Assessment of clinical and virological outcomes of rural and urban populations: COVID-19.

Objective: To assess the clinical and virological status in urban and rural populations. Methods: A cross-sectional study was conducted in a tertiary care hospital, Postgraduate Institute of Medical Sciences, Rohtak for a period of six months. Upper respiratory tract (URT) specimens including nasopharyngeal and oropharyngeal swabs were collected from the patients and their contacts and processed by RT-PCR technique for COVID-19 detection. Further, clinical and virological response in both the population were assessed and compared. Results: A total of 37,724 URT samples were tested, out of which 20,144 (53%) samples were from the rural population and 17,580 (47%) from the urban population. Out of the total samples from urban and rural population, COVID-19 positivity was 13.9% in urban population and 6.2% in rural population. Around 86% patients or contacts were asymptomatic in both the rural and urban population and rests were symptomatic 14%. Among the symptomatic patients, sore throat was seen as the most common presenting symptom (95-100%) followed by fever (80-83%), dry cough (55-61%), nasal discharge (18-23%), and breathlessness (3-5%) in both the rural and urban population. Conclusion: Our outcomes provide novel facts that the COVID-19 epidemic severely affected both rural and urban populations but with few differences. In our study, positivity rate in case of urban population was 13.9% as compared to 6.2% in rural population. There are two foremost facets that contributed variation in positivity in both the population. First, better immune response in rural population as compared to urban population which can be due to the fact that rural people in India are more exposed to various pathogens during their early lifetime thus, improving their immune status. Second, factor could be elevated population densities in urban areas which can contribute to increased infectiousness thus higher positivity rate. In addition, people living in urban population have to commute more for their work and are exposed to more people throughout the day thus, having more possibility to get infection of COVID-19 as compared to the rural population. To the best of our knowledge, there are no studies conducted on COVID-19, among rural population of Haryana. Hence, this study will allow us to fill the gap in knowledge about the variation in contagion spread and immune response in both rural and urban populations.

In-vitro and in-silico evaluation of the anti-chikungunya potential of Psidium guajava leaf extract and their synthesized silver nanoparticles.

Chikungunya is a notorious viral infection, which affects a large segment of world populations in absence of vaccines and antivirals. The current study evaluates of anti-chikungunya activities of Psidium guajava leaves extract and their green synthesized silver nanoparticles. Green synthesized nanoparticles were well characterized for their size and stability by dynamic light scattering (DLS), zeta potential, scanning electron microscopy (SEM) and their functional groups were analyzed by FTIR. Maximum non-toxic doses (MNTD) of extracts and nanoparticles were analysed by using Vero cell-lines. Anti-chikungunya activities of extracts and nano-particles were determined on Vero cells and their effects on cell viability were measured by MTT assay. The P. guajava nano-particles and extracts revealed the anti-chikungunya activities in the Vero cell. The cells viability was increased by 40% and 60% as compared to the virus control, when these cells were treated with MNTD of P. guajava nano-particles and extracts, respectively. To know the reason for antiviral activity, molecular docking of phytochemicals was done against a replication essential cysteine protease (nsP2) of Chikungunya. It was found that phytochemicals; Longifollen and Quercetin showed the minimum binding energy with nsP2. P. guajava extracts can be exploited to develop an effective anti-chikungunya agent. In the absence of CHIKV vaccines and antivirals, P. guajava may be used to develop rapid, responsive, specific, and cost-effective anti-chikungunya agents. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-021-00685-4.

Zika virus: an emerging challenge to public health worldwide.

Zika virus (ZIKV) is a mosquito-borne virus that was first isolated from Zika forest, Uganda, in 1947. Since its inception, major and minor outbreaks have been documented from several parts of world. Aedes spp. mosquitoes are the primary vectors of ZIKV, but the virus can also be transmitted through sexual practices, materno-fetal transmission, and blood transfusion. The clinical presentations of symptomatic ZIKV infections are similar to dengue and chikungunya, including fever, headache, arthralgia, retro-orbital pain, conjunctivitis, and rash. ZIKV often causes mild illness in the majority of cases, but in some instances, it is linked with congenital microcephaly and autoimmune disorders like Guillain-Barré syndrome. The recent Indian ZIKV outbreak suggests that the virus is circulating in the South East Asian region and may cause new outbreaks in future. At present, no specific vaccines or antivirals are available to treat ZIKV, so management and control of ZIKV infections rely mostly on preventive measures.

Phylogenetic analysis of the hemagglutinin gene of influenza A(H1N1)pdm09 and A(H3N2) virus isolates from Haryana, India.

Influenza A viruses are highly adaptable and are the main pathogen behind winter time morbidity. The present study reports the molecular and phylogenetic characterization of A(H1N1)pdm09 and H3N2 isolates from Haryana, India during 2015 influenza outbreak. A total of 144 nasopharyngeal samples were collected from Post Graduate Institute of Medical Sciences, Rohtak, Haryana, India form September 2014 to February 2016. The samples were screened for influenza A subtypes; A(H1N1)pdm09 and H3N2 by using real-time RT-PCR. Virus isolation and hemagglutinin gene sequencing studies were performed for selected positive samples. Out of 24 (16.6%) Influenza A positive samples, 13 (54.2%) and 11 (45.8%) were subtyped into A(H1N1)pdm09 and H3N2, respectively by real-time RT-PCR. Genetic analysis of A(H1N1)pdm09 isolates revealed the presence of key mutations (P100S, S202T and S220T) in HA gene as compare to reference strain A/California/07/2009 and these isolates were grouped in clade 6B.1 and 6B.2. All A(H3N2) isolates were clustered in clade 3C.2a and revealed specific amino acid substitutions of N161S and P214S in their HA genes in comparison to the reference strain A/Texas/50/2012. The HA gene sequences of all isolates showed 97-98% of nucleotide sequence similarity with their respective reference strains. Influenza A(H1N1)pdm09 and H3N2 isolates were drifted significantly from their respective vaccines strains of 2015-2016 and were more closely related to recommended vaccine strains for flu season 2017-2018. The study supports the need of routine influenza surveillance and continuous monitoring of the genetic changes in the major antigenic sites of these viruses.

Green synthesis of silver nanoparticles from medicinal plants and evaluation of their antiviral potential against chikungunya virus.

The exploration of nanoscale materials for their therapeutic potential against emerging and re-emerging infections has been increased in recent years. Silver nanoparticles (AgNPs) are known to possess antimicrobial activities against different pathogens including viruses and provide an excellent opportunity to develop new antivirals. The present study focused on biological synthesis of AgNPs from Andrographis paniculata, Phyllanthus niruri, and Tinospora cordifolia and evaluation of their antiviral properties against chikungunya virus. Synthesized plants AgNPs were characterized to assess their formation, morphology, and stability. The cytotoxicity assays in Vero cells revealed that A. paniculata AgNPs were most cytotoxic with maximum non-toxic dose (MNTD) value of 31.25 µg/mL followed by P. niruri (MNTD, 125 µg/mL) and T. cordifolia AgNPs (MNTD, 250 µg/mL). In vitro antiviral assay of AgNPs based on degree of inhibition of cytopathic effect (CPE) showed that A. paniculata AgNPs were most effective, followed by T. cordifolia and P. niruri AgNPs. The results of antiviral assay were confirmed by cell viability test using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) dye, which revealed that A. paniculata AgNPs inhibited the virus to a maximum extent. The cell viability of CHIKV-infected cells significantly increased from 25.69% to 80.76 and 66.8%, when treated with A. paniculata AgNPs at MNTD and ½MNTD, respectively. These results indicated that use of plants AgNPs as antiviral agents is feasible and could provide alternative treatment options against viral diseases which have no specific antiviral or vaccines available yet.

Applicability of molecular assays for detection and typing of herpes simplex viruses in encephalitis cases.

Herpes simplex viruses (HSVs) cause a latent infection in humans which is mainly associated with characteristic cold sores or fever blisters and genital blisters. Large segments of the world population are suffering from the HSV infection and early diagnosis as well as treatments are needed to avoid further complications. HSV surveillance is very sparse, especially from developing countries including India. The aim of the present study is to develop and evaluate molecular assays for rapid detection and typing of HSV. In the present study, viral DNA was extracted from cerebro-spinal fluid from HSV suspected encephalitis patients. The conventional multiplex PCR for HSV-1 and HSV-2 was optimized and their comparative analysis was done with Real-Time qPCR for detection and typing of HSV. Out of 137 clinical samples, eleven samples (8.03%) were diagnosed as HSV positive by Real-Time qPCR while ten (7.3%) by conventional multiplex PCR which were further typed as subtyping HSV-1 (nine) and HSV-2 (two). Real-Time qPCR is highly sensitive and able to detect 9.4 × 101 to 3.1 × 106 copies/ml of HSV DNA. Conventional PCR was found to be having 99.21% specificity with 100% sensitivity. The positive predictive value was 90.91% whereas negative predictive value was 100%. Logistic regression indicates blisters with pain and skin rash as the most significant symptoms associated with HSV infection. The present study could be applied for rapid, specific, sensitive and cost-effective diagnosis of HSV-1 and HSV-2 thereby helpful in better patient management through early detection and treatment of HSV.