Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of viable but non-culturable Vibrio cholerae O1

<i>Vibrio cholerae</i>, an important waterborne pathogen, is a rod-shaped bacterium that naturally exists in aquatic environments. When conditions are unfavorable for growth, the bacterium can undergo morphological and physiological changes to assume a coccoid morphology. This stage in its life cycle is referred to as viable but non-culturable (VBNC) since VBNC cells do not grow on conventional bacteriological culture media. The current study compared polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to detect and identify VBNC <i>V. cholerae</i>. Because it is difficult to detect and identify VBNC <i>V. cholerae</i>, the results of the current study are useful in showing LAMP to be more sensitive and rapid than PCR in detecting and identifying non-culturable, coccoid forms of <i>V. cholerae</i>. Furthermore, the LAMP method is effective in detecting and identifying very low numbers of coccoid VBNC <i>V. cholerae</i> in environmental water samples, with the added benefit of being inexpensive to perform.

Quantification of Brownian motion under presence of flow using particle diffusometry

Particle diffusometry (PD), a quantification method for the Brownian motion, is performed by recording temporally sequential images and using correlation analysis to obtain an ensemble diffusion coefficient for all particles captured in the imaging region (Clayton et al., 2017). PD is proven to be successful in the detection of the waterborne pathogen V. cholerae in environmental samples using different imaging techniques, including an inverted fluorescence microscope as well as a handheld hardware device operated with a smartphone (Clayton et al., 2019; Moehling et al., 2020). Although we intend to use PD to calculate diffusion coefficients in quiescent fluid, oftentimes unintentional fluid flows occur, creating measurement error when calculating the diffusion coefficient. In previous work, recordings under the presence of flow were discarded to avoid incorrect measurements of the sample.

The ability of Aliarcobacter butzleri strains isolated from foods of animal origin in Costa Rica to form biofilm

Aliarcobacter butzleri is a zoonotic emerging food and waterborne pathogen widely distributed in nature. It is present in food processing environments and can easily be spread through the food industry because of its ability to form biofilm. The aim of this work was to determine the ability of strains isolated in Costa Rica from different food matrixes of animal origin to form biofilm. Thirty-eight A. butzleri strains previously isolated and identified from animal origin products were analyzed using the method described by Stepmovic et al. (2000), in three culture broths, brain heart infusion broth, Boer broth and Houf broth. Results showed that 67% of poultry origin strains, 62.5% of meat origin strains and just 8% of milk origin strains showed ability to form biofilm. The findings of this study confirm the adherence ability of A. butzleri to form biofilm, a characteristic that can promote dispersion and cross contamination along food industry processing lines.

Population genomics provides insights into the evolution and adaptation to humans of the waterborne pathogen Mycobacterium kansasii

Mycobacterium kansasii can cause serious pulmonary disease. It belongs to a group of closely-related species of non-tuberculous mycobacteria known as the M. kansasii complex (MKC). Here, we report a population genomics analysis of 358 MKC isolates from worldwide water and clinical sources. We find that recombination, likely mediated by distributive conjugative transfer, has contributed to speciation and on-going diversification of the MKC. Our analyses support municipal water as a main source of MKC infections. Furthermore, nearly 80% of the MKC infections are due to closely-related M. kansasii strains, forming a main cluster that apparently originated in the 1900s and subsequently expanded globally. Bioinformatic analyses indicate that several genes involved in metabolism (e.g., maintenance of the methylcitrate cycle), ESX-I secretion, metal ion homeostasis and cell surface remodelling may have contributed to M. kansasii’s success and its ongoing adaptation to the human host.

First Experimental Evidence for the Presence of Potentially Toxic Vibrio cholerae in Snails, and Virulence, Cross-Resistance and Genetic Diversity of the Bacterium in 36 Species of Aquatic Food Animals

Vibrio cholerae is the most common waterborne pathogen that can cause pandemic cholera in humans. Continuous monitoring of V. cholerae contamination in aquatic products is crucial for assuring food safety. In this study, we determined the virulence, cross-resistance between antibiotics and heavy metals, and genetic diversity of V. cholerae isolates from 36 species of aquatic food animals, nearly two-thirds of which have not been previously detected. None of the V. cholerae isolates (n = 203) harbored the cholera toxin genes ctxAB (0.0%). However, isolates carrying virulence genes tcpA (0.98%), ace (0.5%), and zot (0.5%) were discovered, which originated from the snail Cipangopaludina chinensis. High occurrences were observed for virulence-associated genes, including hapA (73.4%), rtxCABD (68.0–41.9%), tlh (54.2%), and hlyA (37.9%). Resistance to moxfloxacin (74.9%) was most predominant resistance among the isolates, followed by ampicillin (59.1%) and rifampicin (32.5%). Approximately 58.6% of the isolates displayed multidrug resistant phenotypes. Meanwhile, high percentages of the isolates tolerated the heavy metals Hg2+ (67.0%), Pb2+ (57.6%), and Zn2+ (57.6%). Distinct virulence and cross-resistance profiles were discovered among the V. cholerae isolates in 13 species of aquatic food animals. The ERIC-PCR-based genome fingerprinting of the 203 V. cholerae isolates revealed 170 ERIC-genotypes, which demonstrated considerable genomic variation among the isolates. Overall, the results of this study provide useful data to fill gaps for policy and research related to the risk assessment of V. cholerae contamination in aquatic products.

Microbial Assay of the Waterborne Pathogen on Supplied Drinking Water in Gopalganj Area, Bangladesh and its Future Effect

Around 60% of water constitute the human body if about 2% of the body’s water content drop anyone dehydration must have a remarkable effect. Studies show that the loss of body water can break many aspects of brain function. About Eight 8-ounce (237-mL) glasses of water always consult by the physician for a sound person. But unfortunately, If this drinking water becomes impure it affects to develop the various vital dysfunction in our body. Infrequent bowel actions and trouble passing footrest, brain associate dysfunction along with familiar waterborne infections such as looseness, polio, diarrhoea and meningitis are the common conditions that arise for the water-borne pathogen in water. Gopalganj is a town in Gopalgonj District belonging to the Dhaka Division of Bangladesh. According to the census(cf) of 2011-03-15, 53778 individuals are living here from the divergent profession. For geographical reasons the most of the area specially Gopalganj town along with Gobra, Ghonapra, Tungipara where most government institutions are located the groundwater is not suitable for drinking as Iron and Arsenic are the supremacy amount. That’s why this is the common scenery of these areas to use drinking water jar for the purpose of safe water. For this reason here have already found about 10-15 drinking water jar supplier company. In this research, we performed the microbiological assay to observe the presence or absence of water-born pathogen on these drinking water containing the water jar along with the morphological characteristics of these pathogens. In point of fact, we found that there is 3 to 6 type of microbes are present in this water. Among these pathogens, there are the supremacy of gram-positive bacteria than gram-negative bacteria along with some moulds co-exist on these microbiomes.

The detection efficiency of digital PCR for the virulence genes of waterborne pathogenic bacteria

Waterborne pathogens are the primary concern for the safe reuse of wastewater. Although digital PCR (dPCR) is considered promising for absolutely quantitating genes, the detection efficiency of dPCR is affected by many factors. This study tested 8 virulence genes of pathogenic bacteria on a control plasmid and reclaimed water samples with reported primer-probe sets and designed ones on quantitative PCR (qPCR) and dPCR. Probe efficiency, data analysis, and PCR inhibition, were found to affect the detection efficiency of dPCR. Firstly, poor probe quality, which is determined by probe quenching and activation efficiencies, was the main cause of PCR failure. Secondly, even if the PCR was successful, the probe quality and signal intensity could still affect the quantitative process. Manual analysis of dPCR data on the weak signal intensity would significantly reduce errors. And lastly, the sensitivity of PCR inhibition was lower in dPCR than qPCR, but inhibition still existed. The dPCR produced various detection efficiencies for different targets in one sample indicating inconstant inhibitory effects. Dilution was still the proper approach to overcome inhibition, but decreased detection limit. More studies are required to ensure accurate waterborne pathogen quantitation by dPCR.