Diversity of integron- and culture-associated antibiotic resistance genes in freshwater floc

Clinically important antibiotic resistance genes were detected in culturable bacteria and class 1 integron gene cassettes recovered from suspended floc, a significant aquatic repository for microorganisms and trace elements, across freshwater systems variably impacted by anthropogenic activities. Antibiotic resistance gene cassettes in floc total community DNA differed appreciably in number and type from genes detected in bacteria cultured from floc. The number of floc antibiotic resistance gene cassette types detected across sites was positively correlated with total (the sum of Ag, As, Cu, and Pb) trace element concentrations in aqueous solution and in a component of floc readily accessible to bacteria. In particular, concentrations of Cu and Pb in the floc component were positively correlated with floc resistance gene cassette diversity. Collectively, these results identify suspended floc as an important reservoir, distinct from bulk water and bed sediment, for antibiotic resistance in aquatic environments ranging from heavily impacted urban sites to remote areas of nature reserves and indicate that trace elements, particularly Cu and Pb, are geochemical markers of resistance diversity in this environmental reservoir. The increase in contamination of global water supplies suggests that aquatic environments will become an even more important reservoir of clinically important antibiotic resistance in the future.     

High Diversity of Extended-Spectrum Beta-Lactamase-Producing Bacteria in an Urban River Sediment Habitat

Antibiotic-resistant bacteria (ARB) have been surveyed widely in water bodies, but few studies have determined the diversity of ARB in sediment, which is the most taxon-abundant habitat in aquatic environments. We isolated 56 extended-spectrum β-lactamase (ESBL)-producing bacteria from a single sediment sample taken from an urban river in China. All strains were confirmed for ESBL-producing capability by both the clavulanic acid combination disc method and MIC determination. Of the isolated strains, 39 were classified as Enterobacteriaceae (consisting of the genera Escherichia, Klebsiella, Serratia, and Aeromonas) by 16S rRNA gene sequencing and biochemical analysis. The present study identifies, for the first time, ESBL-producing strains from the families Brucellaceae and Moraxellaceae. The bla_CTX-M gene was the most dominant of the ESBL genes (45 strains), while the bla_TEM gene was the second-most dominant (22 strains). A total of five types of bla_CTX-M fragments were identified, with both known and novel sequences. A library of bla_CTX-M cloned from the sediment DNA showed an even higher diversity of bla_CTX-M sequences. The discovery of highly diverse ESBL-producing bacteria and ESBL genes, particularly bla_CTX, in urban river sediment raises alarms for potential dissemination of ARB in communities through river environments.Antibiotic-resistant bacteria (ARB) have been found widely in aquatic environments (1, 18, 24). ARB in rivers may originate from anthropogenic sources, such as hospital, municipal, and aquaculture effluents (3, 16, 23); in addition, they could occur naturally, since many acquired resistance mechanisms originated in producers of antibiotics, such as actinomycetes (12). Both anthropogenic and naturally occurring ARB in water environments may compromise human health, since people may be infected by ARB through drinking water, aquatic products, and direct contact with water bodies. Moreover, the ARB may transfer the antibiotic resistance genes to other pathogens through horizontal gene transfer (1).Sediment has the highest microbial diversity in water environments. The species richness and abundance of the sediment community are comparable to those of soil and are orders of magnitude higher than those of the planktonic community in the upper water layer (10, 20). It is reasonable to deduce that a wide variety of ARB might exist in sediment environments, as they are taxon-rich habitats, particularly in sediments receiving wastewater. Nevertheless, despite extensive studies surveying ARB in water columns, the diversity of antibiotic resistance in sediment environments has seldom been investigated.The present study therefore focused on an urban river sediment environment as a model, concentrating on the diversity of extended-spectrum β-lactamase (ESBL)-producing bacteria. ESBL-producing organisms have been emerging both in nosocomial and in community settings since the 1980s (4, 13, 14). In aquatic environments, ESBL-producing bacteria have been found in sewage and water samples (11, 13, 16), but their diversity in sediment habitats has never been analyzed before. In the present study, the diversity of ESBL-producing organisms was studied by both culture-dependent and culture-independent methods. A wide variety of ESBL-producing bacteria were isolated using a vast array of different nutrient media. The ESBL-producing abilities of the isolated strains were determined by both the clavulanic acid combination disc method and MIC determination. ESBL genes were identified and characterized by PCR and sequencing. A clone library of the bla_CTX-M gene, which was the most abundant type of ESBL gene found in the isolated bacteria, was cloned directly from the sediment and sequenced. The data revealed a diverse community of ESBL-producing bacteria that exists in urban aquatic sediment.     

Vertical distribution of diazotrophic bacterial community associated with temperature and oxygen gradients in a subtropical reservoir

Nitrogen-fixing microorganisms play important roles in the structure and function of aquatic ecosystems. However, the diversity and distribution of diazotrophic bacteria along the lake depth continuum are so far poorly understood. In this study, we investigated the dynamic variations of diazotrophs in a subtropical deep reservoir during the stratified period. We applied an in-depth biomolecular approach (DGGE, clone libraries, and quantitative real-time PCR) to explore the nitrogenase (nifH) gene diversity and abundance. The diazotrophic community shifted between the oxic/anoxic interface and the nifH diversity increased with depth. The Cyanobacteria, affiliated to the toxic bloom-forming Cylindrospermopsis raciborskii, were the dominant diazotrophic cluster in the surface waters, whereas diazotrophic Alphaproteobacteria were dominant in the bottom waters. The relationships between microbial and environmental factors clearly demonstrated that the temperature gradient and the oxygen concentration affect the heterogeneity of the diazotrophic community, thereby influencing the entire aquatic nitrogen cycle.     

Molecular diversity of the methanotrophic bacteria communities associated with disused tin-mining ponds in Kampar,Perak, Malaysia

In a previous study, notable differences of several physicochemical properties, as well as the community structure of ammonia oxidizing bacteria as judged by 16S rRNA gene analysis, were observed among several disused tin-mining ponds located in the town of Kampar, Malaysia. These variations were associated with the presence of aquatic vegetation as well as past secondary activities that occurred at the ponds. Here, methane oxidizing bacteria (MOB), which are direct participants in the nutrient cycles of aquatic environments and biological indicators of environmental variations, have been characterised via analysis of pmoA functional genes in the same environments. The MOB communities associated with disused tin-mining ponds that were exposed to varying secondary activities were examined in comparison to those in ponds that were left to nature. Comparing the sequence and phylogenetic analysis of the pmoA clone libraries at the different ponds (idle, lotus-cultivated and post-aquaculture), we found pmoA genes indicating the presence of type I and type II MOB at all study sites, but type Ib sequences affiliated with the Methylococcus/Methylocaldum lineage were most ubiquitous (46.7 % of clones). Based on rarefaction analysis and diversity indices, the disused mining pond with lotus culture was observed to harbor the highest richness of MOB. However, varying secondary activity or sample type did not show a strong variation in community patterns as compared to the ammonia oxidizers in our previous study.     

Antimicrobial and Antibiofilm Activity of Mauriporin,a Multifunctional Scorpion Venom Peptide

Many pathogenic free living and biofilm forming bacterial organisms can cause serious infections to humans that could consequently have devastating effects on human health. A significant number of these microbial organisms are resistant to almost all known conventional antibiotics and the ability of some these strains to form sessile communities of biofilms increases the resistance ability of bacteria to antibiotic treatment. Global research is currently focused on finding novel therapies to counteract the threat of bacterial and biofilm infections rather than using conventional antibiotics. Mauriporin, a novel cationic α-helical peptide identified from the venom derived cDNA library of the scorpion Androctonus mauritanicus was reported to display selective cytotoxic and anti-proliferative activity against prostate cancer cell lines. In the present study, we investigated the antimicrobial and antibiofilm activities of Mauriporin. Our results show that Mauriporin displays potent antimicrobial activities against a range of Gram-positive and Gram-negative planktonic bacteria with MIC values in the range 5 µM to 10 µM. Mauriporin was also able to prevent Pseudomonas aeruginosa biofilm formation while showing weak hemolytic activity towards human erythrocytes. Studies on the mechanism of action of Mauriporin revealed that the peptide is probably inducing bacterial cell death through membrane permeabilization determined by the release of β-galactosidase enzyme from peptide treated Escherichia coli cells. Moreover, DNA binding studies found that Mauriporin can cause potent binding to intracellular DNA. All these results indicate that Mauriporin has a considerable potential for therapeutic application as a novel drug candidate for eradicating bacterial infections.     

Macrozoobenthic Community and Assessment of Aquatic Ecosystem Health of three Waterbodies of East Calcutta Wetlands,India

The present study was aimed to understand the diversity, abundance and distribution of macrozoobenthic community of three selected fishponds differing in sewage intake and culture practices of East Calcutta Wetlands of Kolkata, India, as well as the aquatic ecosystem health of these waterbodies considering benthos as the best indicator of pollution. One year seasonal samplings were done to study the diversity and distribution of macrozoobenthos along with 12 water quality parameters to reflect the ecological conditions and aquatic ecosystem health of these waterbodies through biodiversity indices and statistical analysis (SPSS 10). A total of 27 species of macrozoobenthic organisms belonging to 18 families under four phyla comprising eight major groups viz. Polychaeta, Oligochaeta, Hirudinea, Diptera, Odonata, Gastropoda, Bivalvia and Pisces have been recorded. The diversity of macrozoobenthic fauna of these three waterbodies revealed that Ruby wetland had the highest representation of 24 species followed by Sukantanagar pond (17 species) and Sukantanagar bheri (16 species). The result of Pearson’s correlation coefficient showed that amongst the collected macrozoobenthos the gastropod species, Bellamya bengalensis and Lymnaea acuminata were wetland site-wise significant. The total benthic population and biomass have positive correlation with water transparency and organic enrichment of the waterbody. Shannon Weiner and Margalef’s indices have revealed higher values for Ruby wetland. Biodiversity indices and BMWP Score System designated to benthic organisms, suggested Ruby wetland is healthier than the other two waterbodies, differing in sewage intake and culture practices.     

Ecological determinants of methylmercury bioaccumulation in benthic invertebrates of polar desert lakes

We investigated concentrations of monomethylmercury (MMHg) at the base of benthic food webs in six lakes from polar desert (biologically poor and low annual precipitation) on Cornwallis Island (Nunavut, Canada, ~75°N latitude). Anthropogenic mercury emissions reach the Arctic by long-range atmospheric transport, and information is lacking on processes controlling MMHg entry into these simple lake food webs, despite their importance in determining transfer to lake-dwelling Arctic char. We examined the influences of diet (using carbon and nitrogen stable isotopes), water depth, and taxonomic composition on MMHg bioaccumulation in benthic invertebrates (Chironomidae and Trichoptera). We also estimated MMHg biomagnification between benthic algae and invertebrates. Similar MMHg concentrations of chironomid larvae in nearshore and offshore zones suggest that benthic MMHg exposure was homogeneous within the lakes. Chironomid δ¹³C values were also similar in both depth zones, suggesting that diet items with highly negative δ¹³C, specifically methanogenic bacteria and planktonic organic matter, were not important food (and therefore mercury) sources for profundal larvae. MMHg concentrations were significantly different among two subfamilies of chironomids (Diamesinae, Chironominae) and Trichoptera. Higher MMHg concentrations in Diamesinae were likely related to predation on other chironomids. We found high MMHg biomagnification between benthic algae and chironomid larvae compared with literature estimates for aquatic ecosystems at lower latitudes; thus, benthic processes may affect the sensitivity of polar desert lakes to mercury. Information on benthic MMHg exposure is important for evaluating and tracking impacts of atmospheric mercury deposition and environmental change in this remote High Arctic environment.     

Decreased thermal niche breadth as a trade-off of antibiotic resistance

Evolutionary theory predicts that adaptations, including antibiotic resistance, should come with associated fitness costs; yet, many resistance mutations seemingly contradict this prediction by inducing no growth rate deficit. However, most growth assays comparing sensitive and resistant strains have been performed under a narrow range of environmental conditions, which do not reflect the variety of contexts that a pathogenic bacterium might encounter when causing infection. We hypothesized that reduced niche breadth, defined as diminished growth across a diversity of environments, can be a cost of antibiotic resistance. Specifically, we test whether chloramphenicol-resistant Escherichia coli incur disproportionate growth deficits in novel thermal conditions. Here we show that chloramphenicol-resistant bacteria have greater fitness costs at novel temperatures than their antibiotic-sensitive ancestors. In several cases, we observed no resistance cost in growth rate at the historic temperature but saw diminished growth at warmer and colder temperatures. These results were consistent across various genetic mechanisms of resistance. Thus, we propose that decreased thermal niche breadth is an under-documented fitness cost of antibiotic resistance. Furthermore, these results demonstrate that the cost of antibiotic resistance shifts rapidly as the environment changes; these context-dependent resistance costs should select for the rapid gain and loss of resistance as an evolutionary strategy.Subject terms: Bacterial evolution, Microbial ecology, Antibiotics     

Bacterial diversity assessment in soil of an active Brazilian copper mine using high-throughput sequencing of 16S rDNA amplicons

Mining activities pose severe environmental risks worldwide, generating extreme pH conditions and high concentrations of heavy metals, which can have major impacts on the survival of organisms. In this work, pyrosequencing of the V3 region of the 16S rDNA was used to analyze the bacterial communities in soil samples from a Brazilian copper mine. For the analysis, soil samples were collected from the slopes (geotechnical structures) and the surrounding drainage of the Sossego mine (comprising the Sossego and Sequeirinho deposits). The results revealed complex bacterial diversity, and there was no influence of deposit geographic location on the composition of the communities. However, the environment type played an important role in bacterial community divergence; the composition and frequency of OTUs in the slope samples were different from those of the surrounding drainage samples, and Acidobacteria, Chloroflexi, Firmicutes, and Gammaproteobacteria were responsible for the observed difference. Chemical analysis indicated that both types of sample presented a high metal content, while the amounts of organic matter and water were higher in the surrounding drainage samples. Non-metric multidimensional scaling (N-MDS) analysis identified organic matter and water as important distinguishing factors between the bacterial communities from the two types of mine environment. Although habitat-specific OTUs were found in both environments, they were more abundant in the surrounding drainage samples (around 50 %), and contributed to the higher bacterial diversity found in this habitat. The slope samples were dominated by a smaller number of phyla, especially Firmicutes. The bacterial communities from the slope and surrounding drainage samples were different in structure and composition, and the organic matter and water present in these environments contributed to the observed differences.     

Phylogenetic and functional prokaryotic diversity in the Hoito-Gol mesothermal mineral spring (Eastern Sayan Mountains,Buryat Republic)

High-throughput sequencing was used for comparative analysis of microbial communities of the water and mat from the Hoito-Gol mesothermal mineral sulfide spring (Eastern Sayan Mountains, Buryat Republic). Activity of microbial communities was determined. While both spring biotopes were dominated by members of three bacterial phyla—Proteobacteria, Bacteroidetes, and Firmicutes—they differed drastically in the composition of predominant phylotypes (at the genus level). In the water, the organisms widespread in aquatic environments were predominant, mostly aerobic chemoorganotrophs of the genera Acinetobacter, Pedobacter, and Flavobacterium. In the microbial mat, the organisms actively involved in the sulfur cycle predominated, including sulfur-reducing bacteria Sulfurospirillum, sulfate-reducing deltaproteobacteria, sulfuroxidizing chemoautotrophic bacteria, anoxygenic phototrophic bacteria of the phyla Chloroflexi and Chlorobi, as well as purple bacteria belonging to the α-, ß-, and γ-Proteobacteria. Microbial mats of the spring exhibited higher phylogenetic diversity compared to high-temperature mats containing photosynthetic microorganisms.     

Unusual pathogenic bacterium isolated from microbial communities of bioaerosols at Chilean Patagonian lakes

Bioaerosols are transported from warm regions and lower latitudes of the planet to colder regions and higher latitudes, such as the Chilean Patagonia. The role of bioaerosols deposition in remote lake ecosystems is a potentially important process, but it has not yet been fully studied. The aim of this study was to detect and characterize potentially pathogenic viable microorganisms in bioaerosols in a pristine area. Samples were collected from the air, at three remote lakes in the Chilean Patagonia, using a sterile filtration system equipped with 0.2-μm-pore-size nitrocellulose filters. The bacterial community present in bioaerosols was studied using scanning electron microscopy (SEM) and denaturing gradient gel electrophoresis. Isolates were identified and characterized for phenotypic and 16S rDNA analysis and antibiotic resistance. SEM observations of samples from each lake showed the presence of bacteria with different morphologies, and after culturing, the identification results revealed that they were strains of Acinetobacter, Alcaligenes, Edwarsiella, Pseudomonas, Burklolderia, Moraxella, Sphingomonas and CDC NO-1. CDC NO-1, uncommonly isolated worldwide, stands out from the rest of the isolates because it is a rarely found bacterium so far associated with dog and cat bites and was found at two out of three pristine lakes studied (Alto and Verde). This work demonstrates, for the first time, the presence of CDC NO-1, a clinically important Gram-negative microorganism, in bioaerosols and first report of CDC-NO1 isolation in Chile. Besides its presence in remote lakes, its antibiotic resistance is worth mentioning.     

Isolation of a newLegionella species from thermally altered waters

Legionella-like bacteria were isolated from thermally altered water in Pennsylvania and Minnesota. These organisms were isolated from tissues of infected guinea pigs after intraperitoneal inoculation of water concentrates. While the cultural and morphological characteristics indicated the isolates wereLegionella, they did not react with antibodies prepared against known species of the genus. Antisera prepared against one of the isolates reacted maximally with the other ten isolates. Fluorescent antibody analysis of water concentrates from geographically disparate sites indicated that the environmental distribution is broad and that concentrations of the Oak Ridge strain ofLegionella were similar to those of serogroups 1, 2, 3, and 4 ofLegionella pneumophila.     

Bacteriophage Diversity in the North Sea

In recent years interest in bacteriophages in aquatic environments has increased. Electron microscopy studies have revealed high numbers of phage particles (10⁴ to 10⁷ particles per ml) in the marine environment. However, the ecological role of these bacteriophages is still unknown, and the role of the phages in the control of bacterioplankton by lysis and the potential for gene transfer are disputed. Even the basic questions of the genetic relationships of the phages and the diversity of phage-host systems in aquatic environments have not been answered. We investigated the diversity of 22 phage-host systems after 85 phages were collected at one station near a German island, Helgoland, located in the North Sea. The relationships among the phages were determined by electron microscopy, DNA-DNA hybridization, and host range studies. On the basis of morphology, 11 phages were assigned to the virus family Myoviridae, 7 phages were assigned to the family Siphoviridae, and 4 phages were assigned to the family Podoviridae. DNA-DNA hybridization confirmed that there was no DNA homology between phages belonging to different families. We found that the 22 marine bacteriophages belonged to 13 different species. The host bacteria were differentiated by morphological and physiological tests and by 16S ribosomal DNA sequencing. All of the bacteria were gram negative, facultatively anaerobic, motile, and coccoid. The 16S rRNA sequences of the bacteria exhibited high levels of similarity (98 to 99%) with the sequences of organisms belonging to the genus Pseudoalteromonas, which belongs to the γ subdivision of the class Proteobacteria.The marine bacterial community is responsible for a considerable portion of primary production and regeneration of nutrients in the microbial loop and is associated with a great variety of marine bacteriophages (5, 12). These phages are capable of infecting a large portion of the bacterioplankton (32, 34). It is assumed that as part of the marine food web, bacteriophages play important quantitative and qualitative roles in controlling marine bacterial populations (8, 24, 34, 39, 45). The phenotypic diversity and genotypic diversity of the phage populations are related to the interaction between phages and their host organisms, which provides a tool for understanding the interaction itself (13). To estimate the influence of marine bacteriophages on the diversity of bacterioplankton, we investigated phage diversity. The virus species concept proposed by Murphy et al. (37) delineates seven different families of bacteriophages based on morphological criteria and provides criteria for new phage species based on several traits, such as DNA homologies, serological data, protein profiles, and host ranges.In this paper, we describe the diversity and genetic relationships of marine phages based on investigations of 22 representatives from 85 phage-host systems (35, 36) collected between 1988 and 1992 from waters around an island, Helgoland, located in the North Sea. All of the phages were virulent and formed plaques on their host bacteria. We assigned the phages to different virus families, species, and strains based on morphology, DNA homology, and host range. Furthermore, we characterized the phenotypic and genotypic features of the host bacteria.     

Toxicity of zinc to schistosoma Mansoni Cercariae in a Chemically Defined Water Medium

Time-until-death studies were run on cercariae of Schistosoma mansoni in 15 concentrations of zinc (from zinc sulfate) ranging from 57.6 ppm to 1 ppb. A chemically defined water medium was used for the dilution and control medium, and a partially enclosed slide chamber was perfected for the detailed observation of test cercariae. In concentrations of less than 57.6 ppm, zinc was found to have little lethal effect on cercariae during that period in which these larvae were most likely to be infective (0–6 hours) after emergence. Zinc concentration of 1 ppb killed all test cercariae within 46 hours at 21–22°C.At the highest concentration tested, 57.6 ppm zinc immobilized all test organisms within 6 hours at 21–2°C. The average time for 100 percent mortality for the control was 49 hours. The cercaricidal effects of zinc were shown to be insignificant in concentrations that would not be detrimental to other aquatic organisms.     

New antibiotics produced by Bacillus subtilis strains

Two Bacillus subtilis strains isolated from the fruiting body of a basidiomycete fungus Pholiota squarrosa exhibited a broad range of antibacterial activity, including those against methicillin-resistant Staphylococcus aureus INA 00761 (MRSA) and Leuconostoc mesenteroides VKPM B-4177 resistant to glycopeptide antibiotics, as well as antifungal activity. The strains were identified as belonging to the “B. subtilis” complex based on their morphological and physiological characteristics, as well as by sequencing the 16S rRNA gene fragments. Both strains (INA 01085 and INA 01086) produced insignificant amounts of polyene antibiotics (hexaene and pentaene, respectively). Strain INA 01086 also produced a cyclic polypeptide antibiotic containing Asp, Gly, Leu, Pro, Tyr, Thr, Trp, and Phe, while the antibiotic of strain INA 01085 contained, apart from these, two unidentified nonproteinaceous amino acids. Both polypeptide antibiotics were new compounds efficient against gram-positive bacteria and able to override the natural bacterial antibiotic resistance.     

Toxicity of gold nanoparticles for plants in experimental aquatic system

Increased production and use of nanomaterials can lead to new types of pollution of the environment, including aquatic ecosystems. Pollution of the aqueous environment with nanoparticles can be a new type of pollution of the environment. This requires a more detailed study of the biological effects during exposure of nanoparticles on aquatic organisms. The interactions of gold nanoparticles (Au) with aquatic macrophytes Ceratophyllum demersum have been studied. Aquatic microcosms with these plants were used. Gold nanoparticles (Au) were added to the aqueous medium of C. demersum macrophyte containing microcosms. The state of the plants was then analyzed. Phytotoxicity of Au nanoparticles for aquatic macrophytes was shown for the first time. A new method of phytotoxicity detection was suggested and successfully approved. Phytotoxicity at a concentration of Au (in the form of nanoparticles) of 6 × 10^?6 M-1.8 × 10^?5 M was shown.     

Determination of total antibiotic resistance in waterborne bacteria in rivers and streams in Northern Ireland: Can antibiotic-resistant bacteria be an indicator of ecological change?

Antibiotic resistance (ABR) has now become a major public health issue. Relatively little studies have been published on the incidence of ABR in environmental isolates circulating within the community. A study was performed to determine the diversity of total ABR (intrinsic + acquired resistance) in waterborne bacteria. Surface water from 12 waterways, including 11 rivers/steams and 1 lake, were examined for the presence of ABR phenotypes, using a direct antibiotic susceptibility assay and demonstrated the presence of ABR (in increasing order of resistance), to the following 19 agents: amikacin (17%), gentamicin (17%), ciprofloxacin (33%), colistin (42%), linezolid (42%), tobramycin (42%), vancomycin (42%), ertapenem (67%), erythromycin (75%), meropenem (75%), rifampin (75%), teichoplanin (75%), tetracycline (75%), trimethoprim (75%), fusidic acid (83%), aztreonam (92%), clindamycin (92%), penicillin (92%) and cefoxitin (100%). Multiple resistance to the major classes of antibiotics was noted, which varied from one to six classes, with a mean resistance to 3.7 major antibiotics classes, with diminishing antibacterial effectiveness in the following order: aminoglycosides > fluoroquinolones > glycopeptides > macrolides > tetracyclines > β-lactams. Overall, these data indicate that waterborne bacteria are an important source of ABR determinants and contribute to the mass balance of ABR in the environment, and may be used as an indicator of ecological change in water ecosystems. The waterborne ABR organisms may potentially act as donors in pathogens, which may acquire these through horizontal gene transfer or other genetic exchange events, thus leading to clinically significant cases in both animal and human health. Therefore, environmental bacteria should not be regarded as being devoid of ABR determinants, simply because they are physically removed from clinical settings. Such bacteria have natural intrinsic resistance, as well as having the ability to acquire determinants from agricultural run-off and human wastewater discharge, which may contain ABR organisms, as well as sublethal concentrations of metabolically active antibiotic. The tracking of such organisms to their source may help determine the source of fecal pollution in aquatic ecosystems.     

Metal tolerance assisted antibiotic susceptibility profiling in <Emphasis Type="Italic">Comamonas acidovorans</Emphasis>

Metal ions are known selective agents for antibiotic resistance and frequently accumulate in natural environments due to the anthropogenic activities. However, the action of metals that cause the antibiotic resistance is not known for all bacteria. The present work is aimed to investigate the co-selection of metals and antibiotic resistance in Comamonas acidovorans. Tolerance profile of 16 metals revealed that the strain could tolerate high concentrations of toxic metals i.e., Cr (710 ppm), As (380 ppm), Cd (320 ppm), Pb (305 ppm) and Hg (205 ppm). Additionally, metal tolerant phenotypes were subjected to antibiotic resistance profiling; wherein several metal tolerant phenotypes (Cr 1.35-fold; Co-1.33 fold; Mn-1.29 fold) were resistant, while other metal tolerant phenotypes (Mg 1.32-fold; Hg 1.29-fold; Cu 1.28-fold) were susceptible than control phenotype. Metal accumulation may alter the metabolism of C. acidovorans that activates or inactivates the genes responsible for antibiotic resistance, resulting in the resistance and/or susceptibility pattern observed in metal resistant phenotypes.     

Protozoan Predation Is Differentially Affected by Motility of Enteric Pathogens in Water vs. Sediments

Survival of enteric bacteria in aquatic habitats varies depending upon species, strain, and environmental pressures, but the mechanisms governing their fate are poorly understood. Although predation by protozoa is a known, top-down control mechanism on bacterial populations, its influence on the survival of fecal-derived pathogens has not been systematically studied. We hypothesized that motility, a variable trait among pathogens, can influence predation rates and bacterial survival. We compared the survival of two motile pathogens of fecal origin by culturing Escherichia coli O157 and Salmonella enterica Typhimurium. Each species had a motile and non-motile counterpart and was cultured in outdoor microcosms with protozoan predators (Tetrahymena pyriformis) present or absent. Motility had a significant, positive effect on S. enterica levels in water and sediment in the presence or absence of predators. In contrast, motility had a significant negative effect on E. coli O157 levels in sediment, but did not affect water column levels. The presence/absence of protozoa consistently accounted for a greater proportion of the variability in bacterial levels (>95 %) than in bacterial motility (<4 %) in the water column. In sediments, however, motility was more important than predation for both bacteria. Calculations of total CFU/microcosm showed decreasing bacterial concentrations over time under all conditions except for S. enterica in the absence of predation, which increased ～0.5–1.0 log over 5 days. These findings underscore the complexity of predicting the survival of enteric microorganisms in aquatic habitats, which has implications for the accuracy of risk assessment and modeling of water quality.