The present study tracked the city-wide dynamics of severe acute respiratory syndrome-corona virus 2 ribonucleic acids (SARS-CoV-2 RNA) in the wastewater from nine different wastewater treatment plants (WWTPs) in Jaipur during the second wave of COVID-19 out-break in India. A total of 164 samples were collected weekly between February 19th and June 8th, 2021. SARS-CoV-2 was detected in 47.2% (52/110) influent samples and 37% (20/54) effluent samples. The increasing percentage of positive influent samples correlated with the city’s increasing active clinical cases during the second wave of COVID-19 in Jaipur. Furthermore, wastewater-based epidemiology (WBE) evidence clearly showed early detection of about 20 days (9/9 samples reported positive on April 20th, 2021) before the maximum cases and maximum deaths reported in the city on May 8th, 2021. The present study further observed the presence of SARS-CoV-2 RNA in treated effluents at the time window of maximum active cases in the city even after tertiary disinfection treatments of ultraviolet (UV) and chlorine (Cl 2 ) disinfection. The average genome concentration in the effluents and removal efficacy of six commonly used treatments, activated sludge process + chlorine disinfection (ASP + Cl 2 ), moving bed biofilm reactor (MBBR) with ultraviolet radiations disinfection (MBBR + UV), MBBR + chlorine (Cl 2 ), sequencing batch reactor (SBR), and SBR + Cl 2 , were compared with removal efficacy of SBR + Cl 2 (81.2%) > MBBR + UV (68.8%) > SBR (57.1%) > ASP (50%) > MBBR + Cl 2 (36.4%). The study observed the trends and prevalence of four genes (E, RdRp, N, and ORF1ab gene) based on two different kits and found that prevalence of N > ORF1ab > RdRp > E gene suggested that the effective genome concentration should be calculated based on the presence/absence of multiple genes. Hence, it is imperative to say that using a combination of different detection genes (E, N, RdRp, & ORF1ab genes) increases the sensitivity in WBE. Graphical abstract

The present study tracked the city-wide dynamics of severe acute respiratory syndrome-corona virus 2 (SARS-CoV-2) RNA in the wastewater from nine different wastewater treatment plants (WWTPs) in Jaipur during second wave of COVID-19 out-break in India. A total of 164 samples were collected weekly between February 19th and June 8th, 2021. SARS-CoV-2 was detected in 47.2% (52/110) influent samples and 37% (20/54) effluent samples. The increasing percentage of positive influent samples correlated with the city’s increasing active clinical cases during the second wave of COVID-19 in Jaipur. Furthermore, WBE based evidence clearly showed early detection of about 20 days (9/9 samples reported positive on April 20th, 2021) prior to the maximum cases & maximum deaths reported in the city on May 8th, 2021. The present study further observed the presence of SARS-CoV-2 RNA in treated effluents at the time window of maximum active cases in the city even after tertiary disinfection treatments of UV & Chlorine. The average genome concentration in the effluents and removal efficacy of six commonly used treatments; Activated Sludge Treatment + Chlorine disinfection (ASP + Cl2), Moving Bed Biofilm Reactor (MBBR) with Ultraviolet radiations disinfection (MBBR + UV), MBBR + Chlorine (Cl2), Sequencing Batch Reactor (SBR) and SBR + Cl2 were compared with removal efficacy of SBR + Cl2 (81.2%)> MBBR + UV (68.8%) > SBR (57.1%) > ASP (50%) > MBBR + Cl2(36.4%). The study observed the trends & prevalence of four genes (E, RdRp, N, and ORF1ab gene) based on two different kits and found that prevalence of N> ORF1ab >RdRp> E gene, suggested that the effective genome concentration should be calculated based on the presence/absence of multiple genes. Hence, it is imperative to say that using a combination of different detection genes (E, N, RdRp & ORF1ab genes) reduce false positives in WBE.Graphical AbstractHighlightsSuccessful application of WBE with prediction of 14-20 days for COVID-19 in JaipurA comparison of SARS-CoV-2 RNA removal efficacy of 9 WWTPs was investigatedSBR showed better performance than MBBR with SARS-CoV-2 RNA removal from wastewaterPresence of SARS-CoV-2 in effluents even after UV and Chlorine disinfectionUsing a combination of different detection genes reduce false positives in WBE

An alarming increase in Candidiasis infections has been observed worldwide as well as in India. This increase is attributed to increase in immune-compromised individuals and an increase in plethora of species causing the disease. The emergence of drug resistance of isolates has further worsened the situation. Candida isolates causing infections were obtained from patients and evaluated for their macroscopic and microscopic characteristics by colony morphology and staining procedures. The patients’ demography was also analyzed to identify any correlation between the isolates. Doubling time was determined to analyze growth characteristics of these isolates. Isolates were also characterized biochemically for their ability to assimilate carbon and nitrogen sources and for fermentative capabilities. Additionally, drug sensitivity profiles of these isolates were analyzed towards Azoles. Demographic analysis of the isolates suggested that all age groups were affected by Candida via both albicans and non-albican species. The infections were not gender biased but majority of isolates were obtained from urine samples suggesting Candida as an important species causing urino-genital infections. Macroscopic and microscopic analysis showed cream- colored circular colonies with smooth surface and entire margins with cells in single and budding stage. Biochemical characterization showed sucrose to be the most metabolizable sugar with maximum fermentative capacity with glucose. Surprisingly, very low nitrogen assimilation capacity was observed with all the nitrogen sources tested (nitrate, urea and glycine). Drug sensitivity towards Azoles suggested almost 50% and 90% isolates were resistant to Fluconazole and Itraconazole respectively, the most common Azoles used against fungal infections in the current scenario. The results obtained from the study suggested differential characteristics of the isolates towards various parameters thereby indicating the relevance of isolate characterization, for appropriate control and prevention of the disease.

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Purpose: An alarming increase in Candidiasis infections has been observed worldwide and in India. This increase is attributed to increase in immune-compromised individuals and an increase in plethora of species causing the disease. The emergence of drug resistance of isolates has further worsened the situation. Materials & Methods: Candida isolates causing infections were obtained from patients and evaluated for their macroscopic and microscopic characteristics by colony morphology and staining procedures. The patients demography was also analyzed to identify any correlation between the isolates. Doubling time was determined to analyze growth characteristics of these isolates. Isolates were also characterized biochemically for their ability to assimilate carbon and nitrogen sources and for fermentative capabilities. Additionally, drug sensitivity profiles of these isolates were analyzed towards Azoles. Results: Demographic analysis of the isolates suggested that all age groups were affected by Candida via both albicans and non-albican species. The infections were not gender biased but majority isolates were obtained from urine samples suggesting Candida as an important species causing urine-genital infections. Macroscopic and microscopic analysis showed cream colored circular colonies with smooth surface and entire margins with cells in single and budding stage. The doubling time ranged between 40 mins to 180 mins with 81 mins being the average. Biochemical characterization showed sucrose to be the most metabolizable sugar with maximum fermentative capacity with glucose. Surprisingly, very low nitrogen assimilation capacity was observed with all the nitrogen sources tested (nitrate, urea and glycine). Drug sensitivity towards Azoles suggested almost 50% and 90% isolates were resistant to Fluconazole and Itraconazole respectively, the most common Azoles used against fungal infections in the current scenario. Conclusions: The results obtained from the study suggested differential characteristics of the isolates towards various parameters thereby indicating the relevance of isolate characterization, for appropriate control and prevention of the disease.