Isolation, identification and characterization of fluoride resistant bacteria: possible role in bioremediation

Microorganisms found in industrial effluents and near the sites of the contamination can be used to indicate pollution and detoxify the contaminated water resources. Emergence of xenobiotic resistant bacteria among them might be potential application in bioremediation. The objective of this study was to isolate and characterize fluoride resistant bacteria from soil and water samples of different regions of India. Five isolates were recovered from different samples which were found to be fluoride resistant. Two of them effectively reduced the fluoride from their media. Through the current study it can be predicted that fluoride pollution results in selective pressure that leads to the development of fluoride resistant among bacterial populations, probably through the mechanism which involved high affinity anion binding compounds called ionophores. Resistant microbes may play a bioremediative role by transforming and concentrating these anions so that they are less available and less dangerous.     

Assessment of heavy metal contamination and Hg-resistant bacteria in surface water from different regions of Delhi,India

The present study aims to monitor the surface water quality of different regions in Delhi (India). With many physical and chemical properties, all samples had a high load of pollution in which Najafgarh drain (Nd) exhibited maximum and laboratory tap water (Ltw) minimum contamination. Water samples contained notable amounts of heavy metals including Cr, Cd, As, Cu, Pb and Hg. A total of 88 Hg-resistant bacteria were isolated from all the regions except Ltw. Among all the samples, the density of Hg-resistant bacteria was highest in sample of Nd and their morphotype heterogeneity was highest in sample collected from river Yamuna nearby Kashmiri gate (Kg). Different strains showed different patterns of resistance to different heavy metals and antibiotics. Multiple antibiotic resistance (MAR) indices were high in two samples, the highest reported in a sample taken from river Yamuna nearby Majnu ka tila (Mkt) (0.34). The 12.5% and 24.45% isolates showed β- and α-hemolytic natures, respectively that might be of pathogenic concern. In this account, high concentrations of heavy metals and their resistant bacteria in surface water have severely damaged the quality of water and their resources and produced high risk to the associated life forms.     

The fate of stormwater-associated bacteria in constructed wetland and water pollution control pond systems

The performances of a constructed wetland and a water pollution control pond were compared in terms of their abilities to reduce stormwater bacterial loads to recreational waters. Concentrations of thermotolerant coliforms, enterococci and heterotrophic bacteria were determined in inflow and outflow samples collected from each system over a 6-month period. Bacterial removal was significantly less effective in the water pollution control pond than in the constructed wetland. This was attributed to the inability of the pond system to retain the fine clay particles (< 2 microm) to which the bacteria were predominantly adsorbed. Sediment microcosm survival studies showed that the persistence of thermotolerant coliforms was greater in the pond sediments than in the wetland sediments, and that predation was a major factor influencing bacterial survival. The key to greater bacterial longevity in the pond sediments appeared to be the adsorption of bacteria to fine particles, which protected them from predators. These observations may significantly affect the choice of treatment system for effective stormwater management.     

Air and water qualities around small ruminant houses in Central Java - Indonesia

There is a general concern that livestock can have a profound effect on the environment, also in smallholder production systems. This paper presented the impact of small ruminants on the quality of air and water in and around small ruminant houses. In total, 27 small ruminant houses from the three agro-ecological zones, lowlands, middle zone and uplands, in the Province of Yogyakarta, Indonesia, were monitored to investigate gas emissions and water pollution. Air samples were taken by using a gas catching tool filled with absorbents for gases. The gases mainly consisted of ammonia, nitrogen oxides, dihydrogen sulphide, sulphur dioxide and hydrocarbons. Groundwater samples were collected from the farmers’ wells, adjacent to the small ruminants houses. The main water chemical parameters analysed were pH, turbidity, iron, fluoride, calcium carbonate, chloride, manganese, nitrate, nitrite, dihydrogen sulphide, and organic matter, while the bacterial indicators were faecal coliform and total coliform. Gas concentrations in the air were highest inside the houses, while around the houses their concentrations decreased rapidly on both the tailwind and headwind side. The concentrations of the gases were below the admissible levels assessed by the local government, except hydrocarbons in the middle zone, which was probably related to the storage of the manure inside the houses. The impact of small ruminants on water pollution was much greater than on air pollution. Some of the physico-chemical parameters showed high concentrations and nearly reached the admissible limit. The water sources had very high levels of faecal coliform bacteria and total coliform bacteria, two groups of bacteria used as indicators for water contamination caused by manure. It can be concluded that, in Central Java, management and housing of small ruminants close to the family quarters causes little environmental problems; however, attention should be paid to water quality.     

F+ RNA coliphage typing for microbial source tracking in surface waters

AIMS: The utility of coliphages to detect and track faecal pollution was evaluated using South Carolina surface waters that exceeded State faecal coliform standards. METHODS AND RESULTS: Coliphages were isolated from 117 surface water samples by single agar layer (SAL) and enrichment presence/absence (EP/A) methods. Confirmed F+ RNA coliphages were typed for microbial source tracking using a library-independent approach. Concentrations of somatic coliphages using 37 and 44.5 degrees C incubation temperatures were found to be significantly different and the higher temperature may be more specific for faecal contamination. The EP/A technique detected coliphages infecting Escherichia coli Famp in 38 (66%) of the 58 surface water samples negative for F+ coliphages by the SAL method. However, coliphages isolated by EP/A were found to be less representative of coliphage diversity within a sample. Among the 2939 coliphage isolates tested from surface water and known source samples, 813 (28%) were found to be F+ RNA. The majority (94%) of surface water F+ RNA coliphage isolates typed as group I. Group II and/or III viruses were identified from 14 surface water stations, the majority of which were downstream of wastewater discharges. These sites were likely contaminated by human-source faecal pollution. CONCLUSIONS: The results suggest that faecal contamination in surface waters can be detected and source identifications aided by coliphage analyses. SIGNIFICANCE AND IMPACT OF THE STUDY: This study supports the premise that coliphage typing can provide useful, but not absolute, information to distinguish human from animal sources of faecal pollution. Furthermore, the comparison of coliphage isolation methods detailed in this study should provide valuable information to those wishing to incorporate coliphage detection into water quality assessments.     

Tracking human sewage microbiome in a municipal wastewater treatment plant

Human sewage pollution is a major threat to public health because sewage always comes with pathogens. Human sewage is usually received and treated by wastewater treatment plants (WWTPs) to control pathogenic risks and ameliorate environmental health. However, untreated sewage that flows into water environments may cause serious waterborne diseases, as reported in India and Bangladesh. To examine the fate of the human sewage microbiome in a local municipal WWTP of Hong Kong, we used massively parallel sequencing of 16S rRNA gene to systematically profile microbial communities in samples from three sections (i.e., influent, activated sludge, and effluent) obtained monthly throughout 1 year. The results indicated that: (1) influent sewage bacterial profile reflected the human microbiome; (2) human gut bacterial community was the dominant force shaping influent sewage bacterial profile; (3) most human sewage bacteria could be effectively removed by the WWTP; (4) a total of 75 genera were profiled as potentially pathogenic bacteria, most of which were still present in the effluent although at a very low level; (5) a grouped pattern of bacterial community was observed among the same section samples but a dispersed pattern was found among the different section samples; and (6) activated sludge was less affected by the influent sewage bacteria, but it showed a significant impact on the effluent bacteria. All of these findings provide novel insights toward a mechanistic understanding of the fate of human sewage microbiome in the WWTP.     

Percentage of gelatinolytic bacteria among heterotrophic bacteria as indicator of water quality

The relationship between the physiological group of gelatinolytic bacteria and the abundance of heterotrophic bacteria in freshwater ecosystems was described, based on analysis of 1082 different freshwater samples collected in Croatia. Percentages of gelatinolytic bacteria among the population of heterotrophic bacteria showed a significant negative correlation with the abundance of heterotrophic bacteria. The relation between the physiological group (gelatinolytic bacteria) and heterotrophic bacteria can be considered to be an indicator of the pollution degree of freshwaters. A high relative content of gelatinolytic bacteria (> 76%) always indicates the colony-forming units of heterotrophic bacteria < 1000/mL, which corresponds to the high water quality; gelatinolytic bacteria < 11% indicate polluted waters. Isolated strains of aerobically grown gelatinolytic bacteria were Gram-negative rod-shaped or Gram-positive endospore-forming rod-shaped cells.     

Investigation of N-acyl homoserine lactone (AHL) molecule production in Gram-negative bacteria isolated from cooling tower water and biofilm samples

In this study, 99 Gram-negative rod bacteria were isolated from cooling tower water, and biofilm samples were examined for cell-to-cell signaling systems, N-acyl homoserine lactone (AHL) signal molecule types, and biofilm formation capacity. Four of 39 (10 %) strains isolated from water samples and 14 of 60 (23 %) strains isolated from biofilm samples were found to be producing a variety of AHL signal molecules. It was determined that the AHL signal molecule production ability and the biofilm formation capacity of sessile bacteria is higher than planktonic bacteria, and there was a statistically significant difference between the AHL signal molecule production of these two groups (p?<?0.05). In addition, it was found that bacteria belonging to the same species isolated from cooling tower water and biofilm samples produced different types of AHL signal molecules and that there were different types of AHL signal molecules in an AHL extract of bacteria. In the present study, it was observed that different isolates of the same strains did not produce the same AHLs or did not produce AHL molecules, and bacteria known as AHL producers did not produce AHL. These findings suggest that detection of signal molecules in bacteria isolated from cooling towers may contribute to prevention of biofilm formation, elimination of communication among bacteria in water systems, and blockage of quorum-sensing controlled virulence of these bacteria.     

Microbiological quality of water and dialysate in a haemodialysis unit in Ponta Grossa-PR, Brazil

AIMS: The objective of the study was to determine the microbiological quality of samples of water and dialysate in a haemodialysis unit. METHODS AND RESULTS: Seventy-two samples each of water and dialysate were collected during November 2003 to April 2004. The following microbiological analyses were performed: test for total and faecal coliforms, which produced negative results for all the samples; counts of total heterotrophic bacteria, where three samples of water and two of dialysate showed levels higher than those permitted by national standards; and endotoxin assay, which revealed high quantities only in samples of water that preceded reverse osmosis. Nonfermenting Gram-negative bacteria were identified in 54 samples of dialysate and in 26 samples of water. The test for adhesion to an inert surface showed that various bacteria were capable of forming biofilms. Twenty-seven per cent of the bacteria were resistant to sodium hypochlorite at 500 ppm for 10-min contact time. Sixty per cent of the isolates were resistant to three or more antibiotics. CONCLUSIONS: Water and dialysate can be a source of infection for patients who need haemodialysis. SIGNIFICANCE AND IMPACT OF THE STUDY: An adequate system for water treatment, disinfection of the haemodialysis system and microbiological monitoring of the water and dialysate are necessary to reduce bacteraemia and pyrogenia outbreaks.     

FURTHER OBSERVATIONS ON THE BACTERIAL CONTENT OF WATER IN WATERCRESS BEDS

SUMMARY: Observations on two watercress beds in Kent have confirmed previous findings (Jones & Baker, 1955) that even in comparatively well-protected beds the coli-aerogenes content of the water may be high in summer. They have also shown that in spite of circumstances which would lead one to expect pollution, as in the case of one bed, the coli-aerogenes content can be low in winter. The season appears to influence the content in the water of coli-aerogenes bacteria, pectate liquefying bacteria and those capable of growing at 37°, but in the case of those growing at 22° other factors can play a more important part, as higher counts were not uncommon in winter. Seasonal effects were less marked with inlet samples; they were less influenced by atmospheric temperature, and little extraneous contamination occurred. Numbers increased as the sampling point moved from inlet to outlet.     

Detection of Legionella species in reclaimed water and air with the EnviroAmp Legionella PCR kit and direct fluorescent antibody staining.

Reclaimed water is an important resource for areas with inadequate water supplies. However, there have been few studies on the variety of microorganisms found in this type of water, since typically reclaimed water is examined only for the presence of coliform bacteria. Many microorganisms, including the legionellae, are known to be more resistant to chlorine than are coliform bacteria. Previously, we detected > 10(3) Legionella cells per ml in primary and secondary sewage effluents and observed no significant reduction in population numbers throughout the treatment process. In this study, we detected Legionella spp. in chlorinated effluent by using an EnviroAmp Legionella PCR kit and direct fluorescent antibody (DFA) staining. However, we were not able to isolate Legionella spp. from either natural or seeded reclaimed water samples. This suggests that the Legionella spp. detected by the PCR and DFA methods may be injured or viable but nonculturable after exposure to the high residual chlorine levels typically found in this type of water source. The numbers of coliform bacteria were low (< 2 cells per 100 ml) in most reclaimed water samples and were not correlated with the presence or absence of Legionella spp. We also collected air samples from above a secondary aeration basin and analyzed them by using the PCR, DFA, and plate culture methods. Legionella spp. were detected in the air obtained from above the secondary basin with all three methods. We concluded that the PCR was superior to the culture and DFA methods for detecting Legionella spp. in environmental water samples.     

Antibiotic resistance genes in the bacteriophage DNA fraction of environmental samples

Antibiotic resistance is an increasing global problem resulting from the pressure of antibiotic usage, greater mobility of the population, and industrialization. Many antibiotic resistance genes are believed to have originated in microorganisms in the environment, and to have been transferred to other bacteria through mobile genetic elements. Among others, β-lactam antibiotics show clinical efficacy and low toxicity, and they are thus widely used as antimicrobials. Resistance to β-lactam antibiotics is conferred by β-lactamase genes and penicillin-binding proteins, which are chromosomal- or plasmid-encoded, although there is little information available on the contribution of other mobile genetic elements, such as phages. This study is focused on three genes that confer resistance to β-lactam antibiotics, namely two β-lactamase genes (blaTEM and blaCTX-M9) and one encoding a penicillin-binding protein (mecA) in bacteriophage DNA isolated from environmental water samples. The three genes were quantified in the DNA isolated from bacteriophages collected from 30 urban sewage and river water samples, using quantitative PCR amplification. All three genes were detected in the DNA of phages from all the samples tested, in some cases reaching 104 gene copies (GC) of blaTEM or 102 GC of blaCTX-M and mecA. These values are consistent with the amount of fecal pollution in the sample, except for mecA, which showed a higher number of copies in river water samples than in urban sewage. The bla genes from phage DNA were transferred by electroporation to sensitive host bacteria, which became resistant to ampicillin. blaTEM and blaCTX were detected in the DNA of the resistant clones after transfection. This study indicates that phages are reservoirs of resistance genes in the environment.     

Assessment of the Bioaccessibility of Trace Elements in Cat’s Claw Teas by In Vitro Simulated Gastrointestinal Digestion Using FAAS

Beer is a widely consumed drink throughout the world, and because its manufacture involves the use of water, beer can be, in some cases, a source of fluorides. For this reason, the objective of this study was to determine the concentration of fluorides in 50 samples of beers from different sources sold in two different types of container (aluminum can and glass bottle). The possible significant differences between the different types of packaging and the intake of fluoride from the consumption of these beers were evaluated. The concentration of fluoride in beers has been determined using the potentiometric method of fluoride determination by standard addition. The concentration of fluoride ranged between 0.06 and 1.77 mg/L. In general, the concentration was below 1 mg/L, except for three beer samples from Ireland and the USA, whose concentration was over 1.5 mg/L. No significant differences were found between the types of packaging. The contribution of fluoride to the diet from beer consumption is not high (<27%); however, it is necessary to warn consumers whenever they are in areas of high concentrations of fluoride in the water supply.     

Tolerance to Various Toxicants by Marine Bacteria Highly Resistant to Mercury

Bacteria highly resistant to mercury isolated from seawater and sediment samples were tested for growth in the presence of different heavy metals, pesticides, phenol, formaldehyde, formic acid, and trichloroethane to investigate their potential for growth in the presence of a variety of toxic xenobiotics. We hypothesized that bacteria resistant to high concentrations of mercury would have potential capacities to tolerate or possibly degrade a variety of toxic materials and thus would be important in environmental pollution bioremediation. The mercury-resistant bacteria were found to belong to Pseudomonas, Proteus, Xanthomonas, Alteromonas, Aeromonas, and Enterobacteriaceae. All these environmental bacterial strains tolerant to mercury used in this study were capable of growth at a far higher concentration (50 ppm) of mercury than previously reported. Likewise, their ability to grow in the presence of toxic xenobiotics, either singly or in combination, was superior to that of bacteria incapable of growth in media containing 5 ppm mercury. Plasmid-curing assays done in this study ascertained that resistance to mercury antibiotics, and toxic xenobiotics is mediated by chromosomally borne genes and/or transposable elements rather than by plasmids.     

Tolerance of soil bacterial complexes to salt shock

Investigations showed that bacteria present in soil are resistant to one-day exposure to a saturated solution of ammonium nitrate and can well develop when transferred to laboratory nutrient media. The evaluated number of bacteria in NH4NO3-treated soil samples was nearly the same as in native soil samples, while was 1.5-2.5 times smaller in the former than in the latter case when microbial succession in the soil samples was initiated by wetting them. Bacteria (particularly gram-negative ones) occurring at the early stages of succession were the most sensitive to salt stress. Bacteria in soil were found to be much more resistant to salt stress than the same bacteria isolated in pure cultures.     

Toolbox Approaches Using Molecular Markers and 16S rRNA Gene Amplicon Data Sets for Identification of Fecal Pollution in Surface Water

In this study, host-associated molecular markers and bacterial 16S rRNA gene community analysis using high-throughput sequencing were used to identify the sources of fecal pollution in environmental waters in Brisbane, Australia. A total of 92 fecal and composite wastewater samples were collected from different host groups (cat, cattle, dog, horse, human, and kangaroo), and 18 water samples were collected from six sites (BR1 to BR6) along the Brisbane River in Queensland, Australia. Bacterial communities in the fecal, wastewater, and river water samples were sequenced. Water samples were also tested for the presence of bird-associated (GFD), cattle-associated (CowM3), horse-associated, and human-associated (HF183) molecular markers, to provide multiple lines of evidence regarding the possible presence of fecal pollution associated with specific hosts. Among the 18 water samples tested, 83%, 33%, 17%, and 17% were real-time PCR positive for the GFD, HF183, CowM3, and horse markers, respectively. Among the potential sources of fecal pollution in water samples from the river, DNA sequencing tended to show relatively small contributions from wastewater treatment plants (up to 13% of sequence reads). Contributions from other animal sources were rarely detected and were very small (<3% of sequence reads). Source contributions determined via sequence analysis versus detection of molecular markers showed variable agreement. A lack of relationships among fecal indicator bacteria, host-associated molecular markers, and 16S rRNA gene community analysis data was also observed. Nonetheless, we show that bacterial community and host-associated molecular marker analyses can be combined to identify potential sources of fecal pollution in an urban river. This study is a proof of concept, and based on the results, we recommend using bacterial community analysis (where possible) along with PCR detection or quantification of host-associated molecular markers to provide information on the sources of fecal pollution in waterways.     

Structure of bacterial communities in diverse freshwater habitats

The structures and dynamics of bacterial communities from raw source water, groundwater, and drinking water before and after filtration were studied in four seasons of a year, with culture-independent methods. Genomic DNA from water samples was analyzed by the polymerase chain reaction?- denaturing gradient gel electrophoresis system and by cloning of the 16S rRNA gene. Water samples exhibited complex denaturing gradient gel electrophoresis genetic profiles composed of many bands, corresponding to a great variety of bacterial taxa. The bacterial communities of different seasons from the four sampling sites clustered into two major groups: (i) water before and after filtration, and (ii) source water and groundwater. Phylogenetic analyses of the clones from the autumn sampling revealed 13 phyla, 19 classes, and 155 operational taxonomic units. Of the clones, 66% showed less than 97% similarities to known bacterial species. Representatives of the phyla Proteobacteria, Bacteroidetes, and Actinobacteria were found at all four sampling sites. Species belonging to the phylum Firmicutes were an important component of the microbial community in filtered water. Representatives of Enterobacteriaceae were not detected, indicating the absence of fecal pollution in the drinking water. Differences were found in the bacterial populations that were sampled from the same sites in different seasons. Each water habitat had a unique bacterial profile. Drinking water harbors diverse and dynamic microbial communities, part of which may be active and resilient to chlorine disinfection. This study provides, for the first time, basic data for uncultivable drinking water bacteria in Israel.     

Survival and transport of bacteria in egg washwater.

An evaluation of methods for monitoring the quality of water used to wash eggs at grading stations was undertaken to improve maintenance of bacterial viability during overnight sample transport. Bacterial content of samples at analysis would then better reflect conditions at the time eggs were washed. The interactive effects of temperature and the highly alkaline water conditions upon viability were the subjects of this study. Nine transport methods were examined for their efficacy in recovering total and coliform bacteria from recycled water used to wash eggs, and these were compared with samples analyzed at two commercial egg grading stations. Samples were shipped under test to the laboratory for analysis the following day. The survival of Staphylococcus aureus and Escherichia coli was also examined, but in a synthetic washwater matrix under various combinations of temperature (6 to 32 degrees C) and pH (9.5 to 10.5) to determine whether there was likely to be a different response to variations in transport treatment among gram-positive and -negative bacteria. S. aureus was much more resistant to the lethal effects of high pH and moderate temperature than E. coli. These results indicated that samples of high pH should be held (transported) at less than or equal to 13 degrees C to optimize bacterial survival. Considering cost, ease of manipulation, and the ability to protect both coliforms and the bacterial population as a whole, the method of choice for transport of industrial samples was the direct addition of washwater to containers in which powdered KH2PO4 and Na2S2O3 had been placed to yield final concentrations, when dissolved, of 0.2 and 0.05% (wt/vol), respectively.     

Survival and transport of bacteria in egg washwater

An evaluation of methods for monitoring the quality of water used to wash eggs at grading stations was undertaken to improve maintenance of bacterial viability during overnight sample transport. Bacterial content of samples at analysis would then better reflect conditions at the time eggs were washed. The interactive effects of temperature and the highly alkaline water conditions upon viability were the subjects of this study. Nine transport methods were examined for their efficacy in recovering total and coliform bacteria from recycled water used to wash eggs, and these were compared with samples analyzed at two commercial egg grading stations. Samples were shipped under test to the laboratory for analysis the following day. The survival of Staphylococcus aureus and Escherichia coli was also examined, but in a synthetic washwater matrix under various combinations of temperature (6 to 32 degrees C) and pH (9.5 to 10.5) to determine whether there was likely to be a different response to variations in transport treatment among gram-positive and -negative bacteria. S. aureus was much more resistant to the lethal effects of high pH and moderate temperature than E. coli. These results indicated that samples of high pH should be held (transported) at less than or equal to 13 degrees C to optimize bacterial survival. Considering cost, ease of manipulation, and the ability to protect both coliforms and the bacterial population as a whole, the method of choice for transport of industrial samples was the direct addition of washwater to containers in which powdered KH2PO4 and Na2S2O3 had been placed to yield final concentrations, when dissolved, of 0.2 and 0.05% (wt/vol), respectively.     

Microbiological investigation on black crusts from open-air stone monuments of Bologna (Italy)

Samples of black crusts taken from open-air stone monuments in Bologna dating back to between the twelfth and the nineteenth century were considered. The microbiological test procedures isolated from a high number of chemoorganotrophic bacteria, each of the samples although it was expected to find chemo-lithotrophic (specially sulfur-oxidizing and nitrifying) bacteria on account of the great amounts of sulphur, carbon, nitrogen dioxides and ammonia in the air. Sporogenous and asporogenous species were found among the isolated bacteria: the first were recognized as Bacillus subtilis, Bacillus brevis, Bacillus licheniformis, Bacillus mycoides and Bacillus megaterium; the latter were Corynebacterium glutamicum, Actinomyces sp., Alcaligenes denitrificans subsp. denitrificans, Flavobacterium breve, Acinetobacter calcoaceticus, Enterobacter cloacae, Pseudomonas stutzeri. These species were differently associated in the samples studied with a frequency of more than sixty-four per cent. This finding leads us to believe that their presence in the black crusts is not occasional, but the result of an ecologically significant process of colonization. Although behavioural tests (solubilization of calcium carbonate, of calcium sulphate and marble dust) did not reveal lithic activity among the bacteria studied, one can presume that their behaviour in nature - as a more or less complex microbial consortium - can be totally different and exert a decomposing action with regard to several minerals, most of them acidify the media in which they develop.