PLOS Neglected Tropical Diseases

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ISSN / EISSN : 1935-2735 / 1935-2735
Published by: Public Library of Science (PLoS) (10.1371)
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Kaori Terasaki, Birte Kalveram, , Terry Juelich, Jennifer K. Smith, Lihong Zhang, ,
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009785

Abstract:
Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus with a wide host range including ruminants and humans. RVFV outbreaks have had devastating effects on public health and the livestock industry in African countries. However, there is no approved RVFV vaccine for human use in non-endemic countries and no FDA-approved antiviral drug for RVFV treatment. The RVFV 78kDa protein (P78), which is a membrane glycoprotein, plays a role in virus dissemination in the mosquito host, but its biological role in mammalian hosts remains unknown. We generated an attenuated RVFV MP-12 strain-derived P78-High virus and a virulent ZH501 strain-derived ZH501-P78-High virus, both of which expressed a higher level of P78 and carried higher levels of P78 in the virion compared to their parental viruses. We also generated another MP-12-derived mutant virus (P78-KO virus) that does not express P78. MP-12 and P78-KO virus replicated to similar levels in fibroblast cell lines and Huh7 cells, while P78-High virus replicated better than MP-12 in Vero E6 cells, fibroblast cell lines, and Huh7 cells. Notably, P78-High virus and P78-KO virus replicated less efficiently and more efficiently, respectively, than MP-12 in macrophage cell lines. ZH501-P78-High virus also replicated poorly in macrophage cell lines. Our data further suggest that inefficient binding of P78-High virus to the cells led to inefficient virus internalization, low virus infectivity and reduced virus replication in a macrophage cell line. P78-High virus and P78-KO virus showed lower and higher virulence than MP-12, respectively, in young mice. ZH501-P78-High virus also exhibited lower virulence than ZH501 in mice. These data suggest that high levels of P78 expression attenuate RVFV virulence by preventing efficient virus replication in macrophages. Genetic alteration leading to increased P78 expression may serve as a novel strategy for the attenuation of RVFV virulence and generation of safe RVFV vaccines.
Fatima Zohra Hamlili, Fatou Thiam, Maureen Laroche, Adama Zan Diarra, , Papa Mouhamadou Gaye, Cheikh Binetou Fall, Babacar Faye, Cheikh Sokhna, Doudou Sow, et al.
PLoS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009725

Abstract:
Freshwater snails of the genera Biomphalaria, Bulinus, and Oncomelania are intermediate hosts of schistosomes that cause human schistosomiasis, one of the most significant infectious neglected diseases in the world. Identification of freshwater snails is usually based on morphology and, potentially, DNA-based methods, but these have many drawbacks that hamper their use. MALDI-TOF MS has revolutionised clinical microbiology and has emerged in the medical entomology field. This study aims to evaluate MALDI-TOF MS profiling for the identification of both frozen and ethanol-stored snail species using protein extracts from different body parts. A total of 530 field specimens belonging to nine species (Biomphalaria pfeifferi, Bulinus forskalii, Bulinus senegalensis, Bulinus truncatus, Bulinus globosus, Bellamya unicolor, Cleopatra bulimoides, Lymnaea natalensis, Melanoides tuberculata) and 89 laboratory-reared specimens, including three species (Bi. pfeifferi, Bu. forskalii, Bu. truncatus) were used for this study. For frozen snails, the feet of 127 field and 74 laboratory-reared specimens were used to validate the optimised MALDI-TOF MS protocol. The spectral analysis yielded intra-species reproducibility and inter-species specificity which resulted in the correct identification of all the specimens in blind queries, with log-score values greater than 1.7. In a second step, we demonstrated that MALDI-TOF MS could also be used to identify ethanol-stored snails using proteins extracted from the foot using a specific database including a large number of ethanol preserved specimens. This study shows for the first time that MALDI-TOF MS is a reliable tool for the rapid identification of frozen and ethanol-stored freshwater snails without any malacological expertise.
, Heather N. White, , Oliver W. Stringer, Vicente Y. Belizario Jr, , Eugenio L. de Hostos, Tala De Los Santos
PLoS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009782

Abstract:
Background Soil-transmitted helminths (STHs) are parasitic nematodes that inhabit the human intestine. They affect more than 1.5 billion people worldwide, causing physical and cognitive impairment in children. The global strategy to control STH infection includes periodic mass drug administration (MDA) based on the results of diagnostic testing among populations at risk, but the current microscopy method for detecting infection has diminished sensitivity as the intensity of infection decreases. Thus, improved diagnostic tools are needed to support decision-making for STH control programs. Methodology We developed a nucleic acid amplification test based on recombinase polymerase amplification (RPA) technology to detect STH in stool. We designed primers and probes for each of the four STH species, optimized the assay, and then verified its performance using clinical stool samples. Principal findings Each RPA assay was as sensitive as a real-time polymerase chain reaction (PCR) assay in detecting copies of cloned target DNA sequences. The RPA assay amplified the target in DNA extracted from human stool samples that were positive for STH based on the Kato-Katz method, with no cross-reactivity of the non-target genomic DNA. When tested with clinical stool samples from patients with infections of light, moderate, and heavy intensity, the RPA assays demonstrated performance comparable to that of real-time PCR, with better results than Kato-Katz. This new rapid, sensitive and field-deployable method for detecting STH infections can help STH control programs achieve their goals. Conclusions Semi-quantitation of target by RPA assay is possible and is comparable to real-time PCR. With proper instrumentation, RPA assays can provide robust, semi-quantification of STH DNA targets as an alternative field-deployable indicator to counts of helminth eggs for assessing infection intensity.
, Geofrey Ssentamu, Enock Matovu, Kevin Kamanyi Marucha, Francisco Aresta-Branco, Claudia Helbig,
PLoS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009738

Abstract:
Most researchers who study unicellular eukaryotes work with an extremely limited number of laboratory-adapted isolates that were obtained from the field decades ago, but the effects of passage in laboratory rodents, and adaptation to in vitro culture, have been little studied. For example, the vast majority of studies of Trypanosoma brucei biology have concentrated on just two strains, Lister 427 and EATRO1125, which were taken from the field over half a century ago and have since have undergone innumerable passages in rodents and culture. We here describe two new Trypanosoma brucei brucei strains. MAK65 and MAK98, which have undergone only 3 rodent passages since isolation from Ugandan cattle. High-coverage sequencing revealed that adaptation of the parasites to culture was accompanied by changes in gene copy numbers. T. brucei has so far been considered to be uniformly diploid, but we also found trisomy of chromosome 5 not only in one Lister 427 culture, but also in the MAK98 field isolate. Trisomy of chromosome 6, and increased copies of other chromosome segments, were also seen in established cultured lines. The two new T. brucei strains should be useful to researchers interested in trypanosome differentiation and pathogenicity. Initial results suggested that the two strains have differing infection patterns in rodents. MAK65 is uniformly diploid and grew more reproducibly in bloodstream-form culture than MAK98.
José Wandilson Barboza Duarte Júnior, Elaine Hellen Nunes Chagas, Ana Carolina Silva Serra, Lizandra Caroline Dos Santos Souto, Edvaldo Tavares Da Penha Júnior, Renato Da Silva Bandeira, , Hanna Gabriela Da Silva Oliveira, Thaymis Kiara Santos Sousa, Cinthia Távora De Albuquerque Lopes, et al.
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0008792

Abstract:
The present study reports the occurrence of rotavirus A (RVA), rotavirus D (RVD), rotavirus F (RVF), rotavirus G (RVG), and picobirnavirus (PBV) in fecal specimens of wild (n = 22), and exotic birds (n = 1) from different cities of Pará state. These animals were hospitalized at Veterinary Hospital of the Federal University of Pará, Brazil, in a period from January 2018 to June 2019. The animals exhibited different clinical signs, such as diarrhea, malnutrition, dehydration, and fractures. The results showed 39.1% (9/23) of positivity for RVA by RT-qPCR. Among these, one sample (1/9) for the NSP3 gene of T2 genotype was characterized. About 88.9% (8/9) for the VP7 gene belonging to G1, G3 equine like and G6 genotypes, and 55.5% (5/9) for the VP4 gene of P[2] genotype were obtained. In the current study, approximately 4.5% of the samples (1/23) revealed coinfection for the RVA, RVD and RVF groups. Furthermore, picobirnavirus (PBV) was detected in one of the 23 samples tested, and was classified in the Genogroup I. The findings represent the first report of RVA, RVD, RVF, RVG, and PBV genotypes in wild birds in Brazil, and due to wide distribution it can implies potential impacts of RVs, and PBVs on avian health, and other animals contributing to construction of new knowledge, and care perspectives.
Kaki Tsang, Gilles de Wildt, Upendo Mwingira,
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009727

Abstract:
Background Despite aspects of the SAFE strategy for reducing trachoma in Tanzania have been somewhat successful, the disease still persists in marginalised communities even with repeated trachoma control interventions. This study aims to understand the facilitators and barriers associated with implementing trachoma control programmes in these communities, from the perspective of non-governmental organisations (NGOs). Methods Participants were the representatives of NGOs who had knowledge and experience in the implementation of trachoma control programmes. Data was collected using in-depth, semi-structured interviews guided by a topic guide, which was updated after each interview using a constant comparative method. Interviews were audio-recorded and then transcribed verbatim. Thematic analysis was done inductively. Codes were generated from the transcripts and then clustered into themes. Findings The context within marginalised communities often acted as a perceived barrier to successful implementation of control programmes. This included poor environmental cleanliness, lack of trust, poor disease knowledge and traditional lifestyles. Community values could either be a facilitator or a barrier, depending on the scenario. The anatomical location of the disease and the poor understanding of the disease progression also served as barriers. Considerations affecting decision-making among NGO’s include financial feasibility, community needs and whether the quality of the intervention could be improved. NGOs felt that the collaboration and the opportunity to learn from other organisations were beneficial aspects of having different actors. However, this also resulted in variability in the effectiveness of interventions between districts. Conclusion NGOs should focus on behaviour change and health education that is tailored to marginalised communities and seek innovative ways to implement trachoma intervention programmes whilst being minimally intrusive to the traditional way of life. Partners should also implement ways to ensure high quality programmes are being provided, by increasing staff accountability and compensating volunteers fairly.
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009731

Abstract:
Snakebite is a major public health problem in Eswatini and serious envenomations can be responsible for considerable morbidity and mortality if not treated correctly. Antivenom should be administered in hospital in case of adverse reactions and any delays due to distance, transport, costs, antivenom availability and cultural beliefs can be critical. Myths and superstition surround snakes, with illness from snakebite considered a supernatural phenomenon best treated by traditional medicine since healers can explore causes through communication with the ancestors. Traditional consultations can cause significant delays and the remedies may cause further complications. Four rural focus group discussions were held in varying geographical regions to establish why people may choose traditional medicine following snakebite. The study revealed four themes, with no apparent gender bias. These were ‘beliefs and traditions’, ‘logistical issues’, ‘lack of knowledge’ and ‘parallel systems’. All snakes are feared, regardless of geographical variations in species distribution. Deep-seated cultural beliefs were the most important reason for choosing traditional medicine, the success of which is largely attributed to the ‘placebo effect’ and positive expectations. Collaboration and integration of the allopathic and traditional systems assisted by the regulation of healers and their methods could improve future treatment success. The plight of victims could be further improved with more education, lower costs and improved allopathic facilities.
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009644

Abstract:
Background Trachoma is a worldwide infectious disease causing blindness. Trachoma continued as a public health problem in Ethiopia due to a lack of sanitation and inadequate prevention strategies. This study aimed to identify the impact of water supply and sanitation intervention on preventing active trachoma among children. Methods Systematic literature searches were performed from 4 international databases. The search involved articles published from January 1995 up to March 2019. The Cochran Q and I2 statistical tests were used to check heterogeneity among the studies. A random-effect meta-analysis was employed to determine the pooled estimates with a 95% confidence interval (CI). Data analysis was performed using the CMA V.3 and RevMan 5 software program, and the result of the systematic review was reported using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Findings Out of 211 studies screened for the analysis, only 29 studies were finally included in this systematic review and meta-analysis. The result revealed factors that are significantly associated with increased odds of active trachoma. Accordingly, households with no access to toilet facilities (odds ratio [OR]: 2.04, 95% CI: 1.75–2.38), no access to improved water (OR: 1.58, 95% CI: 1.27–1.96), and do not practice regular face washing for children (OR: 4.19, 95% CI: 3.02–5.81) have shown increased odds of active trachoma. Besides, the results show a higher prevalence of active trachoma among children who did not wash their faces with soap and frequently. Conclusions The study found strong evidence that lack of access to water, sanitation, and hygiene (WASH) was associated with increased prevalence of active trachoma among children. Therefore, a comprehensive and partnership-oriented program is needed to tackle the problem, but further study will be required to strengthen its implementation.
Guilherme Kemeron Maciel Salazar, Joseir Saturnino Cristino, Alexandre Vilhena Silva-Neto, Altair Seabra Farias, João Arthur Alcântara, , , , , André Sachett, et al.
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009758

Abstract:
In the Brazilian Amazon, long distances, low healthcare coverage, common use of ineffective or deleterious self-care practices, and resistance to seeking medical assistance have an impact on access to antivenom treatment. This study aimed to estimate snakebite underreporting, and analyze barriers that prevent victims from obtaining healthcare in communities located in 15 municipalities on the banks of the Solimões, Juruá and Purus Rivers, in the remote Western Brazilian Amazon. Information on the participants’ demographics, previous snakebites, access to healthcare, time taken to reach medical assistance, use of self-care practices, and the reason for not accessing healthcare were collected through semi-structured interviews. In the case of deaths, information was collected by interviewing parents, relatives or acquaintances. A total of 172 participants who reported having suffered snakebites during their lifetime were interviewed. A total of 73 different treatment procedures was reported by 65.1% of the participants. Participants living in different river basins share few self-care procedures that use traditional medicine, and 91 (52.9%) participants reported that they had access to healthcare. Living in communities along the Juruá River [OR = 12.6 (95% CI = 3.2–49.7; p<0.001)] and the use of traditional medicine [OR = 11.6 (95% CI = 3.4–39.8; p<0.001)] were variables that were independently associated to the lack of access to healthcare. The main reasons for not accessing healthcare were the pprioritization of traditional treatments (70.4%), and the failure to recognize the situation as being potentially severe (50.6%). Four deaths from complications arising from the snakebite were reported, and three of these were from communities on the banks of the Juruá River. Only one of these received medical assistance. We found an unexpectedly high underreporting of snakebite cases and associated deaths. Snakebite victims utilized three main different healing systems: 1) self-care using miscellaneous techniques; 2) official medical healthcare generally combined with traditional practices; and 3) self-care using traditional practices combined with Western medicines. To mitigate snakebite burden in the Brazilian Amazon, an innovative intervention that would optimize timely delivery of care, including antivenom distribution among existing community healthcare centers, is needed.
Antonios Kolimenakis, Sabine Heinz, Michael Lowery Wilson, , , Antonios Michaelakis, , ,
PLOS Neglected Tropical Diseases, Volume 15; https://doi.org/10.1371/journal.pntd.0009631

Abstract:
Background This systematic review aims to assess how different urbanisation patterns related to rapid urban growth, unplanned expansion, and human population density affect the establishment and distribution of Aedes aegypti and Aedes albopictus and create favourable conditions for the spread of dengue, chikungunya, and Zika viruses. Methods and findings Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic review was conducted using the PubMed, Virtual Health Library, Cochrane, WHO Library Database (WHOLIS), Google Scholar, and and the Institutional Repository for Information Sharing (IRIS) databases. From a total of 523 identified studies, 86 were selected for further analysis, and 29 were finally analysed after applying all inclusion and exclusion criteria. The main explanatory variables used to associate urbanisation with epidemiological/entomological outcomes were the following: human population density, urban growth, artificial geographical space, urban construction, and urban density. Associated with the lack of a global definition of urbanisation, several studies provided their own definitions, which represents one of the study’s limitations. Results were based on 8 ecological studies/models, 8 entomological surveillance studies, 7 epidemiological surveillance studies, and 6 studies consisting of spatial and predictive models. According to their focus, studies were categorised into 2 main subgroups, namely “Aedes ecology” and “transmission dynamics.” There was a consistent association between urbanisation and the distribution and density of Aedes mosquitoes in 14 of the studies and a strong relationship between vector abundance and disease transmission in 18 studies. Human population density of more than 1,000 inhabitants per square kilometer was associated with increased levels of arboviral diseases in 15 of the studies. Conclusions The use of different methods in the included studies highlights the interplay of multiple factors linking urbanisation with ecological, entomological, and epidemiological parameters and the need to consider a variety of these factors for designing effective public health approaches.
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