Dataset of assembly and annotation of the mitogenomes of Triatoma dimidiata and Triatoma huehuetenanguensis captured from Yucatán, México.

Triatoma dimidiata is a species complex, and its members are responsible for the transmission of Trypanosoma cruzi, the causative agent of Chagas disease. We present the assembly and annotation of the mitogenome of the Triatoma dimidiata (Latreille, 1811) and Triatoma huehuetenanguensis Lima-Cordón & Justi, 2019. The mitochondrial genomes were successfully sequenced using the Illumina Nextseq 500 platform, 2×75 cycles, and 5 million reads per sample. Contigs were assembled and annotated using the reference genomes of T. dimidiata and T. huehuetenanguensis available in Genbank (NC_002609 and NC_050325.1, respectively). The mitogenomes of T. dimidiata have lengths of 17,008 bp, while those of T. huehuetenanguensis are 15,910 bp and 15,909 bp. The genome comprises 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region. The mitogenomes will be valuable to scholars and students focused on integrative taxonomy, phylogeography, and evolutionary studies of the Triatoma dimidiata complex and the transmission of Chagas diseases.

Congenital Transmission of Trypanosoma cruzi in Naturally Infected Dogs.

Background: Congenital transmission (CT) of Trypanosoma cruzi in dogs has not been clearly demonstrated, even though dogs are important reservoirs of this agent. Materials and Methods: Seventeen late pregnant dogs seropositive for T. cruzi were selected, and a total of 84 fetuses were obtained. Blood and heart tissues from the fetuses and dams, and placental tissue from dam were collected. All tissues were analyzed by quantitative polymerase chain reaction (qPCR) for T. cruzi DNA (TcDNA) and inflammatory infiltrate and pathology by histological examination. CT was determined when physical, histological, or molecular evidence of T. cruzi was detected in blood or tissues of the fetuses. Results: A general transmission frequency of 59% was found, and 0.20 ± 0.24 of fetuses per litter were infected. Dams that were qPCR positive for TcDNA in cardiac tissue or blood displayed a transmission frequency of 100% and 67%, respectively. The highest parasite burden was noted in dams that were positive for TcDNA in both blood (82E-01 ± 1.54E-01) and cardiac (5.28E+03 ± 8.85E+03) tissues. In fetuses, higher parasitic burden in blood and cardiac tissue was found in those carried by dams that were seropositive and qPCR positive for TcDNA in cardiac tissue and blood. No amastigote nests were recorded in the cardiac tissue of fetuses in the histopathological studies, but typical lesions of T. cruzi infection were identified in all fetuses where CT occurred. Conclusions: CT of T. cruzi occurred at a high frequency in naturally infected pregnant dogs from the endemic areas.

Mosquito Fauna in the Mangroves of Yucatan, Mexico, and Identification of Alphavirus RNA.

The surveillance of arboviruses in mangrove mosquitoes is a neglected topic in Mexico. The Yucatan State is part of a peninsula and, therefore, is rich in mangroves along its coast. The purpose of the study was to identify alphavirus in the mosquito fauna of mangroves. Mosquitoes were captured in mangrove settings in seven communities in Yucatan between June 2019 and August 2021. From 1900 to 2200 h and from 0500 to 0800 h, mosquitoes were captured with a backpack-mounted aspirator. In total, 3,167 female mosquitoes of five genera and nine species were captured. Aedes taeniorhynchus and Anopheles crucians were the most abundant mosquitoes collected. Mosquitoes were sorted into 210 pools and tested by reverse transcription-polymerase chain reaction for alphavirus ribonucleic acid (RNA). Alphavirus RNA was found in Ae. taeniorhynchus, An. pseudopunctipennis, and An. crucians collected in the Celestun Mangrove. The community is part of the Ria Celestun Biosphere Reserve, and the presence arbovirus-infected mosquitoes could pose a health risk to residents and visitors alike in the area.

Mites associated with budgerigars Melopsittacus undulatus (Psittaciformes: Psittacidae) and the first report of Ornithonyssus bursa (Mesostigmata: Macronyssidae) in Mexico.

Background: Hematophagous mites affect numerous bird species, causing severe injuries to the budgerigars. Some species can cause dermatitis in humans. Aims: The purpose was to morphologically identify the mites related to budgerigars (Melopsittacus undulatus) and their nests in Yucatan, Mexico. Methods: In May 2022, a private budgerigar hatchery was visited and mites were collected from the bodies of the birds and their nests. The morphological traits of the mites were confirmed by scanning electron microscopy. Results: Four of 30 birds showed severe clinical signs of mite infestation. The Budgerigars revealed lesions in the cere, nostrils, eyelids, beak, and paws. The bird's skin showed signs of dryness and beige coloring. The birds with severe damage also presented anorexia and had deformed paws and beaks. The parasitosis was caused by the "burrowing mites," Knemidocoptes pilae. The burrowing mites and the Grallacheles bakeri were recovered and identified from paw scabs. To eliminate mites, a topical application of Ivermectin was administered to the necks of the birds. The dose was a single, which has a residuality of 21 days. Two drops (0.115 mg/ml) of ivermectin were applied to each bird. A gradual reduction in crusted lesions due to mite mortality was noted. The "tropical fowl mite" Ornithonyssus bursa was identified in the nests, which represents the first record in Mexico. Conclusions: Three species of mites were discovered in a single budgerigar hatchery. This emphasizes the importance of deworming birds and keeping a clean environment in their cages to reduce the potential for parasitic mite infestation.

New State Record for Culex rejector and Cx. restrictor in Yucatan, Mexico.

Several known mosquito species occur in the Mexican state of Yucatan, including competent and suspected vectors responsible for transmitting zoonotic arboviruses. Between February and July 2022, mosquito collections were carried out in several forested areas in Yucatan. As part of the ongoing monitoring, we present the first reports of Culex (Microculex) rejector and Cx. (Anoedioporpa) restrictor. Another 14 species were identified during the monitoring: Aedes albopictus, Ae. bimaculatus, Ae. tormentor, Ae. cozumelensis, Anopheles albimanus, Cx. coronator s.l., Cx. erraticus, Cx. lactator, Cx. salinarius, Coquillettidia venezuelensis, Limatus durhamii, Psorophora ciliata, Toxorhynchites theobaldi, and Wyeomyia mitchellii. Currently, the mosquito fauna in Yucatan consists of 65 species. The subgenera Microculex and Anoedioporpa had not been documented in Yucatan State prior to the current investigation.

Impacto indirecto de COVID-19 en las tasas de incidencia de enfermedades transmitidas por vector en México / Indirect impact of COVID-19 on the incidence rates of vector-borne diseases in Mexico

Resumen Introducción: La distribución espacial y temporal de la infección por SARS-CoV-2 sobrepasa las áreas endémicas de enfermedades transmitidas por vector (ETV), cuya vigilancia en México ha cambiado sustancialmente a partir del primer caso confirmado de COVID-19. Objetivos: Estimar y comparar las tasas de incidencia de las ETV antes y después de la introducción del SARS-CoV-2 en México. Métodos: Estudio retrospectivo de casos de ETV de 2014 a 2021. Las tasas de incidencia de cada ETV en el periodo previo (2014-2019) y posterior (2020-2021) a la introducción del SARS-CoV-2 en México fueron calculadas y comparadas. Resultados: Antes de la introducción del SARS-CoV-2, las tasas de incidencia de las ETV fueron altas y posterior a la introducción del coronavirus hubo un descenso en los índices epidemiológicos; sin embargo, solo se identificó diferencia estadística significativa en la tasa de incidencia de la malaria (p ≤ 0.05) y otras rickettsias (p ≤ 0.05). Conclusiones: Algunas medidas para reducir los casos de COVID-19, como el distanciamiento social, el confinamiento domiciliario, la reducción en el aforo en el transporte público y el trabajo en casa, probablemente contribuyeron a disminuir temporalmente el número de casos de las ETV; sin embargo, puede haber rebrote de las ETV en el futuro cercano.

Abstract Introduction: SARS-CoV-2 infection spatial and temporal distribution overlaps with endemic areas of vector-borne diseases (VBD), whose surveillance in Mexico has substantially changed since the first COVID-19 confirmed case. Objectives: To estimate and compare the incidence rates of VBDs before and after the introduction of SARS-CoV-2 in Mexico. Methods: Retrospective study of VBD cases from 2014 to 2021. The incidence rates of each VBD in the period before (2014-2019) and after (2020-2021) the introduction of SARS-CoV-2 in Mexico were calculated and compared. Results: Before the introduction of SARS-CoV-2, the incidence rates of VBDs were high and after the introduction of coronavirus there was a decrease in epidemiological indices; however, there was only statistically significant difference in the incidence rate of malaria (p ≤ 0.05) and other rickettsiae (p ≤ 0.05). Conclusions: Some measures to reduce COVID-19 cases, such as social distancing, home confinement, reductions in public transport and working at home (home office), probably temporarily decreased the number of VBD cases; however, there may be a resurgence of VBDs in the near future.

Cluster Analysis of Dengue Morbidity and Mortality in Mexico from 2007 to 2020: Implications for the Probable Case Definition.

Dengue cases and deaths occur frequently in Mexico, although the trend is not uniform across the country. We performed a Spatio-temporal analysis of dengue cases and deaths in Mexico from 2007 to 2020, and clustered states according to whether there was a low, moderate, or high risk of dengue. A total of 501,600 confirmed dengue cases were registered from 2007 to 2020, with 378,122 cases classified as dengue fever (DF) and 123,478 cases classified as dengue hemorrhagic fever (DHF). For each confirmed case, there were 4.68 probable cases. There were 1,230 dengue deaths, with highest numbers reported in 2009, 2012, 2013, and 2019. The number of deaths had a significant correlation (P ≤ 0.01) with DF (r = 0.82), DHF (r = 0.94), and probable dengue cases (r = 0.84). States were clustered using Machine Learning technique according to select indices associated with dengue. Cluster 1 (low risk) primarily contained states in the northwest, northcentral, and east. Cluster 2 (moderate risk) includes states in the northeast. Cluster 3 (high risk) mostly contained coastal states in the southeast, southwest, and west. The generation of the clusters was supported by the Kruskal-Wallis test. A significant difference was found in the incidence, mortality rates, and case-fatality rates of dengue among the clusters (P ≤ 0.01). Notably, cluster 3 contributed 71.4% of the confirmed cases and 89.2% of the deaths. Public health and vector control strategies designed to mitigate the burden of dengue in Mexico should consider the states in cluster 3 as high priority areas.

Combined use of real-time PCR and serological techniques for improved surveillance of chronic and acute American trypanosomiasis in dogs and their owners from an endemic rural area of Neotropical Mexico.

In this study, the prevalence of T. cruzi infection was estimated in dogs and their owners from a rural community in Mexico using serological techniques for chronic infection cases, qPCR for acute phase cases, and a combination of both techniques to detect chronic and acute infections. Eighty-nine blood samples were collected from owners and their dogs for obtaining serum and parasite DNA. Prevalence was calculated using (i) positive cases detected in a serological test (ELISA and Western blot), (ii) positive cases detected in a qPCR test, and (iii) positive cases detected by both techniques. Sensitivity, specificity, and predictive values were determined individually for serology, qPCR and for both techniques used simultaneously. The prevalence observed varied: for serology, 25.8% of the dogs and 7.9% of the owners were seropositive, while for qPCR 29.2% of the dogs and 10.1% of the owners were identified as positive. Combination of serological and molecular techniques resulted in a prevalence of 38.2% for dogs and 12.4% for their owners. The sensitivity, specificity and predictive values calculated for both techniques improved when both techniques were used simultaneously (sensitivity of 92.4% and specificity of 100% for infected dogs and sensitivity of 93.4% and specificity of 100% for infected owners). Combined use of serological tests and qPCR allowed identifying a greater number of positive cases in dogs and their owners. This strategy may help implement adequate and timely epidemiological surveillance of American trypanosomiasis in order to prevent the appearance of new cases of Trypanosoma cruzi infections in endemic zones.

Indirect impact of COVID-19 on the incidence rates of vector-borne diseases in Mexico.

INTRODUCTION: SARS-CoV-2 infection spatial and temporal distribution overlaps with endemic areas of vector-borne diseases (VBD), whose surveillance in Mexico has substantially changed since the first COVID-19 confirmed case. OBJECTIVES: To estimate and compare the incidence rates of VBDs before and after the introduction of SARS-CoV-2 in Mexico. METHODS: Retrospective study of VBD cases from 2014 to 2021. The incidence rates of each VBD in the period before (2014-2019) and after (2020-2021) the introduction of SARS-CoV-2 in Mexico were calculated and compared. RESULTS: Before the introduction of SARS-CoV-2, the incidence rates of VBDs were high and after the introduction of coronavirus there was a decrease in epidemiological indices; however, there was only statistically significant difference in the incidence rate of malaria (p ≤ 0.05) and other rickettsiae (p ≤ 0.05). CONCLUSIONS: Some measures to reduce COVID-19 cases, such as social distancing, home confinement, reductions in public transport and working at home (home office), probably temporarily decreased the number of VBD cases; however, there may be a resurgence of VBDs in the near future.

INTRODUCCIÓN: La distribución espacial y temporal de la infección por SARS-CoV-2 sobrepasa las áreas endémicas de enfermedades transmitidas por vector (ETV), cuya vigilancia en México ha cambiado sustancialmente a partir del primer caso confirmado de COVID-19. OBJETIVOS: Estimar y comparar las tasas de incidencia de las ETV antes y después de la introducción del SARS-CoV-2 en México. MÉTODOS: Estudio retrospectivo de casos de ETV de 2014 a 2021. Las tasas de incidencia de cada ETV en el periodo previo (2014-2019) y posterior (2020-2021) a la introducción del SARS-CoV-2 en México fueron calculadas y comparadas. RESULTADOS: Antes de la introducción del SARS-CoV-2, las tasas de incidencia de las ETV fueron altas y posterior a la introducción del coronavirus hubo un descenso en los índices epidemiológicos; sin embargo, solo se identificó diferencia estadística significativa en la tasa de incidencia de la malaria (p ≤ 0.05) y otras rickettsias (p ≤ 0.05). CONCLUSIONES: Algunas medidas para reducir los casos de COVID-19, como el distanciamiento social, el confinamiento domiciliario, la reducción en el aforo en el transporte público y el trabajo en casa, probablemente contribuyeron a disminuir temporalmente el número de casos de las ETV; sin embargo, puede haber rebrote de las ETV en el futuro cercano.

An Updated Review of the Invasive Aedes albopictus in the Americas; Geographical Distribution, Host Feeding Patterns, Arbovirus Infection, and the Potential for Vertical Transmission of Dengue Virus.

Aedes (Stegomyia) albopictus is a mosquito native to Southeast Asia. Currently, it has a wide distribution in America, where natural infection with arboviruses of medical and veterinary importance has been reported. In spite of their importance in the transmission of endemic arbovirus, the basic information of parameters affecting their vectorial capacity is poorly investigated. The aim of the work was to update the distribution range of Ae. albopictus in the Americas, review the blood-feeding patterns, and compare the minimum infection rate (MIR) of the Dengue virus (DENV) between studies of vertical and horizontal transmission. The current distribution of Ae. albopictus encompasses 21 countries in the Americas. An extensive review has been conducted for the blood-feeding patterns of Ae. albopictus. The results suggest that the mosquito is capable of feeding on 16 species of mammals and five species of avian. Humans, dogs, and rats are the most common hosts. Eight arboviruses with the potential to infect humans and animals have been isolated in Ae. albopictus. In the United States of America (USA), Eastern equine encephalitis virus, Keystone virus, La Crosse Virus, West Nile virus, and Cache Valley virus were isolated in the Asian mosquito. In Brazil, Mexico, Colombia, and Costa Rica, DENV (all serotypes) has been frequently identified in field-caught Ae. albopictus. Overall, the estimated MIR in Ae. albopictus infected with DENV is similar between horizontal (10.95) and vertical transmission (8.28). However, in vertical transmission, there is a difference in the MIR values if the DENV is identified from larvae or adults (males and females emerged from a collection of eggs or larvae). MIR estimated from larvae is 14.04 and MIR estimated in adults is 4.04. In conclusion, it has to be highlighted that Ae. albopictus is an invasive mosquito with wide phenotypic plasticity to adapt to broad and new areas, it is highly efficient to transmit the DENV horizontally and vertically, it can participate in the inter-endemic transmission of the dengue disease, and it can spread zoonotic arboviruses across forest and urban settings.

Prevalence and Factors Associated with Ectoparasite Infestations in Dogs from the State of Tabasco, Mexico.

This study was carried out to identify the ectoparasites that infest owned dogs in the state of Tabasco, Mexico. In total, 1,302 dogs were sampled in the 5 ecological regions of Tabasco. The dog owners were surveyed to identify the factors associated with infestations. Ectoparasites were identified using taxonomic keys. Eleven species of ectoparasites were observed. General prevalence was 26.65%. Rhipicephalus sanguineus and Ctenocephalides felis were the most prevalent and abundant ectoparasites. The most important factors associated with ectoparasite infestations in the studied dogs were living outdoors, being a non-purebred, having short hair, being dark-haired, and having a body condition <3. Ectoparasite studies such as the one presented herein generate important information to create control programs focused on decreasing infestations in companion animals and thus the likelihood of zoonotic transmission of pathogens.

Evidence of Zika Virus Infection in Pigs and Mosquitoes, Mexico.

Evidence suggests that pigs seroconvert after experimental exposure to Zika virus and are potential sentinels. We demonstrate that pigs are also susceptible to natural Zika virus infection, shown by the presence of antibodies in domestic pigs in Yucatan, Mexico. Zika virus RNA was detected in 5 species of mosquitoes collected inside pigpens.

Identification of parasitic arthropods collected from domestic and wild animals in Yucatan, Mexico.

The morphological characterization of ectoparasites from domestic and wild animals is crucial to distinguish those that may be involved in the transmission of zoonotic pathogens. The objectives of this work were to identify parasitic arthropods collected from several hosts in Yucatan State, Mexico and to determine the prevalence of the parasite Trypanosoma cruzi in sylvatic specimens of Triatoma dimidiata. Morphological traits and scanning electron microscopy were used to confirm some parasitic identities. In total, 834 parasitic arthropods of various taxa were identified. Amblyomma americanum, Amblyomma maculatum, Amblyomma parvum, Dermacentor variabilis, and Rhipicephalus sanguineus were identified from dogs, horses, sheep, and deer. The flea Ctenocephalides felis was identified from dogs and the human flea, Pulex irritans was found on horses and sheep. The Neotropical deer ked Lipoptena mazamae was identified from brown brocket deer Mazama pandora. The chewing lice Chelopistes meleagridis, Menacanthus stramineus, Menopon gallinae, Myrsidea sp. and Lipeurus caponis were identified from poultry (Meleagris gallopavo and Gallus gallus). Other chewing lice, Tyranniphilopterus sp., Columbicola columbae, and Physconelloides eurysema were identified from wild birds Pachyramphus aglaiae, Patagioenas flavirostris and Zenaida asiatica, respectively. Trypanosoma cruzi was present in 52.5% of sylvatic adult T. dimidiata. Several of these findings represent new records of ectoparasites for Yucatan and new distribution areas in Mexico. The implications for human and animal health are discussed.

Four Species of Under-reported Parasitic Arthropods in Mexico and their Potential Role as Vectors of Pathogens.

Numerous arthropod taxa are important in human and veterinary medicine. The salivary secretions and feces of arthropods can cause allergic reactions in host vertebrates or harbor pathogens. Also, bites can be a risk factor for secondary infections. Documenting the diversity of arthropods of medical and veterinary importance remains an important aspect of disease control and prevention. We provide new records of ectoparasitic arthropods from Mexico that are of potential medical or veterinary relevance. Scanning electron microscopy along with amplification and sequencing of a fragment of the mitochondrial gene (16S rRNA) was used to confirm some species identities. We report the cat louse Felicola subrostratus from cats and the chewing louse Heterodoxus spiniger from dogs, which are common ectoparasites but largely not reported in Mexico. The chigger Eutrombicula alfreddugesi is common on wild lizards (Squamata). For the first time, E. alfreddugesi is reported on Hemidactylus frenatus (common house gecko). This reptile has a close relationship with humans and its chiggers can cause dermatitis (i.e., trombiculiasis) or transmit pathogens. In addition, the common bed bug Cimex lectularius is reported for the first time in the state of Yucatan, an atypical area for its natural distribution. Phylogenetic analysis revealed that Cimex lectularius from Yucatan is closely related to genetic sequences of Cimex lectularius from China. Knowing the regional distribution of arthropods allows the design and implementation of prevention strategies for those that have potential roles as reservoirs or vectors.

Dirofilaria immitis proteins recognized by antibodies from individuals living with microfilaremic dogs.

INTRODUCTION: Dirofilaria immitis is a nematode that affects human health in several countries of the world. This study was conducted to examine whether serum samples from the owners of microfilaremic dogs present immunoreactivity to parasite proteins. METHODOLOGY: Eight serum samples from the owners of microfilaremic dogs were examined. Total proteins were extracted from adult worms and 12% SDS-PAGE was performed. The gel was electroblotted to a nitrocellulose membrane, and a Western blot (WB) was performed. Reactive bands of 22, 33, 39, 49, and 63 kDa in WB were excised from the gel and analyzed by mass spectrometry (MS). RESULTS: The MS results showed the presence of 10 different proteins of D. immitis recognized by the human serum samples. CONCLUSIONS: These results indicate that in endemic areas of D. immitis, owners of infected dogs recognize specific proteins of the parasite, suggesting a possible infection.

Rickettsia parkeri (Rickettsiales: Rickettsiaceae) detected in Amblyomma maculatum ticks collected on dogs in Tabasco, Mexico.

The present study was carried out to identify Rickettsia species with zoonotic potential in ticks collected from dogs in a rural area in Tabasco, Mexico. In total 197 Amblyomma maculatum ticks were collected from 40 domestic dogs. The collected specimens were pooled and subjected to DNA extraction. A fragment (380 bp) of citrate synthase gene (gltA) was amplified by polymerase chain reaction (PCR) using universal primers for Rickettsia. A second PCR was later performed to amplify a fragment (420 bp) of the outer membrane protein B gene (ompB). The PCR products were purified, sequenced and compared using the basic local alignment search tool (BLAST). Twenty out of 40 (50%) tick pools assayed were positive for rickettsial DNA using both primer pairs. The consensus sequence obtained from the ompB gene fragments showed 99.5-100% of identity with strains of Rickettsia parkeri. This study provides the first molecular evidence of the presence of R. parkeri in A. maculatum ticks infesting domestic dogs from southeastern Mexico. Close contact between dogs and humans should lead to consider the infection caused by this species of Rickettsia among the differential diagnoses for people of Tabasco, Mexico, who show acute febrile syndrome associated to inoculation eschar and have a clinical history of tick exposure.

Entomological and virological surveillance for dengue virus in churches in Merida, Mexico.

This study was designed to assess whether churches in endemic dengue districts in Merida, Mexico provide suitable breeding habitats for mosquitoes and are potential sites for dengue virus (DENV) transmission. Churches were inspected for immature and adult mosquitoes once every week from November 2015 to October 2016. A total of 10,997 immatures of five species were collected. The most abundant species were Aedes aegypti (6,051) and Culex quinquefasciatus (3,018). The most common source of immature Ae. aegypti were buckets followed by disposable containers. Adult collections yielded 21,226 mosquitoes of nine species. The most common species were Cx. quinquefasciatus (15,215) and Ae. aegypti (3,902). Aedes aegypti were found all year long. Female Ae. aegypti (1,380) were sorted into pools (166) and assayed for flavivirus RNA by RT-PCR and Sanger sequencing. Two pools were positive for DENV (DENV-1 and 2). In conclusion, we demonstrated that some churches in Merida are infested with mosquitoes all year long and they potentially serve as sites for DENV transmission and should therefore be considered for inclusion in mosquito and arboviruses control and surveillance efforts.

Complete Genome Sequence of Houston Virus, a Newly Discovered Mosquito-Specific Virus Isolated from Culex quinquefasciatus in Mexico.

We fully sequenced the genome of Houston virus, a recently discovered mosquito-associated virus belonging to the newly established family Mesoniviridae. The isolate was recovered from Culex quinquefasciatus in southern Mexico, which shows that the geographic range of Houston virus is not restricted to the United States in North America.

Survey of mosquito-borne flaviviruses in the Cuitzmala River Basin, Mexico: do they circulate in rodents and bats?

BACKGROUND: RNA viruses commonly infect bats and rodents, including mosquito-borne flaviviruses (MBFV) that affect human and animal health. Serological evidence suggests past interactions between these two mammalian orders with dengue viruses (DENV), West Nile virus (WNV), and yellow fever virus (YFV). Although in Mexico there are reports of these viruses in both host groups, we know little about their endemic cycles or persistence in time and space. METHODS: Rodents and bats were captured at the Cuitzmala River Basin on the Pacific coast of Jalisco state, Mexico, where MBFV, such as DENV, have been reported in both humans and bats. Samples were taken during January, June, and October 2014, at locations adjacent to the river. Tissue samples were collected from both bats and rodents and serum samples from rodents only. Highly sensitive serological and molecular assays were used to search for current and past evidence of viral circulation. RESULTS: One thousand nine hundred forty-eight individuals were captured belonging to 21 bat and 14 rodent species. Seven hundred sixty-nine liver and 764 spleen samples were analysed by means of a specific molecular protocol used to detect flaviviruses. Additionally, 708 serum samples from rodents were examined in order to demonstrate previous exposure to dengue virus serotype 2 (which circulates in the region). There were no positive results with any diagnostic test. DISCUSSION: To our knowledge, this is the first survey of rodents and only the second survey of bats from the Pacific Coast of Mexico in a search for MBFV. We obtained negative results from all samples. We validated our laboratory tests with negative and positive controls. Our findings are consistent with other empirical and experimental studies in which these mammalian hosts may not replicate mosquito-borne flaviviruses or present low prevalence. CONCLUSIONS: True-negative results are essential for the construction of distribution models and are necessary to identify potential areas at risk. Negative results should not be interpreted as the local absence of MBFV in the region. On the contrary, we need to establish a long-term surveillance programme to find MBFV presence in the mosquito trophic networks, identifying the potential role of rodents and bats in viral dynamics.