Epidemiological Field Studies Research Articles

Quantifying salivary iodine biomarker using total reflection X-ray fluorescence (TXRF): A method for assessing and monitoring iodine status simulated by Monte Carlo

Iodine stands as a vital trace element essential crucial for synthesizing thyroid hormones, which are essential for regulating metabolism and tissue growth. The “gold standard” for assessing iodine status in a population quantifies the iodine content in urine samples and is based on Inductively Coupled Plasma - Mass Spectrometry (ICP-MS). Such method, or methods involving the readings of the Sandell-Kolthoff (SK) reaction, require complex sample preparations and the use of several toxic chemical elements. However, recent studies have shown a strong correlation between urinary iodine content (UIC) and iodine present in saliva. Using Monte Carlo simulations, this paper presents a TXRF-based method to quantify salivary iodine biomarker for assessing and monitoring iodine status. In the theoretical analysis, mono and polychromatic (filtered) excitation sources are considered. The detection and quantification limits of iodine, as well as the calibration curves for mass recovery, used the strong I Kα1 and I Kα2 lines. It was found that the optimal X-ray excitation source energy for iodine detection is 50 keV monochromatic X-rays. In contrast, using simulated spectra from a tungsten anode target, the best result was found at 70 kVp (peak voltage), after filtering the white beam with a copper absorber. Detection limits of iodine are 0.121 ng and 2.292 ng, for mono and polychromatic sources, respectively. The theoretical results of this paper show that TXRF could be a fast, simple and accurate alternative to ICP-MS or SK reaction-based methods. Of clinical significance, quantifying salivary iodine using TXRF to assess and monitor iodine status could be particularly helpful to patients undertaking 131I treatment. In addition, it could be used in epidemiological field studies to determine iodine deficiency/excess in children and pregnant women.

Evaluacija performansi Veri-Q hemoglobinskog merača za procenu hemoglobina i hematokrita na mestu pružanja pomoći

Abstract: Introduction: Haemoglobin (Hb) and packed cell volume (PCV) are tests used in the assessment of anaemia. The Veri-Q haemoglobin meter is a new device in the point-of-care market used for the quantitative measurement of haemoglobin and packed cell volume. This study aimed to evaluate the performance of the Veri-Q haemoglobin meter for the assessment of haemoglobin and packed cell volume. Materials and Methods: Haemoglobin and packed cell volume estimations of one hundred and eleven participants were determined using the Veri-Q Red haemoglobin meter and statistically compared with those obtained from the conventional method (gold standard). Results: One hundred and eleven undergraduate students participated in this study, of whom 72 (64.9%) were females and 39 (35.1%) were males. The mean haemoglobin values calculated using the Veri-Q haemoglobin meter (11.97 ± 1.95 g/dl) were significantly lower than the values obtained with the conventional method (12.35 ± 1.66 g/dl) (t = 4.7549; p = 0.0001). Similar results were obtained with packed cell volume estimation. The performance indices of the Veri-Q haemoglobin meter were as follows: sensitivity (75.41% for Hb and 77.05% for PCV), specificity (28.0% for Hb and PCV), positive predictive value (PPV) (56.1% for Hb and 56.63% for PCV), negative predictive value (48.28% for Hb and 50% for PCV). The prevalence of anaemia in this study was 54.05%. Positive Likelihood Ratio (1.05 for Hb and 1.07 for PCV), Negative Likelihood Ratio (0.88 for Hb and 0.82 for PCV), Accuracy (54.05% for Hb and 54.95% for PCV). The Receiver Operative Characteristics (ROC) Area Under Curve (AUC) was 0.996 for Hb and 0.984 for PCV. A significant but weak positive correlation was found to exist between haemoglobin estimations using conventional and Veri-Q methods (r = 0.3013, P = 0.01); PCV (R = 0.2512, P = 0.07). The sensitivity of the device can be described as moderate while the specificity is low. The accuracy of the device is just average. Conclusion: The Veri-Q haemoglobin meter demonstrates an average level of accuracy and a high AUC, making it potentially useful for field epidemiological studies.

An outbreak of trichophytic tinea capitis in a group of schoolchildren in a rural area of the department of Cauca, Colombia

Introduction. Tinea capitis is a mycosis of keratinized tissue, which affects the scalp and may cause alopecia, pruritus, and desquamation. This type of mycosis is more frequent in school-age children, and it may represent a public health problem; the main etiological agents reported for Colombia are zoophilic dermatophytes. Objective. To characterize an outbreak of Tinea capitis in 32 children from a rural school in the department of Cauca. Materials and methods. We conducted an epidemiological field study using a structured survey to characterize sociodemographic aspects and predisposing factors for this mycosis. We collected samples of affected scalp scales and hair for mycological studies. The children and the general population received recommendations, about these mycoses’ prevention, from Cauca’s health authorities and the local hospital. The parents verbally approved the informed consent. Results. The etiological agent isolated in 63% of the collected samples was Trichophyton tonsurans, an anthropophilic dermatophyte, and the main predisposing factor was sharing razors (87.5%). Conclusions. Ideally, mycological studies define the etiological agent to propose therapeutics and recommendations in agreement with management guidelines. Implementation of multidisciplinary measures to control the outbreak and educate the population is required.

Microenvironment Tracker (MicroTrac) model to estimate time-location of individuals for air pollution exposure assessments: model evaluation using smartphone data.

A critical aspect of air pollution exposure assessments is determining the time spent in various microenvironments (ME), which can have substantially different pollutant concentrations. We previously developed and evaluated a ME classification model, called Microenvironment Tracker (MicroTrac), to estimate time of day and duration spent in eight MEs (indoors and outdoors at home, work, school; inside vehicles; other locations) based on input data from global positioning system (GPS) loggers. In this study, we extended MicroTrac and evaluated the ability of using geolocation data from smartphones to determine the time spent in the MEs. We performed a panel study, and the MicroTrac estimates based on data from smartphones and GPS loggers were compared to 37 days of diary data across five participants. The MEs were correctly classified for 98.1% and 98.3% of the time spent by the participants using smartphones and GPS loggers, respectively. Our study demonstrates the extended capability of using ubiquitous smartphone data with MicroTrac to help reduce time-location uncertainty in air pollution exposure models for epidemiologic and exposure field studies.

Triplex Immunostrip Assay for Rapid Diagnosis of Tobacco Mosaic Virus, Tobacco Vein Banding Mosaic Virus, and Potato Virus Y.

Mixed virus infection has increasingly become a problem in the production of Solanaceae crops in recent years; therefore, a fast and accurate detection method is needed. In this study, a novel triplex immunostrip assay was developed for the simultaneous detection of tobacco mosaic virus (TMV), tobacco vein banding mosaic virus (TVBMV), and potato virus Y (PVY). The limits of detection of this novel immunostrip reached 200 ppb (ng/ml), 1 ppm (µg/ml), and 2 ppm for TMV, PVY, and TVBMV particles, respectively. Importantly, no cross-reactivity was observed among TMV, TVBMV, and PVY or to a nontarget virus. When the assay was applied to suspected virus-infected tobacco, tomato, and potato samples collected from fields in Southwest China, samples of single or mixed virus infection were successfully identified. In conclusion, the triplex immunostrip assay provides a fast and easy to use on-site detection method for field epidemiological studies of TMV, TVBMV, and PVY, and for managing diseases that are caused by them.

Pesticide exposure and cortical brain activation among farmworkers in Costa Rica

BackgroundPrevious epidemiological studies have reported associations of pesticide exposure with poor cognitive function and behavioral problems. However, these findings have relied primarily on neuropsychological assessments. Questions remain about the neurobiological effects of pesticide exposure, specifically where in the brain pesticides exert their effects and whether compensatory mechanisms in the brain may have masked pesticide-related associations in studies that relied purely on neuropsychological measures. MethodsWe conducted a functional neuroimaging study in 48 farmworkers from Zarcero County, Costa Rica, in 2016. We measured concentrations of 13 insecticide, fungicide, or herbicide metabolites or parent compounds in urine samples collected during two study visits (approximately 3–5 weeks apart). We assessed cortical brain activation in the prefrontal cortex during tasks of working memory, attention, and cognitive flexibility using functional near-infrared spectroscopy (fNIRS). We estimated associations of pesticide exposure with cortical brain activation using multivariable linear regression models adjusted for age and education level. ResultsWe found that higher concentrations of insecticide metabolites were associated with reduced activation in the prefrontal cortex during a working memory task. For example, 3,5,6-trichloro-2-pyridinol (TCPy; a metabolite of the organophosphate chlorpyrifos) was associated with reduced activation in the left dorsolateral prefrontal cortex (β = −2.3; 95% CI: −3.9, −0.7 per two-fold increase in TCPy). Similarly, 3-phenoxybenzoic acid (3-PBA; a metabolite of pyrethroid insecticides) was associated with bilateral reduced activation in the dorsolateral prefrontal cortices (β = −3.1; 95% CI: −5.0, −1.2 and −2.3; 95% CI: −4.5, −0.2 per two-fold increase in 3-PBA for left and right cortices, respectively). These associations were similar, though weaker, for the attention and cognitive flexibility tasks. We observed null associations of fungicide and herbicide biomarker concentrations with cortical brain activation during the three tasks that were administered. ConclusionOur findings suggest that organophosphate and pyrethroid insecticides may impact cortical brain activation in the prefrontal cortex – neural dynamics that could potentially underlie previously reported associations with cognitive and behavioral function. Furthermore, our study demonstrates the feasibility and utility of fNIRS in epidemiological field studies.

Prevalence of pvmrp1 Polymorphisms and Its Contribution to Antimalarial Response.

As more sporadic cases of chloroquine resistance occur (CQR) in Plasmodium vivax (P. vivax) malaria, molecular markers have become an important tool to monitor the introduction and spread of drug resistance. P. vivax multidrug resistance-associated protein 1 (PvMRP1), as one of the members of the ATP-binding cassette (ABC) transporters, may modulate this phenotype. In this study, we investigated the gene mutations and copy number variations (CNVs) in the pvmrp1 in 102 P. vivax isolates from China, the Republic of Korea (ROK), Myanmar, Papua New Guinea (PNG), Pakistan, the Democratic People’s Republic of Korea (PRK), and Cambodia. And we also obtained 72 available global pvmrp1 sequences deposited in the PlasmoDB database to investigate the genetic diversity, haplotype diversity, natural selection, and population structure of pvmrp1. In total, 29 single nucleotide polymorphisms reflected in 23 non-synonymous, five synonymous mutations and one gene deletion were identified, and CNVs were found in 2.9% of the isolates. Combined with the antimalarial drug susceptibility observed in the previous in vitro assays, except the prevalence of S354N between the two CQ sensitivity categories revealed a significant difference, no genetic mutations or CNVs associated with drug sensitivity were found. The genetic polymorphism analysis of 166 isolates worldwide found that the overall nucleotide diversity (π) of pvmrp1 was 0.0011, with 46 haplotypes identified (Hd = 0.9290). The ratio of non-synonymous to synonymous mutations (dn/ds = 0.5536) and the neutrality tests statistic Fu and Li’s D* test (Fu and Li’s D* = −3.9871, p < 0.02) suggests that pvmrp1 had evolved under a purifying selection. Due to geographical differences, genetic differentiation levels of pvmrp1 in different regions were different to some extent. Overall, this study provides a new idea for finding CQR molecular monitoring of P. vivax and provides more sequences of pvmrp1 in Asia for subsequent research. However, further validation is still needed through laboratory and epidemiological field studies of P. vivax samples from more regions.

Application and reflection of laboratory-based monitoring in early warning of infectious diseases

Despite the fact that our cognition towards infectious disease prevention, the advanced technology and the economic status of the whole society has made a great progress in the last decade, the outbreak of COVID-19 pneumonia has again enabled the public to acquire more about super-challenges of infectious diseases, epidemics and the relevant preventive measurements. In order to identify the epidemic signals in early stage or even before the onset of epidemic, the data research and utilization of a series of factors related to the occurrence and transmission of infectious diseases have played a significant role in research of prevention and control during the whole period of surveillance and early warning. Laboratory-based monitoring for the etiology has always been an important part of infectious disease warning system due to pathogens as the direct cause of such diseases. China has initially established a laboratory-based monitoring and early warning system for bacterial infectious diseases based on the Chinese Pathogen Identification Network with an aim to identify pathogens, outbreaks and sources. This network has played an essential role in early detection, tracking and precise prevention and control of bacterial infectious diseases, such as plague, cholera, and epidemic cerebrospinal meningitis. This issue focuses on the function of laboratory-based monitoring during the period of early warning, prevention, and control of bacterial infectious diseases, and conducted a wide range of researches based on the analysis of the epidemic and outbreak isolates, together with field epidemiological studies and normal monitoring systems. All of these could illustrate the effect of laboratory surveillance in the infectious disease risk assessment and epidemic investigation. At the same time, we have put forward our review and expectation of scenarios about laboratory-based monitoring and early warning technologies to provide innovative thoughts for promoting a leapfrog development of infectious disease monitoring and early warning system in China.

Incidence of Malnutrition in Outpatient Department of Kerbala Teaching Hospital with some Associated Factors

Measurement of mid-upper arm circumference provides a simple and reliable tool for screening nutritional status and also enables rapid assessment of large populations in epidemiological field study.

Establishment of a Blocking ELISA Detection Method for Against African Swine Fever Virus p30 Antibody.

African swine fever (ASF) is a highly lethal hemorrhagic viral disease of domestic pigs caused by African swine fever virus (ASFV). A sensitive and reliable serological diagnostic assay is required, so laboratories can effectively and quickly detect ASFV infection. The p30 protein is abundantly expressed early in cells and has excellent antigenicity. Therefore, this study aimed to produce and characterize p30 monoclonal antibodies with an ultimate goal of developing a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for ASFV antibody detection. Three monoclonal antibodies against p30 protein that were expressed in E. coli were generated, and their characterizations were investigated. Furthermore, a blocking ELISA based on a monoclonal antibody was developed. To evaluate the performance of the assay, 186 sera samples (88 negative and 98 positive samples) were analyzed and a receiver-operating characteristic (ROC) analysis was applied to determine the cutoff value. Based on the ROC analysis, the area under the curve (AUC) was 0.997 (95% confidence interval: 99.2 to 100%). Besides, a diagnostic sensitivity of 97.96% (95% confidence interval: 92.82 to 99.75%) and a specificity of 98.96% (95% confidence interval: 93.83 to 99.97%) were achieved when the cutoff value was set to 38.38%. Moreover, the coefficients of inter- and intra-batches were <10%, indicating the good repeatability of the method. The maximum dilution of positive standard serum detected by this ELISA method was 1:512. The blocking ELISA was able to detect seroconversion in two out of five pigs at 10 Dpi and the p30 response increasing trend through the time course of the study (0–20 Dpi). In conclusion, the p30 mAb-based blocking ELISA developed in this study demonstrated a high repeatability with maximized diagnostic sensitivity and specificity. The assay could be a useful tool for field surveillance and epidemiological studies in swine herd.

The first reported outbreak of an undetermined species of human infection with spotted fever group Rickettsia in Lu'an, China

ObjectiveOn May 2, 2017, an outbreak of unexplained fever with rashes was reported in Lu'an, China. In this study, we aimed to identify the possible pathogens, epidemiological characteristics, and risk factors of this outbreak. MethodsWe conducted descriptive field epidemiological studies. Blood samples were tested using an indirect immunofluorescence assay for Rickettsia rickettsii antibody, and nested polymerase chain reaction and gene sequencing assays were performed. ResultsWe recruited 39 cases who had symptomatic onset from April 20 to June 8, 2017. Among these, 9 were suspected cases, 18 were probable cases, and 12 were confirmed cases. No one died. The main clinical manifestations were fever (100%), rash (100%), fatigue (97.3%), myalgia (83.8%), and anorexia (83.8%). None of the patients died. Thirty-seven patients who were treated with antibiotics during hospitalization showed significant improvement. The cases were distributed across 14 townships in 2 counties. The median age was 59 (43.0–81.0) years, of which 93.3% had a history of tea picking (28/30), and 77.3% (17/22) had a history of tick bites. The mean incubation period was 5.0 days (2.0–13.0 days). Serum IgG titers were higher in convalescent patients than in the general population (p = 0.016). Phylogenetic analysis revealed that the ompA sequences of Rickettsia sp. Lu'an-2018 had an 86.8%–99.0% sequence identity with the 23 strains of Rickettsia found worldwide. ConclusionsThis was the first reported outbreak of an undetermined species of a human infection with the spotted fever group of Rickettsia in China, which might be caused by ticks biting local residents when picking tea.

Development of a dual-labeled, hydrolysis probe-based, real-time quantitative PCR assay for detection of both genotypes of duck circovirus-1 (DuCV-1) and DuCV-2.

In this study, we developed a real-time quantitative polymerase chain reaction (qPCR) assay based on a dual-labeled hydrolysis probe to simultaneously detect both duck circovirus (DuCV) 1 and DuCV-2. The reproducibility, sensitivity and specificity of the primer set and probe were evaluated using other duck pathogens. The detection limit was 20 copies per µL. The intra-assay coefficients of variation (CVs) were ≤ 0.73% and the inter-assay CVs were ≤ 1.89%. No cross-reaction occurred with other duck pathogens. In addition, the qPCR assay was successfully applied to the simultaneous detection of DuCV-1 and DuCV-2 in clinical field samples. Therefore, this assay will be useful for laboratory diagnosis and epidemiological field studies of DuCV.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11262-021-01862-9.

Challenges in conducting epidemiological field studies evaluating associations between heat stress and renal health among workers in informal sectors: experiences from India

Well-designed Environmental Epidemiological Studies (EES) play a crucial role in quantifying the influence of environmental exposures and their associated risks on health in the wider population. They provide critical research evidence for identifying and developing interventions to avert adverse health consequences from those exposures. However, uncertainty and variability inherent to any field based EES could hinder the nature and magnitude of association between an exposure and health outcome. This is particularly pronounced in resource limited settings and resource-tight research projects. The present study evaluated the association between occupational heat stress and renal health among informal work sectors in India which had some significant challenges. Informal workers exposed to chronic occupational heat stress had significantly higher adverse renal health outcomes than the unexposed workers. Our field challenges included gaining access and permissions to conduct the study, participant recalls bias and attrition, accurately estimating exposures, confounding from causes of both exposure and disease, and to a large extent tight-funding. Though opportunities are abundant, we must ensure field conditions are optimized to attain study objectives. A keen understanding and sensitivity towards the cultural and work settings is essential for successful project completion. Based on our experiences, we provide strategies to adopt to improve fieldwork and provide recommendations to help overcome the field challenges and achieve better results for future EES studies in developing country settings.

Discriminating the eight genotypes of the porcine circovirus type 2 with TaqMan-based real-time PCR

BackgroundThe porcine circovirus type 2 (PCV2) is divided into eight genotypes including the previously described genotypes PCV2a to PCV2f and the two new genotypes PCV2g and PCV2h. PCV2 genotyping has become an important task in molecular epidemiology and to advance research on the prophylaxis and pathogenesis of PCV2 associated diseases. Standard genotyping of PCV2 is based on the sequencing of the viral genome or at least of the open reading frame 2. Although, the circovirus genome is small, classical sequencing is time consuming, expensive, less sensitive and less compatible with mass testing compared with modern real-time PCR assays. Here we report about a new PCV2 genotyping method using qPCR.MethodsBased on the analysis of several hundred PCV2 full genome sequences, we identified PCV2 genotype specific sequences or single-nucleotide polymorphisms. We designed six TaqMan PCR assays that are specific for single genotypes PCV2a to PCV2f and two qPCRs targeting two genotypes simultaneously (PCV2g/PCV2d and PCV2h/PCV2c). To improve specific binding of oligonucleotide primers and TaqMan probes, we used locked nucleic acid technology. We evaluated amplification efficiency, diagnostic sensitivity and tested assay specificity for the respective genotypes.ResultsAll eight PCV2 genotype specific qPCRs demonstrated appropriate amplification efficiencies between 91 and 97%. Testing samples from an epidemiological field study demonstrated a diagnostic sensitivity of the respective genotype specific qPCR that was comparable to a highly sensitive pan-PCV2 qPCR system. Genotype specificity of most qPCRs was excellent. Limited unspecific signals were obtained when a high viral load of PCV2b was tested with qPCRs targeting PCV2d or PCV2g. The same was true for the PCV2a specific qPCR when high copy numbers of PCV2d were tested. The qPCR targeting PCV2h/PCV2c showed some minor cross-reaction with PCV2d, PCV2f and PCV2g.ConclusionGenotyping of PCV2 is important for routine diagnosis as well as for epidemiological studies. The introduced genotyping qPCR system is ideal for mass testing and should be a valuable complement to PCV2 sequencing, especially in the case of simultaneous infections with multiple PCV2 genotypes, subclinically infected animals or research studies that require large sample numbers.

Determining the serotype composition of mixed samples of pneumococcus using whole-genome sequencing.

Serotyping of Streptococcus pneumoniae is a critical tool in the surveillance of the pathogen and in the development and evaluation of vaccines. Whole-genome DNA sequencing and analysis is becoming increasingly common and is an effective method for pneumococcal serotype identification of pure isolates. However, because of the complexities of the pneumococcal capsular loci, current analysis software requires samples to be pure (or nearly pure) and only contain a single pneumococcal serotype. We introduce a new software tool called SeroCall, which can identify and quantitate the serotypes present in samples, even when several serotypes are present. The sample preparation, library preparation and sequencing follow standard laboratory protocols. The software runs as fast as or faster than existing identification tools on typical computing servers and is freely available under an open source licence at https://github.com/knightjimr/serocall. Using samples with known concentrations of different serotypes as well as blinded samples, we were able to accurately quantify the abundance of different serotypes of pneumococcus in mixed cultures, with 100 % accuracy for detecting the major serotype and up to 86 % accuracy for detecting minor serotypes. We were also able to track changes in serotype frequency over time in an experimental setting. This approach could be applied in both epidemiological field studies of pneumococcal colonization and experimental laboratory studies, and could provide a cheaper and more efficient method for serotyping than alternative approaches.

The Research Protocol of SENIOR Project-Psychiatric Services and Support System for Serious Mental Illness in Old Age, Kerala, India.

Background:India has the second-largest population of elderly in the world. Serious mental illness (SMI) is a subset of the mental disorders that result in significant functional impairment and is usually long term. Persons with SMI face several challenges in their old age that are different from the issues faced by younger people with SMI. Understanding the problems faced by elderly individuals suffering from SMI is fundamental for planning programs to address them. The SENIOR (Support Systems Evaluation of Neuropsychiatric Illness in Old age) project is a study aimed at evaluating the problems faced in obtaining mental health care by elderly persons having SMI in the Kerala state of India.Aim:To describe the scientific methodology of the SENIOR project.Methods:This study employs mixed-methods cross-sectional design among a minimum sample of 768 SMI patients identified through cluster sampling from three districts, and Focus Group Discussion among mental health program officials.Discussion:This paper presents a methodological model to assist researchers in future field epidemiological studies on mental illness. Assessing service needs and barriers to access for the most vulnerable among the mentally ill will help the policymakers make evidence-based decisions to improve their quality of life.

Increased tropospheric ozone levels enhance pathogen infection levels of amphibians

As a result of anthropogenic activities, changes to the chemistry of Earth's atmosphere pose a threat to ecosystem health and biodiversity. One such change is the increase in tropospheric ozone (O3), which is particularly severe in the Mediterranean basin area, where the levels of this pollutant are chronically high during spring and summer time. Within this region, Mediterranean mountain ecosystems are hot spots for biodiversity which may be especially vulnerable to changes in O3 levels. Declines in montane amphibian populations have been recorded worldwide, including the Mediterranean basin. A significant driver of these declines is the emerging infection disease, chytridiomycosis, caused by the aquatic fungus Batrachochytrium dendrobatidis (Bd). Chytridiomycosis has negatively affected populations of several amphibian species in the Spanish Central Range, including in the Sierra Guadarrama, and interactions with other biotic and abiotic factors are an important part of these declines. However, there is little evidence or knowledge of whether tropospheric O3 levels may be another factor in the outbreaks of this disease. To test the hypothesis that O3 levels are another interactive driver of Bd infection dynamics, two different approaches were followed: 1) an experimental study in open top chambers was used to quantify the aspects of how Bd infection progressed throughout the metamorphic process under four different O3 levels; and 2) a field epidemiological study was used to analyse the relationship between the Bd infection load in the Sierra de Guadarrama and tropospheric O3 levels during a 9 year period. Our results suggest that high O3 levels significantly delayed the rate of development of tadpoles and increased Bd infection, providing empirical evidence of two new separate ways that may explain population declines of montane amphibians.

Highly sensitive serological approaches for Pepino mosaic virus detection.

Pepino mosaic virus (PepMV) causes severe disease in tomato and other Solanaceous crops around globe. To effectively study and manage this viral disease, researchers need new, sensitive, and high-throughput approaches for viral detection. In this study, we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies (mAbs). A panel of highly specific and sensitive murine mAbs (15B2, 8H6, 23D11, 20D9, 3A6, and 8E3) could be produced through cell fusion, antibody selection, and cell cloning. Using the mAbs as the detection antibodies, we established double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), Dot-ELISA, and Tissue print-ELISA for detecting PepMV infection in tomato plants. Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1 310 720 and 1:20 480 (weight/volume ratio (w/v), g/mL), respectively. Among the three methods developed, the Tissue print-ELISA was found to be the most practical detection technique. Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing, demonstrating all three serological methods are reliable and effective for monitoring PepMV. Anti-PepMV mAbs and the newly developed DAS-ELISA, Dot-ELISA, and Tissue print-ELISA can benefit PepMV detection and field epidemiological study, and management of this viral disease, which is already widespread in tomato plants in Yunnan Province of China.

Clinical, Epidemiological and Experimental Approaches to Assess Adverse Health Outcomes of Indoor Biomass Smoke Exposure: Conclusions from An Indo-Swedish Workshop in Mysuru, January 2020

This report summarizes the outcome of a workshop held in Mysuru, India in January 2020 addressing the adverse health effects of exposure to biomass smoke (BMS). The aim of the workshop was to identify uncertainties and gaps in knowledge and possible methods to address them in the Mysuru study on Determinants of Health in Rural Adults (MUDHRA) cohort. Specific aims were to discuss the possibility to improve and introduce new screening methods for exposure and effect, logistic limitations and other potential obstacles, and plausible strategies to overcome these in future studies. Field visits were included in the workshop prior to discussing these issues. The workshop concluded that multi-disciplinary approaches to perform: (a) indoor and personalized exposure assessment; (b) clinical and epidemiological field studies among children, adolescents, and adults; (c) controlled exposure experiments using physiologically relevant in vitro and in vivo models to understand molecular patho-mechanisms are warranted to dissect BMS-induced adverse health effects. It was perceived that assessment of dietary exposure (like phytochemical index) may serve as an important indicator for understanding potential protective mechanisms. Well trained field teams and close collaboration with the participating hospital were identified as the key requirements to successfully carry out the study objectives.

Insights into the evolutionary history and epidemiological characteristics of the emerging lineage 1 porcine reproductive and respiratory syndrome viruses in China.

The newly emerged lineage 1 porcine reproductive and respiratory syndrome viruses (PRRSVs) (especially the NADC30-like and NADC34-like viruses) have posed a direct threat to the Chinese pig industry since 2013. The phylogenetic, epidemic, and recombinant properties of these viruses have not yet systematically analysed in China. This report presents regular surveillance and field epidemiological studies for PRRSV across China from 2007 to 2019. From over 4,000 detected clinical samples, 70 open reading frame five sequences and four complete genomes of lineage 1 viruses were successfully obtained. Combined with global data, we conducted an extensive and systematic molecular phylogeny analysis using a maximum likelihood tree. The Chinese lineage 1 viruses were clustered, and their temporal and spatial distribution was further explored. Multiple viral introductions of lineage 1 virus from the United States to China were detected, and some became endemic in China. There are three sub-lineage 1 clusters: lineage 1.5 (NADC34-like), lineage 1.6 and New Intro cluster (NADC30-like). These viruses show high genetic diversity and a wide distribution in China, with Henan Province showing the highest diversity. Moreover, Chinese lineage 1 viruses have developed an endemic NADC30-like cluster. The demographic feature of this cluster showed a more or less constant population expansion history with a recent decreasing trend. Moreover, the genome recombination of Chinese lineage 1 with two dominant clusters (Chinese HP-PRRSVs: lineage 8.7 and VR2332-like: lineage 5.1) was frequently detected, both of which have commercial vaccine strains available. Furthermore, recombination hotspots were discovered near NSP9 and ORF2-4 regions of the genome. Overall, these findings provide important insights into the evolution and geographical diversity of Chinese lineage 1 PRRSV. These results will facilitate the development of programmes for the control and prevention of the emerging lineage 1 viruses in China.