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Dípteros ectoparásitos hematófagos: ¿reservorios potenciales del virus del dengue? / Ectoparasitic hematophagous dipters: potential reservoirs of dengue virus?

Gac Med Mex; 153(Supl. 2): S82-S90, 2017.
Artigo em Espanhol | MEDLINE | ID: mdl-29099114


Recently, the presence of antibodies and dengue virus (DV) RNA in neotropical wild mammals, including Desmodus rotundus, was reported. In a previous study, DV was also found in a high percentage (39.6%) of ectoparasitic hematophagous dipters specifics of these hematophagous bats. In order to verify the susceptibility of these ectoparasites to DV, in this work experimental infections with VD2 of organs explants of Strebla wiedemanni and of Melophagus ovinus were performed using C6/36 cells as control. Viral titers (UFP/mL) were determined at 0, 48 and 96 hrs pi. Infected organs were observed by electron microscopy and under the confocal microscopy indirect immunofluorescence (IIF) using specific conjugates against DV. The infected organs of both species of ectoparasites replicated DV at titers similar to those obtained with the C6/36 cell line (≥10 UFP/mL). Electron microscopy and IIF showed DV replication in the digestive tract, tracheoles, reproductive organs of males but not in females, and milk glands (MG) of both species. In the fatty bodies of the MG of M. ovinus, zones with a high affinity for the DV were observed. In this work the susceptibility of S. wiedemanni and M. ovinus to DV was demonstrated and consequently the probable role of this ectoparasites as wild reservoirs of DV.

"An Uncommon Complication of Dengue".

J Stroke Cerebrovasc Dis; 2017 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-29103866


Intracranial hemorrhage is an uncommon complication of dengue fever, which is caused by a flavivirus and transmitted via Aedes mosquito. We present here bilateral cerebellar bleed because of dengue virus infection.

Surveillance of intensity level and geographical spreading of dengue outbreak among males and females in Punjab, Pakistan: A case study of 2011.

J Infect Public Health; 2017 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-29103928


BACKGROUND: Dengue fever is viral disease which spreads due to the bite of the Aedes aegypti mosquito. In recent years, it has affected around 40% population of the world. Its endemic flow has led to a large disease burden, in terms of human and financial resources. METHODS: Geographical Information Systems (GIS) are normally used to develop epidemiological thematic maps. This study explores the patterns and hotspots, associated with the catastrophic outbreak of dengue, in Punjab, in 2011. The ArcView software was used to analyze the data reported by the district hospitals of Punjab. Twenty-one-thousand cases were reported from March to December 2011, with 300 causalities. RESULTS AND CONCLUSION: This research reveals that from among the total 37 epidemiological weeks, the maximum impact was observed between weeks 22 and 27. The geographical flow and hotspots associated with dengue have been shown through thematic maps. A positive correlation between the risk for dengue and age was observed. The findings of this research can help health officials and decision-makers alert the public about future outbreaks and take preventive measures to considerably reduce the mortality and morbidity associated with the disease.

Differences in activation and tissue homing markers of natural killer cell subsets during acute dengue infection.

Immunology; 2017 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-29105052


Dengue virus (DENV) infection is considered as one of the most important mosquito-borne diseases. It causes a spectrum of illness that could be due to qualitative and/or quantitative difference(s) of the natural killer (NK)-cell responses during acute DENV infection. This view prompted us to perform a detailed phenotypic comparative characterization of NK cell subsets from DENV-infected patients with dengue fever (DF), dengue hemorrhagic fever (DHF) and healthy controls. The activation/differentiation molecules, CD69 and CD57 and a variety of tissue homing molecules were analyzed on the CD56 CD16 and CD56 CD16 NK cells. While there was no increase in the frequency of the total NK cells during DENV infection as compared with the healthy subjects, there was a significant increase in the frequency of the CD56 CD16 subset and the frequency of CD69 expression by both NK cell subsets during the febrile phase of infection. We also found an increase in the frequencies of cells expressing CD69 and CD57 in the CD56 CD16 subset than those in the CD56 CD16 subset. Moreover, while the CD56 CD16 subset contained a high frequency of cells expressing skin-homing markers, the CD56 CD16 subset contained a high frequency of cells expressing bone marrow and lymph node trafficking markers. Interestingly, no differences of these NK cell subsets were noted in samples from DF versus DHF patients. These findings suggest that activation and differentiation and the patterns of tissue homing molecules of the two major NK cell subsets are different and these might play a critical role in the immune response against acute DENV infection. This article is protected by copyright. All rights reserved.

Interplay between Keratinocytes and Myeloid Cells Drives Dengue Virus Spread in Human Skin.

J Invest Dermatol; 2017 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-29106931


The skin is the site of dengue virus (DENV) transmission following the bite of an infected mosquito, but the contribution of individual cell types within skin to infection is unknown. We studied the dynamics of DENV infection in human skin explants using quantitative in situ imaging. DENV replicated primarily in the epidermis and induced a transient interferon-α response. DENV infected a wide range of cells, including Langerhans cells, macrophages, dermal dendritic cells (DC), mast cells, fibroblasts and lymphatic endothelium, but keratinocytes were the earliest targets of infection and made up 60% of infected cells over time. Virus inoculation led to recruitment and infection of Langerhans cells, macrophages and dermal DC, and these cells emigrated from skin in increased numbers as a result of infection. DENV induced expression of proinflammatory cytokines and chemokines by infected keratinocytes. Blocking keratinocyte-derived IL-1ß alone reduced infection of Langerhans cells, macrophages and dermal DC by 75 to 90% and reduced the overall number of infected cells in dermis by 65%. These data show that the innate response of infected keratinocytes attracts virus-permissive myeloid cells that inadvertently spread DENV infection. Our findings highlight a previously undescribed role for keratinocytes and their interplay with myeloid cells in dengue.

Relevance of transportation to correlations among criticality, physical means of propagation, and distribution of dengue fever cases in the state of Bahia.

Sci Total Environ; 2017 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-29107376


Dengue infection is a public health problem with a complex distribution. The physical means of propagation and the dynamics of diffusion of the disease between municipalities need to be analysed to direct efficient public policies to prevent dengue infection. The present study presents correlations of occurrences of reported cases of dengue infection among municipalities, self-organized criticality (SOC), and transportation between areas, identifying the municipalities that play an important role in the diffusion of dengue across the state of Bahia, Brazil. The significant correlation found between the correlation network and the SOC demonstrates that the pattern of intramunicipal diffusion of dengue is coupled to the pattern of synchronisation between the municipalities. Transportation emerges as influential in the dynamics of diffusion of epidemics by acting on the aforementioned variables.

XBP1-Mediated BiP/GRP78 Upregulation Copes with Oxidative Stress in Mosquito Cells during Dengue 2 Virus Infection.

Biomed Res Int; 2017: 3519158, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29098151


Dengue viruses (DENVs) cause dengue fever which is an important mosquito-borne disease in tropical areas. Generally, DENV does not cause cellular damage in mosquito cells. However, alterations in cytosolic calcium ions ([Ca ]cyt) and the mitochondrial membrane potential (MMP), as well as accumulated reactive oxygen species (ROS), including superoxide anions (O ) and hydrogen peroxide (H O ), can be detected in C6/36 cells with DENV2 infection. Evident upregulation of BiP/GRP78 also appeared at 24 h postinfection in DENV2-infected C6/36 cells. As expression of BiP/GRP78 mRNA was reduced when the transcription factor X-box-binding protein-1 (XBP1) was knocked down in C6/36 cells, it demonstrated that BiP/GRP78 is the target gene regulated by the XBP1 signal pathway. We further demonstrated that the expression and splicing activity of XBP1 were upregulated in parallel with DENV2 infection in C6/36 cells. In C6/36 cells with BiP/GRP78 overexpression, oxidative stress indicators including [Ca ]cyt, MMP, O , and H O were all pushed back to normal. Taken together, DENV2 activates XBP1 at earlier stage of infection, followed by upregulating BiP/GRP78 in mosquito cells. This regulatory pathway contributes a cascade in relation to oxidative stress alleviation. The finding provides insights into elucidating how mosquitoes can healthily serve as a vector of arboviruses in nature.

Antibody-dependent enhancement of severe dengue disease in humans.

Science; 2017 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-29097492


For dengue viruses (DENV1-4), a specific range of antibody titer has been shown to enhance viral replication in vitro and severe disease in animal models. Although suspected, such antibody-dependent enhancement (ADE) of severe disease has not been shown to occur in humans. Using multiple statistical approaches to study a long-term pediatric cohort in Nicaragua, we show that risk of severe dengue disease is highest within a narrow range of pre-existing anti-DENV antibody titers. By contrast, we observe protection from all symptomatic dengue disease at high antibody titers. Thus, immune correlates of severe dengue must be evaluated separately from correlates of protection against symptomatic disease. These results have implications for studies of dengue pathogenesis and for vaccine development, because enhancement, not just lack of protection, is of concern.

Computer-aided analysis of phytochemicals as potential dengue virus inhibitors based on molecular docking, ADMET and DFT studies.

J Vector Borne Dis; 54(3): 255-262, 2017 Jul-Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29097641


BACKGROUND & OBJECTIVES: Dengue fever, caused by dengue virus (DENV), has become a serious threat to human lives. Phytochemicals are known to have great potential to eradicate viral, bacterial and fungal-borne diseases in human beings. This study was aimed at in silico drug development against nonstructural protein 4B (NS4B) of dengue virus 4 (DENV4). METHODS: A total of 2750 phytochemicals from different medicinal plants were selected for this study. These plants grow naturally in the climate of Pakistan and India and have been used for the treatment of various pathologies in human for long-time. The ADMET studies, molecular docking and density functional theory (DFT) based analysis were carried out to determine the potential inhibitory properties of these phytochemicals. RESULTS: The ADMET analysis and docking results revealed nine phytochemicals, i.e. Silymarin, Flavobion, Derrisin, Isosilybin, Mundulinol, Silydianin, Isopomiferin, Narlumicine and Oxysanguinarine to have potential inhibitory properties against DENV and can be considered for additional in vitro and in vivo studies to assess their inhibitory effects against DENV replication. They exhibited binding affinity ≥ -8 kcal/mol against DENV4-NS4B. Furthermore, DFT based analysis revealed high reactivity for these nine phytochemicals in the binding pocket of DENV4-NS4B, based on ELUMO, EHOMO and band energy gap. INTERPRETATION & CONCLUSION: Five out of nine phytochemicals are reported for the first time as novel DENV inhibitors. These included three phytochemicals from Silybum marianum, i.e. Derrisin, Mundulinol, Isopomiferin, and two phytochemicals from Fumaria indica, i.e. Narlumicine and Oxysanguinarine. However, all the nine phytochemicals can be considered for in vitro and in vivo analysis for the development of potential DENV inhibitors.

The bacterium Wolbachia exploits host innate immunity to establish a symbiotic relationship with the dengue vector mosquito Aedes aegypti.

ISME J; 2017 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-29099491


A host's immune system plays a central role in shaping the composition of the microbiota and, in return, resident microbes influence immune responses. Symbiotic associations of the maternally transmitted bacterium Wolbachia occur with a wide range of arthropods. It is, however, absent from the dengue and Zika vector mosquito Aedes aegypti in nature. When Wolbachia is artificially forced to form symbiosis with this new mosquito host, it boosts the basal immune response and enhances the mosquito's resistance to pathogens, including dengue, Zika virus and malaria parasites. The mechanisms involved in establishing a symbiotic relationship between Wolbachia and A. aegypti, and the long-term outcomes of this interaction, are not well understood. Here, we have demonstrated that both the immune deficiency (IMD) and Toll pathways are activated by the Wolbachia strain wAlbB upon its introduction into A. aegypti. Silencing the Toll and IMD pathways via RNA interference reduces the wAlbB load. Notably, wAlbB induces peptidoglycan recognition protein (PGRP)-LE expression in the carcass of A. aegypti, and its silencing results in a reduction of symbiont load. Using transgenic mosquitoes with stage-specific induction of the IMD and Toll pathways, we have shown that elevated wAlbB infection in these mosquitoes is maintained via maternal transmission. These results indicate that host innate immunity is utilized to establish and promote host-microbial symbiosis. Our results will facilitate a long-term projection of the stability of the Wolbachia-A. aegypti mosquito system that is being developed to control dengue and Zika virus transmission to humans.The ISME Journal advance online publication, 3 November 2017; doi:10.1038/ismej.2017.174.

Ability to serologically confirm recent Zika virus infection in areas with varying past incidence of dengue virus infection - United States and territories, 2016.

J Clin Microbiol; 2017 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-29093104


Cross-reactivity within flavivirus antibody assays, produced by shared epitopes in the envelope proteins, can complicate serological diagnosis of Zika virus (ZIKAV) infection. We assessed the utility of the plaque reduction neutralization test (PRNT) to confirm recent ZIKAV infections and rule out misleading positive IgM results in areas with varying past dengue virus (DENV) infection incidence. We reviewed PRNT results of sera collected for diagnosis of ZIKAV infection from January 1 through August 31, 2016 with positive ZIKAV IgM results and ZIKAV and DENV PRNT performed. PRNT result interpretations included ZIKAV, unspecified flavivirus, DENV infection, or negative. For this analysis, ZIKAV IgM was considered false-positive for samples interpreted as DENV infection or negative. In US states, 208 (27%) of 759 IgM positives were confirmed as ZIKAV, compared to 11 (21%) of 52 in the US Virgin Islands (USVI), 15 (15%) of 103 in American Samoa, and 13 (11%) of 123 in Puerto Rico. In American Samoa and Puerto Rico, more than 80% of IgM positives were unspecified flavivirus infections. The false-positivity rate was 27% in US states, 18% in USVI, 2% in American Samoa, and 6% in Puerto Rico. In US states, PRNT provided a virus-specific diagnosis or ruled out infection in the majority of IgM positive samples. Almost a third of ZIKAV IgM positive results did not confirm; therefore, providers and patients must understand that IgM results are preliminary. In territories with historically higher DENV transmission, PRNT usually could not differentiate between ZIKAV and DENV infections.

Adherence to Clinical Practice Guidelines (CPG) management of dengue infection in adults (revised 2nd edition).

PLoS One; 12(11): e0184559, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29095822


The Malaysian Dengue Clinical Practice Guidelines (CPG) have been developed to provide evidence-based guidance in the management of dengue infections. The use of these guidelines is essential to ensure its recommendations are being practiced. However, the adherence to the guidelines for management of dengue (revised 2nd edition) by healthcare providers still remains unknown. Therefore, the aim of this study was to evaluate the proportion among healthcare providers that adhere to this Dengue CPG. A retrospective cohort study of dengue cases registered from 1 January 2014 to 1 June 2015 was conducted in public hospitals and health clinics in Selangor, Putrajaya and Kuala Lumpur. Adherence to the CPG recommendations were recorded by reviewing patients' case notes. Overall proportion of adherence in clinical components of the recommendation were (7.1 to 100.0% versus 7.7 to 73.8%) in history taking, (6.7 to 100.0% versus 12.3 to 60.0%) in physical examinations, (18.4 to 100.0% versus 23.1 to 83.2%) in assessment of warning signs, (0.6 to 100.0% versus 12.3 to 87.7%) in assessment of haemodynamic status, (60.0 to 100.0% versus 27.7 to 40.0%) in diagnosis, (46.6 to 80.0% versus 52.3%) in case notifications, (73.2 to 100.0% versus 89.2 to 96.9%) in performing specific laboratory investigations and (7.9 to 100.0% versus 21.5%) in monitoring, for outpatient versus inpatient, respectively. Adherence trends were demonstrated to be higher in hospital settings compared to outpatient settings. Adherence to this Dengue CPG varies widely with overall good clinical outcomes observed.

A young female presenting with unilateral sacroiliitis following dengue virus infection: a case report.

J Med Case Rep; 11(1): 307, 2017 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-29089045


BACKGROUND: Dengue is a common arthropod-borne viral infection in Sri Lanka which is spread by the mosquitos of the genus Aedes. The clinical features of dengue include high-grade fever associated with arthralgia and myalgia. However, dengue virus is not considered an arthritogenic virus. We report a case of a previously healthy young female who presented with imaging-confirmed right-sided sacroiliitis 10 days after developing dengue fever. This is the first reported case that shows a possible link between dengue infection and development of arthritis. CASE PRESENTATION: A 14-year-old Sri Lankan female presented to our medical unit with right buttock and hip pain of 3 weeks' duration. She had serologically confirmed dengue infection 10 days prior to the onset of buttock pain. A clinical examination revealed features of right sacroiliitis. An X-ray of her sacroiliac joint showed joint space widening and reactive bone changes. Magnetic resonance imaging of her pelvis and sacroiliac joint confirmed the diagnosis of acute sacroiliitis. She had an erythrocyte sedimentation rate of 110 mm first hour with a normal C-reactive protein. Her human leukocyte antigen-B27, rheumatoid factor, antinuclear antibody, chikungunya antibody, hepatitis serology, Brucella serology, and tuberculin skin test were negative. She was treated with nonsteroidal anti-inflammatory drugs and showed gradual improvement. CONCLUSIONS: After excluding possible causes for sacroiliitis, we postulated that sacroiliitis in the index case could have been caused or triggered by dengue virus infection. However there is a possibility that the sacroiliitis merely coincided with the dengue virus infection. This case illustrates the possibility that dengue virus could have a link with the development of arthritis in the same manner as other arthritogenic viruses; possible mechanisms for this include direct invasion of the synovium and the joint tissue by the virus, immune complex formation and deposition in the joint tissue, and immune dysregulation. Further studies are needed in this field to gain more knowledge, as dengue infection is highly prevalent in Sri Lanka.

Involvement of caspase-4 in IL-1 beta production and pyroptosis in human macrophages during dengue virus infection.

Immunobiology; 2017 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-29113699


Caspase-4 physically interacts with caspase-1 and is believed to be a proinflammatory caspase that can induce the inflammatory form of programmed cell death (pyroptosis) and the release of mature interleukin (IL)-1ß. However, the function of caspase-4 in dengue virus infection is not yet fully understood. We examined the function of caspase-4 in IL-1ß production and pyroptosis during dengue virus serotype-2 (DENV-2) infection in human macrophages. In this study, DENV-2 infection increased IL-1ß protein level with activated caspase-4 activity. Using primary macrophages, we observed that caspase-4 induces activation of caspase-1 and secretion of IL-1ß in response to DENV-2 infection, without the need for secondary signals to stimulate the assembly of the inflammasome. These findings indicate that the regulation of caspase-1 activity by capsase-4 could represent a unique mechanism. Our data suggest that caspase-4 is upstream of caspase-1 in the pathway that regulates pyroptosis and IL-1ß synthesis in macrophages during DENV-2 infection.


Lancet; 390(10106): 1941, 2017 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-29115222

The impact of temperature and Wolbachia infection on vector competence of potential dengue vectors Aedes aegypti and Aedes albopictus in the transmission of dengue virus serotype 1 in southern Taiwan.

Parasit Vectors; 10(1): 551, 2017 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-29116011


BACKGROUND: We evaluated the impact of temperature and Wolbachia infection on vector competence of the local Aedes aegypti and Ae. albopictus populations of southern Taiwan in the laboratory. RESULTS: After oral infection with dengue serotype 1 virus (DENV-1), female mosquitoes were incubated at temperatures of 10, 16, 22, 28 and 34 °C. Subsequently, salivary gland, head, and thorax-abdomen samples were analyzed for their virus titer at 0, 5, 10, 15, 20, 25 and 30 days post-infection (dpi) by real-time RT-PCR. The results showed that Ae. aegypti survived significantly longer and that dengue viral genome levels in the thorax-abdomen (10 -10 PFU equivalents/ml) and salivary gland samples (10 -10 PFU equivalents/ml) were significantly higher at high temperature (28-34 °C). The survival of Ae. albopictus was significantly better at 16 or 28 °C, but the virus titers from thorax-abdomen (10 -10 PFU equivalents/ml) and salivary gland samples (10 -10 PFU equivalents/ml) were significantly higher at 22-28 °C. Within viable temperature ranges, the viruses were detectable after 10 dpi in salivary glands and head tissues in Ae. aegypti and after 5-10 dpi in Ae. albopictus. Vector competence was measured in Ae. albopictus with and without Wolbachia at 28 °C. Wolbachia-infected mosquitoes survived significantly better and carried lower virus titers than Wolbachia-free mosquitoes. Wolbachia coinfections (92.8-97.2%) with wAlbA and wAlbB strains were commonly found in a wild population of Ae. albopictus. CONCLUSIONS: In southern Taiwan, Ae. aegypti is the main vector of dengue and Ae. albopictus has a non-significant role in the transmission of dengue virus due to the high prevalence of Wolbachia infection in the local mosquito population of southern Taiwan.

Epitopes based drug design for dengue virus envelope protein: A computational approach.

Comput Biol Chem; 71: 152-160, 2017 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-29096381


Dengue virus (DENV) has emerged as a rapidly spreading epidemic throughout the tropical and subtropical regions around the globe. No suitable drug has been designed yet to fight against DENV, therefore, the need for safe and effective antiviral drug has become imperative. The envelope protein of DENV is responsible for mediating the fusion process between viral and host membranes. This work reports an in silico approach to target B and T cell epitopes for dengue envelope protein inhibition. A conserved region "QHGTI" in B and T cell epitopes of dengue envelope glycoprotein was confirmed to be valid for targeting by visualizing its interactions with the host cell membrane TIM-1 protein which acts as a receptor for serotype 2 and 3. A reverse pharmacophore mapping approach was used to generate a seven featured pharmacophore model on the basis of predicted epitope. This pharmacophore model as a 3D query was used to virtually screen a chemical compounds dataset "Chembridge". A total of 1010 compounds mapped on the developed pharmacophore model. These retrieved hits were subjected to filtering via Lipinski's rule of five, as a result 442 molecules were shortlisted for further assessment using molecular docking. Finally, 14 hits of different structural properties having interactions with the active site residues of dengue envelope glycoprotein were selected as lead candidates. These structurally diverse lead candidates have strong likelihood to act as further starting structures in the development of novel and potential drugs for the treatment of dengue fever.

Dengue vaccine supplies under endemic and epidemic conditions in three dengue-endemic countries: Colombia, Thailand, and Vietnam.

Vaccine; 2017 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-29110932


BACKGROUND: Dengue fever has been a major public health concern in Colombia, Thailand, and Vietnam. Unlike other infectious diseases, dengue vaccines had not been available for a long time, causing difficulties to control the disease. However, the first live attenuated, tetravalent dengue vaccine (CYD-TDV) became available in 2016 and has been already licensed in some dengue-endemic countries. Because several second-generation dengue vaccines are also in the pipeline, it is critical to understand the efficient allocation of dengue vaccines considering the geographical variation of the disease. METHODS: The Climate Risk Factor (CRF) index was created using the climate and non-climate factors in the three countries. A random-coefficient negative binomial model was chosen to validate the relationship between the CRF index and dengue incidence proxy. Given the statistical significance of the CRF index, high risk areas for dengue fever were identified at the 5 km by 5 km resolution and used to estimate vaccination coverage rates and the number of doses required for various types of vaccination scenarios by country. RESULTS AND CONCLUSIONS: Based upon a three-dose scheme, the estimated number of vaccines required for routine vaccination targeting 9 years old ranged from 1 to 2.6 million doses across the countries during the first year of introduction. A one-off catch-up campaign targeting the age group of 10-17 year olds would require 8 to 18 million additional doses. Routine vaccination (with or without a catch-up campaign) covered 63%, 90%, and 91% of the targeted age group populations in Colombia, Thailand, and Vietnam respectively. Given that many dengue-endemic countries face limited resources and that the costs for mass vaccination campaigns may not be trivial, the findings of this study can guide the decision makers in the three countries regarding the efficient distribution of vaccines by identifying populations at high risk at 5 km by 5 km resolution.
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