Antibodies against medically relevant arthropod-borne viruses in the ubiquitous African rodent Mastomys natalensis.

Over the past decades, the number of arthropod-borne virus (arbovirus) outbreaks has increased worldwide. Knowledge regarding the sylvatic cycle (i.e., non-human hosts/environment) of arboviruses is limited, particularly in Africa, and the main hosts for virus maintenance are unknown. Previous studies have shown the presence of antibodies against certain arboviruses (i.e., chikungunya-, dengue-, and Zika virus) in African non-human primates and bats. We hypothesize that small mammals, specifically rodents, may function as amplifying hosts in anthropogenic environments. The detection of RNA of most arboviruses is complicated by the viruses' short viremic period within their hosts. An alternative to determine arbovirus hosts is by detecting antibodies, which can persist several months. Therefore, we developed a high-throughput multiplex immunoassay to detect antibodies against 15 medically relevant arboviruses. We used this assay to assess approximately 1,300 blood samples of the multimammate mouse, Mastomys natalensis from Tanzania. In 24% of the samples, we detected antibodies against at least one of the tested arboviruses, with high seroprevalences of antibodies reacting against dengue virus serotype one (7.6%) and two (8.4%), and chikungunya virus (6%). Seroprevalence was higher in females and increased with age, which could be explained by inherent immunity and behavioral differences between sexes, and the increased chance of exposure to an arbovirus with age. We evaluated whether antibodies against multiple arboviruses co-occur more often than randomly and found that this may be true for some members of the Flaviviridae and Togaviridae. In conclusion, the development of an assay against a wide diversity of medically relevant arboviruses enabled the analysis of a large sample collection of one of the most abundant African small mammals. Our findings highlight that Mastomys natalensis is involved in the transmission cycle of multiple arboviruses and provide a solid foundation to better understand the role of this ubiquitous rodent in arbovirus outbreaks.

Behavioral Changes of Some Arboviral Vectors in Zika Forest: A Concern for Emerging and Re-Emerging Diseases in Uganda.

Background: The increasing reports on emerging/re-emerging arboviral disease outbreaks or epidemics in Sub-Saharan Africa have been impacted by factors, including the changing climate plus human activities that have resulted in land cover changes. These factors influence the prevalence, incidence, behavior, and distribution of vectors and vector-borne diseases. In this study, we assessed the potential effect of land cover changes on the distribution and oviposition behavior of some arboviral vectors in Zika forest, Uganda, which has decreased by an estimated 7 hectares since 1952 due to an increase in anthropogenic activities in the forest and its periphery. Materials and Methods: Immature mosquitoes were collected using bamboo pots and placed at various levels of a steel tower in the forest and at different intervals from the forest periphery to areas among human dwellings. Collections were conducted for 20 months. Results and Conclusion: Inside the forest, 22,280 mosquitoes were collected belonging to four arboviral vectors: Aedes aegypti, Aedes africanus, Aedes apicoargenteus, and Aedes cumminsii. When compared with similar studies conducted in the forest in 1964, there was a change from a sylvatic to a tendency of peridomestic behavior in A. africanus, which was now collected among human dwellings. There was an unexpected change in the distribution of A. aegypti, which was not only collected outside the forest as in previous reports but also collected in the forest. Conversely, A. cumminsii originally collected in the forest expanded its ranges with collections outside the forest in this study. Aedes simpsoni maintained its distribution range outside the forest among agricultural sites. We suspect that land cover changes were favorable to most of the arboviral vectors hence enhancing their proliferation and habitat range. This potentially increases the transmission of arboviral diseases in the area, hence impacting the epidemiology of emerging/remerging diseases in Uganda.

Occurrence and surveillance of Taiwanese bovine arboviruses using hematophagous insects in dairy farms during 2012-2019.

Culicoides-borne viruses are an important arbovirus group causing bovine diseases. During 2012-2019, 2,525 pools consisting of 108,937 specimens of vectors were subjected to PCR detection of bovine arbovirus belonging to Orthobunyavirus, Orbivirus, and Ephemerovirus. Twelve virus RNAs, of which 6, that is, Shuni virus, Shamonda virus, and Sathuperi virus in Orthobunyavirus and Sathuvachari virus and epizootic hemorrhagic disease virus serotypes 4 and 7 in Orbivirus were detected for the first time in the area. Potential vector species were evaluated by the minimum infection rate, and the population abundance of Culicoides oxystoma, Culex tritaeniorhynchus, and Anopheles sinensis indicated that they were the main potential vector species in dairy farms in Taiwan.

Arboviruses in Free-Ranging Birds and Hematophagous Arthropods (Diptera, Nematocera) from Forest Remnants and Urbanized Areas of an Environmental Protection Area in the Amazon Biome.

The rapid and disorderly urbanization in the Amazon has resulted in the insertion of forest fragments into cities, causing the circulation of arboviruses, which can involve hematophagous arthropods and free-ranging birds in the transmission cycles in urban environments. This study aimed to evaluate the circulation of arboviruses in free-ranging birds and hematophagous arthropods captured in an Environmental Protection Area in the Belem metropolitan area, Brazil. Birds were captured using mist nets, and hematophagous arthropods were collected using a human protected attraction technique and light traps. The birds' sera were subjected to a hemagglutination inhibition test to detect antibodies against 29 arbovirus antigens. Arthropod macerates were inoculated into C6/36 and VERO cell cultures to attempt viral isolation and were tested using indirect immunofluorescence, subsequent genetic sequencing and submitted for phylogenetic analysis. Four bird sera were positive for arbovirus, and one batch of Psorophora ferox was positive for Flavivirus on viral isolation and indirect immunofluorescence. In addition, the Ilheus virus was detected in the sequencing and phylogenetic analysis. The presence of antibodies in sera from free-ranging birds and the isolation of Ilheus virus in Psorophora ferox indicate the circulation of arboviruses in forest remnants in the urban center of Belem.

North American Arboviruses and White-Tailed Deer (Odocoileus virginianus): Associated Diseases and Role in Transmission.

Background: Arboviral disease is of increasing concern to human and animal health professionals as emerging and re-emerging arboviruses are more frequently recognized. Wildlife species are known to play a role in the transmission and maintenance of arboviruses and infections can result in morbidity and mortality in wildlife hosts. Materials and Methods: In this review, we detail existing evidence of white-tailed deer (Odocoileus virginianus) as an important host to a diverse collection of arboviruses and evaluate the utility of this species as a resource to better understand the epidemiology of related viral diseases. Results: Relevant veterinary and zoonotic viral pathogens endemic to North America include epizootic hemorrhagic disease virus, bluetongue virus, orthobunyaviruses, vesicular stomatitis virus, Eastern equine encephalitis virus, West Nile virus, and Powassan virus. Exotic viral pathogens that may infect white-tailed deer are also identified with an emphasis on zoonotic disease risks. The utility of this species is attributed to the high degree of contact with humans and domestic livestock and evidence of preferential feeding by various insect vectors. Conclusions: There is mounting evidence that white-tailed deer are a useful, widely available source of information regarding arboviral circulation, and that surveillance and monitoring of deer populations would be of value to the understanding of certain viral transmission dynamics, with implications for improving human and domestic animal health.

Alphavirus Identification in Neotropical Bats.

Alphaviruses (Togaviridae) are arthropod-borne viruses responsible for several emerging diseases, maintained in nature through transmission between hematophagous arthropod vectors and susceptible vertebrate hosts. Although bats harbor many species of viruses, their role as reservoir hosts in emergent zoonoses has been verified only in a few cases. With bats being the second most diverse order of mammals, their implication in arbovirus infections needs to be elucidated. Reports on arbovirus infections in bats are scarce, especially in South American indigenous species. In this work, we report the genomic detection and identification of two different alphaviruses in oral swabs from bats captured in Northern Uruguay. Phylogenetic analysis identified Río Negro virus (RNV) in two different species: Tadarida brasiliensis (n = 6) and Myotis spp. (n = 1) and eastern equine encephalitis virus (EEEV) in Myotis spp. (n = 2). Previous studies of our group identified RNV and EEEV in mosquitoes and horse serology, suggesting that they may be circulating in enzootic cycles in our country. Our findings reveal that bats can be infected by these arboviruses and that chiropterans could participate in the viral natural cycle as virus amplifiers or dead-end hosts. Further studies are warranted to elucidate the role of these mammals in the biological cycle of these alphaviruses in Uruguay.

Late effects of Beauveria bassiana on larval stages of Aedes aegypti Linneo, 1762 (Diptera: Culicidae) / Efeitos tardios de Beauveria bassiana nos estágios larvais de Aedes aegypti Linneo, 1762 (Diptera: Culicidae)

Aedes aegypti is a culicide that has gained relevance over the years due to its ability to transmit various viruses that cause diseases in humans that all the years cause high mortality rates in the world population. The main problem is that Ae. aegypti has managed to establish and maintain a close relationship with humans and their habitat, which is why the search for alternatives to control vector populations becomes imperative. The objective of the present work was to study the effects of two Beauveria bassiana strains on Aedes aegypti. Third instar larvae of Ae. aegypti in 250 mL plastic containers were inoculated with the GHA and NB3 strains at different concentrations (1.5 × 104, 1.5× 105, 1.5 × 106 and 1.5 × 107 conidia/mL). The NB3 strain presented highest mortality values with 63% in the highest concentration i.e., 1.5 × 107, while for the GHA strain the highest mortality value was 30.7% at the same concentration. The results showed significant difference in mortality with respect to the strain and days post treatment (P = 0.0001), but not with respect to the conidial concentration (P = 0.634). The average mortality of larvae per day for the NB3 for different concentrations ranged from 20 to 25 larvae per day, while for the GHA daily mortality ranged from 5 to 12 larvae. In post-treatment mortality, the highest mortality was recorded in the third stage larvae for the NB3, while for GHA the highest percentage mortality was observed in individuals who managed to reach the adult state. The findings of the current research depicted the noteworthy role of B. bassiana for the management of an important vector of human disease.

O Aedes aegypti é um culicida que vem ganhando relevância ao longo dos anos devido à sua capacidade de transmitir diversos vírus causadores de doenças em humanos que ao longo dos anos ocasionam altas taxas de mortalidade na população mundial. O principal problema é que Ae. aegypti tem conseguido estabelecer e manter uma relação próxima com o homem e seu habitat, por isso a busca por alternativas para o controle das populações de vetores torna-se imperativa. O objetivo do presente trabalho foi estudar os efeitos de duas cepas de Beauveria bassiana sobre Ae. aegypti. Larvas de terceiro instar de Ae. aegypti em recipientes plásticos de 250 mL foram inoculados com as cepas GHA e NB3 em diferentes concentrações (1.5 × 104, 1.5 × 105, 1.5 × 106 e 1.5 × 107 conídios/mL). A cepa NB3 apresentou os maiores valores de mortalidade com 63% na concentração mais alta, ou seja, 1.5 × 107, enquanto para a cepa GHA o maior valor de mortalidade foi 30.7% na mesma concentração. Os resultados mostraram diferença significativa na mortalidade com relação à cepa e dias pós-tratamento (P = 0.0001), mas não com relação à concentração de conídios (P = 0.634). A mortalidade média de larvas por dia para o NB3 para diferentes concentrações variou de 20 a 25 larvas por dia, enquanto para o GHA a mortalidade diária variou de 5 a 12 larvas. Na mortalidade pós-tratamento, a maior mortalidade foi registrada nas larvas de terceiro estágio para o NB3, enquanto para o GHA o maior percentual de mortalidade foi observado em indivíduos que conseguiram atingir o estado adulto. Os resultados da pesquisa atual retratam o papel notável de B. bassiana no manejo de um importante vetor de doenças humanas.

Mosquito-borne arboviruses in Uganda: history, transmission and burden.

Mosquito-transmitted arboviruses constitute a large proportion of emerging infectious diseases that are both a public health problem and a threat to animal populations. Many such viruses were identified in East Africa, a region where they remain important and from where new arboviruses may emerge. We set out to describe and review the relevant mosquito-borne viruses that have been identified specifically in Uganda. We focused on the discovery, burden, mode of transmission, animal hosts and clinical manifestation of those previously involved in disease outbreaks. A search for mosquito-borne arboviruses detected in Uganda was conducted using search terms 'Arboviruses in Uganda' and 'Mosquitoes and Viruses in Uganda' in PubMed and Google Scholar in 2020. Twenty-four mosquito-borne viruses from different animal hosts, humans and mosquitoes were documented. The majority of these were from family Peribunyaviridae, followed by Flaviviridae, Togaviridae, Phenuiviridae and only one each from family Rhabdoviridae and Reoviridae. Sixteen (66.7%) of the viruses were associated with febrile illnesses. Ten (41.7%) of them were first described locally in Uganda. Six of these are a public threat as they have been previously associated with disease outbreaks either within or outside Uganda. Historically, there is a high burden and endemicity of arboviruses in Uganda. Given the many diverse mosquito species known in the country, there is also a likelihood of many undescribed mosquito-borne viruses. Next generation diagnostic platforms have great potential to identify new viruses. Indeed, four novel viruses, two of which were from humans (Ntwetwe and Nyangole viruses) and two from mosquitoes (Kibale and Mburo viruses) were identified in the last decade using next generation sequencing. Given the unbiased approach of detection of viruses by this technology, its use will undoubtedly be critically important in the characterization of mosquito viromes which in turn will inform other diagnostic efforts.

Mosquito-borne arboviruses in Uganda: history, transmission and burden.

Mosquito-transmitted arboviruses constitute a large proportion of emerging infectious diseases that are both a public health problem and a threat to animal populations. Many such viruses were identified in East Africa, a region where they remain important and from where new arboviruses may emerge. We set out to describe and review the relevant mosquito-borne viruses that have been identified specifically in Uganda. We focused on the discovery, burden, mode of transmission, animal hosts and clinical manifestation of those previously involved in disease outbreaks. A search for mosquito-borne arboviruses detected in Uganda was conducted using search terms 'Arboviruses in Uganda' and 'Mosquitoes and Viruses in Uganda' in PubMed and Google Scholar in 2020. Twenty-four mosquito-borne viruses from different animal hosts, humans and mosquitoes were documented. The majority of these were from family Peribunyaviridae, followed by Flaviviridae, Togaviridae, Phenuiviridae and only one each from family Rhabdoviridae and Reoviridae. Sixteen (66.7 %) of the viruses were associated with febrile illnesses. Ten (41.7 %) of them were first described locally in Uganda. Six of these are a public threat as they have been previously associated with disease outbreaks either within or outside Uganda. Historically, there is a high burden and endemicity of arboviruses in Uganda. Given the many diverse mosquito species known in the country, there is also a likelihood of many undescribed mosquito-borne viruses. New generation diagnostic platforms have great potential to identify new viruses. Indeed, four novel viruses, two of which were from humans (Ntwetwe and Nyangole viruses) and two from mosquitoes (Kibale and Mburo viruses) including the 2010 yellow fever virus (YFV) outbreak were identified in the last decade using next generation sequencing. Given the unbiased approach of detection of viruses by this technology, its use will undoubtedly be critically important in the characterization of mosquito viromes which in turn will inform other diagnostic efforts.

Sindbis virus infection of mosquito species in the wetlands of northwestern Iran and modeling the probable ecological niches of SINV vectors in the country.

Sindbis virus (SINV) and Chikungunya virus (CHIKV) are among the most widely spread mosquito-borne viruses worldwide. Due to the key role of mosquitoes in the transmission cycle of vector-borne diseases, models such as Maximum Entropy (MaxEnt) have been used in recent years to predict the environmental suitability and ecological niches of mosquito vectors. Infection of three mosquito species (Anopheles maculipennis s.l., Culex tritaeniorhynchus, and Culiseta longiareolata) with CHIKV has recently been reported in Iran. However, given the importance of vector-borne diseases in the country, there is a need for extensive studies on the infection of mosquitoes with CHIKV and SINV in different areas of the country. Accordingly, the current research was conducted to investigate the infection of mosquitoes with the two aforementioned viruses in the northwestern part of Iran and also to model the ecological niches of the vectors of these mosquito-borne viruses in the country. In the current study, 4639 mosquito specimens, consisting of 2515 adults and 2124 larvae, were collected from the wetlands of West Azerbaijan Province and identified. Ten species belonging to four genera were identified in this study. The specimens were allocated to 149 pools for the determination of infection with CHIKV and SINV. The amplification pattern of five pools comprising two mosquito species (Culex pipiens complex and Cx. Theileri) corresponded to the reference strain of SINV, and the isolates were sequenced to confirm the presence of SINV genome. No cases of CHIKV infection were found among the 149 examined mosquito pools. Data on the distribution of Cx. Pipiens complex and Cx. Theileri were mapped using ArcMap 10.5. Prediction maps of the presence probability for these species revealed that they are most likely to be found in and spread to the north, northwest, south, and southeastern areas of the country and in areas with abundant water resources. For the first time in Iran, our study investigated the presence probability of SINV vectors using ecological niche modeling. Ecological niche profiling showed that the most suitable habitats for Cx. pipiens are mainly concentrated in the north and northwestern parts of the country, whereas Cx. theileri is mostly located in the northwest and western regions. However, there were some other areas of low suitability for these two species in the country. Further studies in a broader geographical area with more species of mosquitos and the determination of infection with other mosquito-borne viruses can provide a clear understanding of the potential spread of mosquito-borne diseases in various regions of Iran.

Development and lifespan of Culicoides obsoletus s.s. (Meigen) and other livestock-associated species reared at different temperatures under laboratory conditions.

Culicoides Latreille (Diptera: Ceratopogonidae) transmit arboviruses affecting wild and domestic ruminants such as bluetongue (BTV) and Schmallenberg virus (SBV). The sub-adult development and lifespan of Culicoides obsoletus s.s. (Meigen), Culicoides circumscriptus Kieffer and Culicoides paolae Boorman were examined at three different temperatures under laboratory conditions. Insects were collected from field between spring and autumn 2015 in two livestock farms located in Majorca (Spain). Gravid females were held individually at 18, 25 or 30 °C. Low temperatures increased the adult lifespan, time to oviposit and rate of development, whereas high temperatures increased the number of eggs, successful pupation and adult emergence as well as the larvae growth rate. The results showed that C. obsoletus s.s. have optimum development at 18 °C, whereas the optimal rearing temperature for C. circumscriptus and C. paolae was under warmer conditions of 25-30 °C. Variations in temperature/humidity and assays with different materials and substrates for oviposition should be considered in future studies. Understanding the requirements of the different species of Culicoides optimizing the results should be of special interest for predicting environmental change effects on these species, in addition to determining the rearing conditions for candidate Culicoides vectors.

Vertebrate-Aedes aegypti and Culex quinquefasciatus (Diptera)-arbovirus transmission networks: Non-human feeding revealed by meta-barcoding and next-generation sequencing.

BACKGROUND: Aedes aegypti mosquito-borne viruses including Zika (ZIKV), dengue (DENV), yellow fever (YFV), and chikungunya (CHIKV) have emerged and re-emerged globally, resulting in an elevated burden of human disease. Aedes aegypti is found worldwide in tropical, sub-tropical, and temperate areas. The characterization of mosquito blood meals is essential to understand the transmission dynamics of mosquito-vectored pathogens. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report Ae. aegypti and Culex quinquefasciatus host feeding patterns and arbovirus transmission in Northern Mexico using a metabarcoding-like approach with next-generation deep sequencing technology. A total of 145 Ae. aegypti yielded a blood meal analysis result with 107 (73.8%) for a single vertebrate species and 38 (26.2%) for two or more. Among the single host blood meals for Ae. aegypti, 28.0% were from humans, 54.2% from dogs, 16.8% from cats, and 1.0% from tortoises. Among those with more than one species present, 65.9% were from humans and dogs. For Cx. quinquefasciatus, 388 individuals yielded information with 326 (84%) being from a single host and 63 (16.2%) being from two or more hosts. Of the single species blood meals, 77.9% were from dogs, 6.1% from chickens, 3.1% from house sparrows, 2.4% from humans, while the remaining 10.5% derived from other 12 host species. Among those which had fed on more than one species, 11% were from dogs and humans, and 89% of other host species combinations. Forage ratio analysis revealed dog as the most over-utilized host by Ae. aegypti (= 4.3) and Cx. quinquefasciatus (= 5.6) and the human blood index at 39% and 4%, respectively. A total of 2,941 host-seeking female Ae. aegypti and 3,536 Cx. quinquefasciatus mosquitoes were collected in the surveyed area. Of these, 118 Ae. aegypti pools and 37 Cx. quinquefasciatus pools were screened for seven arboviruses (ZIKV, DENV 1-4, CHIKV, and West Nile virus (WNV)) using qRT-PCR and none were positive (point prevalence = 0%). The 95%-exact upper limit confidence interval was 0.07% and 0.17% for Ae. aegypti and Cx. quinquefasciatus, respectively. CONCLUSIONS/SIGNIFICANCE: The low human blood feeding rate in Ae. aegypti, high rate of feeding on mammals by Cx. quinquefasciatus, and the potential risk to transmission dynamics of arboviruses in highly urbanized areas of Northern Mexico is discussed.

Laboratory transmission potential of British mosquitoes for equine arboviruses.

BACKGROUND: There has been no evidence of transmission of mosquito-borne arboviruses of equine or human health concern to date in the UK. However, in recent years there have been a number of outbreaks of viral diseases spread by vectors in Europe. These events, in conjunction with increasing rates of globalisation and climate change, have led to concern over the future risk of mosquito-borne viral disease outbreaks in northern Europe and have highlighted the importance of being prepared for potential disease outbreaks. Here we assess several UK mosquito species for their potential to transmit arboviruses important for both equine and human health, as measured by the presence of viral RNA in saliva at different time points after taking an infective blood meal. RESULTS: The following wild-caught British mosquitoes were evaluated for their potential as vectors of zoonotic equine arboviruses: Ochlerotatus detritus for Venezuelan equine encephalitis virus (VEEV) and Ross River virus (RRV), and Culiseta annulata and Culex pipiens for Japanese encephalitis virus (JEV). Production of RNA in saliva was demonstrated at varying efficiencies for all mosquito-virus pairs. Ochlerotatus detritus was more permissive for production of RRV RNA in saliva than VEEV RNA. For RRV, 27.3% of mosquitoes expectorated viral RNA at 7 days post-infection when incubated at 21 °C and 50% at 24 °C. Strikingly, 72% of Cx. pipiens produced JEV RNA in saliva after 21 days at 18 °C. For some mosquito-virus pairs, infection and salivary RNA titres reduced over time, suggesting unstable infection dynamics. CONCLUSIONS: This study adds to the number of Palaearctic mosquito species that demonstrate expectoration of viral RNA, for arboviruses of importance to human and equine health. This work adds to evidence that native mosquito species should be investigated further for their potential to vector zoonotic mosquito-borne arboviral disease of equines in northern Europe. The evidence that Cx. pipiens is potentially an efficient laboratory vector of JEV at temperatures as low as 18 °C warrants further investigation, as this mosquito is abundant in cooler regions of Europe and is considered an important vector for West Nile Virus, which has a comparable transmission ecology.

Sloths host Anhanga virus-related phleboviruses across large distances in time and space.

Sloths are genetically and physiologically divergent mammals. Phleboviruses are major arthropod-borne viruses (arboviruses) causing disease in humans and other animals globally. Sloths host arboviruses, but virus detections are scarce. A phlebovirus termed Anhanga virus (ANHV) was isolated from a Brazilian Linnaeus's two-toed sloth (Choloepus didactylus) in 1962. Here, we investigated the presence of phleboviruses in sera sampled in 2014 from 74 Hoffmann's two-toed (Choloepus hoffmanni, n = 65) and three-toed (Bradypus variegatus, n = 9) sloths in Costa Rica by broadly reactive RT-PCR. A clinically healthy adult Hoffmann's two-toed sloth was infected with a phlebovirus. Viral load in this animal was high at 8.5 × 107  RNA copies/ml. The full coding sequence of the virus was determined by deep sequencing. Phylogenetic analyses and sequence distance comparisons revealed that the new sloth virus, likely representing a new phlebovirus species, provisionally named Penshurt virus (PEHV), was most closely related to ANHV, with amino acid identities of 93.1%, 84.6%, 94.7% and 89.0% in the translated L, M, N and NSs genes, respectively. Significantly more non-synonymous mutations relative to ANHV occurred in the M gene encoding the viral glycoproteins and in the NSs gene encoding a putative interferon antagonist compared to L and N genes. This was compatible with viral adaptation to different sloth species and with micro-evolutionary processes associated with immune evasion during the genealogy of sloth-associated phleboviruses. However, gene-wide mean dN/dS ratios were low at 0.02-0.15 and no sites showed significant evidence for positive selection, pointing to comparable selection pressures within sloth-associated viruses and genetically related phleboviruses infecting hosts other than sloths. The detection of a new phlebovirus closely-related to ANHV, in sloths from Costa Rica fifty years after and more than 3,000 km away from the isolation of ANHV confirmed the host associations of ANHV-related phleboviruses with the two extant species of two-toed sloths.

Infection of Western Gray Kangaroos (Macropus fuliginosus) with Australian Arboviruses Associated with Human Infection.

More than 75 arboviruses (arthropod-borne viruses) have been identified in Australia. While Alfuy virus (ALFV), Barmah Forest virus (BFV), Edge Hill virus (EHV), Kokobera virus (KOKV), Murray Valley encephalitis virus (MVEV), Sindbis virus (SINV), Ross River virus (RRV), Stratford virus (STRV), and West Nile virus strain Kunjin (KUNV) have been associated with human infection, there remains a paucity of data regarding their respective transmission cycles and any potential nonhuman vertebrate hosts. It is likely that these viruses are maintained in zoonotic cycles involving native animals rather than solely by human-to-human transmission. A serosurvey (n = 100) was undertaken to determine the prevalence of neutralizing antibodies against a panel of Australian arboviruses in western gray kangaroos (Macropus fuliginosus) obtained from 11 locations in the midwest to southwest of Western Australia. Neutralizing antibodies against RRV were detected in 25%, against BFV in 14%, and antibodies to both viruses in 34% of serum samples. The prevalence of antibodies against these two viruses was the same in males and females, but higher in adult than in subadult kangaroos (p < 0.05). Twenty-one percent of samples had neutralizing antibodies against any one or more of the flaviviruses ALFV, EHV, KOKV, MVEV, and STRV. No neutralizing antibodies against SINV and KUNV were detected. If this sample of kangaroo sera was representative of the broader Australian population of macropods, it suggests that they are common hosts for RRV and BFV. The absence or low seroprevalence of antibodies against the remaining arboviruses suggests that they are not prevalent in the region or that kangaroos are not commonly infected with them. The detection of neutralizing antibodies to MVEV requires further investigation as this virus has not been identified previously so far south in Western Australia.

Serological evidence of arboviruses and coccidia infecting horses in the Amazonian region of Brazil.

BACKGROUND: Arboviruses and protozoans can cause neurologic disorders in horses. In Brazilian Amazon, several horses presenting signs compatible with disorders caused by these infectious agents have been observed. OBJECTIVE: To contribute to the knowledge of this epidemiological picture, we sought to construct a serological diagnostic panel for neurotrophic infectious agents in local horses. MATERIAL AND METHODS: A total of 213 blood samples from horses were collected from 29 farms in three municipalities. Samples were evaluated and considered positive when they met the following criteria: titers ≥ 1:80 with the indirect fluorescent antibody test (IFAT) for apicomplexan protozoans; positive recombinant enzyme-linked immunosorbent assay (ELISA) with subsequent titers ≥ 1:10 by the PRNt for viruses; and detection under direct microscopic examination for Trypanosoma evansi. RESULTS: No horses were found to be infected by T. evansi, and only two were infected Toxoplasma gondii and/or Neospora spp. The highest protozoan infection rate was observed for Sarcocystis neurona (40.3%; n = 86/213). Among the positive ELISA samples tested by the plaque reduction neutralization test (PRNT90), 92% (n = 76/83) were positive for St Louis Encephalitis virus, 43% (n = 6/14) were positive for West Nile virus and 33% (n = 16/48) were positive for Mayaro virus. Eighteen percent (n = 39/213) of horses were co-infected by S. neurona and at least one arbovirus, particularly SLEV and/or MAYV. CONCLUSION: Samples positive for SLEV associated with S. neurona, including samples from horses that had recovered from neurological signs were frequent, and must be considered when investigating the possible causes of neurological diseases in South Roraima horses.

Sylvatic cycles of arboviruses in non-human primates.

Arboviruses infecting people primarily exist in urban transmission cycles involving urban mosquitoes in densely populated tropical regions. For dengue, chikungunya, Zika and yellow fever viruses, sylvatic (forest) transmission cycles also exist in some regions and involve non-human primates and forest-dwelling mosquitoes. Here we review the investigation methods and available data on sylvatic cycles involving non-human primates and dengue, chikungunya, Zika and yellow fever viruses in Africa, dengue viruses in Asia and yellow fever virus in the Americas. We also present current putative data that Mayaro, o'nyong'nyong, Oropouche, Spondweni and Lumbo viruses exist in sylvatic cycles.

Post-Epidemic Distribution of Schmallenberg Virus in Culicoides Arbovirus Vectors in Poland.

Pooled samples of female and male Culicoides midges (5146 and 332 pools, respectively) that corresponded to a total number of 124,957 specimens were collected between 2013-2017 in the vicinity of cattle barns that were distributed throughout Poland were analyzed for the presence of Schmallenberg virus (SBV) RNA. Sixty-six pools tested positive (1.2%) with mean Ct value of 34.95. The maximum likelihood estimated infection rate (MLE) was calculated at 0.53 per 1000 individuals; however, it peaked in 2016 with MLE of 3.7. Viral RNA was detected in C. obsoletus/scoticus complex, C. punctatus, and C. pulicaris pools. Moreover, viral material was present in nulliparous (virgin) Culicoides females (MLE 0.27) and for the first time reported in males (MLE 0.34), which suggests the possibility of transovarial route of SBV or virus RNA transmission, as both do not fed on host blood. The accuracy of targeted versus random SBV surveillance in Culicoides vectors was compared. The relationship between infection rate (expressed as minimum infection rate; MIR), in addition to MLE, was compared with the density of virus infected midges (DIM). In conclusion, the SBV infection rate in the vector was significantly higher in 2016 as compared to other surveillance years; this is consistent with the simultaneous increase in SBV seroprevalence (seroconversion) in ruminants during the same year.

Arbovirus surveillance using FTATM cards in modified CO2 -baited encephalitis virus surveillance traps in the Northern Territory, Australia.

In 2016, modified CO2 -baited encephalitis virus surveillance (EVS) traps were evaluated for flavivirus surveillance in the Northern Territory, Australia. The traps were fitted with honey-soaked nucleic acid preservation cards (FTATM ) for mosquitoes to expectorate virus while feeding on the cards. Cards were tested for the presence of selected arboviruses, with two cards testing positive for Kunjin virus and Alfuy, while sentinel chickens tested in parallel also showed Kunjin virus activity at the same time. The results from the cards and vector mosquito feeding rates indicate that CO2 -baited EVS traps coupled with honey-baited FTATM cards are an effective tool for broad-scale arbovirus surveillance.

Culicoides Biting Midges-Underestimated Vectors for Arboviruses of Public Health and Veterinary Importance.

Culicoides biting midges, small hematophagous dipterans, are the demonstrated or putative vectors of multiple arboviruses of veterinary and public health importance. Despite its relevance in disease spread, the ceratopogonid genus Culicoides is still a largely neglected group of species, predominantly because the major human-affecting arboviruses are considered to be transmitted by mosquitoes. However, when a pathogen is detected in a certain vector species, a thorough search for further vectors often remains undone and, therefore, the relevant vector species may remain unknown. Furthermore, for many hematophagous arthropods, true vector competence is often merely suspected and not experimentally proven. Therefore, we aim to illuminate the general impact of Culicoides biting midges and to summarize the knowledge about biting midge-borne disease agents using the order Bunyavirales, the largest and most diverse group of RNA viruses, as an example. When considering only viruses evidentially transmitted by Culicoides midges, the Simbu serogroup (genus Orthobunyavirus) is presumably the most important group within the virus order. Its members are of great veterinary importance, as a variety of simbuviruses, e.g., the species Akabane orthobunyavirus or Schmallenberg orthobunyavirus, induces severe congenital infections in pregnant animals. The major zoonotic representative of this serogroup occurs in South and Central America and causes the so-called Oropouche fever, an acute febrile illness in humans.