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1.
Artículo en Inglés | MEDLINE | ID: mdl-32072409

RESUMEN

Mosquitoes are principal vector of several vector-borne diseases affecting human beings leading to thousands of deaths per year and responsible for transmitting diseases like malaria, dengue, chikungunya, yellow fever, Zika virus, Japanese encephalitis, and lymphatic filariasis. In the present study, we evaluated the different solvent extracts of mangrove Avicennia marina for their toxicity against larvae of three major mosquito vectors, as well as selected microbial pathogens. The larvicidal mortality of third instars was observed after 24 h. Highest larval mortality was found for the acetone extract of A. marina against Culex quinquefasciatus (LC50 = 0.197 mg/ml; LC90 = 1.5011 mg/ml), Anopheles stephensi (LC50 = 0.176 mg/ml; LC90 = 3.6290 mg/ml), and Aedes aegypti (LC50 = 0.164 mg/ml; LC90 = 4.3554 mg/ml). GC-MS analysis of acetone extract revealed 5 peaks, i.e., 1-hexyl-2-nitrocyclohexane (3.229%), eicosanoic acid (40.582%), cis-9-hexadecenal (70.54%), oleic acid (4.646%), and di-N-decylsulfone (5.136%). Parallel to larvicidal assay, sub-lethal dosage acetone extracts severely affected the enzyme regulations (α,ß-carboxylesterase, GST and CYP450) of third instars. Larval and pupal durations increased in all treatment sub-lethal dosage (0.127, 0.151, 0.177, and 0.197 mg/ml), whereas egg hatchability and means of fecundity decreased compared to control. The survival rate was reduced statistically in Cx. quinquefasciatus (χ2 = 23.77, df = 1, P = 0.001) in all the treatment dosages as compared to the control. Antimicrobial activity assays showed significant growth inhibition post treatment with acetone and methanol extracts against Salmonella typhimurium, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus pneumoniae, Escherichia coli, and Shigella flexneri. Overall, these results indicated the potential employment of A. marina extracts as a source of natural mosquitocidal and antimicrobial compounds of green-based environment.

4.
Lancet Infect Dis ; 20(2): 172, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32006508
5.
Int J Infect Dis ; 2020 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-32004691

RESUMEN

BACKGROUND: Yellow fever (YF) is a viral hemorrhagic disease caused by an arbovirus from the Flaviviridae family. Data on the clinical profile of severe YF in intensive care units (ICUs) are scarce. This study aimed to evaluate factors associated with YF mortality in patients admitted to a Brazilian ICU during the YF outbreaks of 2017 and 2018. METHODS: This was a longitudinal cohort case series study that included YF patients admitted to the ICU. Demographics, clinical and laboratory data were analyzed. Cox regression identified independent predictors of death risk. RESULTS: A total of 114 patients were studied. The median age was 48 years, and 92.1% were males. In univariate analysis, jaundice, leukopenia, bradycardia, prothrombin time, expressed as a ratio to the international normalized ratio-(PT-INR), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, lactate, arterial pH and bicarbonate, Acute Physiology and Chronic Health Evaluation II (APACHE II) and Simplified Acute Physiology Score 3 (SAPS 3) severity scores, transfusion of fresh frozen plasma, acute renal failure (Acute Kidney Injury Network stage III (AKIN III)), hemodialysis, cumulative fluid balance at 72 hours of ICU, vasopressor use, seizures and grade IV encephalopathy were significantly associated with mortality. In multivariate analysis, factors independently associated with YF mortality were PT-INR, APACHE II, and grade IV hepatic encephalopathy. CONCLUSIONS: In the large outbreak in Brazil, factors independently associated with death risk in YF were: PT-INR, APACHE II, and grade IV hepatic encephalopathy. Early identification of patients with YF mortality risk factors may be very usefull. Once these patients with a poor prognosis have been identified, proper management should be promptly implemented.

6.
J Med Entomol ; 2020 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-32006427

RESUMEN

Effective suppression of container-inhabiting Asian Tiger [Aedes albopictus (Skuse)] (Diptera: Culicidae) and yellow fever [Aedes aegypti (L.)] (Diptera: Culicidae) mosquitoes presents one of the most intractable problems for modern mosquito control. Traditional tools often fail to control populations of these mosquito species, and are prohibitively expensive or have negative environmental impacts. Novel approaches and tools are urgently needed for integrated container-inhabiting mosquito management programs. One of the most promising techniques is autodissemination. We present the results of a long-term large-scale study conducted in a temperate urbanized environment representing typical Ae. albopictus habitats. Three treatment sites with autodissemination stations and three nearby reference sites were monitored for eggs, immature, and adult mosquitoes over a period of 3 yr from 2014 to 2016. Elevated larval and pupal mortality of 12-19% on average was the most notable outcome in sentinel cups of the treatment sites. The number of eggs in the treatment sites was significantly reduced in 2014, but not in 2015 or 2016. Adult populations remained similar in treatment and reference sites throughout the study. The impact of autodissemination on mosquito populations was lower than reported by previous investigations. Technical and logistical problems associated with wider coverage and working in multiple urban neighborhoods contributed to reduced efficacy. Incorporating autodissemination with routine mosquito control operations and commercializing this methodology for general public use will require further research on combining this tool with other novel or conventional technologies.

7.
Artículo en Inglés | MEDLINE | ID: mdl-32049261

RESUMEN

Aedes aegypti is associated with epidemic diseases in Brazil, such as urban yellow fever, dengue, and more recently, chikungunya and Zika viruses infections. More information about Ae. aegypti infestation is fundamental to virological surveillance in order to ensure the effectiveness of control measures in use. Thus, the present study aims to identify and compare infestation and infectivity of Ae. aegypti females in Macapa city, Amapa State (Amazon region), Brazil, between the epidemiological weeks 2017/02 and 2018/20. A total number of 303 Ae. aegypti females were collected at 21 fixed collection points, 171 at the 10 collection points in the Marabaixo neighborhood and 132 at the 11 collection points in the Central neighborhood. Among the collected samples, only two were positive for dengue virus, with a 2.08% (2/96 pools) infectivity rate for Marabaixo. The difference between the medians of Ae. aegypti females captured in Central and Marabaixo sites was not statistically significant. The findings indicate similar mosquito infestation levels between the neighborhoods, and a low-level of mosquito infectivity, although dengue virus was found only in Marabaixo. Virological surveillance of Ae. aegypti was important to identify sites of infection and determine possible routes of transmission to enable health surveillance teams to adopt preventive strategies where infected mosquitoes are present and act faster.

8.
J Med Entomol ; 2020 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-32020186

RESUMEN

Since the last yellow fever (YF) outbreak was detected in Argentina in 2009, vector surveillance and studies of arbovirus infections are carried out intermittently specifically in areas where nonhuman primates of the Alouatta genus are present. We report in these areas of Corrientes province the detection of Haemagogus leucocelaenus (Dyar and Shannon) (Diptera: Culicidae) and Sabethes albiprivus (Theobald) (Diptera: Culicidae), both species involved in the forest YF cycle, and also the presence of Aedes albopictus (Skuse) in new areas in Argentina, which represents the southernmost citation for this species in South America. Aedes albopictus, a mosquito species native to Asia, was reported for the first time in Argentina in 1998, in Misiones province. Since then, no other report has indicated the extension of the distribution of this mosquito. This report shows the importance of performing continual entomological and arboviruses surveillance and highlights the impact that could result from the expansion of Ae. albopictus across Argentina.

9.
J Med Entomol ; 2020 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-32020196

RESUMEN

Aedes aegypti L. (Diptera: Culicidae) is one of the most medically important mosquito species, due to its ability to spread viruses of yellow fever, dengue fever, and Zika in humans. In this study, the insecticidal activity of 17 plant essential oils was evaluated via topical application against two strains of Ae. aegypti mosquito, Orlando (insecticide-susceptible) and Puerto Rico (pyrethroid-resistant). Initial screens with the Orlando strain showed that cucumber seed oil (2017 sample) was the most toxic, followed by sandalwood and thyme oil. When the essential oils were mixed with permethrin, they failed to show any significant synergism of insecticidal activity. Sandalwood and thyme oils displayed consistently high mortality against the resistant Puerto Rico strain, with low resistance ratios of 2.1 and 1.4, respectively. In contrast, cucumber seed oil showed significantly less activity against Puerto Rico mosquitoes, with a resistance ratio of 45. Bioactivity-guided fractionation of the 2017 sample of cucumber seed oil sample via flash column chromatography produced 11 fractions, and gas-chromatography/mass spectrometry analysis revealed that the three active fractions were contaminated with 0.33, 0.36, and 0.33% of chlorpyrifos-methyl, an organophosphorus insecticide, whereas inactive fractions did not show any trace of it. These results suggested that the insecticidal activity of cucumber seed oil was probably due to the presence of the insecticide, later confirmed with a clean batch of cucumber seed oil obtained in 2018, which showed negligible insecticidal activity. These findings demonstrate clearly the need for essential oil analysis to confirm purity before any claims are made about pesticidal potency.

10.
J Insect Physiol ; 121: 104019, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32032591

RESUMEN

Aedes aegypti is the primary vector of several arboviruses that impact human health including the dengue, Zika, and yellow fever viruses. The potential of Ae. aegypti females to transmit viruses is enhanced by mating-induced behavioral and physiological changes that increase female host-seeking behaviors, blood-feeding frequency and longevity. The mating-induced changes are due to female receipt of male seminal fluid proteins (SFPs) during copulation. SFPs also inhibit female re-mating-re-mating incidence is significantly reduced in the initial hours after mating and nearly absent after 24 h. Males, however, are not limited in the number of females they can inseminate and are able to mate with multiple females in succession. As successive mating depletes SFPs, we examined parameters of fertility and re-mating incidence in females after mating with recently mated males. Males of two Ae. aegypti strains (Colombian and Thai) were mated five consecutive times and fecundity, resulting larvae and hatch percentage in each female of the mating sequence was assessed. In both strains, we found that males can mate three times in succession without impacting fertility in their mates. However, significant declines in fecundity, resulting larvae, and hatch percentage were observed after a third mating. Male size influenced female fecundity and fertility as mates of small males showed further reductions compared to mates of big males after mating consecutively. Seven days after the consecutive mating assays, the re-mating rate of females mated fifth in succession was significantly increased (Colombian strain: 33%; Thai strain: 48%) compared to females mated first (0% in both strains). Re-mating incidence was further increased in small, Thai strain males where 82% of fifth mated females re-mated compared to 0% of first mated females. Finally, we show that regardless of male size, mates of experimental males were similarly fertile to mates of control males when mated for a sixth time 48 h after the consecutive mating assays, showing that males recover fertility after 2 d. Our results show that male sexual history influences fertility and re-mating incidence of Ae. aegypti females.

11.
Vaccine ; 38(9): 2172-2182, 2020 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-32008879

RESUMEN

Ever since its development in the 1930's, the live-attenuated Yellow Fever virus vaccine YF-17D has been highly effective. Despite the increasing knowledge on the immune biology of the YF-17D vaccine, most studies have focused only on a few types of immune cells and pathways or mainly on the primary adaptive immune response to YF-17D vaccination. Here, we examined humoral, innate and adaptive cellular responses in a longitudinal YF-17D vaccination study in Switzerland, comparing both primary and booster vaccination. In contrast to the strong innate and adaptive immune response to the primary vaccination, we find that the response to boosting is much reduced. Our data show an inverse association of neutralizing antibodies at baseline with vaccine virus replication and with the immune response upon boosting. These results suggest that booster vaccination may not have major immunological effects when neutralizing antibodies are present. Importantly, our study population was healthy adults in a non-endemic country and ultimately booster vaccine requirement must be assessed based on additional epidemiological and public health considerations in endemic areas.

12.
Front Immunol ; 11: 16, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32038660

RESUMEN

West Nile (WN) virus infection of humans is frequently asymptomatic, but can also lead to WN fever or neuroinvasive disease. CD4 T cells and B cells are critical in the defense against WN virus, and neutralizing antibodies, which are directed against the viral glycoprotein E, are an accepted correlate of protection. For the efficient production of these antibodies, B cells interact directly with CD4 helper T cells that recognize peptides from E or the two other structural proteins (capsid-C and membrane-prM/M) of the virus. However, the specific protein sites yielding such helper epitopes remain unknown. Here, we explored the CD4 T cell response in humans after WN virus infection using a comprehensive library of overlapping peptides covering all three structural proteins. By measuring T cell responses in 29 individuals with either WN virus disease or asymptomatic infection, we showed that CD4 T cells focus on peptides in specific structural elements of C and at the exposed surface of the pre- and postfusion forms of the E protein. Our data indicate that these immunodominant epitopes are recognized in the context of multiple different HLA molecules. Furthermore, we observed that immunodominant antigen regions are structurally conserved and similarly targeted in other mosquito-borne flaviviruses, including dengue, yellow fever, and Zika viruses. Together, these findings indicate a strong impact of virion protein structure on epitope selection and antigenicity, which is an important issue to consider in future vaccine design.

13.
Acta Trop ; : 105398, 2020 Feb 14.
Artículo en Inglés | MEDLINE | ID: mdl-32068030

RESUMEN

Mosquito-borne diseases affect millions of individuals worldwide; the area of endemic transmission has been increasing due to several factors linked to globalization, urban sprawl, and climate change. The Aedes aegypti mosquito plays a central role in the dissemination of dengue, Zika, chikungunya, and urban yellow fever. Current preventive measures include mosquito control programs; however, identifying high-risk areas for mosquito infestation over a large geographic region based only on field surveys is labor-intensive and time-consuming. Thus, the objective of this study was to assess the potential of remote satellite images (WorldView) for determining land features associated with Ae. aegypti adult infestations in São José do Rio Preto/SP, Brazil. We used data from 60 adult mosquito traps distributed along four summers; the remote sensing images were classified by land cover types using a supervised classification method. We modeled the number of Ae. aegypti using a Poisson probability distribution with a geostatistical approach. The models were constructed in a Bayesian context using the Integrated nested Laplace Approximations and Stochastic Partial Differential Equation method. We showed that an infestation of Ae. aegypti adult mosquitoes was positively associated with the presence of asbestos roofing and roof slabs. This may be related to several other factors, such as socioeconomic or environmental factors. The usage of asbestos roofing may be more prevalent in socioeconomically poor areas, while roof slabs may retain rainwater and contribute to the generation of temporary mosquito breeding sites. Although preliminary, our results demonstrate the utility of satellite remote sensing in identifying landscape differences in urban environments using a geostatistical approach, and indicated directions for future research. Further analyses including other variables, such as land surface temperature, may reveal more complex relationships between urban mosquito micro-habitats and land cover features.

14.
PLoS Negl Trop Dis ; 14(2): e0008034, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32017766

RESUMEN

BACKGROUND: Zika virus has recently spread to South- and Central America, causing congenital birth defects and neurological complications. Many people at risk are flavivirus pre-immune due to prior infections with other flaviviruses (e.g. dengue virus) or flavivirus vaccinations. Since pre-existing cross-reactive immunity can potentially modulate antibody responses to Zika virus infection and may affect the outcome of disease, we analyzed fine-specificity as well as virus-neutralizing and infection-enhancing activities of antibodies induced by a primary Zika virus infection in flavivirus-naïve as well as yellow fever- and/or tick-borne encephalitis-vaccinated individuals. METHODOLOGY: Antibodies in sera from convalescent Zika patients with and without vaccine-induced immunity were assessed by ELISA with respect to Zika virus-specificity and flavivirus cross-reactivity. Functional analyses included virus neutralization and infection-enhancement. The contribution of IgM and cross-reactive antibodies to these properties was determined by depletion experiments. PRINCIPAL FINDINGS: Pre-existing flavivirus immunity had a strong influence on the antibody response in primary Zika virus infections, resulting in higher titers of broadly flavivirus cross-reactive antibodies and slightly lower levels of Zika virus-specific IgM. Antibody-dependent enhancement (ADE) of Zika virus was mediated by sub-neutralizing concentrations of specific IgG but not by cross-reactive antibodies. This effect was potently counteracted by the presence of neutralizing IgM. Broadly cross-reactive antibodies were able to both neutralize and enhance infection of dengue virus but not Zika virus, indicating a different exposure of conserved sequence elements in the two viruses. CONCLUSIONS: Our data point to an important role of flavivirus-specific IgM during the transient early stages of infection, by contributing substantially to neutralization and by counteracting ADE. In addition, our results highlight structural differences between strains of Zika and dengue viruses that are used for analyzing infection-enhancement by cross-reactive antibodies. These findings underscore the possible impact of specific antibody patterns on flavivirus disease and vaccination efficacy.

15.
BMC Infect Dis ; 20(1): 116, 2020 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-32041533

RESUMEN

BACKGROUND: Yellow fever vaccine exists for over 80 years and is considered to be relatively safe. However, in rare cases it can produce serious neurotropic and viscerotropic complications. We report a case of a patient who presented both viscerotropic and neurological manifestations after yellow fever vaccination. CASE PRESENTATION: We describe the case of a 37 years old man who developed after the yellow fever vaccination a yellow fever vaccine-associated viscerotropic disease followed by acute uveitis. Prolonged detection of yellow fever RNA in blood and urine was consistent with yellow fever vaccine-associated adverse event. The final outcome was good, although with persistent fatigue over a few months. CONCLUSIONS: Even if the yellow fever vaccine is relatively safe, physicians should be aware of its possible serious adverse effects.

16.
BMC Genomics ; 21(1): 143, 2020 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-32041546

RESUMEN

BACKGROUND: Successful mating of female mosquitoes typically occurs once, with the male sperm being stored in the female spermatheca for every subsequent oviposition event. The female spermatheca is responsible for the maintenance, nourishment, and protection of the male sperm against damage during storage. Aedes aegypti is a major vector of arboviruses, including Yellow Fever, Dengue, Chikungunya, and Zika. Vector control is difficult due to this mosquito high reproductive capacity. RESULTS: Following comparative RNA-seq analyses of spermathecae obtained from virgin and inseminated females, eight transcripts were selected based on their putative roles in sperm maintenance and survival, including energy metabolism, chitin components, transcriptional regulation, hormonal signaling, enzymatic activity, antimicrobial activity, and ionic homeostasis. In situ RNA hybridization confirmed tissue-specific expression of the eight transcripts. Following RNA interference (RNAi), observed outcomes varied between targeted transcripts, affecting mosquito survival, egg morphology, fecundity, and sperm motility within the spermathecae. CONCLUSIONS: This study identified spermatheca-specific transcripts associated with sperm storage in Ae. aegypti. Using RNAi we characterized the role of eight spermathecal transcripts on various aspects of female fecundity and offspring survival. RNAi-induced knockdown of transcript AeSigP-66,427, coding for a Na+/Ca2+ protein exchanger, specifically interfered with egg production and reduced sperm motility. Our results bring new insights into the molecular basis of sperm storage and identify potential targets for Ae. aegypti control.

17.
Expert Rev Vaccines ; 2020 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-32077343

RESUMEN

INTRODUCTION: Aedes-borne arboviruses imposes significant economic and public health costs inAreas covered: We performed a systematic review of the economic evidence surrounding aedes-borne arboviruses and strategies to prevent and control these diseases to inform disease control policy decisions and research directions. We searched four databases covering an 18-year period (2000-2018) to identify arboviral disease-related cost of illness studies, cost studies of vector control and prevention strategies, cost-effectiveness analyses and cost-benefit analyses. We identified 74 published studies that revealed substantial global total costs in yellow fever virus and dengue virus ranging from $2.1 billion - 57.3 billion. Cost studies of vector control and surveillance programs are limited, but a few studies found that costs of vector control programs ranged from $5.62to $73.5 million. Cost-effectiveness evidence was limited across Aedes-borne diseases, but generally found targeted dengue vaccination programs cost-effective. This review revealed insufficient economic evidence for vaccine introduction and implementation of surveillance and vector control programs.Expert opinion: Evidence of the economic burden of aedes-borne arboviruses and the economic impact of strategies for arboviral disease prevention and control is critical to inform policy decisions and to secure continued financial support for these preventive and control measures.

18.
Einstein (Sao Paulo) ; 18: eRC5041, 2020.
Artículo en Inglés, Portugués | MEDLINE | ID: mdl-32074223

RESUMEN

The yellow fever is a systemic disease that was under control due to the effective campaigns against the vector and promotion of vaccines programs. However, since 1999, outbreaks appeared because of inefficient control of the vector, and led to the need of amplifying the immunization in large scale against the yellow fever virus, and consequently, raising the risk of adverse reactions to the vaccine. We report a case of previously healthy infant, who was referred to our care service, after 3 days with fever, chills, nausea and vomits, he received support therapy and was discharged from the hospital. After 24 hours of supportive measures, he was discharge. The patient returned to our service with general condition decline, strabismus, inability to control of cervical musculature and reduced force of the legs. The patient vaccine had received all vaccines from the calendar, and he was vaccinated for yellow fever 20 days before symptoms. During the hospitalization, liquor was collected, and ceftriaxone and aciclovir were administered. After negative cultures from the liquor, the antibiotics were suspended. The computed tomography of patient's brain showed no alterations. Research for antibodies against yellow fever was requested, being positive for IgM in the liquor, and confirming the neurotropic disease associated with the yellow fever vaccine. On the fifth day of hospitalization, the patient showed improvement on the strabismus, cervical tonus, and musculature force. On the tenth day of hospitalization, patient showed complete improvement, and his laboratory exams no alterations. Subsequently, patient was discharged. The vaccine against yellow fever is safe, efficient and highly recommended, however it is not completely free from serious adverse reactions, including death.

19.
Parasit Vectors ; 13(1): 91, 2020 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-32075683

RESUMEN

BACKGROUND: The international movement of used tyres is a major factor responsible for global introductions of Aedes invasive mosquitoes (AIMs) (Diptera: Culicidae) that are major disease vectors (e.g. dengue, Zika, chikungunya and yellow fever). Surveillance methods are restricted by expense, availability and efficiency to detect all life stages. Currently, no tested method exists to screen imported used tyres for eggs in diapause, the life stage most at risk from accidental introduction. Here we test the efficiency of adhesive tape as an affordable and readily available material to screen tyres for eggs, testing its effect on hatch rate, larval development, DNA amplification and structural damage on the egg surface. RESULTS: We demonstrated that the properties of adhesive tape can influence pick up of dormant eggs attached to dry surfaces. Tapes with high levels of adhesion, such as duct tape, removed eggs with high levels of efficiency (97% ± 3.14). Egg numbers collected from cleaned used tyres were found to explain larval hatch rate success well, particularly in subsequent larval to adult emergence experiments. The strength of this relationship decreased when we tested dirty tyres. Damage to the exochorion was observed following scanning electron microscopy (SEM), possibly resulting in the high variance in the observed model. We found that five days was the optimal time for eggs to remain on all tested tapes for maximum return on hatch rate success. Tape type did not inhibit amplification of DNA of eggs from three, five or ten days of exposure. Using this DNA, genotyping of AIMs was possible using species-specific markers. CONCLUSIONS: We demonstrated for the first time that adhesive tapes are effective at removing AIM eggs from tyres. We propose that this method could be a standardised tool for surveillance to provide public health authorities and researchers with an additional method to screen tyre cargo. We provide a screening protocol for this purpose. This method has a global applicability and in turn can lead to increased predictability of introductions and improve screening methods at high risk entry points.

20.
Parasit Vectors ; 13(1): 90, 2020 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-32075684

RESUMEN

BACKGROUND: Yellow fever (YF) is a severe, infectious, but non-communicable arboviral hemorrhagic disease. In the last decades, yellow fever virus (YFV) infections have been prevalent in endemic areas in Brazil, affecting human and non-human primate (NHP) populations. Monitoring of NHP infection started in 1999, and reports of epizootic diseases are considered important indicators of viral transmission, particularly in relation to the sylvatic cycle. This study presents the monitoring of YFV by real-time RT-PCR and the epidemiological findings related to the deaths of NHPs in the south-eastern states and in the north-eastern state of Bahia, during the outbreak of YF in Brazil during 2017 and 2018. METHODS: A total of 4198 samples from 2099 NHPs from south-eastern and north-eastern Brazilian states were analyzed by real-time reverse transcription polymerase chain reaction (rtRT-PCR). RESULTS: A total of 4198 samples from 2099 NHPs from south-eastern and north-eastern Brazilian states were collected between 2017 and 2018. The samples were subjected to molecular diagnostics for YFV detection using real-time reverse transcription polymerase chain reaction (rtRT-PCR) techniques. Epizootics were coincident with human YF cases. Furthermore, our results showed that the YF frequency was higher among marmosets (Callithrix sp.) than in previous reports. Viremia in species of the genus Alouatta and Callithrix differed greatly. DISCUSSION: Our results indicate a need for further investigation of the role of Callithrix spp. in the transmission cycles of YFV in Brazil. In particular, YFV transmission was observed in a region where viral circulation has not been recorded for decades and thus vaccination has not been previously recommended. CONCLUSIONS: This highlights the need to straighten epizootic surveillance and evaluate the extent of vaccination programmes in Brazil in previously considered "YFV-free" areas of the country.

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