This study assessed the presence of sialic acid α-2,3 and α-2,6 linked glycan receptors in seven avian species. The respiratory and intestinal tracts of the chicken, common quail, red-legged partridge, turkey, golden pheasant, ostrich, and mallard were tested by means of lectin histochemistry, using the lectins Maackia amurensis agglutinin II and Sambucus nigra agglutinin, which show affinity for α-2,3 and α-2,6 receptors, respectively. Additionally, the pattern of virus attachment (PVA) was evaluated with virus histochemistry, using an avian-origin H4N5 virus and a human-origin seasonal H1N1 virus. There was a great variation of receptor distribution among the tissues and avian species studied. Both α-2,3 and α-2,6 receptors were present in the respiratory and intestinal tracts of the chicken, common quail, red-legged partridge, turkey, and golden pheasant. In ostriches, the expression of the receptor was basically restricted to α-2,3 in both the respiratory and intestinal tracts and in mallards the α-2,6 receptors were absent from the intestinal tract. The results obtained with the lectin histochemistry were, in general, in agreement with the PVA. The differential expression and distribution of α-2,3 and α-2,6 receptors among various avian species might reflect a potentially decisive factor in the emergence of new viral strains.

An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic avian influenza virus (LPAIV) was carried out in red-legged partridges (Alectoris rufa) in order to study clinical signs, gross and microscopic lesions, and viral distribution in tissues and viral shedding. Birds were infected with a HPAIV subtype H7N1 (A/Chicken/Italy/5093/1999) and a LPAIV subtype H7N9 (A/Anas crecca/Spain/1460/2008). Uninoculated birds were included as contacts in both groups. In HPAIV infected birds, the first clinical signs were observed at 3 dpi, and mortality started at 4 dpi, reaching 100% at 8 dpi. The presence of viral antigen in tissues and viral shedding were confirmed by immunohistochemistry and quantitative real time RT-PCR (qRRT-PCR), respectively, in all birds infected with HPAIV. However, neither clinical signs nor histopathological findings were observed in LPAIV infected partridges. In addition, only short-term viral shedding together with seroconversion was detected in some LPAIV inoculated animals. The present study demonstrates that the red-legged partridge is highly susceptible to the H7N1 HPAIV strain, causing severe disease, mortality and abundant viral shedding and thus contributing to the spread of a potential local outbreak of this virus. In contrast, our results concerning H7N9 LPAIV suggest that the red-legged partridge is not a reservoir species for this virus.

Several studies have shown Dengue Virus (DENV) nucleic acids and/or antibodies present in Neotropical wildlife including bats, suggesting that some bat species may be susceptible to DENV infection. Here we aim to elucidate the role of house-roosting bats in the DENV transmission cycle. Bats were sampled in households located in high and low dengue incidence regions during rainy and dry seasons in Costa Rica. We captured 318 bats from 12 different species in 29 households. Necropsies were performed in 205 bats to analyze virus presence in heart, lung, spleen, liver, intestine, kidney, and brain tissue. Histopathology studies from all organs showed no significant findings of disease or infection. Sera were analyzed by PRNT90 for a seroprevalence of 21.2% (51/241), and by PCR for 8.8% (28/318) positive bats for DENV RNA. From these 28 bats, 11 intestine samples were analyzed by RT-PCR. Two intestines were DENV RNA positive for the same dengue serotype detected in blood. Viral isolation from all positive organs or blood was unsuccessful. Additionally, viral load analyses in positive blood samples by qRT-PCR showed virus concentrations under the minimal dose required for mosquito infection. Simultaneously, 651 mosquitoes were collected using EVS-CO2 traps and analyzed for DENV and feeding preferences (bat cytochrome b). Only three mosquitoes were found DENV positive and none was positive for bat cytochrome b. Our results suggest an accidental presence of DENV in bats probably caused from oral ingestion of infected mosquitoes. Phylogenetic analyses suggest also a spillover event from humans to bats. Therefore, we conclude that bats in these urban environments do not sustain DENV amplification, they do not have a role as reservoirs, but function as epidemiological dead end hosts for this virus.

The Kemp's ridley sea turtle (Lepidochelys kempi) is restricted to the warm temperate zone of the North Atlantic Ocean, whereas the olive ridley turtle (L. olivacea) is globally distributed in warm-temperate and tropical seas, including nesting colonies in the North Atlantic that nearly overlap the range of L. kempi. To explain this lopsided distribution, Pritchard (1969) proposed a scenario in which an ancestral taxon was divided into Atlantic and Pacific forms (L. kempi and L. olivacea, respectively) by the Central American land bridge. According to this model, the olive ridley subsequently occupied the Pacific and Indian Oceans and recently colonized the Atlantic Ocean via southern Africa. To assess this biogeographic model, a 470 bp sequence of the mtDNA control region was compared among 89 ridley turtles, including the sole L. kempi nesting population and 7 nesting locations across the range of L. olivacea. These data confirm a fundamental partition between L. olivacea and L. kempi (p = 0.052-0.069), shallow separations within L. olivacea (p = 0.002-0.031), and strong geographic partitioning of mtDNA lineages. The most divergent L. olivacea haplotype is observed in the Indo-West Pacific region, as are the central haplotypes in a parsimony network, implicating this region as the source of the most recent radiation of olive ridley lineage. The most common olive ridley haplotype in Atlantic samples is distinguished from an Indo-West Pacific haplotype by a single nucleotide substitution, and East Pacific samples are distinguished from the same haplotype by two nucleotide substitutions. These shallow separations are consistent with the recent invasion of the Atlantic postulated by Pritchard (1969), and indicate that the East Pacific nesting colonies were also recently colonized from the Indo-West Pacific region. Molecular clock estimates place these invasions within the last 300,000 years.

Abstract. To identify the relationship between landscape use and dengue virus (DENV) occurrence in bats, we investigated the presence of DENV from anthropogenically changed and unaltered landscapes in two Biosphere Reserves: Calakmul (Campeche) and Montes Azules (Chiapas) in southern Mexico. Spleen samples of 146 bats, belonging to 16 species, were tested for four DENV serotypes with standard reverse transcriptase polymerase chain reaction (RT-PCR) protocols. Six bats (4.1%) tested positive for DENV-2: four bats in Calakmul (two Glossophaga soricina, one Artibeus jamaicensis, and one A. lituratus) and two bats in Montes Azules (both A. lituratus). No effect of anthropogenic disturbance on the occurrence of DENV was detected; however, all three RT-PCR-positive bat species are considered abundant species in the Neotropics and well-adapted to disturbed habitats. To our knowledge, this study is the first study conducted in southeastern Mexico to identify DENV-2 in bats by a widely accepted RT-PCR protocol. The role that bats play on DENV's ecology remains undetermined.Dengue fever is an important public health concern in the tropics, 1-4 and ecological and epidemiological studies are needed to assess the role of bats and other mammals in a possible sylvatic maintenance cycle.5 Dengue viruses (DENVs) comprise four antigenically distinct but genetically related serotypes of the Flavivirus genus (Flaviviridae family). 1DENVs are positive-sense single-stranded RNA viruses that cause one of the most common infectious diseases in humans in tropical regions.2 Their transmission includes an urban endemic/epidemic cycle between Aedes aegypti mosquitoes and humans as the reservoir host and a sylvatic enzootic cycle between non-human primates and arboreal mosquitoes of the genus Aedes.3 The urban cycle is well-documented in the Neotropics, with four serotypes reported in urban areas, 1-4 whereas the sylvatic cycle has been shown in West Africa and peninsular Malaysia.5 Thus far, the sylvatic cycle has not been described in the Neotropics. However, in Bolivia, DENV seroconversions among the indigenous Ayoreo people were found in a remote area where Ae. aegypti, the primary vector, was absent. 6 This finding suggests a possible sylvatic cycle involving a different mosquito species or cross-reaction with antibodies to another flavivirus. In French Guiana, all four DENV serotypes have been identified by molecular methods in 92 wild mammals (bats, rodents, and marsupials) in all settings investigated: periurban, rural, and sparsely populated areas.7 This finding suggests that primarily urban DENV strains could infect wildlife in non-urban forested areas. 7 The role of wildlife in DENV transmission remains unknown.Bats are important reservoirs of many viruses, such as rabies viruses, Nipah viruses, and coronaviruses. [8][9][10][11][12] Flaviviridae are the second most frequently reported viral family in the order Chiroptera (13% frequency; second only to rhabdoviruses) 9 ; however, their role in the dynamics of DENVs remains poorly ...

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In order to understand the mechanism of neuroinvasion of a highly pathogenic avian influenza virus (HPAIV) into the central nervous system (CNS) of chickens, specific pathogen free chickens were inoculated with a H7N1 HPAIV. Blood, cerebrospinal fluid (CSF), nasal cavity and brain tissue samples were obtained from 1 to 4 days post-inoculation (dpi) of infected and control chickens. Viral antigen topographical distribution, presence of influenza A virus receptors in the brain, as well as, the role of the olfactory route in virus CNS invasion were studied using different immunohistochemistry techniques. Besides, viral RNA load in CSF and blood was quantified by means of a quantitative real-time reverse transcription-polymerase chain reaction. Viral antigen was observed widely distributed in the CNS, showing bilateral and symmetrical distribution in the nuclei of the diencephalon, mesencephalon and rhombencephalon. Viral RNA was detected in blood and CSF at one dpi, indicating that the virus crosses the blood-CSF-barrier early during infection. This early dissemination is possibly favoured by the presence of Siaα2,3 Gal and Siaα2,6 Gal receptors in brain vascular endothelial cells, and Siaα2,3 Gal receptors in ependymal and choroid plexus cells. No viral antigen was observed in olfactory sensory neurons, while the olfactory bulb showed only weak staining, suggesting that the virus did not use this pathway to enter into the brain. The sequence of virus appearance and the topographical distribution of this H7N1 HPAIV indicate that the viral entry occurs via the haematogenous route, with early and generalized spreading through the CSF.

Over one hundred cases of human rickettsiosis, many fatal, are reported annually across the US‐Mexico transboundary region, representing a likely undercount. Although cases are often attributed to Rickettsia rickettsii, the agent of Rocky Mountain spotted fever, multiple other Rickettsia pathogens are present in North America. We conducted multiple‐host surveillance of domestic, synanthropic, and sylvatic mammals and their ectoparasites to investigate the ecology of Rickettsia species in this region. A total of 499 mammals, including 83 dogs, 23 wild carnivores, five lagomorphs, and 388 rodents were sampled, and 413 fleas and 447 ticks belonging to 15 and 4 species, respectively, were collected during 2017 and 2018. We detected Rickettsia spp. DNA in one blood sample of coyote (Canis latrans), 11 ear tissues of rodents (10.6%), and 79 ectoparasites (9.5%). Of the 64 Rickettsia‐positive fleas, 54 were Echidnophaga gallinacea and 10 were Pulex simulans, while of the 15 ticks, 11 were Rhipicephalus sanguineus s.l. and four Ixodes pacificus. The DNA sequence alignment of gltA and ompB regions revealed one and ten genetic variants of Rickettsia spp., respectively. These variants were clustered in clades of zoonotic species (R. felis, R. massiliae, R. parkeri, R. rickettsii, and R. typhi) and organisms of unknown pathogenic significance (R. asembonensis and Candidatus Rickettsia tarasevichiae). The finding of a coyote infected with R. rickettsii and the multiple zoonotic SFG rickettsial agents in the study area suggest that: 1) wild canids could serve as an amplifying host for RMSF, an alternate host for Rh. sanguineus s.l. ticks, and a means to spread infection and ticks over large areas; and 2) at least some of the human rickettsiosis cases attributed to R. rickettsii could be caused by other Rickettsia species. This study strongly supports the importance of multiple‐host and vector eco‐epidemiological studies and the One Health approach to better understand disease in a RMSF‐epidemic region.