Entomo-virological surveillance of Flavivirus in mosquitoes in Yucatan State, Mexico.

The genus Flavivirus (Family: Flaviviridae) comprises arboviruses with the capacity to infect humans and animals. It also integrates insect-specific viruses. This study aimed to identify Flavivirus in mosquitoes captured in 17 municipalities in Yucatan State, Mexico. The mosquitoes were caught in households from November 2021 to May 2022. A total of 4,321 adult mosquitoes from five species were caught. The most abundant were Culex quinquefasciatus (n = 3,563) and Aedes aegypti (n = 734). For molecular investigations, 600 female mosquitoes were split into groups of 10, mostly for species and site location. Reverse transcriptase polymerase chain reaction (RT-PCR) amplified a region of the NS5 gene to find the Flavivirus ribonucleic acids (RNA). A total of 24 pools that were positive for Flavivirus were detected in Ae. aegypti specimens and subsequently subjected to sequencing using the Sanger method. A total of 12 sequences matched the established quality criteria and were subsequently employed for sequence homology analysis. We found that one sequence corresponded to the Zika virus (ZIKV), and 11 sequences had sequence similarity with Phlebotomus-associated flavivirus (PAFV), an insect-specific virus (ISF). In conclusion, we found ZIKV in the Merida municipality, Yucatan State, which suggests that the virus is silently circulating. Phlebotomus-associated flavivirus is distributed in five municipalities in Yucatan State, Mexico. Future studies could focus on isolating this virus and studying its biological role within Ae. aegypti.

Entomo-virological surveillance of Flavivirus in mosquitoes in Yucatan State, Mexico

ABSTRACT The genus Flavivirus (Family: Flaviviridae) comprises arboviruses with the capacity to infect humans and animals. It also integrates insect-specific viruses. This study aimed to identify Flavivirus in mosquitoes captured in 17 municipalities in Yucatan State, Mexico. The mosquitoes were caught in households from November 2021 to May 2022. A total of 4,321 adult mosquitoes from five species were caught. The most abundant were Culex quinquefasciatus (n = 3,563) and Aedes aegypti (n = 734). For molecular investigations, 600 female mosquitoes were split into groups of 10, mostly for species and site location. Reverse transcriptase polymerase chain reaction (RT-PCR) amplified a region of the NS5 gene to find the Flavivirus ribonucleic acids (RNA). A total of 24 pools that were positive for Flavivirus were detected in Ae. aegypti specimens and subsequently subjected to sequencing using the Sanger method. A total of 12 sequences matched the established quality criteria and were subsequently employed for sequence homology analysis. We found that one sequence corresponded to the Zika virus (ZIKV), and 11 sequences had sequence similarity with Phlebotomus-associated flavivirus (PAFV), an insect-specific virus (ISF). In conclusion, we found ZIKV in the Merida municipality, Yucatan State, which suggests that the virus is silently circulating. Phlebotomus-associated flavivirus is distributed in five municipalities in Yucatan State, Mexico. Future studies could focus on isolating this virus and studying its biological role within Ae. aegypti.

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.

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.

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.

Discovery of a novel Tymoviridae-like virus in mosquitoes from Mexico.

A novel Tymoviridae-like virus, designated Ek Balam virus, was isolated from male Culex quinquefasciatus mosquitoes collected in Yucatan, Mexico. The genome was fully sequenced and shown to have no more than 69% nt sequence identity to its closest known relative. Mosquito cells were permissive to Ek Balam virus replication, but mammalian and avian cells were refractory, suggesting that vertebrates are not involved in the maintenance of the virus in nature.

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.

Detection of novel and recognized RNA viruses in mosquitoes from the Yucatan Peninsula of Mexico using metagenomics and characterization of their in vitro host ranges.

A metagenomics approach was used to detect novel and recognized RNA viruses in mosquitoes from the Yucatan Peninsula of Mexico. A total of 1359 mosquitoes of 7 species and 5 genera (Aedes, Anopheles, Culex, Mansonia and Psorophora) were sorted into 37 pools, homogenized and inoculated onto monolayers of Aedes albopictus (C6/36) cells. A second blind passage was performed and then total RNA was extracted and analysed by RNA-seq. Two novel viruses, designated Uxmal virus and Mayapan virus, were identified. Uxmal virus was isolated from three pools of Aedes (Ochlerotatus) taeniorhynchus and phylogenetic data indicate that it should be classified within the recently proposed taxon Negevirus. Mayapan virus was recovered from two pools of Psorophora ferox and is most closely related to unclassified Nodaviridae-like viruses. Two recognized viruses were also detected: Culex flavivirus (family Flaviviridae) and Houston virus (family Mesoniviridae), with one and two isolates being recovered, respectively. The in vitro host ranges of all four viruses were determined by assessing their replicative abilities in cell lines of avian, human, monkey, hamster, murine, lepidopteran and mosquito (Aedes, Anopheles and Culex) origin, revealing that all viruses possess vertebrate replication-incompetent phenotypes. In conclusion, we report the isolation of both novel and recognized RNA viruses from mosquitoes collected in Mexico, and add to the growing plethora of viruses discovered recently through the use of metagenomics.

Molecular detection of Dirofilaria immitis in dogs and mosquitoes in Tabasco, Mexico.

BACKGROUND & OBJECTIVES: Dirofilaria immitis is a filarial nematode that causes heartworm disease in domestic as well as wild canines and felines; and cutaneous or pulmonary infections in humans. The purpose of the study was to estimate the prevalence of D. immitis in domestic dogs in Tabasco, Mexico and to assay mosquitoes temporally and spatially associated with dogs for evidence of infection. METHODS: Blood was collected from 1050 dogs in 1039 houses during a random household survey performed in 2016 and 2017. Genomic DNA was extracted and assayed by polymerase chain reaction (PCR) using pan-filarial primers and various species-specific primers. Dog owners were interviewed using a structured questionnaire designed to collect information on factors that may impact the occurrence of filarial infection. The association between canine dirofilariasis prevalence and factors likely to impact infection was determined by univariate logistic regression analysis, followed by multivariate binomial logistic regression analysis. Indoor and outdoor resting mosquitoes were collected from houses by manual aspiration. Mosquitoes were identified according to species, homogenized and tested by PCR for filarial nematodes. RESULTS: A total of 84 (8%) dogs were positive for D. immitis DNA, while 3 (0.3%) dogs contained Acanthocheilonema reconditum DNA. Several factors were significantly associated with D. immitis infection. For example, dogs that lived <100 m from a large source of open standing water were significantly more likely (p = 0.002) to become infected with D. immitis than other dogs. Additionally, dogs with infrequent or no anthelmintic treatment were significantly more likely (p = 0.0) to become infected than dogs that were regularly treated. The entomologic investigation yielded 2618 female mosquitoes from 14 species. Four pools of Culex quinquefasciatus were positive for D. immitis DNA and the minimum infection rate, calculated as the number of positive pools per 1000 mosquitoes tested, was 2.9. INTERPRETATION & CONCLUSION: The study identified several factors positively associated with an increased risk of D. immitis infection in domestic dogs in Tabasco and provides evidence that Cx. quinquefasciatus is potentially an important vector in this region. This information can be used by local veterinarians and dog owners to reduce the burden of D. immitis on canine health.