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Comparative genomic and phenotypic characterization of invasive non-typhoidal <i>Salmonella</i> isolates from Siaya, Kenya

PLoS Neglected Tropical Diseases News - 1 February 2021 - 2:00pm

by Jessica Z. Kubicek-Sutherland, Gary Xie, Migun Shakya, Priya K. Dighe, Lindsey L. Jacobs, Hajnalka Daligault, Karen Davenport, Loreen R. Stromberg, Zachary R. Stromberg, Qiuying Cheng, Prakasha Kempaiah, John Michael Ong’echa, Vincent Otieno, Evans Raballah, Samuel Anyona, Collins Ouma, Patrick S. G. Chain, Douglas J. Perkins, Harshini Mukundan, Benjamin H. McMahon, Norman A. Doggett

Non-typhoidal Salmonella (NTS) is a major global health concern that often causes bloodstream infections in areas of the world affected by malnutrition and comorbidities such as HIV and malaria. Developing a strategy to control the emergence and spread of highly invasive and antimicrobial resistant NTS isolates requires a comprehensive analysis of epidemiological factors and molecular pathogenesis. Here, we characterize 11 NTS isolates that caused bloodstream infections in pediatric patients in Siaya, Kenya from 2003–2010. Nine isolates were identified as S. Typhimurium sequence type 313 while the other two were S. Enteritidis. Comprehensive genotypic and phenotypic analyses were performed to compare these isolates to those previously identified in sub-Saharan Africa. We identified a S. Typhimurium isolate referred to as UGA14 that displayed novel plasmid, pseudogene and resistance features as compared to other isolates reported from Africa. Notably, UGA14 is able to ferment both lactose and sucrose due to the acquisition of insertion elements on the pKST313 plasmid. These findings show for the first time the co-evolution of plasmid-mediated lactose and sucrose metabolism along with cephalosporin resistance in NTS further elucidating the evolutionary mechanisms of invasive NTS phenotypes. These results further support the use of combined genomic and phenotypic approaches to detect and characterize atypical NTS isolates in order to advance biosurveillance efforts that inform countermeasures aimed at controlling invasive and antimicrobial resistant NTS.

Pulse-Controlled Amplification–A new powerful tool for on-site diagnostics under resource limited conditions

PLoS Neglected Tropical Diseases News - 29 January 2021 - 2:00pm

by Katharina Müller, Sarah Daßen, Scott Holowachuk, Katrin Zwirglmaier, Joachim Stehr, Federico Buersgens, Lars Ullerich, Kilian Stoecker

Background

Molecular diagnostics has become essential in the identification of many infectious and neglected diseases, and the detection of nucleic acids often serves as the gold standard technique for most infectious agents. However, established techniques like polymerase chain reaction (PCR) are time-consuming laboratory-bound techniques while rapid tests such as Lateral Flow Immunochromatographic tests often lack the required sensitivity and/or specificity.

Methods/Principle findings

Here we present an affordable, highly mobile alternative method for the rapid identification of infectious agents using pulse-controlled amplification (PCA). PCA is a next generation nucleic acid amplification technology that uses rapid energy pulses to heat microcyclers (micro-scale metal heating elements embedded directly in the amplification reaction) for a few microseconds, thus only heating a small fraction of the reaction volume. The heated microcyclers cool off nearly instantaneously, resulting in ultra-fast heating and cooling cycles during which classic amplification of a target sequence takes place. This reduces the overall amplification time by a factor of up to 10, enabling a sample-to-result workflow in just 15 minutes, while running on a small and portable prototype device. In this proof of principle study, we designed a PCA-assay for the detection of Yersinia pestis to demonstrate the efficacy of this technology. The observed detection limits were 434 copies per reaction (purified DNA) and 35 cells per reaction (crude sample) respectively of Yersinia pestis.

Conclusions/Significance

PCA offers fast and decentralized molecular diagnostics and is applicable whenever rapid, on-site detection of infectious agents is needed, even under resource limited conditions. It combines the sensitivity and specificity of PCR with the rapidness and simplicity of hitherto existing rapid tests.

ATG5 is instrumental in the transition from autophagy to apoptosis during the degeneration of tick salivary glands

PLoS Neglected Tropical Diseases News - 29 January 2021 - 2:00pm

by Yanan Wang, Houshuang Zhang, Li Luo, Yongzhi Zhou, Jie Cao, Xuenan Xuan, Hiroshi Suzuki, Jinlin Zhou

Female tick salivary glands undergo rapid degeneration several days post engorgement. This degeneration may be caused by the increased concentration of ecdysone in the hemolymph during the fast feeding period and both autophagy and apoptosis occur. In this work, we first proved autophagy-related gene (ATG) and caspase gene expression peaks during degeneration of the tick salivary glands. We explored the regulatory role of Rhipicephalus haemaphysaloides autophagy-related 5 (RhATG5) in the degeneration of tick salivary glands. During the fast feeding phase, RhATG5 was cleaved and both calcium concentration and the transcription of Rhcalpains increased in the salivary glands. Recombinant RhATG5 was cleaved by μ-calpain only in the presence of calcium; the mutant RhATG5191-199Δ was not cleaved. Treatment with 20-hydroxyecdysone (20E) led to programmed cell death in the salivary glands of unfed ticks in vitro, RhATG8-phosphatidylethanolamine (PE) was upregulated in ticks treated with low concentration of 20E. Conversely, RhATG8-PE decreased and Rhcaspase-7 increased in ticks treated with a high concentration of 20E and transformed autophagy to apoptosis. High concentrations of 20E led to the cleavage of RhATG5. Calcium concentration and expression of Rhcalpains were also upregulated in the tick salivary glands. RNA interference (RNAi) of RhATG5 in vitro inhibited both autophagy and apoptosis of the tick salivary glands. RNAi of RhATG5 in vivo significantly inhibited the normal feeding process. These results demonstrated that high concentrations of 20E led to the cleavage of RhATG5 by increasing the concentration of calcium and stimulated the transition from autophagy to apoptosis.

Overview of snakebite in Brazil: Possible drivers and a tool for risk mapping

PLoS Neglected Tropical Diseases News - 29 January 2021 - 2:00pm

by Maria Cristina Schneider, Kyung-duk Min, Patricia Nájera Hamrick, Lucia R. Montebello, Tani Maria Ranieri, Lucia Mardini, Volney M. Camara, Ronir Raggio Luiz, Bernhard Liese, Myriam Vuckovic, Milton Ozorio Moraes, Nísia Trindade Lima

Snakebite envenoming affects close to 2.7 million people globally every year. In Brazil, snakebites are reported to the Ministry of Health surveillance system and cases receive antivenom free of charge. There is an urgent need to identify higher risk areas for antivenom distribution, and to develop prevention activities. The objective of this study is to provide an overview of the epidemiological situation of snakebite envenoming in Brazil and explore possible drivers; as well as to create a flowchart tool to support decision-makers identify higher risk areas. An ecological-type study was carried out using data by municipality (2013–2017). Study parts: 1) Create a geocoded database and perform a descriptive and cluster analysis; 2) Statistical analysis to measure the association of snakebite and possible environmental and socioeconomic drivers; 3) Develop a flowchart to support decision-makers and the application of this tool in one state (Rio Grande do Sul) as an example. An average of 27,120 snakebite cases per year were reported at the country level. Clusters of municipalities with high numbers of snakebites are mostly found in the Amazon Legal Region. The negative binomial regression model showed association with the snakebite case count: the type of major habitat, tropical or non-tropical; temperature; percentage of urbanization; precipitation; elevation; GDP per capita; a weaker relation with forest loss; and with venomous snake richness. The state where the instrument was applied reported 4,227 snakebites in the period. Most municipalities were considered as medium risk and 56/496 as high risk according to the tool created. Snakebite cases are distributed across the entire country with the highest concentration in the Legal Amazon Region. This creates a complex situation both for better understanding of the association of environmental and socioeconomic factors with snakebites and for the distribution and maintenance of antivenom to remote areas. Research into types of antivenom with a longer shelf life without the need for refrigeration is needed.

Severe hepatobiliary morbidity is associated with <i>Clonorchis sinensis</i> infection: The evidence from a cross-sectional community study

PLoS Neglected Tropical Diseases News - 28 January 2021 - 2:00pm

by Men-Bao Qian, Hong-Mei Li, Zhi-Hua Jiang, Yi-Chao Yang, Ming-Fei Lu, Kang Wei, Si-Liang Wei, Yu Chen, Chang-Hai Zhou, Ying-Dan Chen, Xiao-Nong Zhou

Clonorchis sinensis infection is highly prevalent in Asia. Diverse hepatobiliary morbidity has been documented for C. sinensis infection. This study aimed to assess the association between C. sinensis infection and hepatobiliary morbidity, taking into consideration of the control, confounders and infection intensity. A cross-sectional community survey was implemented in Hengxian county, southeastern China. Helminth infections were detected by fecal examination. Physical examination and abdominal ultrasonography were then conducted. After excluding confounding effects from gender, age and alcohol drinking, quantitative association between C. sinensis infection and hepatobiliary morbidity was assessed, and the effect from infection intensity was also evaluated, through adjusted odds ratio (aOR) and 95% confidence intervals (95% CI). 696 villagers older than 10 years were enrolled. The prevalence and infection intensity of C. sinensis were higher in male, elder people and the individuals consuming alcohol. Light C. sinensis infection was associated with the increase of diarrhoea (aOR: 2.2, 95% CI: 1.1–4.5). C. sinensis infection was associated with the increase of fatty liver (aOR: 2.7, 95% CI: 1.4–5.2), and the effect was similar in different infection intensities. Moderate C. sinensis infection was associated with the increase of gallbladder stone (aOR: 3.0, 95% CI: 1.1–8.6), while moderate and heavy infections with the increase of intrahepatic bile duct dilatation (aOR: 2.2, 95% CI: 1.0–4.9 and aOR: 4.3, 95% CI: 1.9–9.9, respectively). C. sinensis infection had an effect on the development of periductal fibrosis (aOR: 3.2, 95% CI: 2.1–4.9), which showed increasing trend by infection intensity. The length and width of gallbladder in those with C. sinensis infection were enlarged, especially in those over 30 years old. C. sinensis infection is significantly associated with hepatobiliary morbidity. The occurrence of some morbidity was strongly related to the infection intensity. Awareness on harm of clonorchiasis should be raised both for policy-makers and villagers to adopt effective interventions.

Major risk factors and histopathological profile of treatment failure, relapse and chronic patients with anthroponotic cutaneous leishmaniasis: A prospective case-control study on treatment outcome and their medical importance

PLoS Neglected Tropical Diseases News - 28 January 2021 - 2:00pm

by Mehdi Bamorovat, Iraj Sharifi, Shahriar Dabiri, Simin Shamsi Meymandi, Ali Karamoozian, Rezvan Amiri, Amireh Heshmatkhah, Mehdi Borhani Zarandi, Mohammad Reza Aflatoonian, Fatemeh Sharifi, Reza Kheirandish, Saeid Hassanzadeh

Over the last years, there has been a remarkable increase in the number of unresponsive patients with anthroponotic cutaneous leishmaniasis (ACL) reported worldwide. The primary objective of this study was to explore the role of demographic, clinical and environmental risk related-factors in the development of treatment failure, relapse and chronic cases compared to responsive patients with ACL. Moreover, molecular, histopathological and immunohistochemical (IHC) findings between these forms were explored. This work was undertaken as a prospective and case-control study in southeastern Iran. Culture media and nested PCR were used to identify the causative agent. Univariate multinomial and multiple multinomial logistic regression models and the backward elimination stepwise method were applied to analyze the data. A P<0.05 was defined as significant. Also, for different groups, skin punch biopsies were used to study the histopathological and immunohistochemical (IHC) profile. All samples showed that L. tropica was the only etiological agent in all unresponsive and responsive patients with ACL. Data analysis represented that 8 major risk factors including nationality, age groups, occupation, marital status, history of chronic diseases, duration of the lesion, the lesion on face and presence of domestic animals in the house were significantly associated with the induction of unresponsive forms. The histopathological and immunohistochemical findings were different from one form to another. The present findings clearly demonstrated a positive relation between ACL and distinct demographic, clinical and environmental risk determinants. Knowledge of the main risk factors for ACL infection is crucial in improving clinical and public health strategies and monitor such perplexing factors.

Lessons learned for surveillance strategies for trachoma elimination as a public health problem, from the evaluation of approaches utilised by Guinea worm and onchocerciasis programmes: A literature review

PLoS Neglected Tropical Diseases News - 28 January 2021 - 2:00pm

by Laura Senyonjo, Philip Downs, Elena Schmidt, Robin Bailey, Karl Blanchet

Introduction

A number of neglected tropical diseases are targeted for elimination or eradication. An effective surveillance system is critical to determine if these goals have been achieved and maintained. Trachoma has two related but morphologically different presentations that are monitored for elimination, the active infectious form of trachoma and trachomatous trichiasis (TT), the progression of the disease. There are a number of lessons learnt from the Guinea worm surveillance system that are particularly compatible for TT surveillance and the onchocerciasis surveillance system which can provide insights for surveillance of the infectious form of trachoma.

Methods/Principal findings

A literature search of peer-reviewed published papers and grey literature was conducted using PUBMED and Google Scholar for articles relating to dracunculiasis or Guinea worm, onchocerciasis and trachoma, along with surveillance or elimination or eradication. The abstracts of relevant papers were read and inclusion was determined based on specified inclusion and exclusion criteria. The credibility and bias of relevant papers were also critically assessed using published criteria. A total of 41 papers were identified that were eligible for inclusion into the review.The Guinea worm programme is designed around a surveillance-containment strategy and combines both active and passive surveillance approaches, with a focus on village-based surveillance and reporting. Although rumour reporting and a monetary incentive for the identification of confirmed Guinea worm cases have been reported as successful for identifying previously unknown transmission there is little unbiased evidence to support this conclusion. More rigorous evidence through a randomised controlled trial, influenced by motivational factors identified through formative research, would be necessary in order to consider applicability for TT case finding in an elimination setting. The onchocerciasis surveillance strategy focuses on active surveillance through sentinel surveillance of villages and breeding sites. It relies on an entomological component, monitoring infectivity rates of black flies and an epidemiological component, tracking exposure to infection in humans. Challenges have included the introduction of relatively complex diagnostics that are not readily available in onchocerciasis endemic countries and target thresholds, which are practically unattainable with current diagnostic tests. Although there is utility in monitoring for infection and serological markers in trachoma surveillance, it is important that adequate considerations are made to ensure evidence-based and achievable guidelines for their utility are put in place.

Conclusions/Significance

The experiences of both the Guinea worm and onchocerciasis surveillance strategies have very useful lessons for trachoma surveillance, pre- and post-validation. The use of a monetary reward for identification of TT cases and further exploration into the use of infection and serological indicators particularly in a post-validation setting to assist in identifying recrudescence would be of particular relevance. The next step would be a real-world evaluation of their relative applicability for trachoma surveillance.

Effectiveness of miltefosine in cutaneous leishmaniasis caused by <i>Leishmania tropica</i> in Pakistan after antimonial treatment failure or contraindications to first line therapy—A retrospective analysis

PLoS Neglected Tropical Diseases News - 28 January 2021 - 2:00pm

by Suzette Kämink, Boota Masih, Noor Ali, Aman Ullah, Syed Juma Khan, Shakil Ashraf, Tetyana Pylypenko, Martin P. Grobusch, Jena Fernhout, Margriet den Boer, Koert Ritmeijer

Background

Cutaneous leishmaniasis (CL) is a neglected tropical skin disease, caused by Leishmania protozoa. In Pakistan, where CL caused by L. tropica is highly endemic, therapy with pentavalent antimonials is the standard of care, but has significant toxicity when used in systemic therapy, while are no evidence-based safer alternative treatment options for L. tropica. The efficacy of oral miltefosine has not been studied in CL caused by L. tropica. We evaluated effectiveness and tolerability of miltefosine in patients with previous treatment failure or with contraindications to systemic antimonial treatment.

Methods

A retrospective review was conducted of a cohort of CL patients who were treated with a 28-day course of miltefosine between December 2017 and August 2019, in urban Quetta, Pakistan, an area endemic for L. tropica. Descriptive analyses were performed, and effectiveness was assessed by initial response after treatment, and final cure at routine follow up visits, six weeks to three months post-treatment. Tolerability was assessed by routinely reported adverse events.

Results

Of the 76 CL patients in the cohort, 42 (55%) had contraindications to systemic antimonial treatment, and 34 (45%) had failure or relapse after antimonial treatment. Twelve patients defaulted during treatment and 12 patients were lost to follow up. In the remaining 52 patients, final cure rate was 77% (40/52). In those with contraindications to systemic antimonial treatment the final cure rate was 83% (24/29) and in the failure and relapse group 70% (16/23). Twenty-eight patients (40.0%) reported 39 mild to moderate adverse events with the main complaints being nausea (41.0%), general malaise (25.6%), and stomach pain (12.8%).

Conclusion

Results indicate that miltefosine is an effective second line treatment in CL in areas endemic for L. tropica. Prospective studies with systematic follow up are needed to obtain definitive evidence of effectiveness and tolerability, including identification of risk factors for miltefosine treatment failure.

Human T-cell lymphotropic virus type 1 transmission dynamics in rural villages in the democratic republic of the congo with high nonhuman primate exposure

PLoS Neglected Tropical Diseases News - 28 January 2021 - 2:00pm

by Megan Halbrook, Adva Gadoth, Anupama Shankar, HaoQiang Zheng, Ellsworth M. Campbell, Nicole A. Hoff, Jean-Jacques Muyembe, Emile Okitolonda Wemakoy, Anne W. Rimoin, William M. Switzer

The Democratic Republic of the Congo (DRC) has a history of nonhuman primate (NHP) consumption and exposure to simian retroviruses yet little is known about the extent of zoonotic simian retroviral infections in DRC. We examined the prevalence of human T-lymphotropic viruses (HTLV), a retrovirus group of simian origin, in a large population of persons with frequent NHP exposures and a history of simian foamy virus infection. We screened plasma from 3,051 persons living in rural villages in central DRC using HTLV EIA and western blot (WB). PCR amplification of HTLV tax and LTR sequences from buffy coat DNA was used to confirm infection and to measure proviral loads (pVLs). We used phylogenetic analyses of LTR sequences to infer evolutionary histories and potential transmission clusters. Questionnaire data was analyzed in conjunction with serological and molecular data. A relatively high proportion of the study population (5.4%, n = 165) were WB seropositive: 128 HTLV-1-like, 3 HTLV-2-like, and 34 HTLV-positive but untypeable profiles. 85 persons had HTLV indeterminate WB profiles. HTLV seroreactivity was higher in females, wives, heads of households, and increased with age. HTLV-1 LTR sequences from 109 persons clustered strongly with HTLV-1 and STLV-1 subtype B from humans and simians from DRC, with most sequences more closely related to STLV-1 from Allenopithecus nigroviridis (Allen’s swamp monkey). While 18 potential transmission clusters were identified, most were in different households, villages, and health zones. Three HTLV-1-infected persons were co-infected with simian foamy virus. The mean and median percentage of HTLV-1 pVLs were 5.72% and 1.53%, respectively, but were not associated with age, NHP exposure, village, or gender. We document high HTLV prevalence in DRC likely originating from STLV-1. We demonstrate regional spread of HTLV-1 in DRC with pVLs reported to be associated with HTLV disease, supporting local and national public health measures to prevent spread and morbidity.

The entomological impact of passive metofluthrin emanators against indoor <i>Aedes aegypti</i>: A randomized field trial

PLoS Neglected Tropical Diseases News - 26 January 2021 - 2:00pm

by Gregor J. Devine, Gonzalo M. Vazquez-Prokopec, Wilbert Bibiano-Marín, Norma Pavia-Ruz, Azael Che-Mendoza, Anuar Medina-Barreiro, Josue Villegas, Gabriela Gonzalez-Olvera, Mike W. Dunbar, Oselyne Ong, Scott A. Ritchie, Thomas S. Churcher, Oscar D. Kirstein, Pablo Manrique-Saide

Background

In the absence of vaccines or drugs, insecticides are the mainstay of Aedes-borne disease control. Their utility is challenged by the slow deployment of resources, poor community compliance and inadequate household coverage. Novel application methods are required.

Methodology and principal findings

A 10% w/w metofluthrin “emanator” that passively disseminates insecticide from an impregnated net was evaluated in a randomized trial of 200 houses in Mexico. The devices were introduced at a rate of 1 per room and replaced at 3-week intervals. During each of 7 consecutive deployment cycles, indoor resting mosquitoes were sampled using aspirator collections. Assessments of mosquito landing behaviours were made in a subset of houses. Pre-treatment, there were no differences in Aedes aegypti indices between houses recruited to the control and treatment arms. Immediately after metofluthrin deployment, the entomological indices between the trial arms diverged. Averaged across the trial, there were significant reductions in Abundance Rate Ratios for total Ae. aegypti, female abundance and females that contained blood meals (2.5, 2.4 and 2.3-times fewer mosquitoes respectively; P<0.001). Average efficacy was 60.2% for total adults, 58.3% for females, and 57.2% for blood-fed females. The emanators also reduced mosquito landings by 90% from 12.5 to 1.2 per 10-minute sampling period (P<0.05). Homozygous forms of the pyrethroid resistant kdr alleles V410L, V1016L and F1534C were common in the target mosquito population; found in 39%, 24% and 95% of mosquitoes collected during the trial.

Conclusions/Significance

This is the first randomized control trial to evaluate the entomological impact of any volatile pyrethroid on urban Ae. aegypti. It demonstrates that volatile pyrethroids can have a sustained impact on Ae. aegypti population densities and human-vector contact indoors. These effects occur despite the presence of pyrethroid-resistant alleles in the target population. Formulations like these may have considerable utility for public health vector control responses.

Risk factors for antimony treatment failure in American Cutaneous Leishmaniasis in Northwestern-Argentina

PLoS Neglected Tropical Diseases News - 26 January 2021 - 2:00pm

by María F. García-Bustos, Gabriela González-Prieto, Alberto E. Pániz-Mondolfi, Cecilia Parodi, Josefina Beckar, Sibila Monroig, Federico Ramos, María C. Mora, Lourdes A. Delgado-Noguera, Yoshihisa Hashiguchi, Daniela Jaime, Sonia Moreno, Luisa Ruiz-Morales, César G. Lemir, Alejandra Barrio

Background. To date, there is no specific literature available on the determinants for therapeutic failure (TF) with meglumine antimoniate (MA) in Northwestern-Argentina. This study aimed to identify epidemiological, clinical, and treatment-related factors that could be involved in TF. Methodology/Principal Findings. We performed a case-control study. Cases were represented by patients who showed TF after administration of the first course of MA treatment, whereas, controls were determined as patients who evolved towards healing after the first MA cycle received. Crude Odds Ratios and their corresponding 90% confidence intervals (CI) were calculated, and risk factors were then tested by multivariate analysis using logistic binary regression. Three hundred and eighty-four patients with a presumptive diagnosis of ACL were recruited, and 153 with a positive diagnosis were selected. We included in the study 71 patients, who underwent specific treatment with MA, presented complete data on response to treatment, and had a minimum post-treatment follow-up of 6 months in cutaneous leishmaniasis, and 12 months in mucosal leishmaniasis. Of these, 34 (47.9%) presented TF. In the initial analysis, TF was significantly associated with the geographical area of disease acquisition (p = 0.036), the presence of mucosal lesions (p = 0.042), the presence of concomitant skin and mucosal lesions (p = 0.002), and lesion age ≥ 6 months (p = 0.018). Risk factors influencing TF in the final multivariate model included the geographical area where the disease was acquired (adjusted Odd Ratio 8.062; 95% CI 1.914–33.959; p = 0.004), and lesion age ≥ 6 months (adjusted Odd Ratio 10.037; 95% CI 1.383–72.843; p = 0.023). Conclusions/Significance. The results of the present study suggest the existence of some risk factors linked to TF in Northwestern-Argentina, which deserve further investigation. Herein we recorded a high percentage of TF and we described clinical and epidemiological characteristics associated with TF that could be taken into account improving the clinical management of patients.

Transcriptional and immunological analysis of the putative outer membrane protein and vaccine candidate TprL of <i>Treponema pallidum</i>

PLoS Neglected Tropical Diseases News - 26 January 2021 - 2:00pm

by Austin M. Haynes, Mark Fernandez, Emily Romeis, Oriol Mitjà, Kelika A. Konda, Silver K. Vargas, Maria Eguiluz, Carlos F. Caceres, Jeffrey D. Klausner, Lorenzo Giacani

Background

An effective syphilis vaccine should elicit antibodies to Treponema pallidum subsp. pallidum (T. p. pallidum) surface antigens to induce pathogen clearance through opsonophagocytosis. Although the combination of bioinformatics, structural, and functional analyses of T. p. pallidum genes to identify putative outer membrane proteins (OMPs) resulted in a list of potential vaccine candidates, still very little is known about whether and how transcription of these genes is regulated during infection. This knowledge gap is a limitation to vaccine design, as immunity generated to an antigen that can be down-regulated or even silenced at the transcriptional level without affecting virulence would not induce clearance of the pathogen, hence allowing disease progression.

Principal findings

We report here that tp1031, the T. p. pallidum gene encoding the putative OMP and vaccine candidate TprL is differentially expressed in several T. p. pallidum strains, suggesting transcriptional regulation. Experimental identification of the tprL transcriptional start site revealed that a homopolymeric G sequence of varying length resides within the tprL promoter and that its length affects promoter activity compatible with phase variation. Conversely, in the closely related pathogen T. p. subsp. pertenue, the agent of yaws, where a naturally-occurring deletion has eliminated the tprL promoter region, elements necessary for protein synthesis, and part of the gene ORF, tprL transcription level are negligible compared to T. p. pallidum strains. Accordingly, the humoral response to TprL is absent in yaws-infected laboratory animals and patients compared to syphilis-infected subjects.

Conclusion

The ability of T. p. pallidum to stochastically vary tprL expression should be considered in any vaccine development effort that includes this antigen. The role of phase variation in contributing to T. p. pallidum antigenic diversity should be further studied.

Prevalence of simian malaria parasites in macaques of Singapore

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Meizhi Irene Li, Diyar Mailepessov, Indra Vythilingam, Vernon Lee, Patrick Lam, Lee Ching Ng, Cheong Huat Tan

Plasmodium knowlesi is a simian malaria parasite currently recognized as the fifth causative agent of human malaria. Recently, naturally acquired P. cynomolgi infection in humans was also detected in Southeast Asia. The main reservoir of both parasites is the long-tailed and pig-tailed macaques, which are indigenous in this region. Due to increased urbanization and changes in land use, there has been greater proximity and interaction between the long-tailed macaques and the general population in Singapore. As such, this study aims to determine the prevalence of simian malaria parasites in local macaques to assess the risk of zoonosis to the general human population. Screening for the presence of malaria parasites was conducted on blood samples from 660 peridomestic macaques collected between Jan 2008 and Mar 2017, and 379 wild macaques collected between Mar 2009 and Mar 2017, using a Pan-Plasmodium-genus specific PCR. Positive samples were then screened using a simian Plasmodium species-specific nested PCR assay to identify the species of parasites (P. knowlesi, P. coatneyi, P. fieldi, P. cynomolgi, and P. inui) present. All the peridomestic macaques sampled were tested negative for malaria, while 80.5% of the 379 wild macaques were infected. All five simian Plasmodium species were detected; P. cynomolgi being the most prevalent (71.5%), followed by P. knowlesi (47.5%), P. inui (42.0%), P. fieldi (32.5%), and P. coatneyi (28.5%). Co-infection with multiple species of Plasmodium parasites was also observed. The study revealed that Singapore’s wild long-tailed macaques are natural hosts of the five simian malaria parasite species, while no malaria was detected in all peridomestic macaques tested. Therefore, the risk of simian malaria transmission to the general human population is concluded to be low. However, this can be better demonstrated with the incrimination of the vectors of simian malaria parasites in Singapore.

Are <i>Melanoides tuberculata</i> and <i>Tarebia granifera</i> (Gastropoda, Thiaridae), suitable first intermediate hosts of <i>Clonorchis sinensis</i> in Vietnam?

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Hung Manh Nguyen, Hien Hoang Van, Loan Thi Ho, Yulia V. Tatonova, Henry Madsen

Background

Two thiarid snail species, Melanoides tuberculata and Tarebia granifera have been reported as first intermediate hosts of the liver fluke Clonorchis sinensis; however, their role as true first intermediate hosts has not been verified. Thus, the present study aimed to clarify the suitability of these two snail species as first intermediate hosts of C. sinensis. This was accomplished by collecting snails from a highly endemic area for C. sinensis in Vietnam, the Thac Ba reservoir, and identifying shed cercariae using molecular techniques. We also conducted experimental infections of five snail species including M. tuberculata and T. granifera with eggs of C. sinensis.

Methodology/Principal findings

A total of 11,985 snails, representing 10 species were sampled. Five snail species, M. tuberculata, T. granifera, Lymnaea swinhoei, Parafossarulus manchouricus, and Bithynia fuchsiana were found shedding cercariae with an overall prevalence of infection ranging from 0.7% to 11.5%. Seven cercarial types were recorded. Cercariae of C. sinensis were only found in Parafossarulus manchouricus. Using a multiplex PCR approach for detecting C. sinensis infection, the prevalence in P. manchouricus was 4.2%. Additionally, all five snail species were experimentally exposed to C. sinensis eggs, however only P. manchouricus was successfully infected with an infection rate of 7.87%.

Conclusions/Significance

We confirmed that in the Thac Ba reservoir, Vietnam, the two thiarids, M. tuberculata and T. granifera are not suitable first intermediate hosts of C. sinensis. Only P. manchouricus was found infected by C. sinensis in nature, and was the only species that became infected experimentally.

Relevance of oxidative stress in inhibition of eIF2 alpha phosphorylation and stress granules formation during Usutu virus infection

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Ana-Belén Blázquez, Miguel A. Martín-Acebes, Teresa Poderoso, Juan-Carlos Saiz

Usutu virus (USUV) is an African mosquito-borne flavivirus closely related to West Nile, Japanese encephalitis, Zika, and dengue viruses. USUV emerged in 1996 in Europe, where quickly spread across the continent causing a considerable number of bird deaths and varied neurological disorders in humans, including encephalitis, meningoencephalitis, or facial paralysis, thus warning about USUV as a potential health threat. USUV replication takes place on the endoplasmic reticulum (ER) of infected cells, inducing ER stress and resulting in the activation of stress-related cellular pathways collectively known as the integrated stress response (ISR). The alpha subunit of the eukaryotic initiation factor eIF2 (eIF2α), the core factor in this pathway, is phosphorylated by stress activated kinases: protein kinase R (PKR), PKR-like endoplasmic reticulum kinase (PERK), heme-regulated inhibitor kinase (HRI), and general control non-repressed 2 kinase (GCN2). Its phosphorylation results, among others, in the downstream inhibition of translation with accumulation of discrete foci in the cytoplasm termed stress granules (SGs). Our results indicated that USUV infection evades cellular stress response impairing eIF2α phosphorylation and SGs assembly induced by treatment with the HRI activator ArsNa. This protective effect was related with oxidative stress responses in USUV-infected cells. Overall, these results provide new insights into the complex connections between the stress response and flavivirus infection in order to maintain an adequate cellular environment for viral replication.

Quality and composition of Albendazole, Mebendazole and Praziquantel available in Burkina Faso, Côte d’Ivoire, Ghana and Tanzania

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Moritz Seitzer, Sylvia Klapper, Humphrey D. Mazigo, Ulrike Holzgrabe, Andreas Mueller

Background

Even though the international combat against Neglected Tropical Diseases such as schistosomiasis or soil-transmitted helminthiases depends on reliable therapeutics, anthelminthic pharmacovigilance has been neglected on many national African drug markets. Therefore, quality and composition of Albendazole, Mebendazole and Praziquantel locally collected in Burkina Faso, Côte d’Ivoire, Ghana and Tanzania were analysed.

Methods

Samples of 88 different batches were obtained from randomly selected facilities. Sampling took place in Northwest Tanzania, Western Burkina Faso, Southeast Côte d’Ivoire and Southwest Ghana. Visual examination of both packaging and samples was performed according to the WHO ‘Be Aware’ tool. Products were then screened with the GPHF Minilab, consisting of tests of mass uniformity, disintegration times and thin-layer chromatography (TLC). Confirmatory tests were performed according to international pharmacopoeiae, applying assays for dissolution profiles and high-performance liquid chromatography (HPLC).

Findings

Despite minor irregularities, appearance of the products did not hint at falsified medicines. However, 19.6% of the brands collected in Ghana and Tanzania were not officially licensed for sale. Mass uniformity was confirmed in 53 out of 58 brands of tablets. 41 out of 56 products passed disintegration times; 10 out of the 15 failing products did not disintegrate at all. Evaluating TLC results, only 4 out of 83 batches narrowly missed specification limits, 18 batches slightly exceeded them. No more than 46.3% (31 / 67) of the tablets assayed passed the respective pharmaceutical criteria for dissolution. HPLC findings confirmed TLC results despite shifted specification limits: 10 out of 83 tested batches contained less than 90%, none exceeded 110%.

Conclusion

In the four study countries, no falsified anthelminthic medicine was encountered. The active pharmaceutical ingredient was not found to either exceed or fall below specification limits. Galenic characteristics however, especially dissolution profiles, revealed great deficits.

Variable bites and dynamic populations; new insights in <i>Leishmania</i> transmission

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Samuel Carmichael, Ben Powell, Thomas Hoare, Pegine B. Walrad, Jonathan W. Pitchford

Leishmaniasis is a neglected tropical disease which kills an estimated 50,000 people each year, with its deadly impact confined mainly to lower to middle income countries. Leishmania parasites are transmitted to human hosts by sand fly vectors during blood feeding. Recent experimental work shows that transmission is modulated by the patchy landscape of infection in the host’s skin, and the parasite population dynamics within the vector. Here we assimilate these new findings into a simple probabilistic model for disease transmission which replicates recent experimental results, and assesses their relative importance. The results of subsequent simulations, describing random parasite uptake and dynamics across multiple blood meals, show that skin heterogeneity is important for transmission by short-lived flies, but that for longer-lived flies with multiple bites the population dynamics within the vector dominate transmission probability. Our results indicate that efforts to reduce fly lifespan beneath a threshold of around two weeks may be especially helpful in reducing disease transmission.

Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of <i>Mycobacterium bovis</i>

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Thoko Flav Kapalamula, Jeewan Thapa, Mwangala Lonah Akapelwa, Kyoko Hayashida, Stephen V. Gordon, Bernard Mudenda Hang' ombe, Musso Munyeme, Eddie Samuneti Solo, Precious Bwalya, Mirriam Ethel Nyenje, Aki Tamaru, Yasuhiko Suzuki, Chie Nakajima

Bovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of patients as one of the first-line anti-TB drug, pyrazinamide, is ineffective against M. bovis. Currently, there are no rapid, simplified and low-cost diagnostic methods that can be easily integrated for use in many developing countries. Here, we report the development of a loop-mediated isothermal amplification (LAMP) assay for specific identification of M. bovis by targeting the region of difference 4 (RD4), a 12.7 kb genomic region that is deleted solely in M. bovis. The assay's specificity was evaluated using 139 isolates comprising 65 M. bovis isolates, 40 M. tuberculosis isolates, seven M. tuberculosis complex reference strains, 22 non-tuberculous mycobacteria and five other bacteria. The established LAMP detected only M. bovis isolates as positive and no false positives were observed using the other mycobacteria and non-mycobacteria tested. Our LAMP assay detected as low as 10 copies of M. bovis genomic DNA within 40 minutes. The procedure of LAMP is simple with an incubation at a constant temperature. Results are observed with the naked eye by a color change, and there is no need for expensive equipment. The established LAMP can be used for the detection of M. bovis infections in cattle and humans in resource-limited areas.

Malaria in Southern Venezuela: The hottest hotspot in Latin America

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Maria Eugenia Grillet, Jorge E. Moreno, Juan V. Hernández-Villena, Maria F. Vincenti-González, Oscar Noya, Adriana Tami, Alberto Paniz-Mondolfi, Martin Llewellyn, Rachel Lowe, Ananías A. Escalante, Jan E. Conn

Malaria elimination in Latin America is becoming an elusive goal. Malaria cases reached a historical ~1 million in 2017 and 2018, with Venezuela contributing 53% and 51% of those cases, respectively. Historically, malaria incidence in southern Venezuela has accounted for most of the country's total number of cases. The efficient deployment of disease prevention measures and prediction of disease spread to new regions requires an in-depth understanding of spatial heterogeneity on malaria transmission dynamics. Herein, we characterized the spatial epidemiology of malaria in southern Venezuela from 2007 through 2017 and described the extent to which malaria distribution has changed country-wide over the recent years. We found that disease transmission was focal and more prevalent in the southeast region of southern Venezuela where two persistent hotspots of Plasmodium vivax (76%) and P. falciparum (18%) accounted for ~60% of the total number of cases. Such hotspots are linked to deforestation as a consequence of illegal gold mining activities. Incidence has increased nearly tenfold over the last decade, showing an explosive epidemic growth due to a significant lack of disease control programs. Our findings highlight the importance of spatially oriented interventions to contain the ongoing malaria epidemic in Venezuela. This work also provides baseline epidemiological data to assess cross-border malaria dynamics and advocates for innovative control efforts in the Latin American region.

Sero-prevalence of brucellosis, Q-fever and Rift Valley fever in humans and livestock in Somali Region, Ethiopia

PLoS Neglected Tropical Diseases News - 25 January 2021 - 2:00pm

by Mohammed Ibrahim, Esther Schelling, Jakob Zinsstag, Jan Hattendorf, Emawayish Andargie, Rea Tschopp

Information on zoonotic diseases in humans and livestock are limited in pastoral/agro-pastoral communities in Ethiopia. A multi-stage cross sectional cluster design study was implemented with the aim to establish the seroprevalence of zoonotic diseases including brucellosis, Q-fever and Rift Valley fever (RVF) in humans and livestock in Adadle Woreda of the Somali Region, Ethiopia. Blood samples were collected from humans and livestock and tested by relevant serological tests. For brucellosis, Rose Bengal test (RBT) and indirect ELISA was used for screening and confirmatory diagnosis respectively. Indirect and competitive ELISA were also used for Q-fever and RVF respectively. The individual seropositivity of Q-fever in livestock was 9.6% (95% CI 5.9–15.1) in cattle, 55.7% (95% CI 46.0–65.0) in camels, 48.8% (95% CI 42.5–55.0) in goats, and 28.9% (95% CI 25.0–33.2) in sheep. In humans, seropositivity of Q-fever was 27.0% (95% CI 20.4–34.0), with prevalence in males of 28.9% vs 24.2% in females (OR = 1.3; 95% CI 0.6–2.5). Camel seropositivity of Q-fever was significantly associated with age (OR = 8.1; 95% CI 2.8–23.7). The individual apparent seroprevalence of RVF was 13.2% (95% CI 8.7–18.8) in humans, 17.9% (95% CI 11.0–27.8) in cattle, 42.6% (95% CI 34.8–50.7) in camels, 6.3% (95% CI 3.3–11.6) in goats and 7.4% (95% CI 4.7–11.5) in sheep. Camels had the highest seropositivity of both Q-fever and RVF. Only a weak correlation was observed between human and livestock seropositivity for both Q-fever and RVF. Only cattle and camels were seropositive for brucellosis by iELISA. The individual seroprevalence of brucellosis was 2.8(0.9–6.4) in humans, 1.5% (95% CI 0.2–5.2) in cattle and 0.6% (95% CI 0.0–3.2) in camels. This study showed the importance of zoonoses in Somali Region and is the first published study to describe RVF exposure in humans and livestock in the country. Even though human exposure to RVF virus was reported, public health sector of Somali Region has not given attention to such zoonoses. Collaboration between public and animal health sectors for further investigation on these zoonoses using the One Health concept is indispensable.

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