Increased waterborne blaNDM-1 resistance gene abundances associated with seasonal human pilgrimages to the upper ganges river.

Antibiotic resistance (AR) is often rooted in inappropriate antibiotic use, but poor water quality and inadequate sanitation exacerbate the problem, especially in emerging countries. An example is increasing multi-AR due to mobile carbapenemases, such as NDM-1 protein (coded by blaNDM-1 genes), which can produce extreme drug-resistant phenotypes. In 2010, NDM-1 positive isolates and blaNDM-1 genes were detected in surface waters across Delhi and have since been detected across the urban world. However, little is known about blaNDM-1 levels in more pristine locations, such as the headwaters of the Upper Ganges River. This area is of particular interest because it receives massive numbers of visitors during seasonal pilgrimages in May/June, including visitors from urban India. Here we quantified blaNDM-1 abundances, other AR genes (ARG), and coliform bacteria in sediments and water column samples from seven sites in the Rishikesh-Haridwar region of the Upper Ganges and five sites on the Yamuna River in Delhi to contrast blaNDM-1 levels and water quality conditions between season and region. Water quality in the Yamuna was very poor (e.g., anoxia at all sites), and blaNDM-1 abundances were high across sites in water (5.4 ± 0.4 log(blaNDM-1·mL(-1)); 95% confidence interval) and sediment (6.3 ± 0.7 log(blaNDM-1·mg(-1))) samples from both seasons. In contrast, water column blaNDM-1 abundances were very low across all sites in the Upper Ganges in February (2.1 ± 0.6 log(blaNDM-1·mL(-1))), and water quality was good (e.g., near saturation oxygen). However, per capita blaNDM-1 levels were 20 times greater in June in the Ganges water column relative to February, and blaNDM-1 levels significantly correlated with fecal coliform levels (r = 0.61; p = 0.007). Given that waste management infrastructure is limited in Rishikesh-Haridwar, data imply blaNDM-1 levels are higher in visitor's wastes than local residents, which results in seasonally higher blaNDM-1 levels in the river. Pilgrimage areas without adequate waste treatment are possible "hot spots" for AR transmission, and waste treatment must be improved to reduce broader AR dissemination via exposed returning visitors.

Nitrification in hybrid bioreactors treating simulated domestic wastewater.

AIM: To provide deeper insights into nitrification process within aerobic bioreactors containing supplemental physical support media (hybrid bioreactors). METHODS AND RESULTS: Three bench-scale hybrid bioreactors with different media size and one control bioreactor were operated to assess how biofilm integrity influences microbial community conditions and bioreactor performance. The systems were operated initially at a 5-day hydraulic retention time (HRT), and all reactors displayed efficient nitrification and chemical oxygen demand (COD) removal (>95%). However, when HRT was reduced to 2.5 days, COD removal rates remained high, but nitrification efficiencies declined in all reactors after 19 days. To explain reduced performance, nitrifying bacterial communities (ammonia-oxidizing bacteria, AOB; nitrite-oxidizing bacteria, NOB) were examined in the liquid phase and also on the beads using qPCR, FISH and DGGE. Overall, the presence of the beads in a reactor promoted bacterial abundances and diversity, but as bead size was increased, biofilms with active coupled AOB-NOB activity were less apparent, resulting in incomplete nitrification. CONCLUSIONS: Hybrid bioreactors have potential to sustain effective nitrification at low HRTs, but support media size and configuration type must be optimized to ensure coupled AOB and NOB activity in nitrification. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that AOB and NOB coupling must be accomplished to minimize nitrification failure.

Cryptic striations in the upper mantle revealed by hafnium isotopes in southeast Indian ridge basalts.

The Earth's mantle is isotopically heterogeneous on length scales ranging from centimetres to more than 10(4) kilometres. This heterogeneity originates from partial melt extraction and plate tectonic recycling, whereas stirring during mantle convection tends to reduce it. Here we show that mid-ocean ridge basalts from 2,000 km along the southeast Indian ridge (SEIR) display a bimodal hafnium isotopic distribution. This bimodality reveals the presence of ancient compositional striations (streaks) in the Indian Ocean upper mantle. The number density of the streaks is described by a Poisson distribution, with an average thickness of approximately 40 km. Such a distribution is anticipated for a well-stirred upper mantle, in which heterogeneity is continually introduced by plate tectonic recycling, and redistributed by viscous stretching and convective refolding.

Feasibility tests for treating shampoo and hair colorant wastewaters using anaerobic processes.

Wastes from the personal care product (PCP) industry are often high in biodegradable carbon, which makes them amenable to aerobic biological treatment, although process costs are usually high due to aeration inefficiencies, high electricity demand and production of large amounts of sludge. As such, anaerobic treatment technologies are being considered to lower net energy costs by reducing air use and increasing methane production. To assess the amenability of PCP wastes to anaerobic treatment, methane yields and rates were quantified in different anaerobic reactors treating typical PCP wastes, including wastes from shampoo and hair colorant products. Overall, shampoo wastes were more amenable to methanogenesis with almost double the methane yields compared with colour wastes. To assess relevant microbial guilds, qPCR was performed on reactor biomass samples. Methanosaetaceae abundances were always significantly higher than Methanosarcinaceae and Methanomicrobiales abundances (P < 0.05), and did not differ significantly between waste types. Although colour wastes were less amenable to anaerobic treatment than shampoo wastes, differences cannot be explained by relative microbial abundances and probably result from the presence of inhibiting compounds in hair colorants (e.g., oxidants) at higher levels. Results showed that anaerobic technologies have great potential for treating PCP wastes, but additional work is needed to establish the basis of elevated methane yields and inhibition, especially when colorant wastes are present.

Mercury levels in sediments and mangrove oysters, Crassostrea rizophorae, from the north coast of Villa Clara, Cuba.

Total mercury levels were quantified in sediments and oyster tissues (Crassostrea rizophorae) from the Sagua la Grande River estuary and offshore mangrove keys 19 km downstream of a chlor-alkali plant (CAP) in Villa Clara, Cuba. Relatively elevated total mercury levels were found in sediments from the estuary itself, ranging from 0.507 to 1.81 µg g(-1) dry weight. However, levels were lower in sediments from the keys farther from the estuary. Oyster mercury levels were always acceptable for human consumption, although levels significantly correlated in sediments and oysters across sampling sites (p < 0.05), which suggests that mercury from the CAP is impacting coastal water quality conditions.

Magmatic and amagmatic seafloor generation at the ultraslow-spreading Gakkel ridge, Arctic Ocean.

A high-resolution mapping and sampling study of the Gakkel ridge was accomplished during an international ice-breaker expedition to the high Arctic and North Pole in summer 2001. For this slowest-spreading endmember of the global mid-ocean-ridge system, predictions were that magmatism should progressively diminish as the spreading rate decreases along the ridge, and that hydrothermal activity should be rare. Instead, it was found that magmatic variations are irregular, and that hydrothermal activity is abundant. A 300-kilometre-long central amagmatic zone, where mantle peridotites are emplaced directly in the ridge axis, lies between abundant, continuous volcanism in the west, and large, widely spaced volcanic centres in the east. These observations demonstrate that the extent of mantle melting is not a simple function of spreading rate: mantle temperatures at depth or mantle chemistry (or both) must vary significantly along-axis. Highly punctuated volcanism in the absence of ridge offsets suggests that first-order ridge segmentation is controlled by mantle processes of melting and melt segregation. The strong focusing of magmatic activity coupled with faulting may account for the unexpectedly high levels of hydrothermal activity observed.

Mantle plume helium in submarine basalts from the galapagos platform.

Helium-3/helium-4 ratios in submarine basalt glasses from the Galapagos Archipelago range up to 23 times the atmospheric ratio in the west and southwest. These results indicate the presence of a relatively undegassed mantle plume at the Galápagos hot spot and place Galápagos alongside Hawaii, Iceland, and Samoa as the only localities known to have such high helium-3/helium-4 ratios. Lower ratios across the rest of the Galápagos Archipelago reflect systematic variations in the degree of dilution of the plume by entrainment of depleted material from the asthenosphere. These spatial variations reveal the dynamics of the underlying mantle plume and its interaction with the nearby Galápagos Spreading Center.

Assessment of total mercury levels in Clarias gariepinus from the Sagua la Grande River, Cuba.

Total mercury levels (Thg) were quantified in Clarias gariepinus captured from the Sagua la Grande River (Cuba) in the vicinity of an active chlor-alkali plant, and relationships among place of capture; fish size, weight, and sex; and THg levels were assessed. THg levels ranged from 67 to 375 ng/g ww in collected fish, never exceeding the Cuban recommended maximum limit for fish consumption of 500 ng/g ww. No significant correlation was observed between mercury levels and fish allometric characteristics (p < 0.05); however, levels were significantly higher in fish captured below the chlor-alkali facility, suggesting a connection between mercury bioaccumulation and plant discharges.

Field assessment of oxytetracycline exposure to the freshwater macrophytes Egeria densa Planch. and Ceratophyllum demersum L.

In a microcosm study, two aquatic macrophytes, Egeria densa and Ceratophyllum demersum were exposed to nominal concentrations of 0, 5, 20, 50, and 250 microg/L oxytetracycline (n=3), plus 20 microg/L oxytetracycline amended with additional nitrogen (N) and phosphorus (P). Responses were monitored bi-weekly over a six-week exposure period. Both plant species exhibited a significant decline in growth in the 250 microg/L oxytetracycline and the N- and P-amended units. Decreased light penetration resulting from accumulating oxytetracycline by-products appears to be the primary modifier in the growth of these plants. Increased susceptibility to oxytetracycline exposure was noted in some paired plantings (e.g., E. densa root development), relative to individual plants in these treatments, however, no clear explanation for this response is available. Based on the toxicity data generated in this study, we estimate that current concentrations of oxytetracycline in freshwater environments do not pose a direct risk to E. densa and C. demersum.

Inhibitors of folate biosynthesis. 1. Inhibition of dihydroneopterin aldolase by pteridine derivatives.

2-Amino-6-carboxamido-7,8-dihydropteridin-4-one and 2-amino-6-hydroxymethyl-7,7-dimethyl-7,8-dihydropteridin-4-one have been shown to be good inhibitors of Escherichia coli dihydroneopterin aldolase, an early enzyme of de novo folate biosynthesis.

Inhibition of the mammalian beta-lactamase renal dipeptidase (dehydropeptidase-I) by (Z)-2-(acylamino)-3-substituted-propenoic acids.

The title enzyme deactivates the potent carbapenem antibiotic imipenem in the kidney, producing low antibiotic levels in the urinary tract. A series of (Z)-2-(acylamino)-3-substituted-propenoic acids (3) are specific, competitive inhibitors of the enzyme capable of increasing the urinary concentration of imipenem in vivo. Many of the compounds were prepared in one step from an alpha-keto acid and a primary amide. The optimum R2 groups are 2,2-dimethyl, -dichloro, and -dibromocyclopropyl. With R2 = 2,2-dimethylcyclopropyl (DMCP), a wide variety of R3 groups including alkyl, oxa- and thiaalkyl, and alkyl groups containing acidic, basic, and neutral substituents give effective inhibitors with Ki values of 0.02-1 microM and a range of pharmacokinetic properties. By resolution of enantiomers and X-ray crystallography, the enzyme-inhibitory activity of the DMCP group was found to reside with the 1S isomer. The cysteinyl compound 176 (cilastatin, MK-0791) has the desired pharmacological properties and has been chosen for combination with imipenem.

Benzylated 1,2,3-triazoles as anticoccidiostats.

Substituted 5-amino-4-carbamoyl-1,2,3-triazoles 3a-w were prepared by two synthetic schemes and evaluated in vivo for anticoccidial activity. Both schemes proceeded by brominating appropriately substituted toluenes 4a-s,v to 5a-s,v. In Scheme I, the brominated benzyl analogues 5 were converted to the corresponding benzyl azides 6, which were treated with cyanoacetamide to yield 1-substituted-5-amino-4-carbamoyl-1,2,3-triazoles 3. In Scheme II, the benzyl halides 5 were employed to alkylate the sodium salt of 5-amino-4-carbamoyl-1,2,3-triazole (7). Preliminary screening data against Eimeria acervulina and E. tenella in chickens suggested structural requirements for maximizing activity. Further evaluation against a relatively resistant series of eight Eimeria field isolates revealed L-651,582 (3a) to be a highly effective coccidiostat. However, unacceptable tissue residues precluded further development. Mechanistic studies on this series of 5-amino-4-carbamoyl-1,2,3-triazoles and, in particular, on L-651,582 (3a) revealed that its mode of action does not involve inhibition of IMP dehydrogenase, but probably interferes with host cell calcium entry. In addition, L-651,582 has been found to have antiproliferative activity in several disease models and was recently reported to possess antimetastatic activity in a model of ovarian cancer progression.

Effect of oxygen level on simultaneous nitrogenase and sMMO expression and activity in Methylosinus trichosporium OB3b and its sMMO(C) mutant, PP319: aerotolerant N2 fixation in PP319.

Soluble methane monooxygenase (sMMO) expression and activity were monitored under conditions that either promoted or suppressed the expression of nitrogenase in Methylosinus trichosporium OB3b wild-type (WT) and in its sMMO-constitutive mutant, PP319. Both WT and mutant cultures had reduced sMMO activity and protein levels under elevated O2 conditions (188 microM) compared with low O2 conditions (24 microM). Simultaneous N2 fixation also reduced sMMO activity in both cultures when O2 was low. However, when O2 levels were increased, nitrogenase expression ceased and sMMO activity was reduced by approximately 77% in the WT, whereas sMMO and nitrogenase expression and activity in PP319 were relatively unaffected by the higher O2 levels. Western immunoblot analysis showed that the nitrogenase Fe protein resolved as two components (apparent molecular mass of 30.5 and 32 kDa) in both the WT and PP319 when O2 levels were low. When O2 levels were high, only the 32-kDa form of the Fe protein was present in PP319, whereas neither form was detectable in the WT. Aerotolerant N2 fixation appears to be associated with the 32-kDa Fe protein in M. trichosporium OB3b.

Anticoccidial and tolerance studies in the chicken with two 6-amino-9-(substituted benzyl)purines.

Initial assays of 6-amino-9-(2-chloro-6-fluorobenzyl) purine (MK-302) and 6-amino-9-(2,6-dichlorobenzyl)purine (coded L-628,914) showed potential as anticoccidial agents on the basis of broad-spectrum activity and safety. In battery efficacy studies, dietary levels of 60 to 70 p.p.m. and above MK-302 and 45 to 60 p.p.m. L-628,914 proved to have excellent broad-spectrum anticoccidial activity in chickens given heavy exposure to virulent field isolates of coccidia. Eight-week floor-pen tolerance trials showed that the maximum tolerated diet concentration (MTC) of MK-302 was approximately 95 p.p.m. while the MTC of L-628,914 was approximately 60 p.p.m. Dietary relationships (p.p.m. MK-302:p.p.m. L-628,914 for equivalent effects) derived from the efficacy and tolerance results were 1.2:1 and 1.6:1 respectively and clearly demonstrated a higher therapeutic ratio for MK-302.

Low-energy treatment of colourant wastes using sponge biofilters for the personal care product industry.

Four trickling biofilter designs were assessed as low-energy alternatives to aerobic activated sludge (AS) for the treatment of personal care product industry wastes. The designs included partially submerged packed-media and sponge reactors with and without active aeration. Partial submergence was used to reduce active aeration needs. Simulated colourant wastes (up to COD=12,480 mg/L, TN=128 mg/L) were treated for 201 days, including wastes with elevated oxidant levels. COD and TN removal efficiencies were always >79% and >30% (even without aeration). However, aerated sponge reactors consistently had the highest removal efficiencies, especially for TN (∼60%), and were most tolerant of elevated oxidants. This study shows sponge biofilters have great potential for treating colourant wastes because they achieve high treatment efficiencies and reduce energy use by >40% relative to AS systems.

Anaerobic-aerobic sequencing bioreactors improve energy efficiency for treatment of personal care product industry wastes.

Personal care product (PCP) industry liquid wastes contain shampoo residues, which are usually treated by aerobic activated sludge (AS). Unfortunately, AS is expensive for PCP wastes because of high aeration and energy demands, whereas potentially energy-positive anaerobic designs cannot meet effluent targets. Therefore, combined anaerobic-aerobic systems may be the best solution. Seven treatment systems were assessed in terms of energy and treatment performance for shampoo wastes, including one aerobic, three anaerobic (HUASB, AHR and AnCSTR) and three anaerobic-aerobic reactor designs. COD removals were highest in the HUASB-aerobic (87.9 ± 0.4%) and AHR-aerobic (86.8±0.5%) systems, which used 69.2% and 62.5% less energy than aerobic AS. However, actual methane production rates were low relative to theoretical in the UASB and AHR units (∼10% methane/COD removed) compared with the AnCSTR unit (∼70%). Anaerobic-aerobic sequence reactors show promise for treating shampoo wastes, but optimal designs depend upon whether methane production or COD removal is most important to operations.

A comparative assessment of molecular biological and direct microscopic techniques for assessing aquatic systems.

Molecular techniques have clear value for community characterization; however, almost all previous datasets are based upon non-molecular measurements and it is hard to compare "old" data with "new" data because few correlations have been made. Therefore, the purpose of this evaluation was to simultaneously use molecular and non-molecular methods within the same sampling program to determine how data compare. Three methods were used for characterizing microbial populations in Lake 260 (L260) at Experimental Lakes Area (Ontario, Canada) during a whole-lake exposure study. Methods included whole-cell microscopic counts (for bacteria, cyanobacteria, algae and zooplankton), chlorophyll a, and small sub-unit (ssu)-rRNA hybridization using EUB-338, EUB-785, CYAN-785-a/b, EUCA-1379 and UNIV-1390 gene probes. Strong correlations were found between the EUB-785 probe signal and "bacteria minus cyanobacteria" direct counts, and the EUB-338 probe signal and "bacteria plus cyanobacteria" counts. Furthermore, the difference in probe signal between EUB-338 and EUB-785 (a presumptive signature for cyanobacteria and plastids) correlated with cyanobacterial direct counts and also with chlorophyll a. However, EUCA-1379 probe signal did not correlate with algae counts, and UNIV-1390 probe signal only correlated with total bacteria counts. Results suggest that, although ssu-rRNA methods are fast, reproducible, and specifically detect "viable" organisms, their use may be limited to non-eukaryotic populations unless new probes are developed that are more specific.

Water hyacinths (Eichhornia crassipes) as indicators of heavy metal impact of a large landfill on the Almendares River near Havana, Cuba.

The Almendares River is central to recreational and other activities in Havana, Cuba. However, monitoring indicated significant heavy metal contamination in river sediments, especially below Calle 100, the largest landfill in Havana. This work extended previous sediment studies by determining complementary Cu, Pb, Ni, Cr, Cd, and Zn levels in indigenous water hyacinths (Eichhornia crassipes; EC) above and below the landfill. Pb, Cu, and Zn were significantly elevated in EC roots below the landfill and also correlated with sediment data (p < 0.05), implying elevated levels likely result from landfill activity and might be useful biomonitors as river remediation proceeds.

Fate and effects of enrofloxacin in aquatic systems under different light conditions.

The fate and effects of fluoroquinolone antibacterials (FQ) in the environment is of significance because of apparent increased FQ resistance in environmental and clinical organisms. Here we simultaneously assessed the fate and effects of enrofloxacin (enro), an FQ often used in agriculture, on the chemistry and in situ microbial communities in receiving waters. We added enro to 25 microg/L in nine outdoor mesocosms maintained under three light conditions (in triplicate): full sunlight typical of the upper epilimnion (100% full-light exposure, FLE), partial shading typical of the lower epilimnion (28% FLE), and near-complete shading typical of the hypolimnion (0.5% FLE). Enro disappearance and ciprofloxacin (cipro) formation were monitored over time using LC/MS, and water chemistry and ambient microbial communities (using denaturing gradient gel electrophoresis; DGGE) were characterized. Enro half-lives were 0.8, 3.7, and 72 days for the 100%, 28%, and 0.5% FLE treatments, respectively, creating three distinct FQ exposure scenarios. Although FQ exposures ranged from approximately 6 microg/L for 24 h to approximately 21 microg/L for 30 days, no statistically significant exposure effects were noted in water quality or microbial communities (as indicated by whole-community 16S rDNA DGGE analysis and specific amplification of the QRDR region of gyrase A). Small changes in water chemistry were noted over time; however, changes could not be specifically attributed to FQs. In general, enro addition had minimal effect on water column conditions at the levels and durations used here; however, further investigation is needed to assess effects in aquatic sediments.

Factors affecting competition between type I and type II methanotrophs in two-organism, continuous-flow reactors.

Competition experiments were performed in a continuous-flow reactor using Methylosinus trichosporium OB3b, a type II methanotroph, and Methylomonas albus BG8, a type I methanotroph. The experiments were designed to establish conditions under which type II methanotrophs, which have significant cometabolic potential, prevail over type I strains. The primary determinants of species selection were dissolved methane, copper, and nitrate concentrations. Dissolved oxygen and methanol concentrations played secondary roles. M. trichosporium OB3b proved dominant under copper and nitratelimited conditions. The ratio of M. trichosporium to M. albus in the reactor increased ten-fold in less than 100 hours following the removal of copper from the reactor feed. Numbers of M. albus declined to levels that were below detection limits (<106/ml) under nitrogen-limited conditions. In the latter experiment, the competitive success of M. trichosporiumdepended on the maintenance of an ambient dissolved oxygen level below about 7.5 × 10(-5) M, or 30% of saturation with air. The ability of M. trichosporium to express soluble methane monooxygenase under copper limitation and nitrogenase under nitrate limitation was very significant. M. albus predominated under methane-limited conditions, especially when low levels of methanol were simultaneously added with methane to the reactor. The results imply that nitrogen limitation can be used to select for type II strains such as M. trichosporium OB3b.