Re-emerging Disease Research Articles

Bacteria Emerging As an Opportunistic Pathogen

Due to the re-emergence of infectious diseases, the causative pathogenic microorganisms are becoming the major microbiologic public health threat. It seems to be difficult to control the emergence of new and severe bacterial diseases. However, efforts are continuously being made to identify the main cause to prevent the uncontrolled spread of emerging diseases. This review focuses on emerging bacterial diseases and their causative bacteria and the pathway of pathogenesis.

Comparison of haematological profile in influenza A H1N1 positive and negative patient

Re-emergence of infectious diseases are commonly seen with viral infections among which those caused by Influenza viruses are by nature unstable and unpredictable because of the unique capability of changing their antigenic characteristics by mutation. This study aims to unravel the haematological consequences of Influenza A H1N1 infections and also to compare it with Influenza A H1N1 negative infections. Materials and Methods: Applying the relevant inclusion and exclusion criterion yielded a total hundred patients consisting of sixty-five Influenza A H1N1 positive and thirty-five Influenza A H1N1 negative patients. The CBC profile of all these subjects were obtained from an Automated hematology analyzer which yielded various haematological parameters. Results: The mean hemoglobin was 12.145 gm/dl in H1N1 positive cases and 12.6 gm/dl in H1N1 negative cases. The mean hematocrit value was 36.28% in H1N1 positive cases and 40.65% in H1N1 negative cases. 26 H1N1 positive cases had leucopenia, whereas 4 H1N1 negative cases had leucopenia and 8 H1N1 positive cases had leucocytosis. Lymphopenia was observed in 60% of H1N1 positive cases and in 22.9% H1N1 negative cases. Thrombocytopenia was a feature in 8 H1N1 positive cases and 2 H1N1 negative cases. Conclusions: Several haematological variations are observed in both H1N1 positive cases and H1N1 negative influenza cases, but to different extents. Keywords: H1N1, Influenza, Haematological.

Get comfortable with being uncomfortable: Experiences from diagnostic radiographers a year into the COVID-19 pandemic

Many of us worldwide are living through an unprecedented crisis with the reemergence and rapid spread of coronavirus disease (COVID-19). It may seem like a lifetime to many, but it was only slightly more than a year ago that Singapore reported its first COVID-19 case on January 23, 2020 where, at that point, the virus had yet to be named. As the situation worsened with global outbreaks, the World Health Organization (WHO) eventually declared COVID-19 a pandemic on March 11, 2020 [1].

Compelling Evidence for the Activity of Antiviral Peptides against SARS-CoV-2.

Multiple outbreaks of epidemic and pandemic viral diseases have occurred in the last 20 years, including those caused by Ebola virus, Zika virus, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The emergence or re-emergence of such diseases has revealed the deficiency in our pipeline for the discovery and development of antiviral drugs. One promising solution is the extensive library of antimicrobial peptides (AMPs) produced by all eukaryotic organisms. AMPs are widely known for their activity against bacteria, but many possess additional antifungal, antiparasitic, insecticidal, anticancer, or antiviral activities. AMPs could therefore be suitable as leads for the development of new peptide-based antiviral drugs. Sixty therapeutic peptides had been approved by the end of 2018, with at least another 150 in preclinical or clinical development. Peptides undergoing clinical trials include analogs, mimetics, and natural AMPs. The advantages of AMPs include novel mechanisms of action that hinder the evolution of resistance, low molecular weight, low toxicity toward human cells but high specificity and efficacy, the latter enhanced by the optimization of AMP sequences. In this opinion article, we summarize the evidence supporting the efficacy of antiviral AMPs and discuss their potential to treat emerging viral diseases including COVID-19.

Effectiveness and safety of cladribine in MS: Real-world experience from two tertiary centres

Background: Oral cladribine has been approved for the treatment of relapsing multiple sclerosis (MS) yet real-world evidence regarding its effectiveness and safety remains scarce. Objective: To evaluate efficacy and safety outcomes of MS patients following induction of cladribine. Methods: We evaluated our prospective cohort of cladribine-treated MS patients from two tertiary centres in Germany. Relapses, disability worsening and occurrence of new or enlarging T2-hyperintense magnetic resonance imaging (MRI) lesions were assessed as well as lymphocyte counts and herpes virus infections. Results: Among 270 patients treated with cladribine, we observed a profound reduction of both relapses and new or enlarging MRI lesions. Treatment appeared more efficacious, especially in patients without previous therapy or following platform substances. Patients switching from natalizumab were prone to re-emerging disease activity. Among patients following dimethyl fumarate pre-treatment, severe lymphopenia was common and associated with increased rates of herpes virus manifestations. Conclusion: Overall, we observed an efficacy and safety profile of cladribine consistent with data from the phase 3 clinical trial. However, patients switching from natalizumab experienced suboptimal disease control beyond rebound activity following cessation of natalizumab. Furthermore, dimethyl fumarate pre-treatment was associated with a profound risk of developing severe lymphopenia and subsequent herpes virus infections.

Disease surveillance in England and Wales, April 2021

Disease surveillance in England and Wales, April 2021

Environmental Concerns for Emergence and Reemergence of Novel Viral Infectious Diseases: Implications for SARS-CoV-2 and Other Pathogens

The existing COVID-19 pandemic is an anthropogenic disaster which looked inevitable, because despite repeated natural warnings no attention was paid to them. Recently many viral zoonotic infections – caused by SARS-CoV-1, Ebola and Nipah, to name a few – happened that became public health emergency. The most recently discovered zoonotic SARS-CoV-2 is responsible for the contemporary humanitarian disaster. When a virus undergoes zoonotic transmission, the humans begin to respond urgently to its testing and treatment, giving least attention to the actual cause behind the transmission. To prevent future outbreaks, and possible pandemics, it is indispensable to understand the reasons behind emergence of new infectious diseases in human beings. Some most likely reasons for emergence and reemergence of novel zoonotic infections include deforestation, land use change, wildlife trade, bush-meat consumption and rapid increase in international travel. This article attempts to explore and discuss the factors that trigger emergence and reemergence of novel infectious diseases, with special emphasis on SARS, in human beings.

Retrospective analysis of disease profile and percentage of COVID positivity among patients presenting to paediatric triage of a tertiary care hospital during COVID pandemic lockdown period

Background: Currently, limited reports analyze pediatric patients infected with SARS-Covid-19. We conducted this study with the purpose of assessing the disease profile and percentage of COVID positivity among pediatric patients presenting to our triage area.Methods: A retrospective quantitative observational study was conducted in the pediatric triage area, department of pediatrics, Civil hospital campus, Ahmedabad, India. Patients aged 0-12 years were included with the study period from 1stApril to 30th June, 2020. Patients fulfilling the ICMR testing criteria were considered as suspected COVID patients and tested for COVID. Remaining patients were triaged and managed either on OPD basis or admitted in PICU, NICU or pediatric ward. Also, data of OPD attendance rate and immunization rates of 3 months (April, May and June) of 2019 and 2020 were collected from pediatric OPD and vaccination center.Results: A total of 709 patients presented in the triage out of which 35.5% were neonates, 28.6% were between 1-12 month, 23.2%were between 1-5 years and 12.6% were between 5-12 years. 12 out of 97 suspected patients tested positive for COVID 19. Among COVID positive patients, 50% had uncomplicated illness or mild pneumonia, while 50 % had critical illness and were admitted in COVID ICU. Mortality among non COVID patients was 22.7%, and among COVID patients was 16.6%. A sharp fall was noted in the OPD attendance (10 times) and immunization rates (3 times) this year compared to last year.Conclusions: Incidence of COVID positivity was comparable to the studies done in other countries. Even during the pandemic peak, the major chunk of pediatric admissions still belonged to the non COVID conditions. The drastic fall in immunization rates can have implications in the form of re-emergence of vaccine preventable diseases in the coming times.

Evaluation of Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus Mosquitoes Competence to Oropouche virus Infection.

The emergence of new human viral pathogens and re-emergence of several diseases are of particular concern in the last decades. Oropouche orthobunyavirus (OROV) is an arbovirus endemic to South and Central America tropical regions, responsible to several epidemic events in the last decades. There is little information regarding the ability of OROV to be transmitted by urban/peri-urban mosquitoes, which has limited the predictability of the emergence of permanent urban transmission cycles. Here, we evaluated the ability of OROV to infect, replicate, and be transmitted by three anthropophilic and urban species of mosquitoes, Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus. We show that OROV is able to infect and efficiently replicate when systemically injected in all three species tested, but not when orally ingested. Moreover, we find that, once OROV replication has occurred in the mosquito body, all three species were able to transmit the virus to immunocompromised mice during blood feeding. These data provide evidence that OROV is restricted by the midgut barrier of three major urban mosquito species, but, if this restriction is overcome, could be efficiently transmitted to vertebrate hosts. This poses a great risk for the emergence of permanent urban cycles and geographic expansion of OROV to other continents.

Ecological and Socio-Economic Determinants of Livestock Animal Leptospirosis in the Russian Arctic.

Leptospirosis is a re-emerging zoonotic infectious disease caused by pathogenic bacteria of the genus Leptospira. Regional differences in the disease manifestation and the role of ecological factors, specifically in regions with a subarctic and arctic climate, remain poorly understood. We here explored environmental and socio-economic features associated with leptospirosis cases in livestock animals in the Russian Arctic during 2000–2019. Spatial analysis suggested that the locations of the majority of 808 cases were in “boreal” or “polar” climate regions, with “cropland,” “forest,” “shrubland,” or “settlements” land-cover type, with a predominance of “Polar Moist Cropland on Plain” ecosystem. The cases demonstrated seasonality, with peaks in March, June, and August, corresponding to the livestock pasturing practices. We applied the Forest-based Classification and Regression algorithm to explore the relationships between the cumulative leptospirosis incidence per unit area by municipal districts (G-rate) and a number of socio-economic, landscape, and climatic factors. The model demonstrated satisfactory performance in explaining the observed disease distribution (R2 = 0.82, p < 0.01), with human population density, livestock units density, the proportion of crop area, and budgetary investments into agriculture per unit area being the most influential socio-economic variables. Climatic factors demonstrated a significantly weaker influence, with nearly similar contributions of mean yearly precipitation and air temperature and number of days with above-zero temperatures. Using a projected climate by 2100 according to the RCP8.5 scenario, we predict a climate-related rise of expected disease incidence across most of the study area, with an up to 4.4-fold increase in the G-rate. These results demonstrated the predominant influence of the population and agricultural production factors on the observed increase in leptospirosis cases in livestock animals in the Russian Arctic. These findings may contribute to improvement in the regional system of anti-leptospirosis measures and may be used for further studies of livestock leptospirosis epidemiology at a finer scale.

Phylogenomics of Tick Inward Rectifier Potassium Channels and Their Potential as Targets to Innovate Control Technologies.

This study was conducted to enhance the identification of novel targets to develop acaricides that can be used to advance integrated tick-borne disease management. Drivers for the emergence and re-emergence of tick-borne diseases affecting humans, livestock, and other domestic animals in many parts of the world include the increased abundance and expanded geographic distribution of tick species that vector pathogens. The evolution of resistance to acaricides among some of the most important tick vector species highlights the vulnerability of relying on chemical treatments for tick control to mitigate the health burden of tick-borne diseases. The involvement of inward rectifier potassium (Kir) channels in homeostasis, diuresis, and salivary gland secretion in ticks and other pests identified them as attractive targets to develop novel acaricides. However, few studies exist on the molecular characteristics of Kir channels in ticks. This bioinformatic analysis described Kir channels in 20 species of hard and soft ticks. Summarizing relevant investigations on Kir channel function in invertebrate pests allowed the phylogenomic study of this class of ion channels in ticks. How this information can be adapted to innovate tick control technologies is discussed.

No Such Thing as a Free-Rider? Understanding Drivers of Childhood and Adult Vaccination through a Multicountry Discrete Choice Experiment.

Increased vaccine hesitancy and refusal negatively affects vaccine uptake, leading to the reemergence of vaccine preventable diseases. We aim to quantify the relative importance of factors people consider when making vaccine decisions for themselves, or for their child, with specific attention for underlying motives arising from context, such as required effort (accessibility) and opportunism (free riding on herd immunity). We documented attitudes towards vaccination and performed a discrete choice experiment in 4802 respondents in The United Kingdom, France and Belgium, eliciting preferences for six attributes: (1) vaccine effectiveness, (2) vaccine preventable disease burden, (3) vaccine accessibility in terms of copayment, vaccinator and administrative requirements, (4) frequency of mild vaccine-related side-effects, (5) vaccination coverage in the country’s population and (6) local vaccination coverage in personal networks. We distinguished adults deciding on vaccination for themselves from parents deciding for their youngest child. While all attributes were found to be significant, vaccine effectiveness and accessibility stood out in all (sub)samples, followed by vaccine preventable disease burden. We confirmed that people attach more value to severity of disease compared to its frequency, and discovered that peer influence dominates free-rider motives, especially for the vaccination of children. These behavioral data are insightful for policy and are essential to parameterize dynamic vaccination behavior in simulation models. In contrast to what most game theoretical models assume, social norms dominate free-rider incentives. Policy-makers and healthcare workers should actively communicate on high vaccination coverage, and draw attention to the effectiveness of vaccines while optimizing their practical accessibility.

Qualitative Phytochemical Screening of Selected Medicinal Plants

Phytochemicals have great potency as therapeutic agents. There is continuous and urgent need to discover new therapeutic compounds with diverse chemical structures and novel mechanism of action because there has been an alarming increase in the incidence of new and re-emerging infectious diseases. Hence, the present investigation was carried out to assess the phytoconstituents of leaf extracts of four different medicinal plants viz. Acorus calamus, Senna alata, Solanum torvum and Solanum trilobatum. The leaves of four plants were collected from their wild habitats, washed, air dried and then powdered. The solvent extracts of the respective leaves were prepared using Soxhlet apparatus with acetone, ethanol and petroleum ether. The extracts were subjected to preliminary phytochemical analyses as per standard procedures. The results showed that alkaloids were present in all the four plants analyzed and all the three solvents extracted. Meanwhile, carbohydrates and proteins were absent in the same. It was also indicated that the ethanol extract of Solanum trilobatum leaf showed the presence of 7 compounds among 9 phytochemicals analyzed. The result of this study is encouraging further quantitative estimation and the need for clinical studies to determine the potential effectiveness of particular phytochemical in vivo.&#x0D; Keywords: Preliminary phytochemical analyses, Acorus calamus, Senna alata, Solanum torvum and Solanum trilobatum.

Novel and Re-emerging Zoonotic Viral Diseases in India during Last Two Decades: An Overview

Zoonotic diseases or zoonosis is an infectious disease caused by the transmission of pathogens from animals to humans. Depending on the pathogens, these diseases can be bacterial, viral, or parasitic and the route of transmission of the pathogens can be via reservoirs including birds, bats, pigs, and mosquitoes, eventually infecting humans. Spread among humans arises primarily through person-person contact and fomites, causing major outbreaks (epidemics and pandemics). Some of the major zoonotic outbreaks include plague, influenza, West Nile fever, brucellosis, and rabies. This review focuses on zoonotic viral outbreaks in India over the past 20 years highlighting the spread and severity of the disease, measures to control the infection and prevention, treatment and management of the infectious diseases. New or emerging and re-emerging viral diseases such as coronavirus, Japanese encephalitis, dengue and chikungunya are discussed. These outbreaks have been reported to cause high morbidity and mortality, in addition, pose a major risk to the health security, safety, and economy of the country. Insufficient knowledge of emerging diseases is a major challenge for producing effective anti-viral drugs and vaccines. Understanding the etiology and spread of the disease is essential for preventive measures and the development of efficient treatment strategies.

American Mammals Susceptibility to Dengue According to Geographical, Environmental, and Phylogenetic Distances

Many human emergent and re-emergent diseases have a sylvatic cycle. Yet, little effort has been put into discovering and modeling the wild mammal reservoirs of dengue (DENV), particularly in the Americas. Here, we show a species-level susceptibility prediction to dengue of wild mammals in the Americas as a function of the three most important biodiversity dimensions (ecological, geographical, and phylogenetic spaces), using machine learning protocols. Model predictions showed that different species of bats would be highly susceptible to DENV infections, where susceptibility mostly depended on phylogenetic relationships among hosts and their environmental requirement. Mammal species predicted as highly susceptible coincide with sets of species that have been reported infected in field studies, but it also suggests other species that have not been previously considered or that have been captured in low numbers. Also, the environment (i.e., the distance between the species' optima in bioclimatic dimensions) in combination with geographic and phylogenetic distance is highly relevant in predicting susceptibility to DENV in wild mammals. Our results agree with previous modeling efforts indicating that temperature is an important factor determining DENV transmission, and provide novel insights regarding other relevant factors and the importance of considering wild reservoirs. This modeling framework will aid in the identification of potential DENV reservoirs for future surveillance efforts.

Evaluation of the diagnostic potential of recombinant leptospiral OMP A-like protein (Loa22) and transmembrane (OmpL37) protein in latex agglutination test for serodiagnosis of leptospirosis in animals.

Leptospirosis is a re-emerging zoonotic disease of animals and humans caused by pathogenic Leptospira, which has major public health concerns. The study is aimed to express the recombinant outer membrane protein (OMP) A-like protein (rLoa22) and transmembrane (rOmpL37) protein of Leptospira interrogans serovar Hardjo in the Escherichia coli and their evaluation as a diagnostic antigen in the latex agglutination test (LAT) to detect anti-leptospiral antibodies in the sera of animals. The Loa22 and OmpL37 genes lacking signal peptide coding sequences were individually amplified (522 and 963bp), by polymerase chain reaction, and directionally cloned into a pETite N-His Kan vector for expression. The expressed purified proteins were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblot, which confirmed leptospiral specific reactive protein with a molecular weight of ~19 and 36kDa, respectively. The sensitized latex beads coated with these OM proteins separately were evaluated in LAT using cattle sera of microscopic agglutination test (MAT) confirmed positive (n=53) and negative (n=52) cases of leptospirosis. The rLoa22 LAT and rOmpL37 LAT revealed the relative diagnostic sensitivity of 94·34 and 96·23%, diagnostic specificity of 92·31 and 96·15% and accuracy of 93·33 and 96·19%, with the excellent agreement of Cohen's kappa value of 0·87 and 0·92, respectively. After extensive evaluation, this rapid recombinant protein-based field diagnostic test can be applied as a screening test for the detection of anti-leptospiral antibodies in the sera of animals in the field conditions.

Recurrent neural network ensemble, a new instrument for the prediction of infectious diseases.

Infectious diseases afflict human beings since ancient times. We can classify the infectious disease in two principal types: the emerging diseases, that are caused by new pathogens, and the re-emerging diseases, due to a new spread of a known pathogen. Both types can then be subdivided in natural, accidental or intentional spreads. The risk associated to infectious diseases strongly increased in the last decades, especially because of the globalisation, which leads to a denser and more efficient link between nations, involving that a local infectious may easily spread worldwide, such as the SARS-CoV-2 in 2019–2020. The development of new methods to predict the spread of diseases is crucial. However, sometimes the variables are too many that classical algorithms fail in the prediction. Aim of this work is to investigate the use of an ensemble of recurrent neural networks for disease prediction, using real flu’s data to train and develop an instrument with the capability to determine the future flues. Two different types of study have been conducted. The first study investigates the influence of the neural network architecture, and it has been performed using 12 seasons to train the model and 3 seasons to test it. The second test aims to investigate the number of seasons needed to have a good prediction for future ones. The results demonstrated that this approach could ensure very high performances also with simple architectures. The ensemble approach allows to have information about the uncertainty of the prediction, allowing also to take countermeasures as a function of that value. In the future, the use of this approach may be applied to many other types of disease.

Human and bovine tuberculosis knowledge, attitude and practice (KAP) among cattle owners in Ethiopia

Tuberculosis (TB) is a re-emerging disease occurring worldwide, resulting in multi-billion-dollar loss and human death annually. The situation is worse in developing countries like Ethiopia, where lower knowledge, attitude, and practice (KAP) of the people is poor about the disease. A questionnaire-based cross-sectional study was conducted to assess livestock owners' KAP level towards human and bovine Tuberculosis in Gondar, Ethiopia. A total of 349 study participants were addressed through a face-to-face interview. Descriptive statistics and Pearson's chi-squares analysis were used to analyze the data and observe the association between outcome (KAP level) and predictor variables (sociodemographic characteristics). Out of the 349 respondents, 223 (63.9%) were males, while 126 (36.1%) were females. The KAP measuring interview indicated that 97.4% of the participants are aware of human tuberculosis, while only 84 (24.1%) know about bovine tuberculosis cause and mode of transmission. Inhalation was reported as the main route of transmission for human TB (41.1%) whereas, 50% of the respondent mentioned inhalation, contact, and ingestion of raw animal products as the main route of TB transmission from animal to human. Among those who have heard of bovine tuberculosis, only 56 (66.7%) of respondents consider bovine tuberculosis as a significant threat to public health. The study showed there is a lower KAP on bovine TB among cattle owners in the study area. Therefore, community health education about the impact of the disease, transmission, control, and prevention should be integrated with one health-oriented education and research to eradicate the disease from the country.

Diphtheria: a re-emerging infectious disease

&lt;p class="abstract"&gt;&lt;strong&gt;Background: &lt;/strong&gt;Diphtheria is a fatal bacterial infection which affects the mucous membranes of oropharyngeal and nasal cavity, caused by aerobic gram-positive bacteria Corynebacterium diphtheriae. Clinical diphtheria is on the increase worldwide, mainly affecting developing countries and leading cause of morbidity and mortality. We sought to understand its presentation among patients and also early intervention of the disease. The objective of the study was to study the clinico-demographic profile of cases, the relationship between immunization status and clinical disease, the role of microbiological investigations and to identify complications in diphtheria cases.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Methods:&lt;/strong&gt; This study is a hospital based observational study from September 2019 - September 2020 at a tertiary care centre, S.Nijalingappa Medical College and HSK hospital, Bagalkot, Karnataka. The cases were analysed with respect to demographic details, clinical features, immunization status, microbiological confirmation and complications of diphtheria cases.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Results: &lt;/strong&gt;32 cases were suspected to have diphtheria on clinical examination. The mean age of presentation was 15 years. Fever, sore throat, difficulty in swallowing, neck swelling and pseudomembrane in oral cavity were the common signs and symptoms. Airway compromise, myocarditis and neurological complications were noted. Antidiphtheritic serum (ADS) was tried in all 32 cases and 8 cases had adverse reactions. Case fatality rate was 12.5%.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;Shifting of occurrence of diphtheria in the age group of &amp;gt;5 years suggest the need to improve and strengthen the immunization program specially the booster doses.&lt;/p&gt;

COVID-19 Crisis Creates Opportunity towards Global Monitoring & Surveillance.

The spectrum of emerging new diseases as well as re-emerging old diseases is broadening as infectious agents evolve, adapt, and spread at enormous speeds in response to changing ecosystems. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a recent phenomenon and may take a while to understand its transmission routes from less traveled territories, ranging from fomite exposure routes to wastewater transmission. The critical challenge is how to negotiate with such catastrophic pandemics in high-income countries (HICs ~20% of the global population) and low-and middle-income countries (LMICs ~ 80% of the global population) with a total global population size of approximately eight billion, where practical mass testing and tracing is only a remote possibility, particularly in low-and middle-income countries (LMICs). Keeping in mind the population distribution disparities of high-income countries (HICs) and LMICs and urbanisation trends over recent years, traditional wastewater-based surveillance such as that used to combat polio may help in addressing this challenge. The COVID-19 era differs from any previous pandemics or global health challenges in the sense that there is a great deal of curiosity within the global community to find out everything about this virus, ranging from diagnostics, potential vaccines/therapeutics, and possible routes of transmission. In this regard, the fact that the gut is the common niche for both poliovirus and SARS-CoV-2, and due to the shedding of the virus through faecal material into sewerage systems, the need for long-term wastewater surveillance and developing early warning systems for better preparedness at local and global levels is increasingly apparent. This paper aims to provide an insight into the ongoing COVID-19 crisis, how it can be managed, and what measures are required to deal with a current global international public health concern. Additionally, it shed light on the importance of using wastewater surveillance strategy as an early warning practical tool suitable for massive passive screening, as well as the urgent need for microfluidic technology as a rapid and cost-effective approach tracking SARS-CoV-2 in wastewater.