A Summer of Cyanobacterial Blooms in Belgian Waterbodies: Microcystin Quantification and Molecular Characterizations

In the context of increasing occurrences of toxic cyanobacterial blooms worldwide, their monitoring in Belgium is currently performed by regional environmental agencies (in two of three regions) using different protocols and is restricted to some selected recreational ponds and lakes. Therefore, a global assessment based on the comparison of existing datasets is not possible. For this study, 79 water samples from a monitoring of five lakes in Wallonia and occasional blooms in Flanders and Brussels, including a canal, were analyzed. A Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) method allowed to detect and quantify eight microcystin congeners. The mcyE gene was detected using PCR, while dominant cyanobacterial species were identified using 16S RNA amplification and direct sequencing. The cyanobacterial diversity for two water samples was characterized with amplicon sequencing. Microcystins were detected above limit of quantification (LOQ) in 68 water samples, and the World Health Organization (WHO) recommended guideline value for microcystins in recreational water (24 µg L−1) was surpassed in 18 samples. The microcystin concentrations ranged from 0.11 µg L−1 to 2798.81 µg L−1 total microcystin. For 45 samples, the dominance of the genera Microcystis sp., Dolichospermum sp., Aphanizomenon sp., Cyanobium/Synechococcus sp., Planktothrix sp., Romeria sp., Cyanodictyon sp., and Phormidium sp. was shown. Moreover, the mcyE gene was detected in 75.71% of all the water samples.

Growth and chemotaxis of nematodes reduced upon exposure to Third Fork Creek surface water

Background Third Fork Creek is a historically impaired urban stream that flows through the city of Durham, North Carolina. Caenorhabditis elegans (C. elegans) are non-parasitic, soil and aquatic dwelling nematodes that have been used frequently as a biological and ecotoxicity model. We hypothesize that exposure to Third Fork Creek surface water will inhibit the growth and chemotaxis of C. elegans. Using our ring assay model, nematodes were enticed to cross the water samples to reach a bacterial food source which allowed observation of chemotaxis. The total number of nematodes found in the bacterial food source and the middle of the plate with the water source was recorded for 3 days. Results Our findings suggest a reduction in chemotaxis and growth on day three in nematodes exposed to Third Fork Creek water samples when compared to the control (p value < 0.05). These exploratory data provide meaningful insight to the quality of Third Fork Creek located near a Historically Black University. Conclusions Further studies are necessary to elucidate the concentrations of the water contaminants and implications for human health. The relevance of this study lies within the model C. elegans that has been used in a plethora of human diseases and exposure research but can be utilized as an environmental indicator of water quality impairment.

CRISPR/Cas12a-Assisted Visual Logic-Gate Detection of Pathogenic Microorganisms Based on Water-Soluble DNA-Binding AIEgens

Here, we developed a rapid, visual and double-checked Logic Gate detection platform for detection of pathogenic microorganisms by aggregation-induced emission luminogens (AIEgens) in combination with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated (Cas). DNA light-up AIEgens (1,1,2,2-tetrakis[4-(2-bromo-ethoxy) phenyl]ethene, TTAPE) was non-emissive but the emission was turned on in the presence of large amount of DNA produced by recombinase polymerase amplification (RPA). When CRISPR/Cas12a was added, all long-stranded DNA were cut leading to the emission quenched. Thus, a method that can directly observe the emission changes with the naked eye has been successfully constructed. The detection is speedy within only 20 min, and has strong specificity to the target. The result can be judged by Logic Gate. Only when the output signal is (1,0), does it represent the presence of pathogenic microorganisms in the test object. Finally, the method was applied to the detect pathogenic microorganisms in environmental water samples, which proved that this method has high selectivity, specificity and applicability for the detection of pathogenic microorganisms in environmental water samples.

Can Humidifier Reservoir Bacteria Colonize the Circuit During Mechanical Ventilation?

Background:Although the circuit condensate, an ideal bacterial reservoir, may flow into the humidifier reservoir (HR), no study has investigated if HR-colonized bacteria colonize other circuit locations with airflow. Therefore, the objective of this study was to explore if bacterial growth in the HR leads to bacterial colonization in the ventilator circuit. Methods: A randomized controlled experiment was performed in a public tertiary hospital in Guangdong Province, China. In vitro mechanical ventilation models (n = 60), divided into sterile water samples (n = 30) and broth samples (n = 30), were established. Sterile water was used for humidification in the ventilation models. The sterile water group contained either Acinetobacter baumannii (n = 15) or Pseudomonas aeruginosa (n = 15) in humidifier water. The broth group was similar to the sterile water group, but brain heart infusion broth was added to the HR. After 24, 72, and 168 h of continuous ventilation, bacteria in the humidifier water and at different circuit locations were sampled and cultured, and the results were analyzed by the Chi-square test. The difference in bacterial concentration at the HR outlet was analyzed by the F test, and P < 0.05 was considered statistically significant.Results:Bacterial culture results of the sterile water samples were negative. Bacteria in the humidifier water continued to proliferate in the broth group, and the bacterial concentration at different times was not significantly different (P > 0.05). With prolonged ventilation, the bacterial concentration at the HR outlet increased (P < 0.05). During continuous ventilation, no bacterial growth occurred at 10 cm from the HR outlet and the Y-piece of the ventilator circuit. The bacterial concentration at the HR outlet was higher in the P. aeruginosa group than in the A. baumannii group (P < 0.05).Conclusions:Sterile water in the HR was not conducive to bacterial growth. Although bacteria grew in the HR and could reach the HR outlet, colonization of other circuit locations was unlikely.

Assessment of faecal contamination in selected concrete and earthen ponds stocked with Clarias gariepinus

Background This study was carried out to monitor the levels of faecal pollution markers in catfish (Clarias gariepinus) and their growing waters in selected earthen and concrete ponds. Water and catfish samples were collected weekly in the months of February, March, April, May, June and July, 2019. The concentrations of enteric bacteria in the water and catfish samples were determined using membrane filtration and pour plate methods, respectively. The rate of bioaccumulation of faecal indicator bacteria was obtained by dividing the log concentration of each organism in catfish by the corresponding log concentration in the growing waters. Result The concentration of faecal coliforms in catfish samples from concrete and earthen ponds ranged from 1.41 to 2.28 log10 CFU/100 ml and 1.3 to 2.47 log10 CFU/100 ml respectively and in growing waters from the concrete and earthen ponds; 1.43 to 2.41 log10 CFU/100 ml and 1.50 to 2.80 log10 CFU/100 ml respectively. Faecal coliforms exhibited positive relationships with alkalinity in water samples from the earthen (r = 0.61) and concrete ponds (r = 0.62). Salmonella and faecal coliforms had the highest and least bioaccumulation in catfish raised in earthen pond, respectively, whereas Salmonella and enterococci had the highest and least bioaccumulation in catfish raised in concrete pond, respectively. Faecal coliforms and E. coli had the highest and least counts in water samples from the earthen pond during the dry and wet months, Salmonella and E. coli had the highest and least counts in water samples from the concrete pond during the dry and wet months. Conclusion There were high levels of bacterial faecal pollution markers in water and C. gariepinus from the earthen and concrete ponds. Physicochemical characteristics of the water and seasonality played major roles in the rate of bioaccumulation of the faecal pollution markers in C. gariepinus raised in the earthen and concrete ponds.

Nickel contamination analysis at cost-effective silver printed paper-based electrodes based on carbon black dimethylglyoxime ink as electrode modifier

Electrochemical detection of metal cations at paper-based sensors has been suggested as an attractive alternative to current spectroscopic and chromatographic detection techniques due to the ease of fabrication, disposable nature, and low cost. Herein, a novel carbon black (CB), dimethylglyoxime (DMG) ink is designed as an electrode modifier in conjunction with 3-electrode inkjet-printed paper substrates for use in the adsorptive stripping voltammetric electroanalysis of nickel cations in water samples. The developed method provides a novel, low-cost, rapid, and portable adsorptive stripping detection approach towards metal analysis in the absence of the commonly used toxic metallic films. The study demonstrated a novel approach to nickel detection at paper-based sensors and builds on previous work in the field of paper-based metal analysis by limiting the use of toxic metal films. The device sensitivity is improved by increasing the active surface area, electron transfer kinetics, and catalytic effects associated with non-conductive dimethylglyoxime films through CB nanoparticles for the first time and confirmed by electroanalysis. The first use of the CB-DMG ink allows for the selective preconcentration of analyte at the electrode surface without the use of toxic Mercury or Bismuth metallic films. Compared to similarly reported paper-based sensors, improved limits of detection (48 µg L-1), selectivity, and intermetallic interferences were achieved. The method was applied to the detection of nickel in water samples well below World Health Organization (WHO) standards.