SARS-CoV-2 Burden in Wastewater and its Elimination Using Disinfection.

Background: Pathogenic viruses have been abundant and diverse in wastewater, reflecting the pattern of infection in humans. Human feces, urine, and perhaps other washouts that frequently circulate in sewage systems may contaminate wastewater with SARS-CoV-2. It's crucial to effectively disinfect wastewater since poorly handled wastewater could put the population at risk of infection. Aims: To emphasize the presence and spread of SARS-CoV-2 in sewage (wastewater) through viral shedding from the patients to detect the virus in the population using wastewater-based epidemiology. Also, to effectively manage the transmission of SARS-CoV-2 and reduce the spread of the virus in the population using disinfectants is highlighted. Methods: We evaluated articles from December 2019 to August 2022 that addressed SARS-CoV-2 shedding in wastewater and surveillance through wastewater-based epidemiology. We included the papers on wastewater disinfection for the elimination of SARS-CoV-2. Google Scholar, PubMed, and Research4Life are the three electronic databases from which all of the papers were retrieved. Results: It is possible for viral shedding to get into the wastewater. The enumeration of viral RNA from it can be used to monitor virus circulation in the human community. SARS-CoV-2 can be removed from wastewater by using modern disinfection techniques such as sodium hypochlorite, liquid chlorine, chlorine dioxide, peracetic acid, and ultraviolet light. Conclusion: SARS-CoV-2 burden estimates at the population level can be obtained via longitudinal examination of wastewater, and SARS-CoV-2 can be removed from the wastewater through disinfection.

Extended-spectrum ß-lactamases producing Enterobacteriaceae (ESBL-PE) prevalence in Nepal: A systematic review and meta-analysis.

An alarming increase in the occurrence of extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-PE) has threatened the treatment and management of bacterial infections. This systematic review and meta-analysis aimed to provide a quantitative estimate of the prevalence of ESBL among the members of the Enterobacteriaceae family by analyzing the community-based and clinical studies published between 2011 and 2021 from Nepal and determine if ESBL-PE correlates with multidrug resistance (MDR). The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines were followed for systematic review and meta-analysis and the articles' quality was assessed using the Newcastle-Ottawa scale. Of the 2529 articles screened, 65 articles were systematically reviewed, data extracted, and included in in-depth meta-analysis. The overall pooled prevalence of ESBL-producers in Enterobacteriaceae was 29 % (95 % CI: 26-32 %) with high heterogeneity (I2 = 96 %, p < 0.001). Escherichia coli was the predominant ESBL-producing member of the Enterobacteriaceae family, followed by Citrobacter spp. and Klebsiella spp. The prevalence of ESBL-PE increased from 18.7 % in 2011 to 29.5 % in 2021. A strong positive correlation (r = 0.98) was observed between ESBL production and MDR in Enterobacteriaceae. ESBL-PE isolates showed high resistance to ampicillin, cephalosporins, and amoxicillin-clavulanic acid, and blaCTX-M type was the most reported gene variant among ESBL-PE. In conclusion, this study demonstrated an increased prevalence of ESBL-PE in Nepal over the last decade, and such isolates showed a high level of MDR against the ß-lactams and non-ß-lactam antibiotics. Tackling the rising antibiotic resistance (AR) and MDR in ESBL-PE would require concerted efforts from all stakeholders to institute effective infection control programs in the community and clinical settings.

Multi-drug Resistance, ß-Lactamases Production, and Coexistence of bla NDM-1 and mcr-1 in Escherichia coli Clinical Isolates From a Referral Hospital in Kathmandu, Nepal.

The ability of pathogenic Escherichia coli to produce carbapenemase enzymes is a characteristic that allows them to resist various antibiotics, including last-resort antibiotics like colistin and carbapenem. Our objectives were to identify rapidly developing antibiotic resistance (AR), assess ß-lactamases production, and detect mcr-1 and bla NDM-1 genes in the isolates. A prospective cross-sectional study was carried out in a referral hospital located in Kathmandu from November 2019 to December 2020 using standard laboratory and molecular protocols. Among 77 total E. coli isolates, 64 (83.1%) of them were categorized as MDR. Phenotypically 13 (20.3%) colistin-resistant, 30 (46.9%) ESBL and 8 (12.5%) AmpC producers, and 5 (7.8%) ESBL/AmpC co-producers were distributed among MDR-E. coli. Minimum inhibitory concentrations (MIC) against the majority of MDR isolates were exhibited at 1 g/L. Of these 77 E. coli isolates, 24 (31.2%) were carbapenem-resistant. Among these carbapenem-resistant bacteria, 11 (45.9%) isolates were reported to be colistin-resistant, while 15 (62.5%) and 2 (8.3%) were MBL and KPC producers, respectively. Out of 15 MBL producers, 6 (40%) harbored bla NDM-1, and 8 (61.5%) out of 13 colistin-resistant pathogens possessed mcr-1. The resistance by colistin- and carbapenem were statistically associated (P < .001). However, only 2 (18.2%) of the co-resistant bacteria were found to have both genes. Our study revealed the highly prevalent MDR and the carbapenem-resistant E. coli and emphasized that the pathogens possess a wide range of capabilities to synthesize ß-lactamases. These findings could assist to expand the understanding of AR in terms of enzyme production.

Systematic Review and Meta-analysis on Extended-Spectrum ß-lactamases Producing Klebsiella pneumoniae in Nepal.

Objective: This systematic review and meta-analysis aimed to assess the pool estimates of extended-spectrum ß-lactamases producing K. pneumoniae (ESBL-KP) and study their drug resistance profile by evaluating the studies from Nepal. Methods: A literature search was carried out in PubMed, Google Scholar, and NepJOL to screen all articles on ESBL-KP published between 2011 and 2021 from Nepal. This review was conducted following Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. Relevant data were extracted, and R language 4.2.0 software was used for statistical analysis. Results: The pooled prevalence of K. pneumoniae was 5%, while the pooled prevalence of ESBL and multidrug resistance (MDR) in K. pneumoniae were 23% and 55%, respectively. Imipenem was the drug of choice (in vitro) against ESBL-KP infection. Conclusion: Our analyses showed a high prevalence of ESBL-KP and their high resistance toward commonly used drugs. This study highlights the need for the development of new antibiotics for the management of ESBL-KP infections.

Isolation of Lactococcus garvieae NEP21 from raw cow (Bos indicus) milk in Nepal.

Lactococcus garvieae is an emerging zoonotic pathogen impacting both humans and animals. Infection of this bacterium is known to cause mastitis in cattle, and endocarditis, osteomyelitis, liver abscess, and gastrointestinal problems are reported in immunocompromised and elderly people that regularly consume or handle raw meat, milk, dairy products, and seafood. This study aimed at investigating and detecting lactic acid bacteria in raw cow (Bos indicus) milk samples from a smallholder farm in Nepal. Based on the plate culture, biochemical tests, and molecular sequencing of 16 s ribosomal RNA coding nuclear DNA region followed by phenotypic and genotypic analyses, L. garvieae NEP21 was detected and identified for the first time in Nepal in raw cow milk samples. This finding suggests the prevalence of L. garvieae NEP21 in raw cow milk and recommends further research and surveillance for understanding the extent of its presence in Nepal and globally for informed management of its infection in cattle and humans.

Antibiotic Resistance, Biofilm Formation and Sub-Inhibitory Hydrogen Peroxide Stimulation in Uropathogenic Escherichia coli.

Uropathogenic Escherichia coli (UPEC) is the most prevalent cause of urinary tract infections (UTIs). Biofilm formation and antibiotic resistance could be high among the causative agent. The purpose of this study was to determine antibiotic resistance, biofilm production, and biofilm-associated genes, bcsA and csgD, and sub-inhibitory hydrogen peroxide (H2O2) stimulation in UPEC for biofilm formation. A total of 71 UPEC were collected from a tertiary care hospital in Kathmandu and subjected to identify antibiotic susceptibility using Kirby-Bauer disk diffusion. The biofilm formation was assessed using microtiter culture plate method while pellicle formation was tested by a tube method. In representative 15 isolates based on biofilm-forming ability, bcsA and csgD were screened by conventional polymerase chain reaction, and treated with sub-lethal H2O2. The UPEC were found the most susceptible to meropenem (90.2%), and the least to ampicillin (11.3%) in vitro and 90.1% of them were multi-drug resistant (MDR). Most UPEC harbored biofilm-producing ability (97.2%), and could form pellicle at 37°C. Among representative 15 isolates, csgD was detected only among 10 isolates (66.67%) while bcsA gene was present in 13 isolates (86.67%). This study revealed that level of biofilm production elevated after sub-lethal H2O2 treatment (P = .041). These findings suggested that the pathogens are emerging as MDR. The biofilm production is high and the majority of selected strains contained bcsA and csgD genes. Pellicle formation test was suggestive to be an alternative qualitative method to screen biofilm production in UPEC. The sub-inhibitory concentration of H2O2 may contribute in increasing biofilm formation in UPEC.

Prevalence of metallo-ß-lactamases as a correlate of multidrug resistance among clinical Pseudomonas aeruginosa isolates in Nepal.

Pseudomonas aeruginosa is an opportunistic human pathogen that has developed antibiotic resistance (AR) and causes a range of illnesses, including respiratory pneumonia, gastrointestinal infections, keratitis, otitis media and bacteremia in patients with compromised immune system. The production of metallo-ß-lactamases (MBLs) is one of the major mechanisms of AR in this bacterium with ensuing infections difficult to treat. The main goal of this study was to provide a quantitative estimate of MBLs producing clinical P. aeruginosa isolates among the Nepalese patients and determine if MBL correlates with multi-drug resistance (MDR). Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guideline was followed for meta-analysis of relevant literature using PubMed, Research4Life, and Google Scholar. The prevalence of MBLs in P. aeruginosa from clinical samples was determined using R 4.1.2 for data pooled from studies published until 2021. The meta-analysis of a total of 19 studies selected (of 6038 studies for which titles and abstracts were reviewed) revealed the prevalence of MBLs producing P. aeruginosa (MBL-PA) was 14 % (95 % CI: 0.10-0.19) while MDR isolates among P. aeruginosa was 42 % (95 % CI: 0.30-0.55) in Nepal. Combined Disc Test was predominantly used phenotypic method for confirming MBLs phenotypes among the studies. Sputum was the most common specimen from which MBL-PA was recovered. A significant positive correlation was observed between MDR and MBL production in P. aeruginosa. We conclude that MBL producing strains are widespread among the clinical isolates of P. aeruginosa in Nepal and responsible for emerging MDR strains. It is paramount that antibiotics prescription against the bacterium should be monitored closely and alternative therapeutic modalities against MBL-PA explored.

Understanding COVID-19 Situation in Nepal and Implications for SARS-CoV-2 Transmission and Management.

Background: The pandemic of Coronavirus Disease 2019 (COVID-19), one of the most infectious diseases in the modern history, is caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) and has had a profound health and economic toll, globally. This paper identifies the overall health status associated with COVID-19 pandemic in all 7 provinces of Nepal, a developing country in South Asia, analyzing data from January 2020 to February 2022. It focuses on the SARS-CoV-2 prevalence, transmission through wastewater and other routes, diagnostics, treatment options, and alternative medicines, thereby offering key perspectives for its management. Materials and Methods: Studies regarding coronavirus spanning the 2017 to 2022 period were searched on the web, Nepalese database, and Web of Science. Refined criteria included SARS-CoV-2 in wastewater of Nepal or worldwide. Demographic data (sex, age-group, and geographic location) were also obtained from websites and relevant reports of the Ministry of Health and Population (MOHP) of Nepal, ranging from January 2020 to February 2022. Moreover, trends concerning lockdown, business, and border activities in Nepal between February 2020 and October 2020 were evaluated. The viral dissemination pathways, diagnosis, and available treatment options, including the Ayurvedic medicine, were also examined. Results: Aerosols generated during the hospital, industrial, recreational, and household activities were found to contribute to the propagation of SARS-CoV-2 into environmental wastewater, thereby putting the surrounding communities at risk of infection. When lockdown ended and businesses opened in October 2020, the number of active cases of COVID-19 increased exponentially. Bagmati Province had the highest number of cases (53.84%), while the remaining 6 provinces tallied 46.16%. Kathmandu district had the highest number of COVID-19 cases (138, 319 cases), while Manang district had the smallest number of infections (81 cases). The male population was found to be predominantly infected (58.7%). The most affected age groups were the 31 to 40 years old males (25.92%) and the 21 to 30 years old females (26.85%). Conclusion: The pandemic impacted the public health and economic growth in our study duration. SARS-CoV-2 was prevalent in the wastewater of Nepal. The Terai districts and the megacities were mostly affected by SARS-CoV-2 infections. Working-age groups and males were identified as the highest risk groups. More investigations on the therapeutic and alternative cures are recommended. These findings may guide the researchers and professionals with handling the COVID-19 challenges in developing countries such as Nepal and better prepare for future pandemics.

Prevalence of Multidrug-Resistant Klebsiella pneumoniae Clinical Isolates in Nepal.

BACKGROUND: Multidrug-resistant Klebsiella pneumoniae (MDR-KP) are becoming increasingly common over the world. The focus of this research was to get a quantitative assessment of K. pneumoniae and their multidrug resistance (MDR) profile in Nepal. METHODS: Three electronic databases: PubMed, Google Scholar, and Research4Life were used to search publications specifying K. pneumoniae infections and/or their MDR status from January 2015 to October 2021. Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines was followed for the review, and R language 4.1.1 was used for analysis. Depending upon heterogeneity of data, we used random model for pooled data to examine the prevalence of the organism and the multidrug resistance. RESULTS: Evaluation included 16 studies, and the pooled estimation of K. pneumoniae in total clinical samples was 3% (95% CI; 0.01-0.05). In the meta-analysis, 14 studies were combined for determining the prevalence of K. pneumoniae in total positive clinical isolates which was 16% (95% CI: 0.11-0.20), while from 12 research studies, MDR status in the pathogen was found to be 64% (95% CI, 0.53-0.74). CONCLUSION: The MDR status of K. pneumoniae as well as the prevalence of the bacteria in Nepal was analyzed which showed alarming situation about administration of antibiotics and indicated choosing and developing reliable antibiotic strategies.

Prevalence of Colistin-Resistant Escherichia coli from Poultry in South Asian Developing Countries.

BACKGROUND: Antimicrobial resistance has become a huge problem in animal and public health worldwide. Inadvertent use of antibiotics in poultry has led to the emergence of resistance against many antibiotics, even to last resort of drugs like colistin. AIM: This study aimed to provide uniform quantitative estimate on the percentage prevalence of E. coli as well as to analyze their colistin resistance in poultry in South Asian developing countries. METHODS: Two electronic databases, PubMed and Research4Life, were used to search studies as per the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. The pooled data on the prevalence of E. coli and colistin resistance were analyzed. RESULTS: In the meta-analysis of 9 studies in South Asian region (Nepal, Bangladesh, Pakistan, and India), the pooled prevalence of E. coli was 73% (95% CI, 0.549-0.916). The pooled prevalence of colistin resistance against E. coli from poultry was found to be 28% (95% CI, 0.158-0.438). CONCLUSION: There is high prevalence of E. coli and their resistance rate against colistin in poultry. Therefore, antimicrobials in raising livestock should be supervised.

Asymptomatic SARS-CoV-2 Carriers: A Systematic Review and Meta-Analysis.

Asymptomatic cases of SARS-CoV-2 can be unknown carriers magnifying the transmission of COVID-19. This study appraised the frequency of asymptomatic individuals and estimated occurrence by age group and gender by reviewing the existing published data on asymptomatic people with COVID-19. Three electronic databases, PubMed, Embase, and Web of Science (WoS), were used to search the literature following the guidelines of Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA). The study population for this review included asymptomatic individuals infected with SARS-CoV-2 reported in original articles published up to 30 April 2020. A random effects model was applied to analyze pooled data on the prevalence of asymptomatic cases among all COVID-19 patients and also by age and gender. From the meta-analysis of 16 studies, comprising 2,788 SARS-CoV-2 infected patients, the pooled prevalence according to the random effect size of asymptomatic cases was 48.2% (95% CI, 30-67%). Of the asymptomatic cases, 55.5% (95% CI, 43.6-66.8%) were female and 49.6% (95% CI, 20.5-79.1%) were children. Children and females were more likely to present as asymptomatic COVID-19 cases and could act as unknown carriers of SARS-CoV-2. Symptom-based screening might fail to identify all SARS-CoV-2 infections escalating the threat of global spread and impeding containment. Therefore, a mass surveillance system to track asymptomatic cases is critical, with special attention to females and children.