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Infection of Rift Valley fever virus (RVFV), a highly pathogenic mosquito-borne zoonotic virus, triggers severe inflammatory pathogenesis but the underlying mechanism of inflammation activation is currently unclear. Here, we report that the non-structural protein NSs of RVFV triggers mitochondrial damage to activate the NLRP3 inflammasome leading to viral pathogenesis in vivo. It is found that the host transcription inhibition effect of NSs causes rapid down-regulation of myeloid cell leukemia-1(MCL-1), a pro-survival member of the Bcl-2 (B-cell lymphoma protein 2) protein family. MCL-1 down-regulation led to BAK activation in the mitochondria, which triggered mtROS production and release of oxidized mitochondrial DNA (ox-mtDNA) into the cytosol. Cytosolic ox-mtDNA binds and activates the NLRP3 inflammasome triggering NLRP3-GSDMD pyroptosis in RVFV infected cells. A NSs mutant virus (RVFV-NSsRM) that is compromised in inducing transcription inhibition did not trigger MCL-1 down-regulation nor NLRP3-GSDMD pyroptosis. RVFV infection of the Nlrp3-/- mouse model demonstrated that the RVFV-triggered NLRP3 pyroptosis contributed to RVFV inflammatory pathogenesis and fatal infection in vivo. Infection with the RVFV-NSsRM mutant virus similarly showed alleviated inflammatory pathogenesis and reduced fatality rate. Taken together, these results revealed a mechanism by which a virulence factor activates the mitochondrial MCL-1-BAK axis through inducing host transcription inhibition to trigger NLRP3-dependent inflammatory pathogenesis. Author summary: RVFV is a highly pathogenic mosquito-borne zoonotic virus that infects both ruminants and humans. Humans infected by RVFV can develop acute hepatitis, encephalitis and hemorrhagic fever, which can result in fatal infection in severe cases. RVFV-triggered inflammation is an important driver for severe disease development but the underlying activation mechanism is currently unknown. Here, we found that the major virulence protein NSs of RVFV induces mitochondria damage in infected cells. This is achieved through the transcription inhibition effect of NSs which triggered transcriptional down-regulation of MCL-1 leading to mitochondrial BAK activation. Activated BAK then led to mitochondrial permeabilization and release of ox-mtDNA into the cytosol. The cytoplasmic ox-mtDNA binds and activates NLRP3 inflammasome leading to inflammatory pyroptosis. Using a RVFV infected mouse model and a mutant virus that did not trigger transcription inhibition, we showed that RVFV-triggered NLRP3 activation contributed to viral pathogenesis and fatal infection in vivo. In summary, we showed an important mechanism of RVFV triggered pathogenic inflammation, which may be employed by other viruses that induce transcription inhibition during infection. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a neglected vector-borne disease which is endemic in many countries across Africa and has seen recent geographical expansions into the Arabian Peninsula. RVF can cause severe infections in both animals and humans. RVF infections in livestock can lead to mass fatalities. In humans, the symptoms are nonspecific and can often lead to misdiagnosis. However, a small proportion progresses to haemorrhagic infection with a significantly higher mortality rate. The culmination of this can cause severe socioeconomic impacts. This review aims to identify the main socioeconomic impacts caused by RVF outbreaks as well as existing knowledge gaps. Ninety-three academic and grey papers were selected, covering 19 countries and 10 methodological approaches. A variety of socioeconomic impacts were found across all levels of society: Livestock trade disruptions consequently impacted local food security, local and national economies. Most livestock farmers in endemic countries are subsistence farmers and so rely on their livestock for sustenance and income. RVF outbreaks resulted in a variety of socioeconomic impacts, e.g., the inability to pay for school fees. Main barriers to vaccine uptake in communities were lack of access, funds, interest along with other social aspects. The occupational risks for women (and pregnant women) are largely unknown. To our knowledge, this is the first review on RVF to highlight the clear knowledge gap surrounding the potential gender differences on risks of RVF exposure, as well as differences on occupational health risk in pastoral communities. Further work is required to fill the gaps identified in this review and inform control policies. Author summary: Rift Valley fever (RVF) outbreaks have affected many regions globally with varying severity. The number of reported cases in animals and humans is patchy at best, with some countries not recording animal events. This review focuses on 3 main themes: Health Burden; Economic Impacts; and Prevention and Surveillance. The review explores socioeconomic impacts across local, regional, and national levels. We found women have varying degrees of risk to exposure of RVF; however, more research is required to better understand gender differences. RVF outbreaks tend to begin in livestock and spillover into humans. The complex disease dynamics are poorly understood because of the interactions between animals, humans, and the environment. This cross-boundary dynamics fall outside of the public health remit and into veterinary health. This allows for outbreaks to spread uncontrollably for some time. More research using a One Health Lens is required which explores these complex dynamics with the combination of different social aspects. This review highlights the wider impacts experienced at different levels of society and where there are major gaps in our knowledge. Further analysis into these gaps is required to and will aid policy makers in developing new control measures around RVF. [ABSTRACT FROM AUTHOR]

Background: Coxiella burnetii, the causative agent of Q fever, and Rift Valley fever virus are two under-researched zoonotic pathogens in Ethiopia. Potential outbreaks of these diseases, in light of the high dependency of nomadic pastoralists on their livestock, poses a risk to both human and animal health in addition to risking the pastoralists livelihoods. Our study aimed to determine the seroprevalence and associated risk factors for Q fever and Rift Valley fever in pastoral communities in the Afar region of north-eastern Ethiopia. Methodology/Principal findings: This cross-sectional study screened pastoralists (n = 323) and their livestock (n = 1377) for IgG antibodies to Coxiella burnetii and Rift Valley fever virus. A seroprevalence for Q fever of 25.0% (95%CI 18.6–32.6) was found in pastoralists and 34.3% (95%CI 27.9–41.3) in livestock overall; with 51.9% in goats (95%CI 44.9–58.8), 39.9% in sheep (95%CI 24.6–51.2), 16.3% in camels (95%CI 10.4–24.6) and 8.8% in cattle (95%CI 5.0–15.0). For Rift Valley fever the seroprevalence in pastoralists was 6.1% (95%CI 3.3–11.0) and 3.9% (95%CI 2.6–5.7) in livestock overall; cattle had the highest seroprevalence (8.3%, 95%CI 3.3–19.2), followed by goats (2.7%; 95%CI 1.4–5.1), sheep (2.5%; 95%CI 1.0–5.9) and camels (1.8%; 95%CI 0.4–6.9). Human Q fever seropositivity was found to be associated with goat abortions (OR = 2.11, 95%CI 1.18–3.78, p = 0.011), while Rift Valley fever seropositivity in livestock was found to be associated with cattle abortions (OR = 2.52, 95%CI 1.05–6.08, p = 0.039). Conclusions/Significance: This study provides evidence for a notable exposure to both Q fever and Rift Valley fever in pastoralists and livestock in Afar. The outbreak potential of these pathogens warrants ongoing integrated human and animal surveillance requiring close collaboration of the human and animal health sectors with community representatives following a One Health approach. Author summary: Q fever and Rift Valley fever are two diseases that can affect both humans and animals, causing illness and death. These two diseases can cause large-scale outbreaks, not only affecting the health of individuals and communities but also resulting in a substantial economic loss at the individual, community, regional and national levels through losses in livestock products. We conducted a sero-epidemiological study in the Afar region of north-eastern Ethiopia to determine the presence of these two diseases in nomadic pastoralists and their livestock. Our results indicate that 25.0% of the pastoralists and 34.3% of their animals have had previous exposure to Q fever and 6.1% of the pastoralists and 3.9% of their animals have been exposed to Rift Valley fever. Goats appear to be the most common livestock species associated with Q fever in this region, while cattle are the most common species associated with Rift Valley fever. Our findings highlight the importance of continuous surveillance; additionally, we would recommend the development of rapid response plans for potential outbreaks that integrate the human and animal health sectors. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is an important zoonotic viral disease affecting several species of domestic and wild ruminants, causing major economic losses and dozens of human deaths in various geographical areas of Africa, where it is endemic. Although it is not present in Europe, there is a risk of its introduction and spread linked to globalisation and climate change. At present, the only measure that could help to prevent the disease is vaccination of flocks in areas at risk of RVF. Available live attenuated vaccines are an effective means of controlling the disease, but their use is often questioned due to residual virulence, particularly in susceptible hosts such as pregnant sheep. On the other hand, no vaccine is currently licensed for use in humans. The development of safe and effective vaccines is therefore a major area of research. In previous studies, we selected under selective mutagenic pressure a highly attenuated RVFV 56/74 virus variant called 40Fp8. This virus showed an extremely attenuated phenotype in both wild-type and immunodeficient A129 (IFNARKO) mice, yet was still able to induce protective immunity after a single inoculation, thus supporting its use as a safe, live attenuated vaccine. To further investigate its safety, in this work we have analysed the attenuation level of 40Fp8 in immunosuppressed mice (A129) when administered by the intranasal route, and compared it with other attenuated RVF viruses that are the basis of vaccines in use or in development. Our results show that 40Fp8 has a much higher attenuated level than these other viruses and confirm its potential as a candidate for safe RVF vaccine development. Author summary: Vaccines that use attenuated viruses are highly effective in terms of providing protection, duration, breadth, and quality of the immune response. However, they may pose a risk to immunosuppressed individuals or in certain situations, such as during pregnancy. It is therefore important to analyze the residual virulence of attenuated vaccines to ensure their safety. In this work, we analysed the safety of the RVF virus vaccine variant 40Fp8 developed in our laboratory. We studied its effect on immunodeficient mice and used a more aggressive route of administration such as the intranasal delivery route. To assess the degree of attenuation of 40Fp8, we compared it with other attenuated prototype RVF vaccines. The results clearly demonstrate that 40Fp8 is highly attenuated, and its residual virulence level is extremely low compared to other vaccine viruses tested. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is an encephalitic bunyavirus that can infect neurons in the brain. There are no approved therapeutics that can protect from RVFV encephalitis. Innate immunity, the first line of defense against infection, canonically antagonizes viruses through interferon signaling. We found that interferons did not efficiently protect primary cortical neurons from RVFV, unlike other cell types. To identify alternative neuronal antiviral pathways, we screened innate immune ligands and discovered that the TLR2 ligand Pam3CSK4 inhibited RVFV infection, and other bunyaviruses. Mechanistically, we found that Pam3CSK4 blocks viral fusion, independent of TLR2. In a mouse model of RVFV encephalitis, Pam3CSK4 treatment protected animals from infection and mortality. Overall, Pam3CSK4 is a bunyavirus fusion inhibitor active in primary neurons and the brain, representing a new approach toward the development of treatments for encephalitic bunyavirus infections. Author summary: Rift Valley fever virus (RVFV) is an emerging mosquito-borne pathogen, which has shown the potential to spread geographically, and which can cause severe disease in humans. One of the most serious complications of RVFV infection is invasion into the brain. There are no approved treatments for RVFV infection, or any other bunyavirus that infects the brain. We found that the classically antiviral interferon pathway was not protective in neurons. This led us to screen for new antivirals, and we identified Pam3CSK4, which inhibited RVFV and related bunyaviruses, but not vesicular stomatitis virus. We found that the antiviral activity of Pam3CSK4 was separate from its previously characterized activity as a TLR2 agonist. Instead, Pam3CSK4 blocked RVFV entry at the step of viral membrane fusion. Importantly, we found that Pam3CSK4 prevented RVFV infection in the mouse brain. This work reveals that Pam3CSK4 prevents encephalitic bunyavirus infection in neurons, a vulnerable cell type, and future research should evaluate the therapeutic potential of Pam3CSK4. [ABSTRACT FROM AUTHOR]

Assay validation is an essential component of disease surveillance testing, but can be problematic in settings where access to positive control material is limited and a safety risk for handlers. Here we describe a single non-infectious synthetic control that can help develop and validate the PCR based detection of the viral causes of Crimean-Congo hemorrhagic fever, Ebola virus disease, Lassa fever, Marburg virus disease and Rift Valley fever. We designed non-infectious synthetic DNA oligonucleotide sequences incorporating primer binding sites suitable for five assays, and a T7 promotor site which was used to transcribe the sequence. Transcribed RNA was used as template in a dilution series, extracted and amplified with RT-PCR and RT-qPCR to demonstrate successful recovery and determine limits of detection in a range of laboratory settings. Our results show this approach is adaptable to any diagnostic assay requiring validation of nucleic acid extraction and/or amplification, particularly where sourcing reliable, safe material for positive controls is infeasible. Author summary: The majority of zoonoses originate in wildlife and tend to emerge from biodiverse regions in low to middle income countries, frequently among deprived populations of at-risk people with a lack of access to diagnostic capacity or surveillance. Diseases such as Crimean-Congo hemorrhagic fever, Rift Valley fever, Ebola virus disease, Marburg virus disease and Lassa fever are viral hemorrhagic fevers (VHFs) and rank among the most neglected and serious threats to global public health. This threat is partly due to the severity of disease caused by these pathogens, but also because their geographical distribution is close to human populations with often limited access to medical or diagnostic laboratory services. In our study we describe techniques for PCR based detection of five VHF viruses using a synthetic, multi-target non-infectious positive control. Our work has applications in assay design and optimization, particularly where access to source material is problematic or requires high level biosafety containment, as is the case with VHF viruses. This approach can help learners train in techniques used in nucleic acid extraction, amplification, and sequencing of VHF viruses and can be used for any targets, with potential for multiplexing from a single positive control. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is a viral zoonosis that causes severe disease in ruminants and humans. The nonstructural small (NSs) protein is the primary virulence factor of RVFV that suppresses the host's antiviral innate immune response. Bioinformatic analysis and AlphaFold structural modeling identified four putative LC3-interacting regions (LIR) motifs (NSs 1–4) in the RVFV NSs protein, which suggest that NSs interacts with the host LC3-family proteins. Using, isothermal titration calorimetry, X-ray crystallography, co-immunoprecipitation, and co-localization experiments, the C-terminal LIR motif (NSs4) was confirmed to interact with all six human LC3 proteins. Phenylalanine at position 261 (F261) within NSs4 was found to be critical for the interaction of NSs with LC3, retention of LC3 in the nucleus, as well as the inhibition of autophagy in RVFV infected cells. These results provide mechanistic insights into the ability of RVFV to overcome antiviral autophagy through the interaction of NSs with LC3 proteins. Author summary: Rift Valley fever virus (RVFV) infection can cause severe illnesses including liver disease, encephalitis, and hemorrhagic fever. Unfortunately, there are no FDA-approved vaccines or therapeutics for human use. The NSs protein is the main virulence factor of RVFV and is a potent inhibitor of the host antiviral immune responses. Autophagy has been previously shown to be antiviral in RVFV infected cells, but RVFV can suppress this antiviral pathway by a previously undetermined mechanism. In this manuscript, we demonstrate that NSs interacts with host LC3-family proteins through an LC3 interacting region (LIR) within its C-terminus. This interaction results in LC3 being retained in the nucleus and autophagy inhibition. Our studies provide mechanistic insights by which NSs subverts antiviral autophagy during RVFV infection. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a mosquito-borne viral zoonosis caused by the Rift Valley fever virus (RVFV) that can infect domestic and wild animals. Although the RVFV transmission cycle has been well documented across Africa in savanna ecosystems, little is known about its transmission in tropical rainforest settings, particularly in Central Africa. We therefore conducted a survey in northeastern Gabon to assess RVFV circulation among wild and domestic animals. Among 163 wildlife samples tested using RVFV-specific RT-qPCR, four ruminants belonging to subfamily Cephalophinae were detected positive. The phylogenetic analysis revealed that the four RVFV sequences clustered together with a virus isolated in Namibia within the well-structured Egyptian clade. A cross-sectional survey conducted on sheep, goats and dogs living in villages within the same area determined the IgG RVFV-specific antibody prevalence using cELISA. Out of the 306 small ruminants tested (214 goats, 92 sheep), an overall antibody prevalence of 15.4% (95% CI [11.5–19.9]) was observed with a higher rate in goats than in sheep (20.1% versus 3.3%). RVFV-specific antibodies were detected in a single dog out of the 26 tested. Neither age, sex of domestic animals nor season was found to be significant risk factors of RVFV occurrence. Our findings highlight sylvatic circulation of RVFV for the first time in Gabon. These results stress the need to develop adequate surveillance plan measures to better control the public health threat of RVFV. Author summary: Rift Valley fever (RVF) is a mosquito-borne viral zoonosis caused by the Rift Valley fever virus (RVFV) that can affect wild and domestic animals. Although the RVFV transmission cycle has been well documented across Africa in savanna ecosystems, little is known about its transmission in tropical rainforests, especially in Central Africa. We thus conducted a survey in northeastern Gabon to assess RVFV circulation among wild and domestic animals. In this study, we demonstrated for the first time in Gabon the presence of the RVFV in two wildlife species (Peter's duiker Cephalophus callipygus and the blue duiker Philantomba monticola). In addition, we detected RVFV-specific antibodies in small domestic ruminants (sheep and goats) at an overall antibody prevalence of 15.4%, with a much higher seroprevalence rate in goats than sheep (20.1% versus 3.3%). Furthermore, RVFV-specific antibodies were also observed in a single (hunting) dog out of the 26 tested. These results stress the need to develop adequate surveillance plan measures to better control the public health threat of RVFV. [ABSTRACT FROM AUTHOR]

Background: The escalating risk and contemporary occurrences of arbovirus infections prompt a critical inquiry into the ability of nations to execute efficient surveillance systems capable to detect, prevent and respond to arbovirus outbreaks. Healthcare workers (HCWs) are the major actors in the surveillance of infectious diseases with epidemic potential. The objective of this study was to evaluate the knowledge, attitudes and perceptions of HCWs regarding arboviruses in the public health facilities of Conakry, Guinea. Methods: A cross-sectional survey was conducted during the from December 27, 2022, to January 31, 2023, encompassing from public health facilities in Conakry. The data collection process encompassed various aspects, including the characteristics of health facilities, socio-demographic and professional attributes of HCWs, the information received concerning arboviruses and the sources of information, as well as a series of inquiries designed to evaluate their knowledge, attitudes and perceptions. Subsequently, scores were computed for knowledge, attitude and perception. To identify the factors influencing the knowledge, attitudes, and perceptions of HCWs regarding arboviruses, Decision Tree and Inference Conditional Tree models were used. Results: A total of 352 HCWs participated in the study, comprising 219 from national hospitals, 72 from municipal hospitals and 61 from primary health centers. More than half of the respondents (54.3%) had never received information on arboviruses. Only 1% of the respondents had good knowledge about arboviruses, 95.7% had a negative attitude about arboviruses. Moreover, nearly 60% of the respondents had a moderate perception and 24.1% had a good perception. The analysis revealed significant associations between the knowledge and attitudes of respondents concerning arboviruses and their years of professional experience and age. Conclusion: This study highlights the imperative requirement for comprehensive training targeting HCWs to enhance their capacity for early case detection within healthcare facilities. Additionally, there is a crucial need for analogous studies adopting a mixed-methods approach across all healthcare regions in Guinea. Author summary: Arboviruses, including yellow fever, dengue, Zika, chikungunya, and Rift Valley fever, constitute viral diseases transmitted by blood-sucking arthropod vectors, such as mosquitoes, ticks, and biting midges, to vertebrate hosts. In many countries, particularly within Africa, the prevalence of these arboviruses remains widely underestimated due to the absence of effective surveillance systems, often resulting in the conflation of arboviral infections with other infectious diseases. Nevertheless, sub-Saharan Africa is prone to periodic arbovirus-linked epidemics, propelled by factors such as the expansion of vector habitats, viral genetic mutations, anthropogenic influences, inadequate healthcare services, commercial transportation, and climate fluctuations. While Guinea has predominantly reported suspected cases of yellow fever, widespread arbovirus-related epidemics have yet to be documented. Nevertheless, it is imperative for healthcare workers to be well-informed and prepared for prospective epidemics. The assessment of the knowledge, attitudes, and perceptions of healthcare workers can serve as a catalyst to heighten awareness, bolster surveillance mechanisms, and enhance the capabilities of public health facilities in the prevention and mitigation of potential epidemics. [ABSTRACT FROM AUTHOR]

Background: In Africa, little is known about the role of telephone hotlines in outbreak response. We systematically reviewed the role and effectiveness of hotlines on outbreak response in Africa. Method: We used the Cochrane handbook and searched five databases. The protocol was registered on PROSPERO (CRD42021247141). Medline, Embase, PsycINFO, Global Health and Web of Science were searched from 30 June 2020 to August 2020 for studies on the use of telephone hotlines in outbreak response in Africa published between January 1995 and August 2020. The search was also repeated on 16 September 2022. Data on effectiveness (alerts generated, cases confirmed) were extracted from peer-reviewed studies. Meta-analysis of alerts generated, and proportion of cases confirmed was done using the random effects model. The quality of studies was assessed using the Joanna Briggs Institute (JBI) tools. The heterogeneity and publication bias were assessed using the Galbraith and funnel plots, respectively. Results: Our search yielded 1251 non-duplicate citations that were assessed. 41 full texts were identified, and 21 studies were included in the narrative synthesis, while 12 were included in the meta-analysis. The hotlines were local (seven studies) or national (three studies). A combination of a local and national hotline was used in one study. The hotlines were set up for unusual respiratory events (one study), polio (one study), Ebola (10 studies), COVID-19 (two studies), malaria (one study), influenza-like illnesses (ILI) (one study) and rift valley fever in livestock (one study). Hotlines were mainly used for outbreak surveillance at the local level. A total of 332,323 alerts were generated, and 67,658 met the case definition, corresponding to an overall pooled proportion of alerts generated(sensitivity) of 38% (95%CI: 24–52%). The sensitivity was 41% (95% CI: 24–59%) for local hotlines and 26%(95%CI:5–47%) for national hotlines. Hotlines were also used for surveillance of rift valley fever in livestock (one study) vaccination promotion (one study), death reporting (five studies), rumour tracking and fighting misinformation (two studies) and community engagement (five studies). The studies were of low to moderate quality with high publication bias and heterogeneity(I2 = 99%). The heterogeneity was not explained by the sample size. Conclusion: These data suggest that telephone hotlines can be effective in outbreak disease surveillance in Africa. Further implementation research is needed to scale up telephone hotlines in rural areas. [ABSTRACT FROM AUTHOR]

Crimean-Congo Hemorrhagic fever (CCHF) is an important zoonotic disease transmitted to humans both by tick vectors and contact with fluids from an infected animal or human. Although animals are not symptomatic when infected, they are the main source of human infection. Uganda has reported sporadic human outbreaks of CCHF in various parts of the country since 2013. We designed a nationwide epidemiological study to investigate the burden of CCHF in livestock. A total of 3181 animals were sampled; 1732 cattle (54.4%), 1091 goats (34.3%), and 358 sheep (11.3%) resulting in overall livestock seropositivity of IgG antibodies against CCHF virus (CCHFV) of 31.4% (999/3181). Seropositivity in cattle was 16.9% and in sheep and goats was 48.8%. Adult and juvenile animals had higher seropositivity compared to recently born animals, and seropositivity was higher in female animals (33.5%) compared to male animals (24.1%). Local breeds had higher (36.8%) compared to exotic (2.8%) and cross breeds (19.3%). Animals that had a history of abortion or stillbirth had higher seropositivity compared to those without a history of abortion or stillbirth. CCHFV seropositivity appeared to be generally higher in northern districts of the country, though spatial trends among sampled districts were not examined. A multivariate regression analysis using a generalized linear mixed model showed that animal species, age, sex, region, and elevation were all significantly associated with CCHFV seropositivity after adjusting for the effects of other model predictors. This study shows that CCHFV is actively circulating in Uganda, posing a serious risk for human infection. The results from this study can be used to help target surveillance efforts for early case detection in animals and limit subsequent spillover into humans. [ABSTRACT FROM AUTHOR]

Brucella spp. and Rift Valley fever virus (RVFV) are classified as priority zoonotic agents in Kenya, based on their public health and socioeconomic impact on the country. Data on the pathogen-specific and co-exposure levels is scarce due to limited active surveillance. This study investigated seroprevalence and co-exposure of Brucella spp. and RVFV and associated risk factors among slaughterhouse workers in Isiolo County, northern Kenya. A cross-sectional serosurvey was done in all 19 slaughterhouses in Isiolo County, enrolling 378 participants into the study. The overall seroprevalences for Brucella spp. and RVFV were 40.2% (95% CI: 35.2–45.4) and 18.3% (95% CI: 14.5–22.5), respectively while 10.3% (95% CI 7.4%-13.8%) of individuals were positive for antibodies against both Brucella spp. and RVFV. Virus neutralisation tests (VNT) confirmed anti-RVFV antibodies in 85% of ELISA-positive samples. Our seroprevalence results were comparable to community-level seroprevalences previously reported in the area. Since most of the study participants were not from livestock-keeping households, our findings attribute most of the detected infections to occupational exposure. The high exposure levels indicate slaughterhouse workers are the most at-risk population and there is need for infection, prevention, and control programs among this high-risk group. This is the first VNT confirmation of virus-neutralising antibodies among slaughterhouse workers in Isiolo County and corroborates reports of the area being a high-risk RVFV area as occasioned by previously reported outbreaks. This necessitates sensitization campaigns to enhance awareness of the risks involved and appropriate mitigation measures. Author summary: Brucellosis and Rift Valley fever are zoonotic diseases with significant public health and economic burden in Kenya. We investigated the levels of exposure to Brucella spp. and Rift Valley fever virus among slaughterhouse workers in Isiolo County, northern Kenya. We found high exposure levels that are comparable to those previously reported within the community. Occupational exposure was the most probable source of infection since most of the workers did not keep livestock. Therefore, slaughterhouses could be targeted for sensitization on appropriate infection control measures besides being important points for disease surveillance. [ABSTRACT FROM AUTHOR]

Zika virus (ZIKV) is a Flavivirus responsible for recent epidemics in Pacific Islands and in the Americas. In humans, the consequences of ZIKV infection range from asymptomatic infection to severe neurological disease such as Guillain-Barré syndrome or fetal neurodevelopmental defects, suggesting, among other factors, the influence of host genetic variants. We previously reported similar diverse outcomes of ZIKV infection in mice of the Collaborative Cross (CC), a collection of inbred strains with large genetic diversity. CC071/TauUnc (CC071) was the most susceptible CC strain with severe symptoms and lethality. Notably, CC071 has been recently reported to be also susceptible to other flaviviruses including dengue virus, Powassan virus, West Nile virus, and to Rift Valley fever virus. To identify the genetic origin of this broad susceptibility, we investigated ZIKV replication in mouse embryonic fibroblasts (MEFs) from CC071 and two resistant strains. CC071 showed uncontrolled ZIKV replication associated with delayed induction of type-I interferons (IFN-I). Genetic analysis identified a mutation in the Irf3 gene specific to the CC071 strain which prevents the protein phosphorylation required to activate interferon beta transcription. We demonstrated that this mutation induces the same defective IFN-I response and uncontrolled viral replication in MEFs as an Irf3 knock-out allele. By contrast, we also showed that Irf3 deficiency did not induce the high plasma viral load and clinical severity observed in CC071 mice and that susceptibility alleles at other genes, not associated with the IFN-I response, are required. Our results provide new insight into the in vitro and in vivo roles of Irf3, and into the genetic complexity of host responses to flaviviruses. Author summary: Recent ZIKV outbreaks led to millions of infected people, with rare but severe complications such as Guillain-Barré syndrome and encephalitis in adults suggesting that host genes influence the susceptibility to severe forms of infection. We previously reported the importance of host genes in ZIKV pathogenesis using a panel of genetically diverse mouse strains and identified CC071 as the most susceptible strain. Importantly, this mouse strain has been shown by others to be also susceptible to several other RNA viruses. Through a combination of functional and genetic approaches in a cellular model, we identified a mutation in the Irf3 gene which plays a key role in activating the expression of interferon beta to induce the type I interferon response, the first line of host defense against the virus. This mutation fully explains the high viral replication observed in CC071 cells. However, it was not able to induce the elevated viremia and the disease signs displayed by CC071 ZIKV-infected mice, unraveling the implication of other host genes which are not associated with the type I interferon response. Because of the broad susceptibility of CC071 to multiple viruses, our results have implications beyond ZIKV infection and contribute to shedding light on the plurality of host mechanisms fighting infectious diseases. [ABSTRACT FROM AUTHOR]

Epidemics of Rift Valley fever (RVF), a mosquito-borne zoonotic disease caused by RVF virus, have been linked to exceptionally heavy rainfall and widespread flooding. The disease is endemic in most African countries and pose a major global health risk. Given that the disease was reported in various districts of Tanzania, we hypothesized a lack of knowledge about RVF epidemiology among agropastoral and pastoral communities. The research took place in a period of 7 months, from July, 2021 to January, 2022. The aim of this study was to assess the knowledge, attitudes, and practices (KAP) among the agropastoral and pastoral communities of Ngorongoro district towards RVF. The survey employed a mixed method system, which included 3 focus groups (each comprised 12 individuals), 20 key informant interviews and administration of questionnaire (N = 352) in agropastoral and pastoral community members of Ngorongoro district. The relationship between demographic characteristics and communities' knowledge, attitudes, and practices regarding RVF was observed using a multiple logistic regression model. A total of 352 participants were interviewed, with the majority (67.61%) being male and 32.39% being female, majority (39.5%) attending primary school, and majority (58.2%) being pastoralists. The findings showed that only 36.1%, 38.64% and 16.19% of participants had good knowledge, positive attitude and good practices regarding RVF respectively. Significant demographic factors related with knowledge included: gender (OR = 1.9, CI = 1.03–3.56, P = 0.041), education levels (primary: OR = 3.97, CI = 2–8.16, P = 0.000; secondary: OR = 15.27, CI = 5.5–46.23, P = 0.000 and college: OR = 34. 23, CI = 5.4–67.22, P = 0.000), and locality (Pinyinyi: OR = 0.14, CI = 0.05–0.38, P = 0.000 and Sale: OR = 0.14, CI = 0.04–0.44, P = 0.001). Male participants showed significant positive attitude towards RVF compared to female (OR = 2.37, CI = 1.35–4.17, P = 0.003). Individuals with formal education showed a significant positive attitude toward RVF compared to informal (OR>1, P<0.05). Agropastoral members showed a significant negative attitude toward RVF compared to pastoralists (OR = 0.51, CI = 0.26–0.99, P = 0.048). The calculated RVF prevention practices values were insignificantly (P = 0.853) correlated with knowledge values. The significant correlation between knowledge and attitude, as well as attitude and practice were found (P<0.05). In general, the study revealed poor knowledge, negative attitude and poor practices of communities towards RVF. The lack of regular education programs to make the communities aware of the disease was implicated for these findings. This recommends that provision of health education should be a long-term practice among agropastoral and pastoral communities in order to prevent further RVF outbreaks in Tanzania. Author summary: In the current study, we demonstrated that the KAP for RVF transmission and prevention is low among the pastoral and agropastoral community members in Ngorongoro district, Tanzania. The causes, route of transmission and signs of RVF in both humans and animals were mostly unknown to the respondents. The population was unaware that the high mortality rate of young animals and the miscarriage of pregnant animals were severe signs of RVF. The crucial preventive measures against RVF were not being used, which created a gap between recommended and real practices. Low level of knowledge about the RVF was linked to poor preventive efforts, so raising awareness could launch public awareness in a positive way. We suggest that important authorities should launch public awareness campaigns to help bridge these knowledge gaps and strengthen people's understanding about RVF. It might be necessary to use radio and mobile applications to provide the public with access to such health information. In order to alter behavioral patterns in the study communities to avert RVF occurrences, barriers to change must be identified and eliminated. The livelihoods of agropastoral and pastoralist communities will benefit if these procedures are put into action since they may prevent regular disruptions of livestock market chains that occur whenever RVF outbreaks happen. [ABSTRACT FROM AUTHOR]

Brucellosis, Rift Valley fever (RVF) and Q fever are zoonoses prevalent in many developing countries, causing a high burden on human and animal health. Only a few studies are available on these among agro-pastoralist communities and their livestock in Chad. The objective of our study was to estimate brucellosis, RVF and Q fever seroprevalence among Chadian agro-pastoralist communities and their livestock, and to investigate risk factors for seropositivity. We conducted a multi-stage cross-sectional serological survey in two rural health districts, Yao and Danamadji (966 human and 1041 livestock (cattle, sheep, goat and equine) samples)). The true seroprevalence were calculated applying a Bayesian framework to adjust for imperfect diagnostic test characteristics and accounting for clustering in the study design. Risk factors for each of the zoonotic diseases were estimated using mixed effects logistic regression models. The overall prevalence for brucellosis, Q fever and RVF combined for both regions was estimated at 0.2% [95% credibility Interval: 0–1.1], 49.1% [%CI: 38.9–58.8] and 28.1% [%CI: 23.4–33.3] in humans, and 0.3% [%CI: 0–1.5], 12.8% [%CI: 9.7–16.4] and 10.2% [%CI: 7.6–13.4] in animals. Risk factors correlating significantly with the respective disease seropositivity were sex for human brucellosis, sex and Q fever co-infection for animal brucellosis, age for human Q fever, species and brucellosis co-infection for animal Q fever, age and herd-level animal RVF seroprevalence within the same cluster for human RVF, and cluster-level human RVF seroprevalence within the same cluster for animal RVF. In Danamadji and Yao, Q fever and RVF are notably seroprevalent among agro-pastoralist human and animal communities, while brucellosis appears to have a low prevalence. Correlation between the seroprevalence between humans and animals living in the same communities was detected for RVF, highlighting the interlinkage of human and animal transmissible diseases and of their health, highlighting the importance of a One Health approach. Author summary: Infectious diseases transmitted between humans and animals, called zoonotic diseases, pose a global threat to human and animal health. Furthermore, diseased animals, especially livestock, can compromise the financial resources and livelihood of their owners as these depend on healthy animals for milk or meat production, or for agricultural work purposes. Brucellosis, Q fever and Rift Valley fever are two bacterial and one viral zoonotic disease that were found to be prevalent among many human-animal communities living in close contact, such as it is the case among Chadian agro-pastoralists. Limited data are available on the current status of these diseases in Chad. In this study, the authors investigated the prevalence of these three diseases among humans and their livestock (cattle, sheep, goats, horses and donkeys) by collecting blood samples and conducting serological analyses in two rural regions of Chad, Danamadji and Yao. Results point towards high Q fever and Rift Valley fever seroprevalences (13–49% and 10–28%, respectively), and low prevalence of brucellosis (< 1%), and towards a positive association between human and animal Rift Valley fever seroprevalence. With these findings, the study hopes to support current and future zoonotic disease surveillance and control efforts within the regions. [ABSTRACT FROM AUTHOR]

Uganda has had repeated outbreaks of Rift Valley fever (RVF) since March 2016 when human and livestock cases were reported in Kabale after a long interval. The disease has a complex and poorly described transmission patterns which involves several mosquito vectors and mammalian hosts (including humans). We conducted a national serosurvey in livestock to determine RVF virus (RVFV) seroprevalence, risk factors, and to develop a risk map that could be used to guide risk-based surveillance and control measures. A total of 3,253 animals from 175 herds were sampled. Serum samples collected were screened at the National Animal Disease Diagnostics and Epidemiology Centre (NADDEC) using a competition multispecies anti-RVF IgG ELISA kit. Data obtained were analyzed using a Bayesian model that utilizes integrated nested Laplace approximation (INLA) and stochastic partial differential equation (SPDE) approaches to estimate posterior distributions of model parameters, and account for spatial autocorrelation. Variables considered included animal level factors (age, sex, species) and multiple environmental data including meteorological factors, soil types, and altitude. A risk map was produced by projecting fitted (mean) values, from a final model that had environmental factors onto a spatial grid that covered the entire domain. The overall RVFV seroprevalence was 11.39% (95% confidence interval: 10.35–12.51%). Higher RVFV seroprevalences were observed in older animals compared to the young, and cattle compared to sheep and goats. RVFV seroprevalence was also higher in areas that had (i) lower precipitation seasonality, (ii) haplic planosols, and (iii) lower cattle density. The risk map generated demonstrated that RVF virus was endemic in several regions including those that have not reported clinical outbreaks in the northeastern part of the country. This work has improved our understanding on spatial distribution of RVFV risk in the country as well as RVF burden in livestock. Author summary: Outbreaks of Rift Valley fever occur periodically in Uganda in livestock and humans. An Initial outbreak was reported in Kabale in March 2016 after a long quiescent period. Factors that trigger these outbreaks have not been fully described, but above-normal precipitation and flooding are known to be important causes. This paper presents a study that was conducted in the country to analyse RVF virus seroprevalence and risk factors for exposure in livestock. The study also mapped spatial distribution of RVFV seroprevalence. Data used in the study were generated from a national cross-sectional serosurvey that involved cattle, sheep and goats. Results obtained showed that the national RVFV seroprevalence was 11.39%, with 95% confidence interval of 10.35–12.51%. Cattle had higher RVFV seroprevalence compared to sheep and goats. Environmental factors that were associated with increased seroprevalence were low precipitation seasonality, haplic planosols, and low cattle density. Prevalence maps generated showed that the southwestern, central and parts of the northeastern regions had higher seroprevalence compared to other regions of the country. These can be used to guide the surveillance of the disease in the country. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is a zoonotic arbovirus that has profound impact on domestic ruminants and can also be transmitted to humans via infected animal secretions. Urban areas in endemic regions across Africa have susceptible animal and human hosts, dense vector distributions, and source livestock (often from high risk locations to meet the demand for animal protein). Yet, there has never been a documented urban outbreak of RVF. To understand the likely risk of RVFV introduction to urban communities from their perspective and guide future initiatives, we conducted focus group discussions with slaughterhouse workers, slaughterhouse animal product traders, and livestock owners in Kisumu City and Ukunda Town in Kenya. For added perspective and data triangulation, in-depth interviews were conducted one-on-one with meat inspector veterinarians from selected slaughterhouses. A theoretical framework relevant to introduction, transmission, and potential persistence of RVF in urban areas is presented here. Urban livestock were primarily mentioned as business opportunities, but also had personal sentiment. In addition to slaughtering risks, perceived risk factors including consumption of fresh milk. High risk groups' knowledge and experience with RVFV and other zoonotic diseases impacted their consideration personal risk, with consensus towards lower risk in the urban setting compared to rural areas as determination of health risk was said to primarily rely on hygiene practices rather than the slaughtering process. Groups relied heavily on veterinarians to confirm animal health and meat safety, yet veterinarians reported difficulty in accessing RVFV diagnostics. We also identified vulnerable public health regulations including corruption in meat certification outside of the slaughterhouse system, and blood collected during slaughter being used for food and medicine, which could provide a means for direct RVFV community transmission. These factors, when compounded by diverse urban vector breeding habitats and dense human and animal populations, could create suitable conditions for RVFV to arrive an urban center via a viremic imported animal, transmit to locally owned animals and humans, and potentially adapt to secondary vectors and persist in the urban setting. This explorative qualitative study proposes risk pathways and provides initial insight towards determining how urban areas could adapt control measures and plan future initiatives to better understand urban RVF potential. Author summary: Rift Valley fever is an important mosquito-borne zoonotic virus that can also be transmitted directly to humans via infected livestock. There has never been an urban outbreak of RVF, however, other arboviruses have caused devastating urban outbreaks driven by urban transmission cycles in vectors. This qualitative study aimed to explore and understand the potential risk of Rift Valley fever in the urban context from the perspective of high risk groups. RVF has a complex epidemiology and urban centers of endemic regions may be particularly vulnerable as they import large quantities of livestock for slaughter and milk to meet the high urban demand for animal sourced foods and have dense host and vector populations. This qualitative study provides insights from the opinions and lived experiences of urban high risk groups including slaughterhouse affiliates, livestock owners, and veterinarians. We also demonstrate risks specific to RVFV transmission and regulatory vulnerabilities that would impact the ability to detect RVF disease introduction. The results of this study are intended to guide future initiatives aiming to investigate urban potential of RVF and infer how urban disease ecology could differ from what has been previously demonstrated in rural areas. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a zoonotic arbovirosis which has been reported across Africa including the northernmost edge, South West Indian Ocean islands, and the Arabian Peninsula. The virus is responsible for high abortion rates and mortality in young ruminants, with economic impacts in affected countries. To date, RVF epidemiological mechanisms are not fully understood, due to the multiplicity of implicated vertebrate hosts, vectors, and ecosystems. In this context, mathematical models are useful tools to develop our understanding of complex systems, and mechanistic models are particularly suited to data-scarce settings. Here, we performed a systematic review of mechanistic models studying RVF, to explore their diversity and their contribution to the understanding of this disease epidemiology. Researching Pubmed and Scopus databases (October 2021), we eventually selected 48 papers, presenting overall 49 different models with numerical application to RVF. We categorized models as theoretical, applied, or grey, depending on whether they represented a specific geographical context or not, and whether they relied on an extensive use of data. We discussed their contributions to the understanding of RVF epidemiology, and highlighted that theoretical and applied models are used differently yet meet common objectives. Through the examination of model features, we identified research questions left unexplored across scales, such as the role of animal mobility, as well as the relative contributions of host and vector species to transmission. Importantly, we noted a substantial lack of justification when choosing a functional form for the force of infection. Overall, we showed a great diversity in RVF models, leading to important progress in our comprehension of epidemiological mechanisms. To go further, data gaps must be filled, and modelers need to improve their code accessibility. Author summary: Rift Valley fever (RVF) affects humans and livestock across Africa, South West Indian Ocean islands, and the Arabian Peninsula. This disease is one of the World Health Organization priorities and is caused by a virus which is transmitted by mosquitoes (mainly of Aedes and Culex spp. genera), but also by direct contact from livestock to humans. Mathematical models have been used in the last 20 years to disentangle RVF virus transmission dynamics. These models can further improve our understanding of processes driving outbreaks, test the efficiency of control strategies, or even anticipate possible emergence. Provided with detailed datasets, models can tailor their conclusions to specific geographical contexts and aid in decision-making in the field. This review provides a general overview of mathematical models developed to study RVF virus transmission dynamics. We describe their main results and methodological choices, and identify hurdles to be lifted. To offer innovative animal and public health value, we recommend that future models focus on the relative contribution of host species to transmission, and the role of animal mobility. [ABSTRACT FROM AUTHOR]

Bunyaviruses lack a specific mechanism to ensure the incorporation of a complete set of genome segments into each virion, explaining the generation of incomplete virus particles lacking one or more genome segments. Such incomplete virus particles, which may represent the majority of particles produced, are generally considered to interfere with virus infection and spread. Using the three-segmented arthropod-borne Rift Valley fever virus as a model bunyavirus, we here show that two distinct incomplete virus particle populations unable to spread autonomously are able to efficiently complement each other in both mammalian and insect cells following co-infection. We further show that complementing incomplete virus particles can co-infect mosquitoes, resulting in the reconstitution of infectious virus that is able to disseminate to the mosquito salivary glands. Computational models of infection dynamics predict that incomplete virus particles can positively impact virus spread over a wide range of conditions, with the strongest effect at intermediate multiplicities of infection. Our findings suggest that incomplete particles may play a significant role in within-host spread and between-host transmission, reminiscent of the infection cycle of multipartite viruses. Genome segmentation is assumed to be costly for viruses with non-selective genome packaging, as they produce high numbers of particles with an incomplete genome. However, this study shows that upon co-infection, complementing incomplete bunyavirus particles can reconstitute a complete genome, supporting virus infection and potentially contributing to within-host spread and between-host transmission. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a disease of animals and humans associated with abortions in ruminants and late-gestation miscarriages in women. Here, we use a rat model of congenital RVF to identify tropisms, pathologies, and immune responses in the placenta during vertical transmission. Infection of late-gestation pregnant rats resulted in vertical transmission to the placenta and widespread infection throughout the decidua, basal zone, and labyrinth zone. Some pups from infected dams appeared normal while others had gross signs of teratogenicity including death. Histopathological lesions were detected in placenta from pups regardless of teratogenicity, while teratogenic pups had widespread hemorrhage throughout multiple placenta layers. Teratogenic events were associated with significant increases in placental pro-inflammatory cytokines, type I interferons, and chemokines. RVFV displays a high degree of tropism for all placental tissue layers and the degree of hemorrhage and inflammatory mediator production is highest in placenta from pups with adverse outcomes. Given the potential for RVFV to emerge in new locations and the recent evidence of emerging viruses, like Zika and SARS-CoV-2, to undergo vertical transmission, this study provides essential understanding regarding the mechanisms by which RVFV crosses the placenta barrier. Author summary: Rift Valley fever virus (RVFV) infections cause human health and economical burdens given its ability to induce high rates of abortions in ruminants and possible contributions towards late-term miscarriages in women. In this study, we have identified important structures in the placenta targeted by this emerging bunyavirus. Inflammation was associated with more severe fetal outcomes such as death and fetal deformities. The striking similarities between the pathologies of the placenta in this rodent model of congenital RVF and those observed in naturally infected ruminants highlight the utility of this rodent model. These findings may be further translated towards understanding the mechanisms involved in vertical transmission of RVFV in humans. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is a mosquito-borne pathogen that has caused epidemics involving people and animals across Africa and the Arabian Peninsula. A number of studies have found evidence for the circulation of RVFV among livestock between these epidemics but the population-level incidence of infection during this inter-epidemic period (IEP) is rarely reported. General force of infection (FOI) models were applied to age-adjusted cross-sectional serological data to reconstruct the annual FOI and population-level incidence of RVFV infection among cattle, goats, and sheep in northern Tanzania from 2009 through 2015, a period without reported Rift Valley fever (RVF) cases in people or animals. To evaluate the potential for zoonotic RVFV spillover during this period, the relationship between village-level livestock RVFV FOI and human RVFV seropositivity was quantified using multi-level logistic regression. The predicted average annual incidence was 72 (95% Credible Interval [CrI] 63, 81) RVFV infections per 10,000 animals and 96 (95% CrI 81, 113), 79 (95% CrI 62, 98), and 39 (95% CrI 28, 52) per 10,000 cattle, sheep, and goats, respectively. There was variation in transmission intensity between study villages, with the highest estimated village-level FOI 2.49% (95% CrI 1.89, 3.23) and the lowest 0.12% (95% CrI 0.02, 0.43). The human RVFV seroprevalence was 8.2% (95% Confidence Interval 6.2, 10.9). Human seropositivity was strongly associated with the village-level FOI in livestock, with the odds of seropositivity in an individual person increasing by around 1.2 times (95% CrI 1.1, 1.3) for each additional annual RVFV seroconversion per 1,000 animals. A history of raw milk consumption was also positively associated with human seropositivity. RVFV has circulated at apparently low levels among livestock in northern Tanzania in the period since the last reported epidemic. Although our data do not allow us to confirm human RVFV infections during the IEP, a strong association between human seropositivity and the FOI in cattle, goats, and sheep supports the hypothesis that RVFV circulation among livestock during the IEP poses a risk for undetected zoonotic spillover in northern Tanzania. We provide further evidence for the likely role of raw milk consumption in RVFV transmission from animals to people. Author summary: Rift Valley fever outbreaks are reported in Tanzania every 10 to 15 years. Human and animal cases are not typically reported in the period between outbreaks, but the RVF virus (RVFV) is known to circulate among livestock during these periods. The last officially reported outbreak of RVF in Tanzania was in 2008. The incidence of RVFV infection among livestock in the period since this large epidemic in the country is not known. We used a general force of infection model to quantify the annual incidence of RVFV infection among livestock in northern Tanzania between 2009 and 2015, a period without reported RVF cases in people or animals. Using a sample of over 9,000 animals from 43 villages across northern Tanzania, we estimated a moderately low average annual incidence of 96, 39, and 79 RVFV infections per 10,000 cattle, goats, and sheep, respectively. There was considerable heterogeneity in the intensity of transmission between villages, highlighting important spatial differences in inter-epidemic RVFV transmission among livestock in this region. We also found that human seropositivity was strongly positively associated with the village-level force of RVFV infection among livestock. All people with seropositive results in this study were born before 2008. We therefore cannot be certain that these people were infected in the period since the last outbreak. However, the very strong positive association we find between inter-epidemic circulation of RVFV among livestock and human seropositivity provides support to the hypothesis that zoonotic RVFV spillover may be occuring during the period between large-scale outbreaks in northern Tanzania. Further studies are required to confirm this. We also found a strong positive association between a history of raw milk consumption and human RVFV seropositivity, providing further evidence for the importance of milk as a likely route for zoonotic RVFV transmission. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is a veterinary and human pathogen and is an agent of bioterrorism concern. Currently, RVFV treatment is limited to supportive care, so new drugs to control RVFV infection are urgently needed. RVFV is a member of the order Bunyavirales, whose replication depends on the enzymatic activity of the viral L protein. Screening for RVFV inhibitors among compounds with divalent cation-coordinating motifs similar to known viral nuclease inhibitors identified 47 novel RVFV inhibitors with selective indexes from 1.1–103 and 50% effective concentrations of 1.2–56 μM in Vero cells, primarily α-Hydroxytropolones and N-Hydroxypyridinediones. Inhibitor activity and selective index was validated in the human cell line A549. To evaluate specificity, select compounds were tested against a second Bunyavirus, La Crosse Virus (LACV), and the flavivirus Zika (ZIKV). These data indicate that the α-Hydroxytropolone and N-Hydroxypyridinedione chemotypes should be investigated in the future to determine their mechanism(s) of action allowing further development as therapeutics for RVFV and LACV, and these chemotypes should be evaluated for activity against related pathogens, including Hantaan virus, severe fever with thrombocytopenia syndrome virus, Crimean-Congo hemorrhagic fever virus. [ABSTRACT FROM AUTHOR]

Background: Rift Valley fever (RVF) is a mosquito-borne zoonosis endemic in Africa. With little known of the burden or epidemiology of RVF virus (RVFV) in Cameroon, this study aimed to determine the seroprevalence of RVFV in domestic ruminants of various origins in two markets of Yaoundé, Cameroon. Methodology/Principal findings: The origin of animals randomly sampled at two livestock markets in Yaoundé were recorded and plasma samples collected for competitive and capture Enzyme-linked Immunosorbent Assay (ELISA) to determine the prevalence of Immunoglobulins G (IgG) and Immunoglobulins M (IgM) antibodies. Following ELISA IgM results, a real-time reverse transcription-polymerase chain reaction (qRT-PCR) was performed to detect RVFV RNA. In June-August 2019, February-March 2020, and March-April 2021, 756 plasma samples were collected from 441 cattle, 168 goats, and 147 sheep. RVFV IgG seroprevalence was 25.7% for all animals, 42.2% in cattle, 2.7% in sheep, and 2.4% in goats. However, IgM seroprevalence was low, at 0.9% in all animals, 1.1% in cattle, 1.4% in sheep, and 0% in goats. The seroprevalence rates varied according to the animal's origin with the highest rate (52.6%) in cattle from Sudan. In Cameroon, IgG and IgM rates respectively were 45.1% and 2.8% in the North, 44.8% and 0% in the Adamawa, 38.6% and 1.7% in the Far-North. All IgM positive samples were from Cameroon. In cattle, 2/5 IgM positive samples were also IgG positive, but both IgM positive samples in sheep were IgG negative. Three (42.9%) IgM positive samples were positive for viral RVFV RNA using qRT-PCR but given the high ct values, no amplicon was obtained. Conclusion/Significance: These findings confirm the circulation of RVFV in livestock in Cameroon with prevalence rates varying by location. Despite low IgM seroprevalence rates, RVF outbreaks can occur without being noticed. Further epidemiological studies are needed to have a broad understanding of RVFV transmission in Cameroon. [ABSTRACT FROM AUTHOR]

Background: Co-infection, especially with pathogens of dissimilar genetic makeup, may result in a more devastating impact on the host. Investigations on co-infection with neglected zoonotic pathogens in wildlife are necessary to inform appropriate prevention and control strategies to reduce disease burden in wildlife and the potential transmission of these pathogens between wildlife, livestock and humans. This study assessed co-exposure of various Kenyan wildflife species with Brucella spp, Coxiella burnetii and Rift Valley fever virus (RVFV). Methodology: A total of 363 sera from 16 different wildlife species, most of them (92.6%) herbivores, were analysed by Enzyme-linked immunosorbent assay (ELISA) for IgG antibodies against Brucella spp, C. burnetii and RVFV. Further, 280 of these were tested by PCR to identify Brucella species. Results: Of the 16 wildlife species tested, 15 (93.8%) were seropositive for at least one of the pathogens. Mean seropositivities were 18.9% (95% CI: 15.0–23.3) for RVFV, 13.7% (95% CI: 10.3–17.7) for Brucella spp and 9.1% (95% CI: 6.3–12.5) for C. burnetii. Buffaloes (n = 269) had higher seropositivity for Brucella spp. (17.1%, 95% CI: 13.0–21.7%) and RVFV (23.4%, 95% CI: 18.6–28.6%), while giraffes (n = 36) had the highest seropositivity for C. burnetii (44.4%, 95% CI: 27.9–61.9%). Importantly, 23 of the 93 (24.7%) animals positive for at least one pathogen were co-exposed, with 25.4% (18/71) of the positive buffaloes positive for brucellosis and RVFV. On molecular analysis, Brucella DNA was detected in 46 (19.5%, CI: 14.9–24.7) samples, with 4 (8.6%, 95% CI: 2.2–15.8) being identified as B. melitensis. The Fisher's Exact test indicated that seropositivity varied significantly within the different animal families, with Brucella (p = 0.013), C. burnetii (p = <0.001) and RVFV (p = 0.007). Location was also significantly associated (p = <0.001) with Brucella spp. and C. burnetii seropositivities. Conclusion: Of ~20% of Kenyan wildlife that are seropositive for Brucella spp, C. burnetii and RVFV, almost 25% indicate co-infections with the three pathogens, particularly with Brucella spp and RVFV. Author summary: Infection of an animal with more than one pathogen may result into a more devastating impact. Only few studies have investigated co-infection with multiple pathogens in wildlife, despite their key role as reservoirs of zoonotic diseases. Therefore, there is need for investigations on co-infection with neglected zoonotic pathogens in wildlife to inform prevention and control approaches and reduce disease impact in wildlife and potential transmission of these pathogens between wildlife, livestock, and humans. This study assessed co-exposure of various Kenyan wildlife species with three zoonotic pathogens, including Brucella spp, Coxiella burnetii and Rift Valley fever virus (RVFV). Results from this study revealed widespread, but varied exposure levels to the three individual pathogens within the several wildlife species. Likewise, the study also found the presence of co-exposure with the three pathogens. The findings from this study points to the need for establishment of surveillance and control programmes that target multiple pathogens in the wildlife populations to reduce the risk of transmission of infectious pathogens in wildlife and their zoonotic transmission. [ABSTRACT FROM AUTHOR]

Quantifying the variation of pathogens' life history traits in multiple host systems is crucial to understand their transmission dynamics. It is particularly important for arthropod-borne viruses (arboviruses), which are prone to infecting several species of vertebrate hosts. Here, we focus on how host-pathogen interactions determine the ability of host species to transmit a virus to susceptible vectors upon a potentially infectious contact. Rift Valley fever (RVF) is a viral, vector-borne, zoonotic disease, chosen as a case study. The relative contributions of livestock species to RVFV transmission has not been previously quantified. To estimate their potential to transmit the virus over the course of their infection, we 1) fitted a within-host model to viral RNA and infectious virus measures, obtained daily from infected lambs, calves, and young goats, 2) estimated the relationship between vertebrate host infectious titers and probability to infect mosquitoes, and 3) estimated the net infectiousness of each host species over the duration of their infectious periods, taking into account different survival outcomes for lambs. Our results indicate that the efficiency of viral replication, along with the lifespan of infectious particles, could be sources of heterogeneity between hosts. Given available data on RVFV competent vectors, we found that, for similar infectious titers, infection rates in the Aedes genus were on average higher than in the Culex genus. Consequently, for Aedes-mediated infections, we estimated the net infectiousness of lambs to be 2.93 (median) and 3.65 times higher than that of calves and goats, respectively. In lambs, we estimated the overall infectiousness to be 1.93 times higher in individuals which eventually died from the infection than in those recovering. Beyond infectiousness, the relative contributions of host species to transmission depend on local ecological factors, including relative abundances and vector host-feeding preferences. Quantifying these contributions will ultimately help design efficient, targeted, surveillance and vaccination strategies. Author summary: Viruses spread by mosquitoes present a major threat to animal and public health worldwide. When these pathogenic viruses can infect multiple species, controlling their spread becomes difficult. Rift Valley fever virus (RVFV) is such a virus. It spreads predominantly among ruminant livestock but can also spill over and cause severe disease in humans. Understanding which of these ruminant species are most important for the transmission of RVFV can help for effective control. One piece of this puzzle is to assess how effective infected animals are at transmitting RVFV to mosquitoes. To answer this question, we combine mathematical models with observations from experimental infections in cattle, sheep, and goats and model changes in viremia over time within individuals. We then quantify the relationship between hosts' viremia and the probability to infect mosquitoes. In combining these two analyses, we estimate the overall transmission potential of sheep, when in contact with mosquitoes, to be 3 to 5 times higher than that of goats and cattle. Further, sheep that experience a lethal infection have an even larger overall transmission potential. Once applied at the level of populations, with setting-specific herd composition and exposure to mosquitoes, these results will help unravel species' role in RVF outbreaks. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is an arboviral disease of humans and livestock responsible for severe economic and human health impacts. In humans, RVF spans a variety of clinical manifestations, ranging from an acute flu-like illness to severe forms of disease, including late-onset encephalitis. The large variations in human RVF disease are inadequately represented by current murine models, which overwhelmingly die of early-onset hepatitis. Existing mouse models of RVF encephalitis are either immunosuppressed, display an inconsistent phenotype, or develop encephalitis only when challenged via intranasal or aerosol exposure. In this study, the genetically defined recombinant inbred mouse resource known as the Collaborative Cross (CC) was used to identify mice with additional RVF disease phenotypes when challenged via a peripheral foot-pad route to mimic mosquito-bite exposure. Wild-type Rift Valley fever virus (RVFV) challenge of 20 CC strains revealed three distinct disease phenotypes: early-onset hepatitis, mixed phenotype, and late-onset encephalitis. Strain CC057/Unc, with the most divergent phenotype, which died of late-onset encephalitis at a median of 11 days post-infection, is the first mouse strain to develop consistent encephalitis following peripheral challenge. CC057/Unc mice were directly compared to C57BL/6 mice, which uniformly succumb to hepatitis within 2–4 days of infection. Encephalitic disease in CC057/Unc mice was characterized by high viral RNA loads in brain tissue, accompanied by clearance of viral RNA from the periphery, low ALT levels, lymphopenia, and neutrophilia. In contrast, C57BL/6 mice succumbed from hepatitis at 3 days post-infection with high viral RNA loads in the liver, viremia, high ALT levels, lymphopenia, and thrombocytopenia. The identification of a strain of CC mice as an RVFV encephalitis model will allow for future investigation into the pathogenesis and treatment of RVF encephalitic disease and indicates that genetic background makes a major contribution to RVF disease variation. Author summary: Rift Valley fever virus (RVFV) is a negative sense RNA arbovirus of clinical relevance found throughout Africa and into the Middle East. In humans, RVF disease exhibits various clinical manifestations including acute febrile illness, hepatitis, hemorrhagic fever, and encephalitis. Unfortunately, existing murine models have been limited to studies of acute lethal hepatitis. Here, we identify a mouse model in which all animals survive acute hepatitis and go on to succumb to late onset encephalitis. A key feature of this new experimental system is administration of the virus in the footpad (to mimic mosquito bite), to model a natural route of infection. Importantly in this work, we used the Collaborative Cross recombinant inbred mouse resource to identify this RVFV CNS model. Therefore, our study demonstrates that host genetics can modulate RVF disease manifestations. The identification of this model also enables the field for high throughput, in vivo studies of central nervous system pathogenesis as well as in vivo evaluation of vaccines and therapeutics targeted against RVF encephalitis. [ABSTRACT FROM AUTHOR]

Rift valley fever (RVF) is a mosquito-borne disease of animals and humans. Although RVF outbreaks are usually reported at 5-15-year intervals in sub-Saharan Africa, Zambia has experienced an unusually long inter-epizootic/-epidemic period of more than three decades. However, serological evidence of RVF virus (RVFV) infection in domestic ruminants during this period underscores the need for comprehensive investigation of the mechanisms of virus perpetuation and disease emergence. Mosquitoes (n = 16,778) captured from eight of the ten provinces of Zambia between April 2014 and May 2019 were pooled (n = 961) and screened for RVFV genome by a pan-phlebo RT-PCR assay. Aedes mosquito pools (n = 85) were further screened by nested RT-PCR assay. Sera from sheep (n = 13), goats (n = 259) and wild ungulates (n = 285) were screened for RVFV antibodies by ELISA while genome detection in pooled sera (n = 276) from domestic (n = 248) and wild ungulates (n = 37) was performed by real-time RT-PCR assay. To examine the association between the long inter-epizootic period and climatic variables, we examined El Niño-Southern Oscillation indices, precipitation anomalies, and normalized difference vegetation index. We then derived RVF risk maps by exploring climatic variables that would favor emergence of primary RVFV vectors. While no RVFV genome could be detected in pooled mosquito and serum samples, seroprevalence was significantly high (OR = 8.13, 95% CI [4.63–14.25]) in wild ungulates (33.7%; 96/285) compared to domestic ruminants (5.6%; 16/272). Retrospective analysis of RVF epizootics in Zambia showed a positive correlation between anomalous precipitation (La Niña) and disease emergence. On risk mapping, whilst northern and eastern parts of the country were at high risk, domestic ruminant population density was low (< 21 animals/km2) in these areas compared to low risk areas (>21 animals/km2). Besides evidence of silent circulation of RVFV and the risk of disease emergence in some areas, wildlife may play a role in the maintenance of RVFV in Zambia. Author summary: Rift valley fever (RVF) is an important mosquito borne disease of humans and ruminants. RVF is endemic in sub-Saharan Africa where disease outbreaks are frequently reported in humans and animals. For RVF outbreaks to occur, there has to be presence of the host (animals or humans) and favorable climatic conditions that favor emergence of RVF virus vectors (mosquitoes). Understanding this complex interaction enables us to predict future disease outbreaks and map RVF high risk areas. Above normal rainfall and floods will usually trigger RVF outbreaks. In this study, we conducted surveillance for RVF in mosquitoes, domestic and wild ruminants. We then examined climatic conditions that may have been responsible for past RVF outbreaks and the unusually (>30 years) long inter-epizootic period. Finally, we derived RVF risk maps for Zambia using a combination of climatic variables. We found evidence of RVF infection in both wild and domestic ruminants while there was a positive correlation between past RVF outbreaks and above-normal rainfall. On risk mapping, we found favorable conditions (i.e. high rainfall and risk of flooding) for RVF emergence in northern and eastern parts of the country. Studies like this are important in planning surveillance and disease control programs. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is a highly pathogenic mosquito-borne virus capable of causing hepatitis, encephalitis, blindness, hemorrhagic syndrome, and death in humans and livestock. Upon aerosol infection with RVFV, the brain is a major site of viral replication and tissue damage, yet pathogenesis in this organ has been understudied. Here, we investigated the immune response in the brain of RVFV infected mice. In response to infection, microglia initiated robust transcriptional upregulation of antiviral immune genes, as well as increased levels of activation markers and cytokine secretion that is dependent on mitochondrial antiviral-signaling protein (MAVS) and independent of toll-like receptors 3 and 7. In vivo, Mavs-/- mice displayed enhanced susceptibility to RVFV as determined by increased brain viral burden and higher mortality. Single-cell RNA sequence analysis identified defects in type I interferon and interferon responsive gene expression within microglia in Mavs-/- mice, as well as dysregulated lymphocyte infiltration. The results of this study provide a crucial step towards understanding the precise molecular mechanisms by which RVFV infection is controlled in the brain and will help inform the development of vaccines and antiviral therapies that are effective in preventing encephalitis. Author summary: Rift Valley fever virus causes severe disease in humans and livestock and in some cases can be fatal. There is concern about the use of Rift Valley fever virus as a bioweapon since it can be transmitted through the air, and there are no vaccines or antiviral treatments. Airborne transmission of the virus causes severe inflammation of the brain, yet little is known about the immune response against the virus in this organ. Here, we investigated the immune response in the brain to Rift Valley fever virus following intranasal infection. We determined that microglia, the resident immune cells of the brain, initiate a robust response to Rift Valley fever virus infection and identified a key immune pathway that is critical for the ability of microglia to respond to infection. When this immune pathway is rendered non-functional, mice have a dysregulated response to infection in the brain. This study provides insight into how the immune response can control Rift Valley fever virus infection of the brain. [ABSTRACT FROM AUTHOR]

An important problem for livestock production in Mali is occurrence of several infectious diseases. A particular challenge for control of pathogens that affect different species, especially in a system with mixed herds with cattle, sheep and goats. Therefore, this study aimed to investigate co-exposure with Rift Valley fever virus (RVFV), Coxiella burnetii, bluetongue virus (BTV) and Brucella spp. in different livestock species in mixed herds. With the exception of BTV these pathogens are also zoonotic. A retrospective assessment was carried out on a biobank of sera of cattle and small ruminants collected from Sikasso and Mopti regions. Nine hundred and twelve samples from cattle (n = 304), sheep (n = 318) and goat (n = 290) were screened. Serology tests were conducted using commercial kits as per the protocol of the manufacturers. Sero-prevalence for RVFV was 12.8% (Confidence Interval 95%: 9.3–17.1%); 4.7% (2.7–7.7%) and 3.1% (1.4–5.8%) in cattle, sheep and goat respectively. For Coxiella burnetii, the sero-prevalence was 55.3% (49.5–60.9%), 22.6% (18.2–27.6%), and 16.9% (12.8–21.7%); in cattle, sheep and goat respectively; and for BTV sero-prevalence was 88.8% (84.72–92.13%), 51.6% (45.9–57.2%), 56.2% (50.3–62.0%) in cattle, sheep in goat respectively. Brucella sp. had the lowest sero-prevalence and was only detected in cattle and sheep. Regional differences were observed with sero-prevalence of Coxiella burnetii in sheep and goat and BTV in goat being significantly higher in Sikasso than in Mopti (p<0.001). Evidence of exposure to two pathogens in the same animal was most common for the combination Coxiella burnetii and BTV in cattle (51.6%), followed by sheep (17.0%) and goat (15.5%). Considering the scarcity of disease occurrence and epidemiological data in most sub-saharan countries including Mali, this multi-pathogen survey provides important evidence that cattle, sheep and goat are exposed to pathogens that may negatively impact productivity and pose a risk for public health. The results from this study highlight the urgent need for a better understanding of pathogen diversity and their impact on human and animal health in order to minimize resulting risks. Given that some of the pathogens investigated here are zoonotic, establishment of One-Health surveillance system to monitor disease in animals and people is warranted and intersectoral collaboration is recommended. Author summary: High prevalence of Rift Valley fever virus, Coxiella burnetii and Bluetongue virus were observed in cattle, sheep, and goats. We provided evidence that animals of the same herds have to cope with several pathogens. This study points out to need for important follow up studies to fill further knowledge gaps related to impacts of livestock diseases in Mali, especially zoonoses, and how to address the challenges. [ABSTRACT FROM AUTHOR]

In Mauritania, several mosquito-borne viruses have been reported that can cause devastating diseases in animals and humans. However, monitoring data on their occurrence and local distribution are limited. Rift Valley fever virus (RVFV) is an arthropod-borne virus that causes major outbreaks throughout the African continent and the Arabian Peninsula. The first Rift Valley fever (RVF) epidemic in Mauritania occurred in 1987 and since then the country has been affected by recurrent outbreaks of the disease. To gain information on the occurrence of RVFV as well as other mosquito-borne viruses and their vectors in Mauritania, we collected and examined 4,950 mosquitoes, belonging to four genera and 14 species. The mosquitoes were captured during 2018 in the capital Nouakchott and in southern parts of Mauritania. Evidence of RVFV was found in a mosquito pool of female Anopheles pharoensis mosquitoes collected in December on a farm near the Senegal River. At that time, 37.5% of 16 tested Montbéliarde cattle on the farm showed RVFV-specific IgM antibodies. Additionally, we detected IgM antibodies in 10.7% of 28 indigenous cattle that had been sampled on the same farm one month earlier. To obtain information on potential RVFV reservoir hosts, blood meals captured engorged mosquitoes were analyzed. The mosquitoes mainly fed on humans (urban areas) and cattle (rural areas), but also on small ruminants, donkeys, cats, dogs and straw-colored fruit bats. Results of this study demonstrate the circulation of RVFV in Mauritania and thus the need for further research to investigate the distribution of the virus and its vectors. Furthermore, factors that may contribute to its maintenance should be analyzed more closely. In addition, two mosquito pools containing Aedes aegypti and Culex quinquefasciatus mosquitoes showed evidence of dengue virus (DENV) 2 circulation in the city of Rosso. Further studies are therefore needed to also examine DENV circulation in Mauritania. Author summary: In Mauritania, various mosquito-borne viruses responsible for devastating diseases in livestock and/or humans occur, threatening both human and animal health. One of these viruses is the Rift Valley fever phlebovirus (RVFV), which is widespread throughout the African continent and the Arabian Peninsula. The first outbreak in Mauritania occurred in 1987, and epidemics have recurred since then. However, little is known about the maintenance of RVFV during inter-epidemic periods, in the absence of reported outbreaks, and although there is evidence of an active circulation of the virus during these times, only limited data are available on the abundance of RVFV and its vectors in Mauritania. Therefore, this study aimed to collect and examine mosquitoes in different areas where outbreaks have already occurred. Additionally, blood of collected blood-fed mosquitoes was analyzed to identify feeding patterns and thus wildlife potentially acting as RVFV reservoir hosts. Furthermore, the collected mosquitoes were analyzed for the presence of other neglected viruses in order to participate in an elucidation of the circulation of different viruses within the mosquito population in Mauritania and to be able to better prevent disease outbreaks. [ABSTRACT FROM AUTHOR]

The recent global Zika epidemics have revealed the significant threat that mosquito-borne viruses pose. There are currently no effective vaccines or prophylactics to prevent Zika virus (ZIKV) infection. Limiting exposure to infected mosquitoes is the best way to reduce disease incidence. Recent studies have focused on targeting mosquito reproduction and immune responses to reduce transmission. Previous work has evaluated the effect of insulin signaling on antiviral JAK/STAT and RNAi in vector mosquitoes. Specifically, insulin-fed mosquitoes resulted in reduced virus replication in an RNAi-independent, ERK-mediated JAK/STAT-dependent mechanism. In this work, we demonstrate that targeting insulin signaling through the repurposing of small molecule drugs results in the activation of both RNAi and JAK/STAT antiviral pathways. ZIKV-infected Aedes aegypti were fed blood containing demethylasterriquinone B1 (DMAQ-B1), a potent insulin mimetic, in combination with AKT inhibitor VIII. Activation of this coordinated response additively reduced ZIKV levels in Aedes aegypti. This effect included a quantitatively greater reduction in salivary gland ZIKV levels up to 11 d post-bloodmeal ingestion, relative to single pathway activation. Together, our study indicates the potential for field delivery of these small molecules to substantially reduce virus transmission from mosquito to human. As infections like Zika virus are becoming more burdensome and prevalent, understanding how to control this family of viruses in the insect vector is an important issue in public health. Author summary: Arboviruses pose a significant threat to humans and are an increasing concern as a result of climate change and expanding vector-competent populations. The recent Zika outbreaks demonstrate that mosquito-borne illnesses caused by viral infection remain a prominent and evolving threat that must be actively addressed. As there are currently no post-exposure therapeutics available for Zika virus infection, reducing transmission and, in turn, the likelihood of infection would provide sizeable benefit for human populations most at risk. Here, we show that readily available small molecules can be repurposed to effectively reduce viral replication and likelihood of transmission for a clinically relevant strain of Zika in vector competent mosquitoes. Furthermore, we show how two insulin-mediated canonical antiviral pathways are simultaneously activated in our drug treatment regimen to reduce virus levels in mosquito saliva, through which virus is transmitted to humans. Together, we demonstrate the viability of targeting insulin signaling as a means of reducing the rate of mosquito infection and decreased transmission of Zika virus. [ABSTRACT FROM AUTHOR]

Background: Rift Valley fever (RVF) is an emerging, neglected, mosquito-borne viral zoonosis associated with significant morbidity, mortality and expanding geographical scope. The clinical signs and symptoms in humans are non-specific and case definitions vary. We reviewed and analysed the clinical manifestations of RVF in humans. Methods: In this systematic review and meta-analysis we searched on different dates, the Embase (from 1947 to 13th October 2019), Medline (1946 to 14th October 2019), Global Health (1910 to 15th October 2019), and Web of Science (1970 to 15th October 2019) databases. Studies published in English, reporting frequency of symptoms in humans, and laboratory confirmed RVF were included. Animal studies, studies among asymptomatic volunteers, and single case reports for which a proportion could not be estimated, were excluded. Quality assessment was done using a modified Hoy and Brooks et al tool, data was extracted, and pooled frequency estimates calculated using random effects meta-analysis. Results: Of the 3765 articles retrieved, less than 1% (32 articles) were included in the systematic review and meta-analysis. Nine RVF clinical syndromes were reported including the general febrile, renal, gastrointestinal, hepatic, haemorrhagic, visual, neurological, cardio-pulmonary, and obstetric syndromes. The most common clinical manifestations included fever (81%; 95% Confidence Interval (CI) 69–91; [26 studies, 1286 patients]), renal failure (41%; 23–59; [4, 327]), nausea (38%; 12–67; [6, 325]), jaundice (26%; 16–36; [15, 393]), haemorrhagic disease (26%; 17–36; [16, 277]), partial blindness (24%; 7–45; [11, 225]), encephalitis (21%; 11–33; [4, 327]), cough (4%; 0–17; [4, 11]), and miscarriage (54%) respectively. Death occurred in 21% (95% CI 14–29; [16 studies, 328 patients]) of cases, most of whom were hospitalised. Discussion: This study delineates the complex symptomatology of human RVF disease into syndromes. This approach is likely to improve case definitions and detection rates, impact outbreak control, increase public awareness about RVF, and subsequently inform 'one-health' policies. This study provides a pooled estimate of the proportion of RVF clinical manifestations alongside a narrative description of clinical syndromes. However, most studies reviewed were case series with small sample sizes and enrolled mostly in-patients and out-patients, and captured symptoms either sparsely or using broad category terms. Author summary: Rift Valley fever (RVF) is a neglected, arboviral zoonosis that causes severe and diverse disease manifestations in humans. Currently no licenced vaccines exist for use in humans and there is limited surveillance and estimation of disease burden which in part is due to the inability to concisely define the disease. We searched Medline, Embase, Global Health, and Web of Science for published reports on the clinical manifestations of RVF in humans. Studies published in English, reporting frequency of symptoms in humans, and laboratory confirmed RVF were included. We excluded animal studies, studies among asymptomatic volunteers and single case reports for which a proportion could not be estimated. Pooled symptom frequency estimates were calculated using random effects meta-analysis. This review provides a detailed aggregation of the relative frequency of symptoms, and a description of the RVF clinical manifestations in humans. Previous systematic reviews provided a narrative account and it was difficult to identify the most relevant features of RVF disease in areas where other endemic infections present with similar symptoms. This review will refine the clinical diagnosis, improve case detection, and increase public awareness about RVF presentation in humans. [ABSTRACT FROM AUTHOR]

Uganda established a domestic Viral Hemorrhagic Fever (VHF) testing capacity in 2010 in response to the increasing occurrence of filovirus outbreaks. In July 2018, the neighboring Democratic Republic of Congo (DRC) experienced its 10th Ebola Virus Disease (EVD) outbreak and for the duration of the outbreak, the Ugandan Ministry of Health (MOH) initiated a national EVD preparedness stance. Almost one year later, on 10th June 2019, three family members who had contracted EVD in the DRC crossed into Uganda to seek medical treatment. Samples were collected from all the suspected cases using internationally established biosafety protocols and submitted for VHF diagnostic testing at Uganda Virus Research Institute. All samples were initially tested by RT-PCR for ebolaviruses, marburgviruses, Rift Valley fever (RVF) virus and Crimean-Congo hemorrhagic fever (CCHF) virus. Four people were identified as being positive for Zaire ebolavirus, marking the first report of Zaire ebolavirus in Uganda. In-country Next Generation Sequencing (NGS) and phylogenetic analysis was performed for the first time in Uganda, confirming the outbreak as imported from DRC at two different time point from different clades. This rapid response by the MoH, UVRI and partners led to the control of the outbreak and prevention of secondary virus transmission. Author summary: In the effort to control the on-going COVID-19 pandemic, countries instituted lock downs and closed international borders. But is this the best approach to controlling transboundary infectious diseases? In this publication we demonstrate how we managed to control Ebola Virus Disease (EVD) introduced into Uganda from Democratic Republic of Congo (DRC) in 2019. Once the EVD outbreak was announced in DRC, we intensified cross-border surveillance and instituted acceptable public health control measures following international health regulations to limit onward community and nosocomial transmission in Uganda. Consequently, on July 10th, 2019, three (3) cases crossed into Uganda and were detected at the point of first contact with health facility in Uganda in Kasese district and the fourth case was detected in August 2019 at a point of entry at the Uganda-DRC border by temperature screening. Sequencing of these cases showed that they were independent introductions related to two different clades of the ongoing outbreak in DRC. All the patients died without onward secondary transmissions in Uganda. We invite you to read this publication and learn how this was achieved as it can be used as a model for cross border surveillance to control similar infectious disease outbreaks. [ABSTRACT FROM AUTHOR]

Background: Although vector-borne zoonotic diseases are a major public health threat globally, they are usually neglected, especially among resource-constrained countries, including those in sub-Saharan Africa. This scoping review examined the current knowledge and identified research gaps of vector-borne zoonotic pathogens in Zambia. Methods and findings: Major scientific databases (Web of Science, PubMed, Scopus, Google Scholar, CABI, Scientific Information Database (SID)) were searched for articles describing vector-borne (mosquitoes, ticks, fleas and tsetse flies) zoonotic pathogens in Zambia. Several mosquito-borne arboviruses have been reported including Yellow fever, Ntaya, Mayaro, Dengue, Zika, West Nile, Chikungunya, Sindbis, and Rift Valley fever viruses. Flea-borne zoonotic pathogens reported include Yersinia pestis and Rickettsia felis. Trypanosoma sp. was the only tsetse fly-borne pathogen identified. Further, tick-borne zoonotic pathogens reported included Crimean-Congo Haemorrhagic fever virus, Rickettsia sp., Anaplasma sp., Ehrlichia sp., Borrelia sp., and Coxiella burnetii. Conclusions: This study revealed the presence of many vector-borne zoonotic pathogens circulating in vectors and animals in Zambia. Though reports of human clinical cases were limited, several serological studies provided considerable evidence of zoonotic transmission of vector-borne pathogens in humans. However, the disease burden in humans attributable to vector-borne zoonotic infections could not be ascertained from the available reports and this precludes the formulation of national policies that could help in the control and mitigation of the impact of these diseases in Zambia. Therefore, there is an urgent need to scale-up "One Health" research in emerging and re-emerging infectious diseases to enable the country to prepare for future epidemics, including pandemics. Author summary: Despite vector-borne zoonoses being a major public health threat globally, they are often overlooked, particularly among resource-constrained countries in sub-Saharan Africa, including Zambia. Therefore, we reviewed the current knowledge and identified research gaps of vector-borne zoonotic pathogens in Zambia. We focussed on mosquito-, tick-, flea- and tsetse fly-borne zoonotic pathogens reported in the country. Although we found evidence of circulation of several vector-borne zoonotic pathogens among vectors, animals and humans, clinical cases in humans were rarely reported. This suggests sparse capacity for diagnosis of vector-borne pathogens in healthcare facilities in the country and possibly limited awareness and knowledge of the local epidemiology of these infectious agents. Establishment of facility-based surveillance of vector-borne zoonoses in health facilities could provide valuable insights on morbidity, disease severity, and mortalities associated with infections as well as immune responses. In addition, there is also need for increased genomic surveillance of vector-borne pathogens in vectors and animals and humans for a better understanding of the molecular epidemiology of these diseases in Zambia. Furthermore, vector ecology studies aimed at understanding the drivers of vector abundance, pathogen host range (i.e., including the range of vectors and reservoirs), parasite-host interactions and factors influencing frequency of human-vector contacts should be prioritized. The study revealed the need for Zambia to scale-up One Health research in emerging and re-emerging infectious diseases to enable the country to be better prepared for future epidemics, including pandemics. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a mosquito-borne disease mostly affecting wild and domestic ruminants. It is widespread in Africa, with spillovers in the Arab Peninsula and the southwestern Indian Ocean. Although RVF has been circulating in West Africa for more than 30 years, its epidemiology is still not clearly understood. In 2013, an RVF outbreak hit Senegal in new areas that weren't ever affected before. To assess the extent of the spread of RVF virus, a national serological survey was implemented in young small ruminants (6–18 months old), between November 2014 and January 2015 (after the rainy season) in 139 villages. Additionally, the drivers of this spread were identified. For this purpose, we used a beta-binomial (BB) logistic regression model. An Integrated Nested Laplace Approximation (INLA) approach was used to fit the spatial model. Lower cumulative rainfall, and higher accessibility were both associated with a higher RVFV seroprevalence. The spatial patterns of fitted RVFV seroprevalence pointed densely populated areas of western Senegal as being at higher risk of RVFV infection in small ruminants than rural or southeastern areas. Thus, because slaughtering infected animals and processing their fresh meat is an important RVFV transmission route for humans, more human populations might have been exposed to RVFV during the 2013–2014 outbreak than in previous outbreaks in Senegal. Author summary: An outbreak of Rift Valley fever (RVF)–a zoonotic mosquito-borne viral infection widespread in Africa, was reported in 2013–14 in Senegal. After its end in late 2014, its spatial distribution and drivers were assessed using a national serological survey in small ruminants, thus highlighting the high exposition of humans to RVF virus in urban areas. [ABSTRACT FROM AUTHOR]

Background: Rift Valley fever virus (RVFV) is a lethal threat to humans and livestock in many parts of Africa, the Arabian Peninsula, and the Indian Ocean. This systematic review's objective was to consolidate understanding of RVFV epidemiology during 1999–2021 and highlight knowledge gaps relevant to plans for human vaccine trials. Methodology/Principal findings: The review is registered with PROSPERO (CRD42020221622). Reports of RVFV infection or exposure among humans, animals, and/or vectors in Africa, the Arabian Peninsula, and the Indian Ocean during the period January 1999 to June 2021 were eligible for inclusion. Online databases were searched for publications, and supplemental materials were recovered from official reports and research colleagues. Exposures were classified into five groups: 1) acute human RVF cases, 2) acute animal cases, 3) human RVFV sero-surveys, 4) animal sero-surveys, and 5) arthropod infections. Human risk factors, circulating RVFV lineages, and surveillance methods were also tabulated. In meta-analysis of risks, summary odds ratios were computed using random-effects modeling. 1104 unique human or animal RVFV transmission events were reported in 39 countries during 1999–2021. Outbreaks among humans or animals occurred at rates of 5.8/year and 12.4/year, respectively, with Mauritania, Madagascar, Kenya, South Africa, and Sudan having the most human outbreak years. Men had greater odds of RVFV infection than women, and animal contact, butchering, milking, and handling aborted material were significantly associated with greater odds of exposure. Animal infection risk was linked to location, proximity to water, and exposure to other herds or wildlife. RVFV was detected in a variety of mosquito vectors during interepidemic periods, confirming ongoing transmission. Conclusions/Significance: With broad variability in surveillance, case finding, survey design, and RVFV case confirmation, combined with uncertainty about populations-at-risk, there were inconsistent results from location to location. However, it was evident that RVFV transmission is expanding its range and frequency. Gaps assessment indicated the need to harmonize human and animal surveillance and improve diagnostics and genotyping. Given the frequency of RVFV outbreaks, human vaccination has strong potential to mitigate the impact of this now widely endemic disease. Author summary: Rift Valley fever virus (RVFV) is a globally important mosquito-transmitted zoonosis that is also directly transmissible via aerosolization of body fluids from infected animals. RVFV outbreaks cause mass mortality of young livestock and pregnancy losses in both humans and animals. Severe human cases also result in hemorrhagic fever, encephalitis, and death. Loss of livestock additionally threatens the livelihood of people who depend on animals for income and food. In endemic areas, initiation of RVFV outbreaks is connected to weather events that cause excess rainfall, leading to flooding and subsequent mosquito blooms. However, the natural cycle of RVFV transmission is complex, requiring congregation of susceptible mammalian hosts and mosquito vectors in suitable environments. Several human vaccine candidates are in different stages of development, but none are yet licensed for use in human populations. In this systematic review, we assessed the 1999–2021 frequency and distribution of RVFV outbreaks among humans, animals, and vectors to identify potential locations and population targets for a human RVFV vaccine efficacy trial. It focuses on current understanding of RVFV epidemiology and the identification of gaps that pose critical barriers to controlling expansion of RVFV and implementing new protective measures including human vaccination. [ABSTRACT FROM AUTHOR]

Trypanosoma brucei, the causative agent of human African trypanosomiasis, is highly motile and must be able to move in all three dimensions for reliable cell division. These characteristics make long-term microscopic imaging of live T. brucei cells challenging, which has limited our understanding of important cellular events. To address this issue, we devised an imaging approach that confines cells in small volumes within cast agarose microwells that can be imaged continuously for up to 24 h. Individual T. brucei cells were imaged through multiple rounds of cell division with high spatial and temporal resolution. We developed a strategy that employs in-well "sentinel" cells to monitor potential imaging toxicity during loss-of-function experiments such as small-molecule inhibition and RNAi. Using our approach, we show that the asymmetric daughter cells produced during T. brucei division subsequently divide at different rates, with the old-flagellum daughter cell dividing first. The flagellar detachment phenotype that appears during inhibition of the Polo-like kinase homolog TbPLK occurs in a stepwise fashion, with the new flagellum initially linked by its tip to the old, attached flagellum. We probe the feasibility of a previously proposed "back-up" cytokinetic mechanism and show that cells that initiate this process do not appear to complete cell division. This live-cell imaging method will provide a novel avenue for studying a wide variety of cellular events in trypanosomatids that have previously been inaccessible. Author summary: Trypanosoma brucei causes severe diseases that affect humans and livestock in Sub-Saharan Africa. Efficient strategies for manipulating the T. brucei genome have provided a wealth of information about protein localization and function in diverse cellular processes. However, employing live-cell imaging for phenotypic analysis in T. brucei remains a significant challenge because immobilization of this highly motile parasite rapidly leads to morphologic defects and cell death. While fixed-cell imaging can provide snapshots of cellular events, it cannot provide the direct causal link or precise timing of events that comes from watching a living cell change over time. Our strategy using agarose microwells now allows long-term live cell T. brucei imaging with a simple apparatus that can be adapted for a wide range of experimental conditions. [ABSTRACT FROM AUTHOR]

To evaluate and compare the risk of emerging vector-borne diseases (VBDs), a Model for INTegrated RISK assessment, MINTRISK, was developed to assess the introduction risk of VBDs for new regions in an objective, transparent and repeatable manner. MINTRISK is a web-based calculation tool, that provides semi-quantitative risk scores that can be used for prioritization purposes. Input into MINTRISK is entered by answering questions regarding entry, transmission, establishment, spread, persistence and impact of a selected VBD. Answers can be chosen from qualitative answer categories with accompanying quantitative explanation to ensure consistent answering. The quantitative information is subsequently used as input for the model calculations to estimate the risk for each individual step in the model and for the summarizing output values (rate of introduction; epidemic size; overall risk). The risk assessor can indicate his uncertainty on each answer, and this is accounted for by Monte Carlo simulation. MINTRISK was used to assess the risk of four VBDs (African horse sickness, epizootic haemorrhagic disease, Rift Valley fever, and West Nile fever) for the Netherlands with the aim to prioritise these diseases for preparedness. Results indicated that the overall risk estimate was very high for all evaluated diseases but epizootic haemorrhagic disease. Uncertainty intervals were, however, wide limiting the options for ranking of the diseases. Risk profiles of the VBDs differed. Whereas all diseases were estimated to have a very high economic impact once introduced, the estimated introduction rates differed from low for Rift Valley fever and epizootic haemorrhagic disease to moderate for African horse sickness and very high for West Nile fever. Entry of infected mosquitoes on board of aircraft was deemed the most likely route of introduction for West Nile fever into the Netherlands, followed by entry of infected migratory birds. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) causes morbidity and mortality in humans and domestic ungulates in sub-Saharan Africa, Egypt, and the Arabian Peninsula. Mosquito vectors transmit RVFV between vertebrates by bite, and also vertically to produce infectious progeny. Arrival of RVFV into the United States by infected mosquitoes or humans could result in significant impacts on food security, human health, and wildlife health. Elucidation of the vectors involved in the post-introduction RVFV ecology is paramount to rapid implementation of vector control. We performed vector competence experiments in which field-collected mosquitoes were orally exposed to an epidemic strain of RVFV via infectious blood meals. We targeted floodwater Aedes species known to feed on cattle, and/or deer species (Aedes melanimon Dyar, Aedes increpitus Dyar, Aedes vexans Meigen). Two permanent-water-breeding species were targeted as well: Culiseta inornata Williston of unknown competence considering United States populations, and Culex tarsalis Coquillett as a control species for which transmission efficiency is known. We tested the potential for midgut infection, midgut escape (dissemination), ovarian infection (vertical transmission), and transmission by bite (infectious saliva). Tissues were assayed by plaque assay and RT-qPCR, to quantify infectious virus and confirm virus identity. Tissue infection data were analyzed using a within-host model under a Bayesian framework to determine the probabilities of infection 'outcomes' (midgut-limited infection, disseminated infection, etc.) while estimating barriers to infection between tissues. Permanent-water-breeding mosquitoes (Cx. tarsalis and Cs. inornata) exhibited more efficient horizontal transmission, as well as potential for vertical transmission, which is contrary to the current assumptions of RVFV ecology. Barrier estimates trended higher for Aedes spp., suggesting systemic factors in the differences between these species and Cx. tarsalis and Cs. inornata. These data indicate higher potential for vertical transmission than previously appreciated, and support the consensus of RVFV transmission including a broad range of potential vectors. Author summary: Rift Valley fever virus (RVFV) causes morbidity and mortality in humans and domestic ungulates in sub-Saharan Africa, Egypt, and the Arabian Peninsula. Mosquito vectors transmit RVFV between vertebrates by bite, and also vertically to produce infectious progeny. To inform vector control priorities upon the introduction of RVFV to the United States, we tested the ability of three floodwater Aedes species known to feed on cattle and/or deer in Colorado (Aedes vexans Meigen, Aedes melanimon Dyar, Aedes increpitus Dyar) to transmit RVFV (vector competence). We also tested Culiseta inornata Williston, and Culex tarsalis Coquillett which exhibits high vector competence, and the potential for vertical transmission by testing ovaries. These data were modeled to estimate the potential for virus transmission, based on the infection probabilities of different organs that serve as transmission barriers inside the mosquitoes. The permanent-water-breeders Cs. inornata and Cx. tarsalis exhibited higher efficiency of horizontal transmission as well as potential vertical transmission. Aedes species were less efficient at vertical and horizontal transmission, with high barriers to infection of and dissemination from the midgut. Overall, these data support the transmission of RVFV by a broad range of potential vectors in the United States, posing a major challenge for vector control if this virus is introduced. [ABSTRACT FROM AUTHOR]

Eimeria parasites cause enteric disease in livestock and the closely related Cyclosporacayetanensis causes human disease. Oocysts of these coccidian parasites undergo maturation (sporulation) before becoming infectious. Here, we assessed transcription in maturing oocysts of Eimeria acervulina, a widespread chicken parasite, predicted gene functions, and determined which of these genes also occur in C. cayetanensis. RNA-Sequencing yielded ~2 billion paired-end reads, 92% of which mapped to the E. acervulina genome. The ~6,900 annotated genes underwent temporally-coordinated patterns of gene expression. Fifty-three genes each contributed >1,000 transcripts per million (TPM) throughout the study interval, including cation-transporting ATPases, an oocyst wall protein, a palmitoyltransferase, membrane proteins, and hypothetical proteins. These genes were enriched for 285 gene ontology (GO) terms and 13 genes were ascribed to 17 KEGG pathways, defining housekeeping processes and functions important throughout sporulation. Expression differed in mature and immature oocysts for 40% (2,928) of all genes; of these, nearly two-thirds (1,843) increased their expression over time. Eight genes expressed most in immature oocysts, encoding proteins promoting oocyst maturation and development, were assigned to 37 GO terms and 5 KEGG pathways. Fifty-six genes underwent significant upregulation in mature oocysts, each contributing at least 1,000 TPM. Of these, 40 were annotated by 215 GO assignments and 9 were associated with 18 KEGG pathways, encoding products involved in respiration, carbon fixation, energy utilization, invasion, motility, and stress and detoxification responses. Sporulation orchestrates coordinated changes in the expression of many genes, most especially those governing metabolic activity. Establishing the long-term fate of these transcripts in sporulated oocysts and in senescent and deceased oocysts will further elucidate the biology of coccidian development, and may provide tools to assay infectiousness of parasite cohorts. Moreover, because many of these genes have homologues in C. cayetanensis, they may prove useful as biomarkers for risk. [ABSTRACT FROM AUTHOR]

Globally, arthropod-borne virus (arbovirus) infections continue to pose substantial threats to public health and economic development, especially in developing countries. In Kenya, although arboviral diseases (ADs) are largely endemic, little is known about the factors influencing livestock farmers' knowledge, beliefs, and management (KBM) of the three major ADs: Rift Valley fever (RVF), dengue fever and chikungunya fever. This study evaluates the drivers of livestock farmers' KBM of ADs from a sample of 629 respondents selected using a three-stage sampling procedure in Kenya's three hotspot counties of Baringo, Kwale, and Kilifi. A multivariate fractional probit model was used to assess the factors influencing the intensity of KBM. Only a quarter of the farmers had any knowledge of ADs while over four-fifths of them could not manage any of the three diseases. Access to information (experience and awareness), income, education, religion, and distance to a health facility considerably influenced the intensity of farmers' KBM of ADs in Kenya. Thus, initiatives geared towards improving access to information through massive awareness campaigns are necessary to mitigate behavioral barriers in ADs management among rural communities in Kenya. Author summary: Arboviral infection in humans and animals is on the rise globally due to expansion of vector habitats. Despite the economic and social impact of diseases caused by arboviral infection such as chikungunya, dengue, and Rift Valley fever, little is known in terms of community knowledge, beliefs, and management. Evaluating community knowledge, beliefs, and management practices of arboviral diseases is important for better policy guidance and public health investment. We conducted a survey in Kenya's three hotspot counties of Baringo, Kwale, and Kilifi to understand the factors influencing knowledge, beliefs, and management of arboviral diseases. We found low levels of knowledge and poor managerial skills of arboviral diseases that were largely driven by access to information and asset ownership. Thus, community sensitization through improved access to information is important in increasing awareness and increase the management of arboviral diseases among rural communities in Kenya and other sub-Saharan African countries. [ABSTRACT FROM AUTHOR]

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. Author summary: Rift Valley fever virus (RVFV) is the causative agent of Rift Valley fever, which is primarily endemic in African countries. The virus infects a wide variety of mammalian species, including ruminants and humans, and causes devastating effects on public health and the livestock industry. Because RVFV is transmitted by several mosquito species that are ubiquitous in non-endemic areas, there is a potential risk that the virus will spread outside of the current endemic areas. However, there are no effective therapeutics nor commercially available vaccines for human use. RVFV encodes a 78kDa protein (P78) of unidentified biological function in mammalian cells. We found that a mutant RVFV expressing a higher level of P78 (P78-High virus) showed lower infectivity and less efficient replication than parental RVFV in macrophage cell lines. P78-High virus and a mutant RVFV lacking P78 expression showed lower and higher virulence than the parental virus, respectively, in mice, suggesting that high levels of P78 expression prevent efficient virus replication in macrophages, leading to attenuation of virus virulence. Genetic alteration leading to increased P78 expression may serve as a novel strategy for the generation of attenuated RVFV as vaccine candidates. [ABSTRACT FROM AUTHOR]

The Government of Rwanda reported an outbreak of Rift Valley fever (RVF) in the Eastern Province in 2018. To respond to the outbreak, vaccination and education campaigns about the disease were carried out. Because RVF cases continue to be detected in Rwanda and the disease impacts livelihoods and health, accurate knowledge and communication are imperative. The objectives of this study were to evaluate knowledge and risk perceptions of RVF transmission among livestock farmers in Nyagatare District, Eastern Province, Rwanda, and to compare RVF knowledge, risk perceptions, and farming practices between male and female livestock farmers. This cross-sectional, quantitative study was conducted in selected sectors of Nyagatare District in the Eastern Province of Rwanda in June 2019. A 34-question survey was used to ask about demographics, livestock ownership, risk perceptions about zoonotic diseases and livestock management, RVF knowledge, preferred communication sources and information sharing strategies, and protective strategies for RVF mitigation while working with livestock. Livestock farmers were interviewed at three milk collection centers, two village meeting points, a farm cooperative meeting, and during door-to-door visits in villages. In total, 123 livestock farmers were interviewed. The survey found that most livestock farmers lacked knowledge about epizootic and zoonotic transmission of RVF, more male livestock farmers were familiar with RVF and risk mitigation strategies, and female livestock farmers are not viewed as reliable sources of information. Additionally, most livestock farmers had not vaccinated their animals against RVF despite past vaccination campaigns. Radio was the most popular communication channel. These findings show that RVF knowledge and information sharing are inadequate among livestock farmers in Eastern Province. Therefore, vaccination and education campaigns may need to be reevaluated within the context of these trends in order to prepare for future RVF outbreaks. Author summary: This study was conducted in order to evaluate RVF knowledge and awareness as well as communication and mitigation strategies among livestock farmers in Eastern Province, Rwanda. Rwanda declared an outbreak of RVF in 2018 and cases have continued to be detected. Thus, evaluating the status of knowledge, preventive strategies, and information sharing among livestock farmers is crucial in mitigating future outbreaks. Our team conducted a survey of knowledge, risk perceptions, mitigation strategies, and communication practices among livestock farmers from selected sectors within Nyagatare District and compared them between male and female livestock farmers in order to analyze gender-nuanced differences between these groups. Sectors were chosen for sampling based on their proximity to previous outbreak areas. Survey questionnaire results showed that knowledge and risk perceptions differed between male and female livestock farmers, and that they could be generally improved among all livestock farmers. Female livestock farmers and non-farming community members were viewed as unreliable sources of information which could impact information dissemination. Many farmers also reported that their livestock herds were not vaccinated for the disease despite previous vaccination campaigns. Communication strategies and information sources also differed between male and female livestock farmers, which highlights a need to consider gender in improving RVF vaccination and education campaign coverage. These findings pose implications for future community-based public health interventions as well as policy development for RVF control and mitigating future RVF outbreaks within Rwanda. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV), an arbovirus belonging to the Phlebovirus genus of the Phenuiviridae family, causes the zoonotic and mosquito-borne RVF. The virus, which primarily affects livestock (ruminants and camels) and humans, is at the origin of recent major outbreaks across the African continent (Mauritania, Libya, Sudan), and in the South-Western Indian Ocean (SWIO) islands (Mayotte). In order to be better prepared for upcoming outbreaks, to predict its introduction in RVFV unscathed countries, and to run efficient surveillance programmes, the priority is harmonising and improving the diagnostic capacity of endemic countries and/or countries considered to be at risk of RVF. A serological inter-laboratory proficiency test (PT) was implemented to assess the capacity of veterinary laboratories to detect antibodies against RVFV. A total of 18 laboratories in 13 countries in the Middle East, North Africa, South Africa, and the Indian Ocean participated in the initiative. Two commercial kits and two in-house serological assays for the detection of RVFV specific IgG antibodies were tested. Sixteen of the 18 participating laboratories (88.9%) used commercial kits, the analytical performance of test sensitivity and specificity based on the seroneutralisation test considered as the reference was 100%. The results obtained by the laboratories which used the in-house assay were correct in only one of the two criteria (either sensitivity or specificity). In conclusion, most of the laboratories performed well in detecting RVFV specific IgG antibodies and can therefore be considered to be prepared. Three laboratories in three countries need to improve their detection capacities. Our study demonstrates the importance of conducting regular proficiency tests to evaluate the level of preparedness of countries and of building a network of competent laboratories in terms of laboratory diagnosis to better face future emerging diseases in emergency conditions. [ABSTRACT FROM AUTHOR]

Camels are increasingly becoming the livestock of choice for pastoralists reeling from effects of climate change in semi-arid and arid parts of Kenya. As the population of camels rises, better understanding of their role in the epidemiology of zoonotic diseases in Kenya is a public health priority. Rift Valley fever (RVF), brucellosis and Q fever are three of the top priority diseases in the country but the involvement of camels in the transmission dynamics of these diseases is poorly understood. We analyzed 120 camel serum samples from northern Kenya to establish seropositivity rates of the three pathogens and to characterize the infecting Brucella species using molecular assays. We found seropositivity of 24.2% (95% confidence interval [CI]: 16.5–31.8%) for Brucella, 20.8% (95% CI: 13.6–28.1%) and 14.2% (95% CI: 7.9–20.4%) for Coxiella burnetii and Rift valley fever virus respectively. We found 27.5% (95% CI: 19.5–35.5%) of the animals were seropositive for at least one pathogen and 13.3% (95% CI: 7.2–19.4%) were seropositive for at least two pathogens. B. melitensis was the only Brucella spp. detected. The high sero-positivity rates are indicative of the endemicity of these pathogens among camel populations and the possible role the species has in the epidemiology of zoonotic diseases. Considering the strong association between human infection and contact with livestock for most zoonotic infections in Kenya, there is immediate need to conduct further research to determine the role of camels in transmission of these zoonoses to other livestock species and humans. This information will be useful for designing more effective surveillance systems and intervention measures. Author summary: Dromedary camels are well adapted to the arid and semi-arid environment that makes up about 80% of Kenya's landmass. As such, camels play an important role in the socio-economic wellbeing and food security of pastoralists in the country. However, the species remains relatively neglected in scientific research, one of the main reasons being camels are mostly found in remote, low-income, arid regions of Africa and Asia. We carried out a study to determine the levels of exposure of camels in northern Kenya to Brucella spp., Coxiella burnetii and Rift Valley fever virus, three priority zoonotic pathogens in the country. We found high levels of exposure to the three pathogens, indicating the important role camels might play in the epidemiology of the zoonotic diseases in humans and other livestock. Based on the study findings, we argue for the immediate need for investments in disease surveillance and control strategies for priority zoonotic disease in camels in Kenya. [ABSTRACT FROM AUTHOR]

Rift Valley fever virus (RVFV) is a mosquito-transmitted virus with proven ability to emerge into naïve geographic areas. Limited field evidence suggests that RVFV is transmitted vertically from parent mosquito to offspring, but until now this mechanism has not been confirmed in the laboratory. Furthermore, this transmission mechanism has allowed for the prediction of RVFV epizootics based on rainfall patterns collected from satellite information. However, in spite of the relevance to the initiation of epizootic events, laboratory confirmation of vertical transmission has remained an elusive research aim for thirty-five years. Herein we present preliminary evidence of the vertical transmission of RVFV by Culex tarsalis mosquitoes after oral exposure to RVFV. Progeny from three successive gonotrophic cycles were reared to adults, with infectious RVFV confirmed in each developmental stage. Virus was detected in ovarian tissues of parental mosquitoes 7 days after imbibing an infectious bloodmeal. Infection was confirmed in progeny as early as the first gonotrophic cycle, with infection rates ranging from 2.0–10.0%. Virus titers among progeny were low, which may indicate a host mechanism suppressing replication. Author summary: Rift Valley fever virus (RVFV) represents a significant threat in terms of its ability to emerge into naive geographic areas. Furthermore, RVFV represents a global public health risk due to the ability of many mosquito species to transmit the virus and the ease with which the virus can be transported due to increased globalization. The vertical transmission of RVFV by mosquitoes has long been accepted by the research community due to limited field evidence. However, laboratory confirmation of vertical transmission has remained elusive for thirty-five years. We present the first laboratory evidence of vertical transmission of RVFV in the susceptible North American vector, Culex tarsalis. We present two studies that clearly show 1) the accumulation of RVFV antigen in the ovaries of infected mosquitoes and 2) the transmission of RVFV from parent to offspring immediately following an infectious blood meal. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a vector-borne viral disease of major animal and public health importance. In 2018–19, it caused an epidemic in both livestock and human populations of the island of Mayotte. Using Bayesian modelling approaches, we assessed the spatio-temporal pattern of RVF virus (RVFV) infection in livestock and human populations across the island, and factors shaping it. First, we assessed if (i) livestock movements, (ii) spatial proximity from communes with infected animals, and (iii) livestock density were associated with the temporal sequence of RVFV introduction into Mayotte communes' livestock populations. Second, we assessed whether the rate of human infection was associated with (a) spatial proximity from and (b) livestock density of communes with infected animals. Our analyses showed that the temporal sequence of RVFV introduction into communes' livestock populations was associated with livestock movements and spatial proximity from communes with infected animals, with livestock movements being associated with the best model fit. Moreover, the pattern of human cases was associated with their spatial proximity from communes with infected animals, with the risk of human infection sharply increasing if livestock in the same or close communes were infected. This study highlights the importance of understanding livestock movement networks in informing the design of risk-based RVF surveillance programs. Author summary: Rift Valley fever (RVF) is a vector-borne zoonotic disease, endemic in many sub-Saharan Africa regions with substantial outbreaks. RVF virus (RVFV) is transmitted to animals primarily by the bite of infected mosquitos, whereas direct or indirect contact with infected animals forms the primary route of RVFV transmission to humans. In 2018–19, Mayotte, an archipelago in the Indian Ocean between Madagascar and the coast of Eastern Africa, experienced an RVF epidemic in both livestock and humans. In this study, we investigated factors shaping the spatio-temporal pattern of RVFV infection in livestock and human populations across Mayotte. The diffusion of RVFV through Mayotte's livestock population was associated with livestock movements and, potentially to a lesser extent, spatial proximity from communes with infected animals. Moreover, the pressure of infection on humans was the highest if nearby livestock were infected. This study highlights the value of accounting for the structure of livestock movement networks in the surveillance of zoonotic diseases at the human-animal interface, and the need for One Health approaches. [ABSTRACT FROM AUTHOR]

Rift Valley Fever (RVF) is a zoonotic disease whose outbreak results in heavy economic and public health burdens. In East Africa, RVF is mainly experienced in arid and semi-arid areas predominantly inhabited by the pastoralists. These areas experience sudden, dramatic epidemics of the disease at intervals of approximately 10 years, associated with widespread flooding and the resultant swarms of mosquitoes. Pastoralists' indigenous knowledge and experience of RVF is critical for public health interventions targeting prevention and control of RVF. The study adopted a descriptive cross-sectional design combining both quantitative and qualitative methods of data collection. A total of 204 respondents participated in questionnaire survey and 15 key informants and 4 focus group discussions were interviewed and conducted respectively. In addition, secondary data mainly journal publications, books, policy documents and research reports from conferences and government departments were reviewed. Findings indicated that the Somali pastoralists possess immense knowledge of RVF including signs and symptoms, risk factors, and risk pathways associated with RVF. Ninety eight percent (98%) of respondents identified signs and symptoms such as bloody nose, diarrhea, foul smell and discharge of blood from the orifices which are consistent with RVF. Heavy rains and floods (85%) and sudden emergence of mosquito swarms (91%) were also cited as the major RVF risk factors while mosquito bites (85%), drinking raw milk and blood (78%) and contact with animal fluids during mobility, slaughter and obstetric procedures (77%) were mentioned as the RVF entry risk pathways. Despite this immense knowledge, the study found that the pastoralists did not translate the knowledge into safer health practices because of the deep-seated socio-cultural practices associated with pastoralist production system and religious beliefs. On top of these practices, food preparation and consumption practices such as drinking raw blood and milk and animal ritual sacrifices continue to account for most of the mortality and morbidity cases experienced in humans and animals during RVF outbreaks. This article concludes that pastoralists' indigenous knowledge on RVF has implications on public health delivery approaches. Since the pastoralists' knowledge on RVF was definitive, integrating the community into early warning systems through training on reporting mechanisms and empowering the nomads to use their mobile phone devices to report observable changes in their livestock and environment could prove very effective in providing information for timely mobilization of public health responses. Public health advocacy based on targeted and contextually appropriate health messaging and disseminated through popular communication channels in the community such as the religious leaders and local radio stations would also be needed to reverse the drivers of RVF occurrence in the study area. Author summary: Rift Valley Fever is a viral disease that affects both humans and animals. It is categorized as one of the re-emerging and neglected tropical diseases that mainly affects the poor and marginalized populations that lack access to health services and are readily ignored. Humans usually get RVF through bites from infected mosquitoes. Infections also occur when humans are exposed to the body fluids, or tissues of infected animals. Hence the risk of infection is greatest when slaughtering in the context of traditional sacrificial practices. This is the major reason outbreak of RVF is commonly associated with people whose livelihoods revolve around livestock rearing. In East Africa, RVF is mainly experienced in arid and semi-arid areas predominantly inhabited by the pastoralists. These areas experience epidemics of the disease at intervals of approximately 10 years associated with Elnino events. Understanding the knowledge base of the people in terms of RVF signs and symptoms and risk factors and pathways is important for the adoption of effective prevention and control measures. This study findings suggest that even though the Somali nomads are adept at recognizing RVF, this knowledge has not been translated into appropriate health practices due to the deep-seated socio-cultural practices. Hence, there is need for health authorities to mount locally appropriate public health advocacy campaigns, empower the livestock keepers to report observable changes in livestock and environment using their mobile phone devices and promote cross-disciplinary studies on RVF. [ABSTRACT FROM AUTHOR]

Background: As an invasive mosquito species in the United States, Aedes albopictus is a potential vector of arboviruses including dengue, chikungunya, and Zika, and may also be involved in occasional transmission of other arboviruses such as West Nile, Saint Louis encephalitis, eastern equine encephalitis, and La Crosse viruses. Aedes albopictus feeds on a wide variety of vertebrate hosts, wild and domestic, as well as humans. Methodology/Principal findings: In order to investigate blood feeding patterns of Ae. albopictus, engorged specimens were collected from a variety of habitat types using the Centers for Disease Control and Prevention light traps, Biogents Sentinel 2 traps, and modified Reiter gravid traps in southeast Virginia. Sources of blood meals were determined by the analysis of mitochondrial cytochrome b gene sequences amplified in PCR assays. Our aims were to quantify degrees of Ae. albopictus interactions with vertebrate hosts as sources of blood meals, investigate arboviral infection status, assess the influence of key socioecological conditions on spatial variability in blood feeding, and investigate temporal differences in blood feeding by season. Analysis of 961 engorged specimens of Ae. albopictus sampled between 2017–2019 indicated that 96%, 4%, and less than 1% obtained blood meals from mammalian, reptilian, and avian hosts, respectively. Domestic cats were the most frequently identified (50.5%) hosts followed by Virginia opossums (17.1%), white-tailed deer (12.2%), and humans (7.3%), together representing 87.1% of all identified blood hosts. We found spatial patterns in blood feeding linked to socioecological conditions and seasonal shifts in Ae. albopictus blood feeding with implications for understanding human biting and disease risk. In Suffolk Virginia in areas of lower human development, the likelihood of human blood feeding increased as median household income increased and human blood feeding was more likely early in the season (May-June) compared to later (July-October). Screening of the head and thorax of engorged Ae. albopictus mosquitoes by cell culture and RT-PCR resulted in a single isolate of Potosi virus. Conclusion and significance: Understanding mosquito-host interactions in nature is vital for evaluating vectorial capacity of mosquitoes. These interactions with competent reservoir hosts support transmission, maintenance, and amplification of zoonotic agents of human diseases. Results of our study in conjunction with abundance in urban/suburban settings, virus isolation from field-collected mosquitoes, and vector competence of Ae. albopictus, highlight the potential involvement of this species in the transmission of a number of arboviruses such as dengue, chikungunya, and Zika to humans. Limited interaction with avian hosts suggests that Ae. albopictus is unlikely to serve as a bridge vector of arboviruses such as West Nile and eastern equine encephalitis in the study region, but that possibility cannot be entirely ruled out. Author summary: Native to Southeast Asia, breeding populations of Aedes albopictus were first discovered in Harris County, Texas, in 1985, and as of 2017, seasonal populations of this species have been reported in more than 40 states and the District of Columbia. Aedes albopictus breed readily in natural or man-made environment where stagnant water can accumulate such as gutters, flowerpots, discarded tires, and tree holes. This mosquito species has been implicated in outbreaks of chikungunya, dengue, and Zika viruses and is a competent vector of many arboviruses including West Nile, eastern equine encephalitis, yellow fever, Rift Valley fever, and Japanese encephalitis. Aedes albopictus is regarded as an opportunistic mosquito feeding on a variety of domestic and wild mammals, birds, reptiles and amphibians; however, a preference for human blood meals has been noted in blood meal analysis of field-collected mosquitoes and in laboratory investigations. We studied vector-host interactions of Ae. albopictus in Virginia, United States using molecular methods and identified ten mammalian, three reptilian, and two avian species as blood hosts of this mosquito species. Our study clarifies the host associations of Ae. albopictus and highlights concerns about the potential role of this mosquito species in transmission of emerging and reemerging arboviruses. [ABSTRACT FROM AUTHOR]