embryos by a gradient centrifugation in cesium chloride. The virus had a buoyant density of 1.31 to 1.32 g/ml and a sedimentation coefficient of 148 S. The purified AEV was resistant to treatments with chloroform, acid pH or trypsin. The presence of Mg++ stabilized the virus against heat inactivation (56°C, 1 h). Electron microscopic study showed the virus to be 24 to 32 nm in diameter. The surface structure of the purified virus was not easily discernable. Nevertheless, with uranyl acetate-stained particles, Markham's rotation technique revealed that AEV has five-fold symmetry with 32 or 42 capsomers. Exact classification of AEV awaits characterization of the viral nucleic acid.

Veterinary Microbiology↗
USGS2004

Use of PRD1 bacteriophage in groundwater viral transport, inactivation, and attachment studies

PRD1, an icosahedra-shaped, 62 nm (diameter), double-stranded DNA bacteriophage with an internal membrane, has emerged as an important model virus for studying the manner in which microorganisms are transported through a variety of groundwater environments. The popularity of this phage for use in transport studies involving geologic media is due, in part, to its relative stability over a range of temperatures and low degree of attachment in aquifer sediments. Laboratory and field investigations employing PRD1 are leading to a better understanding of viral attachment and transport behaviors in saturated geologic media and to improved methods for describing mathematically subsurface microbial transport at environmentally significant field scales. Radioisotopic labeling of PRD1 is facilitating additional information about the nature of viral interactions with solid surfaces in geologic media, the importance of iron oxide surfaces, and allowing differentiation between inactivation and attachment in field-scale tracer tests.

FEMS Microbiology Ecology↗
USGS2004

The microbial arsenic cycle in Mono Lake, California

Significant concentrations of dissolved inorganic arsenic can be found in the waters of a number of lakes located in the western USA and in other water bodies around the world. These lakes are often situated in arid, volcanic terrain. The highest concentrations of arsenic occur in hypersaline, closed basin soda lakes and their remnant brines. Although arsenic is a well-known toxicant to eukaryotes and prokaryotes alike, some prokaryotes have evolved biochemical mechanisms to exploit arsenic oxyanions (i.e., arsenate and arsenite); they can use them either as an electron acceptor for anaerobic respiration (arsenate), or as an electron donor (arsenite) to support chemoautotrophic fixation of CO 2 into cell carbon. Unlike in freshwater or marine ecosystems, these processes may assume quantitative significance with respect to the carbon cycle in arsenic-rich soda lakes. For the past several years our research has focused on the occurrence and biogeochemical manifestations of these processes in Mono Lake, a particularly arsenic-rich environment. Herein we review some of our findings concerning the biogeochemical arsenic cycle in this lake, with the hope that it may broaden the understanding of the influence of microorganisms upon the speciation of arsenic in more common, less “extreme” environments, such as drinking water aquifers.

California↗
USGS2012

PhyloChipTM microarray comparison of sampling methods used for coral microbial ecology

Interest in coral microbial ecology has been increasing steadily over the last decade, yet standardized methods of sample collection still have not been defined. Two methods were compared for their ability to sample coral-associated microbial communities: tissue punches and foam swabs, the latter being less invasive and preferred by reef managers. Four colonies of star coral, Montastraea annularis , were sampled in the Dry Tortugas National Park (two healthy and two with white plague disease). The PhyloChip™ G3 microarray was used to assess microbial community structure of amplified 16S rRNA gene sequences. Samples clustered based on methodology rather than coral colony. Punch samples from healthy and diseased corals were distinct. All swab samples clustered closely together with the seawater control and did not group according to the health state of the corals. Although more microbial taxa were detected by the swab method, there is a much larger overlap between the water control and swab samples than punch samples, suggesting some of the additional diversity is due to contamination from water absorbed by the swab. While swabs are useful for noninvasive studies of the coral surface mucus layer, these results show that they are not optimal for studies of coral disease.

Dry Tortugas National Park↗
USGS2015

A real-time, quantitative PCR protocol for assessing the relative parasitemia of Leucocytozoon in waterfowl

Microscopic examination of blood smears can be effective at diagnosing and quantifying hematozoa infections. However, this method requires highly trained observers, is time consuming, and may be inaccurate for detection of infections at low levels of parasitemia. To develop a molecular methodology for identifying and quantifying Leucocytozoon parasite infection in wild waterfowl (Anseriformes), we designed a real-time, quantitative PCR protocol to amplify Leucocytozoon mitochondrial DNA using TaqMan fluorogenic probes and validated our methodology using blood samples collected from waterfowl in interior Alaska during late summer and autumn (n = 105). By comparing our qPCR results to those derived from a widely used nested PCR protocol, we determined that our assay showed high levels of sensitivity (91%) and specificity (100%) in detecting Leucocytozoon DNA from host blood samples. Additionally, results of a linear regression revealed significant correlation between the raw measure of parasitemia produced by our qPCR assay (Ct values) and numbers of parasites observed on blood smears (R2 = 0.694, P = 0.003), indicating that our assay can reliably determine the relative parasitemia levels among samples. This methodology provides a powerful new tool for studies assessing effects of haemosporidian infection in wild avian species.

Journal of Microbiological Methods↗
USGS2016

Multi-laboratory survey of qPCR enterococci analysis method performance in U.S. coastal and inland surface waters

Quantitative polymerase chain reaction (qPCR) has become a frequently used technique for quantifying enterococci in recreational surface waters, but there are several methodological options. Here we evaluated how three method permutations, type of mastermix, sample extract dilution and use of controls in results calculation, affect method reliability among multiple laboratories with respect to sample interference. Multiple samples from each of 22 sites representing an array of habitat types were analyzed using EPA Method 1611 and 1609 reagents with full strength and five-fold diluted extracts. The presence of interference was assessed three ways: using sample processing and PCR amplifications controls; consistency of results across extract dilutions; and relative recovery of target genes from spiked enterococci in water sample compared to control matrices with acceptable recovery defined as 50 to 200%. Method 1609, which is based on an environmental mastermix, was found to be superior to Method 1611, which is based on a universal mastermix. Method 1611 had over a 40% control assay failure rate with undiluted extracts and a 6% failure rate with diluted extracts. Method 1609 failed in only 11% and 3% of undiluted and diluted extracts analyses. Use of sample processing control assay results in the delta–delta Ct method for calculating relative target gene recoveries increased the number of acceptable recovery results. Delta–delta tended to bias recoveries from apparent partially inhibitory samples on the high side which could help in avoiding potential underestimates of enterococci — an important consideration in a public health context. Control assay and delta–delta recovery results were largely consistent across the range of habitats sampled, and among laboratories. The methodological option that best balanced acceptable estimated target gene recoveries with method sensitivity and avoidance of underestimated enterococci densities was Method 1609 without extract dilution and using the delta–delta calculation method. The applicability of this method can be extended by the analysis of diluted extracts to sites where interference is indicated but, particularly in these instances, should be confirmed by augmenting the control assays with analyses for target gene recoveries from spiked target organisms.

Journal of Microbiological Methods↗
USGS2020

Validation of a screening method for the detection of colistin-resistant E. coli containing mcr-1 in feral swine feces

A method was developed and validated for the detection of colistin-resistant Escherichia coli containing mcr-1 in the feces of feral swine. Following optimization of an enrichment method using EC broth supplemented with colistin (1 µg/mL) and vancomycin (8 µg/mL), aliquots derived from 100 feral swine fecal samples were spiked with of one of five different mcr-1 positive E. coli strains (between 100 and 104 CFU/g), for a total of 1,110 samples tested. Enrichments were then screened using a simple boil-prep and a previously developed real-time PCR assay for mcr-1 detection. The sensitivity of the method was determined in swine feces, with mcr-1 E. coli inoculums of 0.1-9.99 CFU/g (n = 340), 10-49.99 CFU/g (n = 170), 50-99 CFU/g (n = 255), 100-149 CFU/g (n = 60), and 200-2,200 CFU/g (n = 175), which were detected with 32%, 72%, 88%, 95%, and 98% accuracy, respectively. Uninoculated controls (n = 100) were negative for mcr-1 following enrichment.

Journal of Microbiological Methods↗
USGS2020

Utilization of multiple microbial tools to evaluate efficacy of restoration strategies to improve recreational water quality at a Lake Michigan Beach (Racine, WI)

Hydro-meteorological conditions facilitate transport of fecal indicator bacteria (FIB) to the nearshore environment, affecting recreational water quality. North Beach (Racine, Wisconsin, United States), is an exemplar public beach site along Lake Michigan, where precipitation-mediated surface runoff, wave encroachment, stormwater and tributary outflow were demonstrated to contribute to beach advisories. Multiple restoration actions, including installation of a stormwater retention wetland, were successfully deployed to improve recreational water quality. Implementation of molecular methods (e.g. human microbial source tracking markers and Escherichia coli ( E. coli ) qPCR) assisted in identifying potential pollution sources and improving public health response time. However, periodic water quality failures still occur. As local beach managers reassess restoration measures in response to climatic changes, use of expanded microbial methods (including bacterial community profiling) may contribute to a better understanding of these dynamic environments. In this 2-year study (2015 and 2019), nearshore/offshore Lake Michigan, stormwater, and tributary samples were collected to determine if, 1) the constructed wetland (~50 m from the shoreline) continued to provide stormwater separation/retention and 2) mixing between onshore sources, Root River and Lake Michigan, was increasing due to rising precipitation/lake levels. Monthly rainfall totals were 1.5× higher in 2019 than 2015, coinciding with a 0.63 m lake-level rise. The prevalence of more intense, onshore winds also increased, facilitating interaction between potential reservoirs of FIB with nearshore water through wind driven waves and lake intrusion, e.g. beach sands and the adjacent Root River. While a strong relationship existed between wet weather wetland and North Beach nearshore E. coli concentrations (all sites), bacterial communities were strikingly different. Conversely, bacterial community overlap existed between the Root River mouth and nearshore/offshore sites. These results suggest the constructed wetland can accommodate the climate-related changes observed in this study. Future restoration activities could be directed towards upstream tributary sources in order to minimize microbial contaminants entering Lake Michigan.

Wisconsin↗
USGS2020

A comparison of E. coli concentration estimates quantified by the EPA and a Michigan laboratory network using EPA Draft Method C

We evaluated data from 10 laboratories that analyzed water samples from 82 recreational water sites across the state of Michigan between 2016 and 2018. Water sample replicates were analyzed by experienced U.S. Environmental Protection Agency (EPA) analysts and Michigan laboratories personnel, many of whom were newly trained, using EPA Draft Method C—a rapid quantitative polymerase chain reaction (qPCR) technique that provides same day Escherichia coli ( E. coli ) concentration results. Beach management decisions (i.e. remain open or issue an advisory or closure) based on E. coli concentration estimates obtained by Michigan labs and by the EPA were compared; the beach management decision agreed in 94% of the samples analyzed. We used the Wilcoxon one-sample signed rank test and nonparametric quantile regression to assess (1) the degree of agreement between E. coli concentrations quantified by Michigan labs versus the EPA and (2) Michigan lab E. coli measurement precision, relative to EPA results, in different years and water body types. The median quantile regression curve for Michigan labs versus EPA approximated the 1:1 line of perfect agreement more closely as years progressed. Similarly, Michigan lab E. coli estimates precision also demonstrated yearly improvements. No meaningful difference was observed in the degree of association between Michigan lab and EPA E. coli concentration estimates for inland lake and Great Lakes samples (median regression curve average slopes 0.93 and 0.95, respectively). Overall, our study shows that properly trained laboratory personnel can perform Draft Method C to a degree comparable with experienced EPA analysts. This allows health departments that oversee recreational water quality monitoring to be confident in qPCR results generated by the local laboratories responsible for analyzing the water samples.

Journal of Microbiological Methods↗
USGS2021

Evaluation of a modified rapid viability-polymerase chain reaction method for Bacillus atrophaeus spores in water matrices

A rapid method that provides information on the viability of organisms is needed to protect public health and ensure that remediation efforts following a release of a biological agent are effective. The rapid viability-polymerase chain reaction (RV-PCR) method combines broth culture and molecular methods to provide results on whether viable organisms are present in less than 15 h. In this study, a modified RV-PCR (mRV-PCR) method was compared to a membrane-filtration culture method for the detection of viable Bacillus spores in water matrices. Samples included small and large volumes of chlorine and non‑chlorine treated tap water. Large volume water samples (up to 100 L), were processed by ultrafiltration using a semi-automated waterborne pathogen concentrator, followed by centrifugation as a secondary concentration technique. The concentrated samples were analyzed by mRV-PCR and culture methods. The overall agreement between the mRV-PCR and culture methods when seed concentrations were greater than 10 spores per sample volume analyzed was 96%. The total time from the start of sample processing to the final sample result for the mRV-PCR method was decreased by approximately 2 h, in comparison to the previously published RV-PCR method because of the incorporation of shorter, more efficient primary and secondary concentration steps and a shorter DNA extraction technique. Overall, this study confirmed that RV-PCR is a promising approach for identifying viable Bacillus spores in small- and large-volume water samples and for producing results in less time than traditional culture methods.

Journal of Microbiological Methods↗
USGS2013

Epifluorescent direct counts of bacteria and viruses from topsoil of various desert dust storm regions

Topsoil from arid regions is the main source of dust clouds that move through the earth's atmosphere, and microbial communities within these soils can survive long-range dispersion. Microbial abundance and chemical composition were analyzed in topsoil from various desert regions. Statistical analyses showed that microbial direct counts were strongly positively correlated with calcium concentrations and negatively correlated with silicon concentrations. While variance between deserts was expected, it was interesting to note differences between sample sites within a given desert region, illustrating the 'patchy' nature of microbial communities in desert environments.

Research in Microbiology↗
USGS2023

The persistence of time: The lifespan of Bacillus anthracis spores in environmental reservoirs

Anthrax is a lethal bacterial zoonosis primarily affecting herbivorous wildlife and livestock. Upon host death Bacillus anthracis vegetative cells form spores capable of surviving for years in soil. Anthrax transmission requires host exposure to large spore doses. Thus, conditions that facilitate higher spore concentrations or promote spore survival will increase the probability that a pathogen reservoir infects future hosts. We investigated abiotic and pathogen genomic variation in relation to spore concentrations in surface soils (0e1 cm depth) at 40 plains zebra (Equus quagga) anthrax carcass sites in Namibia. Specifically, how initial spore concentrations and spore survival were affected by seasonality associated with the timing of host mortality, local soil characteristics, and pathogen genomic variation. Zebras dying of anthrax in wet seasons-the peak season for anthrax in Etosha National Park-had soil spore concentrations 1.36 orders of magnitude higher than those that died in dry seasons. No other variables considered affected spore concentrations, and spore survival rates did not differ among sites. Surface soils at these pathogen reservoirs remained culture positive for a range of 3.8e10.4 years after host death. Future research could evaluate if seasonal patterns in spore concentrations are driven by differences in sporulation success or levels of terminal bacteremia.

Etosha National Park↗
USGS2009

Microbial disease and the coral holobiont

Tropical coral reefs harbour a reservoir of enormous biodiversity that is increasingly threatened by direct human activities and indirect global climate shifts. Emerging coral diseases are one serious threat implicated in extensive reef deterioration through disruption of the integrity of the coral holobiont – a complex symbiosis between the coral animal, endobiotic alga and an array of microorganisms. In this article, we review our current understanding of the role of microorganisms in coral health and disease, and highlight the pressing interdisciplinary research priorities required to elucidate the mechanisms of disease. We advocate an approach that applies knowledge gained from experiences in human and veterinary medicine, integrated into multidisciplinary studies that investigate the interactions between host, agent and environment of a given coral disease. These approaches include robust and precise disease diagnosis, standardised ecological methods and application of rapidly developing DNA, RNA and protein technologies, alongside established histological, microbial ecology and ecological expertise. Such approaches will allow a better understanding of the causes of coral mortality and coral reef declines and help assess potential management options to mitigate their effects in the longer term.

Trends in Microbiology↗