Quantitative detection and reduction of potentially pathogenic bacterial groups of Aeromonas, Arcobacter, Klebsiella pneumoniae species complex, and Mycobacterium in wastewater treatment facilities.

Water quality parameters influence the abundance of pathogenic bacteria. The genera Aeromonas, Arcobacter, Klebsiella, and Mycobacterium are among the representative pathogenic bacteria identified in wastewater. However, information on the correlations between water quality and the abundance of these bacteria, as well as their reduction rate in existing wastewater treatment facilities (WTFs), is lacking. Hence, this study aimed to determine the abundance and reduction rates of these bacterial groups in WTFs. Sixty-eight samples (34 influent and 34 non-disinfected, treated, effluent samples) were collected from nine WTFs in Japan and Thailand. 16S rRNA gene amplicon sequencing analysis revealed the presence of Aeromonas, Arcobacter, and Mycobacterium in all influent wastewater and treated effluent samples. Quantitative real-time polymerase chain reaction (qPCR) was used to quantify the abundance of Aeromonas, Arcobacter, Klebsiella pneumoniae species complex (KpSC), and Mycobacterium. The geometric mean abundances of Aeromonas, Arcobacter, KpSC, and Mycobacterium in the influent wastewater were 1.2 × 104-2.4 × 105, 1.0 × 105-4.5 × 106, 3.6 × 102-4.3 × 104, and 6.9 × 103-5.5 × 104 cells mL-1, respectively, and their average log reduction values were 0.77-2.57, 1.00-3.06, 1.35-3.11, and -0.67-1.57, respectively. Spearman's rank correlation coefficients indicated significant positive or negative correlations between the abundances of the potentially pathogenic bacterial groups and Escherichia coli as well as water quality parameters, namely, chemical/biochemical oxygen demand, total nitrogen, nitrate-nitrogen, nitrite-nitrogen, ammonium-nitrogen, suspended solids, volatile suspended solids, and oxidation-reduction potential. This study provides valuable information on the development and appropriate management of WTFs to produce safe, hygienic water.

Effect of formic acid inflow on microbial properties of the anaerobic granular sludge in a UASB reactor.

In the production of natural rubber, formate or acetate is added to the latex solution to coagulate the rubber; therefore, the wastewater contains high concentrations of organic acids, requiring the application of anaerobic treatment technology. In this study, a two-phase continuous flow experiment using a laboratory-scale upflow anaerobic sludge blanket (UASB) was conducted to investigate the influence of formate inflow on the microbial and physical characteristics of the retained granular sludge. In phase 1, acetate-based wastewater was used as feed, while in phase 2, formate-based wastewater was used as feed. In phase 1, the UASB exhibited high COD removal efficiency (97.2%); in addition, the retained sludge showed increased methane production from acetate and proliferation of acetate-utilizing Methanosaeta species. In phase 2, the UASB performed as well as phase 1, with 98.2% COD removal efficiency. Microbial community structure analysis confirmed that relatives of Methanobacterium formicicum present in the retained sludge were responsible for the degradation of formate in phase 2. However, decreased diameter and slight deterioration of granular sludge settleability were observed. In conclusion, formate inflow has low risk of interference with the process performance of the UASB, but it has negative effects on the physical properties of the granular sludge.

Sediment Microbiota in Response to Circuitry of Sediment Microbial Fuel Cells, Revealed by 16S rRNA Gene Amplicon Sequencing.

Information about sediment microbiota affected by sediment microbial fuel cells (SMFC) is limited. A laboratory-scale SMFC was applied to a eutrophic lake sediment under closed-circuit/open-circuit conditions. We analyzed the prokaryotes in the sediment adhering to the anode material. The archaeal family Methanoperedenaceae was a predominant group under closed-circuit conditions.

Detection of potentially pathogenic Arcobacter spp. in Bangkok canals and the Chao Phraya River.

The management of pathogenic bacteria in waterways is a public health issue. Here, we investigated the concentrations of potentially pathogenic bacteria, Arcobacter spp. and Campylobacter spp., and Escherichia coli, by quantifying species-specific genes in surface water samples from canals and the Chao Phraya River from June 2017 to June 2018 in Bangkok, Thailand. We assessed the relationship between the specific bacterial concentrations, water quality, and seasonal changes. Arcobacter spp. were detected at high density in all samples and showed seasonal fluctuations according to analyses based on 16S rDNA and the invasion gene ciaB. High levels of 16S rDNA and dut gene of E. coli were detected in the polluted drainage canals. A high correlation was observed between E. coli and chemical and biochemical oxygen demand (COD and BOD), suggesting that untreated domestic wastewater was the source of the E. coli. In contrast, Arcobacter spp. were detected with high density even in water samples with relatively low COD, suggesting that Arcobacter spp. are more likely than E. coli to survive in the water environment. The analysis of 16S rDNA and ciaB gene sequence analyses indicated that the Arcobacter spp. isolated from the drainage canals were A. butzleri and A. cryaerophilus.

Complete Genome Sequence of the Cesium-Accumulating Bacterium Rhodococcus qingshengii CS98, Isolated from Soil in Japan.

Rhodococcus qingshengii CS98 is a bacterium isolated from soil in Japan that shows strong cesium-accumulating ability. Here, we report the complete genome sequence of R. qingshengii (6.7 Mb), which may provide useful genetic information supporting its bioremediation features.

Primary production estimated for large lakes and reservoirs in the Mekong River Basin.

Understanding the proximate factors and mechanisms driving primary production in manmade reservoirs is crucial because such production can translate into added fish yields that provide people with food and livelihoods. Furthermore, reservoir fish production could potentially compensate for the loss of fish yields due to habitat fragmentation and alterations caused by damming and impoundment. We monitored primary production, identified environmental factors responsible for its variability, and examined the relationship between primary production and fish production in nine large water bodies of the Lower Mekong Basin for 2 years. The estimated primary production ranged from 40 to 302 g C/m2/y and was generally greater in the wet season than in the dry season. Linear mixed-effects modelling identified the concentration of dissolved inorganic carbon as a significant fixed-effect variable regulating primary production, after variability due to random and fixed effects of water body and seasonality, respectively, were taken into account. Fish yields marginally increased with increasing primary production across the water bodies, with the estimated energy transfer efficiency ranging from 0.004 to 0.009. Dissolved inorganic carbon was partly determined by the lithological composition of the water body catchment, suggesting that the geographic locations of proposed dams determine the magnitude of primary production and hence future fish production.

Performance evaluation of a down-flow hanging sponge (DHS) reactor as a decentralized domestic wastewater treatment system in tropical regions.

In this study, a pilot-scale down-flow hanging sponge (DHS) reactor was operated in the community plant of Bangkok for the treatment of domestic wastewater (COD 285 mg/L, BOD 105 mg/L) collected by separate sewer to evaluate the reactor's feasibility as a decentralized treatment system. The DHS reactor was operated for 600 days at ambient temperatures of 25-30 °C, both with constant flow conditions and with fluctuating flow conditions that simulated wastewater discharge patterns of the community. The results indicate that under constant flow at an HRT of 5 h, the volumetric loading rates of 0.36 kgBOD/m3-sponge/day and 0.16 kgN/m3-sponge/day were the optimum operational conditions of the DHS reactor in order to satisfy the effluent discharge standards. The DHS achieved removal rates of 89, 95, 91 and 90% for COD, BOD, TSS and NH4-N. Under the fluctuating flow condition, improvement of denitrification was confirmed at volumetric loading rates of 0.50 kgBOD/m3-sponge/day and 0.18 kgN/m3-sponge/day. The fluctuating flow of wastewater positively affects retained sludge activities in terms of homogenizing sludge concentration and stimulating oxygen uptake rates. These results suggest that the DHS reactor can be applied as a decentralized treatment system for domestic wastewater with fluctuating flow rates in tropical regions.

Incorporation characteristics of exogenous 15N-labeled thymidine, deoxyadenosine, deoxyguanosine and deoxycytidine into bacterial DNA.

Bacterial production has been often estimated from DNA synthesis rates by using tritium-labeled thymidine. Some bacteria species cannot incorporate extracellular thymidine into their DNA, suggesting their biomass production might be overlooked when using the conventional method. In the present study, to evaluate appropriateness of deoxyribonucleosides for evaluating bacterial production of natural bacterial communities from the viewpoint of DNA synthesis, incorporation rates of four deoxyribonucleosides (thymidine, deoxyadenosine, deoxyguanosine and deoxycytidine) labeled by nitrogen stable isotope (15N) into bacterial DNA were examined in both ocean (Sagami Bay) and freshwater (Lake Kasumigaura) ecosystems in July 2015 and January 2016. In most stations in Sagami Bay and Lake Kasumigaura, we found that incorporation rates of deoxyguanosine were the highest among those of the four deoxyribonucleosides, and the incorporation rate of deoxyguanosine was approximately 2.5 times higher than that of thymidine. Whereas, incorporation rates of deoxyadenosine and deoxycytidine were 0.9 and 0.2 times higher than that of thymidine. These results clearly suggest that the numbers of bacterial species which can incorporate exogenous deoxyguanosine into their DNA are relatively greater as compared to the other deoxyribonucleosides, and measurement of bacterial production using deoxyguanosine more likely reflects larger numbers of bacterial species productions.

The dynamics of pico-sized and bloom-forming cyanobacteria in large water bodies in the Mekong River Basin.

In the face of plans for increased construction of dams and reservoirs in the Mekong River Basin, it is critically important to better understand the primary-producer community of phytoplankton, especially the warm-water cyanobacteria. This is because these algae can serve as the primary source of carbon for higher trophic levels, including fishes, but can also form harmful blooms, threatening local fisheries and environmental and human health. We monitored the dynamics of three cyanobacteria-Synechococcus spp., Microcystis aeruginosa, and Dolichospermum spp.-for two years in nine large lakes and reservoirs in the Mekong River Basin. The densities of these algae were largely system-specific such that their abundance was uniquely determined within individual water bodies. However, after accounting for the system-specific effect, we found that cell densities of Synechococcus spp., M. aeruginosa, and Dolichospermum spp. varied in response to changes in photosynthetically active radiation (PAR), total nitrogen, and water level, respectively. Because both PAR and water level tend to fluctuate concordantly over a wide geographic area, Synechococcus spp., and to a lesser extent Dolichospermum spp., varied synchronously among the water bodies. Sustaining the production of pico-sized primary producers while preventing harmful algal blooms will be a key management goal for the proposed reservoirs in the Mekong Basin.

Dynamics of particulate phosphorus in a shallow eutrophic lake.

We tested the hypothesis that in shallow, eutrophic Lake Kasumigaura, the concentration of particulate phosphorus (PP) is controlled by biogenic P (P in living or dead phytoplankton and bacterial cells), rather than by resuspension of inorganic P in sediment. Increases in wind velocity and turbidity were associated with bottom shear stress exceeding the critical value for the lake (τc=0.15Nm(-2)); this increased turbidity was due to sediment resuspension. However, concentrations of PP; HCl-extractable, reactive P in PP (P-rP); and HCl-extractable, non-reactive P in PP (P-nrP) were not correlated with wind velocity (PP vs. wind velocity: r=0.40, p>0.05). Rather, the P-nrP concentration accounted for approximately 79% of PP, and the concentrations of PP, P-rP, and P-nrP were correlated with the particulate organic carbon (POC) concentration (POC vs. PP: r=0.90, p<0.01; POC vs. P-rP: r=0.82, p<0.01; POC vs. P-nrP: r=0.86, p<0.01). In our (31)P nuclear magnetic resonance spectroscopy results, mononucleotides accounted for the largest proportion among the detected P compound classes. In addition, concentrations of mononucleotides, orthophosphate, and pyrophosphate were significantly higher in samples with high POC concentrations, whereas the DNA-P concentration was not. These results suggest that biogenic P affects PP concentrations more strongly than does sediment resuspension, and the production of biogenic P creates a pool of mononucleotides, a class of easily degradable P, even in shallow, eutrophic Lake Kasumigaura.

Clinical Features of Miller-Fisher Syndrome in Pregnancy.

Miller-Fisher syndrome (MFS) is recognized as a variant of Guillain-Barré syndrome (GBS). MFS is a rare disorder that is characterized by the acute onset of ophthalmoplegia, ataxia, and areflexia/hyporeflexia. MFS has a higher incidence in Asia, where the incidence is estimated to be 18%-26% of GBS compared with 3%-5% in the West. The differential diagnosis of MFS includes Wernicke's encephalopathy (WE) which is characterized by a clinical triad (nystagmus and ophthalmoplegia, mental status changes, and ataxia), myasthenia gravis, and brainstem stroke. The association between MFS and pregnancy has not been reported previously. Here, we describe the clinical features of a pregnant woman in early pregnancy with MFS. This case highlights the fact that it is necessary to establish an accurate diagnosis based on the details from the patient's history on appropriate complementary testing in a pregnant patient with MFS.

Bacterial contribution to dissolved organic matter in eutrophic Lake Kasumigaura, Japan.

Incubation experiments using filtered waters from Lake Kasumigaura were conducted to examine bacterial contribution to a dissolved organic carbon (DOC) pool. Bacterial abundance, bacterial production, concentrations of DOC, total dissolved amino acids (TDAA), and total dissolved neutral sugars (TDNS) were monitored during the experiments. Bacterial production during the first few days was very high (20 to 35 µg C liter(-1) day(-1)), accounting for 40 to 70% of primary production. The total bacterial production accounted for 34 to 55% of the DOC loss during the experiment, indicating high bacterial activities in Lake Kasumigaura. The DOC degradation was only 12 to 15%, whereas the degradation of TDAA and TDNS ranged from 30 to 50%, suggesting the preferential usage of TDAA and TDNS. The contribution of bacterially derived carbon to a DOC pool in Lake Kasumigaura was estimated using d-amino acids as bacterial biomarkers and accounted for 30 to 50% of the lake DOC. These values were much higher than those estimated for the open ocean (20 to 30%). The ratio of bacterially derived carbon to bulk carbon increased slightly with time, suggesting that the bacterially derived carbon is more resistant to microbial degradation than bulk carbon. This is the first study to estimate the bacterial contribution to a DOC pool in freshwater environments. These results indicate that bacteria play even more important roles in carbon cycles in freshwater environments than in open oceans and also suggests that recent increases in recalcitrant DOC in various lakes could be attributed to bacterially derived carbon. The potential differences in bacterial contributions to dissolved organic matter (DOM) between freshwater and marine environments are discussed.

Development of a treatment system for molasses wastewater: the effects of cation inhibition on the anaerobic degradation process.

This study evaluated the process performance of a novel treatment system consisting of an acidification reactor, an upflow staged sludge bed (USSB) reactor, an upflow anaerobic sludge blanket reactor, and an aerobic trickling filter for the treatment of a high-strength molasses wastewater with a chemical oxygen demand (COD) of up to 120,000mg/L. The USSB operating at 35°C was capable of achieving an organic loading rate of 11kgCOD/m(3) day with a methane recovery of 62.4% at an influent COD of 120,000mg/L. The final effluent COD was 4520mg/L. The system was effective with regard to nitrification and sulfur removal. Fifty percent inhibition of the bacterial activity of the retained sludge by the cations was determined at 8gK/L for sucrose degradation, 16gK/L for sulfate reduction, and 12gK/L or 9gNa/L for acetoclastic methane production. Cation inhibition of anaerobic degradation reduced the process performance of the USSB.

Biogenic phosphorus compounds in sediment and suspended particles in a shallow eutrophic lake: a ³¹P-nuclear magnetic resonance (³¹P NMR) study.

Differences in biogenic phosphorus (P) compounds between sediment and suspended particles in aquatic environments are important for understanding the mechanisms of internal P loading, but these differences are still unknown. We used solution-state (31)P-nuclear magnetic resonance spectroscopy ((31)P NMR) with NaOH-ethylenediaminetetraacetic extraction to detect the multiple P compounds in suspended particles and sediment in the eutrophic Lake Kasumigaura, including orthophosphate monoesters, orthophosphate diesters, pyrophosphate, and polyphosphate. We tested the hypothesis that there is a significant difference between these groups in suspended particles and sediment. Biogenic P other than orthophosphate was found in significantly higher proportions in suspended particles (74.3% of total P) than in sediment (25.6%). Orthophosphate monoesters were comparatively more abundant in suspended particles, as indicated by the ratio of orthophosphate diesters to monoesters (average, 0.31 for suspended particles; 1.05 for sediment). The compounds identified as orthophosphate monoesters by (31)P NMR spectroscopy originated mainly from phospholipids (α-glycerophosphate and ß-glycerophosphate) and ribonucleic acid (RNA-P), whereas the orthophosphate diesters included mostly DNA (DNA-P). These results suggest that the dynamics of orthophosphate diesters, the production of DNA-P, or the degradation of phospholipids, play an important role in P cycling in Lake Kasumigaura.

High-rate treatment of molasses wastewater by combination of an acidification reactor and a USSB reactor.

A combination of an acidification reactor and an up-flow staged sludge bed (USSB) reactor was applied for treatment of molasses wastewater containing a large amount of organic compounds and sulfate. The USSB reactor had three gas-solid separators (GSS) along the height of the reactor. The combined system was continuously operated at mesophilic temperature over 400 days. In the acidification reactor, acid formation and sulfate reduction were effectively carried out. The sugars contained in the influent wastewater were mostly acidified into acetate, propionate, and n-butyrate. In addition, 10-30% of influent sulfur was removed from the acidification reactor by means of sulfate reduction followed by stripping of hydrogen sulfide. The USSB achieved a high organic loading rate (OLR) of 30 kgCOD m(-3) day(-1) with 82% COD removal. Vigorous biogas production was observed at a rate of 15 Nm(3) biogas m(-3) reactor day(-1). The produced biogas, including hydrogen sulfide, was removed from the wastewater mostly via the GSS. The GSS provided a moderate superficial biogas flux and low sulfide concentration in the sludge bed, resulting in the prevention of sludge washout and sulfide inhibition of methanogens. By advantages of this feature, the USSB may have been responsible for achieving sufficient retention (approximately 60 gVSS L(-1)) of the granular sludge with high methanogenic activity (0.88 gCOD gVSS(-1) day(-1) for acetate and as high as 2.6 gCOD gVSS(-1) day(-1) for H(2)/CO(2)). Analysis of the microbial community revealed that sugar-degrading acid-forming bacteria proliferated in the sludge of the USSB as well as the acidification reactor at high OLR conditions.

Quantification of microcystis in a eutrophic lake by simple DNA extraction and SYBR green real-time PCR.

A simple extraction method and real-time PCR with SYBR-Green I were combined to monitor Microcystis 16S rRNA gene (16S rDNA) concentrations over a wide range. A Fast DNA SPIN Kit (MP Biomedicals) was used to extract rDNA quantitatively. Real-time PCR amplified Microcystis rDNA for quantification with the forward primer Micro229f, which was newly designed by us and was highly specific for Microcystis, and the reverse universal primer 342r. The method developed here can detect Microcystis at concentrations as low as 3 cells mL(-1). The rDNA concentration and cell count were highly correlated in the range from 1.2×10(4) to 1.1×10(6) copies mL(-1) and from 9.5×10(2) to 1.0×10(5) cells mL(-1), respectively, in a canal where Microcystis algal blooms occur annually. The Microcystis rDNA concentration in Lake Kasumigaura was measured by the application of our method to water samples collected monthly from April 2004 to March 2006. Microcystis was not detected by microscopy from January to June 2005, except in May, but our method detected 1.0×10(3) to 1.0×10(4) copies mL(-1) of Microcystis rDNA during this period. This result clearly showed that our method is useful for clarifying the annual fluctuation in Microcystis concentration, especially when concentrations are low.

Effects of exercise intensity, posture, pressure on the back and ambient temperature on palmar sweating responses due to handgrip exercises in humans.

We have, by using newly developed ratemeters, attempted to examine the effects of exercise intensity, posture, pressure on the skin of the back, and ambient hyperthermic conditions (approximately 30 degrees C) on the 5-s handgrip exercise-mediated responses of active palmar sweating in humans. Thirty-five right-handed male (n=5) and female (n=30) volunteer students (20.2+/-1.3 years old) participated in the present study. Oral explanation of only the isometric handgrip exercise (IHG) caused a rapid and oscillatory response (pre-operational) of active palmar sweating in almost all subjects (10 of 14 subjects). Performing the IHG for 5-s caused a significant increase in active sweating rate (operation-mediated response) in both ipsi- and contra-lateral palmar surfaces of the thumbs of all subjects. The operation-mediated responses of active palmar sweating to the IHG were reproducible, resulting in no habituation. The increase of operation-mediated responses to the IHG was dependent upon exercise intensity (100-25% maximal voluntary contractions). The IHG-mediated ipsi- and contra-lateral responses of active palmar sweating were significantly decreased by changing the body posture from a seated to a supine position or by pressing the skin of the back. Ambient hyperthermic conditions (approximately 30 degrees C) for 60 min also resulted in a significant decrease in the back-pressure-dependent reduction of the operation-mediated responses of active palmar sweating to the IHG. In conclusion, in order to optimize the precision and reproducibility of clinical tests involving palmar sweating responses, it is important that subjects maintain a constant handgrip force and posture and that ambient temperature be kept under normothermic conditions.

Arithmetic calculation, deep inspiration or handgrip exercise-mediated pre-operational active palmar sweating responses in humans.

We examined the effects of repetitive mental stimulation such as arithmetic calculations with sequential subtraction or physical tasks such as handgrip exercise and deep breathing on active palmar sweating responses in humans. Thirty-three healthy, male and female volunteer students (20.4+/-2.1 years) participated in the present study. The responses of active palmar sweating were evaluated by using the newly developed ratemeter. The galvanic skin response (GSR) was also recorded in 10 out of 33 subjects. The oral explanation of the stimulation or tasks caused a rapid and wavy active palmar sweating response. The pre-operational responses of active palmar sweating to the stimulation or tasks were also observed by the GSR recording. The mental stimulation- and physical tasks-mediated pre-operational responses were significantly reduced by the trials. The mental stimulation or physical tasks also caused a rapid and oscillatory response of active palmar sweating during operation of the stimulation or tasks. The operation-mediated responses to physical tasks were observed ipsilaterally and contralaterally. The physical task-mediated responses were also reproducible, resulting in no habituation. In contrast, the operation-mediated responses to mental stimulation were reduced significantly by the trials, resulting in a marked habituation. The findings suggest that the mental stimulation- and physical tasks-mediated pre-operational responses of active palmar sweating obtained by using the newly developed ratemeter will make useful tests for evaluating neuronal activity of limbic system including amygdala, sympathetic sudomotor activity in the palmar skin and functional properties of the palmar sweat glands.