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.

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.

Infection, dissemination, and transmission efficiencies of Zika virus in Aedes aegypti after serial passage in mosquito or mammalian cell lines or alternating passage in both cell types.

BACKGROUND: Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) with an urban transmission cycle that primarily involves humans and Aedes aegypti. Evidence suggests that the evolution of some arboviruses is constrained by their dependency on alternating between disparate (vertebrate and invertebrate) hosts. The goals of this study are to compare the genetic changes that occur in ZIKV after serial passaging in mosquito or vertebrate cell lines or alternate passaging in both cell types and to compare the replication, dissemination, and transmission efficiencies of the cell culture-derived viruses in Ae. aegypti. METHODS: An isolate of ZIKV originally acquired from a febrile patient in Yucatan, Mexico, was serially passaged six times in African green monkey kidney (Vero) cells or Aedes albopictus (C6/36) cells or both cell types by alternating passage. A colony of Ae. aegypti from Yucatan was established, and mosquitoes were challenged with the cell-adapted viruses. Midguts, Malpighian tubules, ovaries, salivary glands, wings/legs and saliva were collected at various times after challenge and tested for evidence of virus infection. RESULTS: Genome sequencing revealed the presence of two non-synonymous substitutions in the premembrane and NS1 regions of the mosquito cell-adapted virus and two non-synonymous substitutions in the capsid and NS2A regions of both the vertebrate cell-adapted and alternate-passaged viruses. Additional genetic changes were identified by intrahost variant frequency analysis. Virus maintained by continuous C6/36 cell passage was significantly more infectious in Ae. aegypti than viruses maintained by alternating passage and consecutive Vero cell passage. CONCLUSIONS: Mosquito cell-adapted ZIKV displayed greater in vivo fitness in Ae. aegypti compared to the other viruses, indicating that obligate cycling between disparate hosts carries a fitness cost. These data increase our understanding of the factors that drive ZIKV adaptation and evolution and underscore the important need to consider the in vivo passage histories of flaviviruses to be evaluated in vector competence studies.

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.

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.

Surveillance for Flaviviruses Near the Mexico-U.S. Border: Co-circulation of Dengue Virus Serotypes 1, 2, and 3 and West Nile Virus in Tamaulipas, Northern Mexico, 2014-2016.

A clinical, serological, and molecular investigation was performed to determine the presence of dengue virus (DENV) and other flaviviruses among residents of the city of Reynosa, Tamaulipas, on the Mexico-U.S. border in 2014-2016. The sample population consisted of 2,355 patients with suspected dengue, in addition to 346 asymptomatic individuals recruited during a household-based epidemiological investigation designed to identify flavivirus seroconversions. Sera were collected from patients with suspected dengue in the acute phase of illness and from asymptomatic individuals at enrollment and every 5-7 months for 19 months. Sera from suspected dengue patients were tested for DENV antigen by enzyme-linked immunosorbent assay (ELISA), and select antigen-positive sera were further tested using a serotype-specific, quantitative reverse transcription-polymerase chain reaction. Sera from the household cohort were tested for flavivirus-reactive antibodies by immunoglobulin (Ig) M and IgG ELISAs using DENV antigen. A total of 418 (17.7%) patients with suspected dengue had laboratory-confirmed DENV infections, including 82 patients who were positive for DENV RNA. The most frequently detected serotype was DENV-1 (61 patients), followed by DENV-2 (16 patients) and DENV-3 (five patients). A total of 217 (62.7%) asymptomatic individuals had flavivirus-reactive antibodies at enrollment, and nine flavivirus-naïve individuals seroconverted. Sera from a subset of dengue patients and household participants, including all those who seroconverted, were further tested by plaque reduction neutralization test, resulting in the detection of antibodies to DENV-1, DENV-2, and West Nile virus. In summary, we provide evidence for the co-circulation of multiple flaviviruses in Reynosa, Tamaulipas, on the Mexico-U.S. border.

Study of Aedes aegypti population with emphasis on the gonotrophic cycle length and identification of arboviruses: implications for vector management in cemeteries.

Aedes aegypti is the vector of the arboviruses causing dengue, chikungunya and zika infections in Mexico. However, its presence in public places has not been fully evaluated. In a cemetery from Merida, Yucatan, Mexico, the productivity of Ae. aegypti, the gonotrophic cycle, and the presence of Ae. aegypti females infected with arboviruses were evaluated. Immature and adult mosquitoes were inspected every two months between April 2016 to June 2017. For the gonotrophic cycle length, the daily pattern of total and parous female ratio was registered and was analyzed using time series analysis. Ae. aegypti females were sorted into pools and assayed for flavivirus RNA by RT-PCR and Sanger sequencing. Aedes aegypti immatures represented 82.86% (8,627/10,411) of the collection. In total, 1,648 Ae. aegypti females were sorted into 166 pools. Two pools were positive; one for dengue virus (DENV-1) and the other for zika virus (ZIKV). The phylogenetic analysis revealed that the DENV-1 is more closely related to isolates from Brazil. While ZIKV is more closely related to the Asian lineage, which were isolates from Guatemala and Mexico. We report some evidence of vertical transmission of DENV-1 in nulliparous females of Ae. aegypti. The gonotrophic cycle was four and three days in the rainy and dry season, respectively. The cemetery of Merida is an important focus of Ae. aegypti proliferation, and these environments may play a role in arboviruses transmission; probably limiting the efficacy of attempts to suppress the presence of mosquitoes in domestic environments.

The Use of Insecticide-Treated Curtains for Control of Aedes aegypti and Dengue Virus Transmission in "Fraccionamiento" Style Houses in México.

Dengue, chikungunya, yellow fever, and Zika viruses transmitted by Aedes aegypti mosquitoes are major public health threats in the tropical and subtropical world. In México, construction of large tracts of "fraccionamientos" high density housing to accommodate population growth and urbanization has provided fertile ground for Ae. aegypti-transmitted viruses. We investigated the utility of pyrethroid-treated window curtains to reduce both the abundance of Ae. aegypti and to prevent dengue virus (DENV) transmission in fraccionamiento housing. Windows and doors of fraccionamiento homes in urban/suburban areas, where Ae. aegypti pyrethroid resistance associated with the Ile1016 knock down resistance (kdr) mutation in the voltage gated sodium channel gene was high, and in rural areas, where kdr resistance was low, were fitted with either insecticide-treated curtains (ITCs) or non-treated curtains (NTCs). The homes were monitored for mosquito abundance and DENV infection. ITCs reduced the indoor abundance of Ae. aegypti and the number of DENV-infected mosquitoes in homes in rural but not in urban/suburban study sites. The presence of non-treated screens also was associated with reduced numbers of mosquitoes in homes. "Super-infested" homes, yielding more than 50 mosquitoes, including DENV-infected mosquitoes, provide a significant public health risk to occupants, visitors, and people in neighboring homes.

Arbovirus Surveillance near the Mexico-U.S. Border: Isolation and Sequence Analysis of Chikungunya Virus from Patients with Dengue-like Symptoms in Reynosa, Tamaulipas.

A total of 1,090 residents of the city of Reynosa, Tamaulipas, on the Mexico-U.S. border presented at hospitals and clinics of the Secretariat of Health, Mexico, in 2015 with symptoms characteristic of dengue. Dengue virus (DENV) antigen was detected by enzyme-linked immunosorbent assay in acute sera from 134 (12.3%) patients. Sera from select patients (N = 34) were also tested for chikungunya virus (CHIKV) RNA by quantitative reverse transcription-polymerase chain reaction. Thirteen (38.2%) patients, including five DENV antigen-positive patients, were positive. Sera from three CHIKV RNA-positive patients were further assayed by virus isolation in cell culture and CHIKV was recovered on each occasion. The genome of one isolate and structural genes of the other two isolates were sequenced. In conclusion, we present evidence of CHIKV and DENV coinfections in patients who live near the Mexico-U.S. border and provide the first genome sequence of a CHIKV isolate from northern Mexico.

Chikungunya Virus in Febrile Humans and Aedes aegypti Mosquitoes, Yucatan, Mexico.

Chikungunya virus (CHIKV) was isolated from 12 febrile humans in Yucatan, Mexico, in 2015. One patient was co-infected with dengue virus type 1. Two additional CHIKV isolates were obtained from Aedes aegypti mosquitoes collected in the homes of patients. Phylogenetic analysis showed that the CHIKV isolates belong to the Asian lineage.

Detection of hepatitis C virus coinfection in patients with dengue diagnosis.

Coinfection produced by dengue virus (DENV) and hepatitis C virus (HCV) is a serious problem of public health in Mexico, as they both circulate in tropical zones and may lead to masking or complicating symptoms. In this research, we detected active coinfected patients by HCV residing in the endemic city of Mérida, Yucatán, Mexico, with positive diagnosis to dengue during the acute phase. We performed a retrospective analysis of 240 serum samples from dengue patients. The IgM-ELISA serological test was used for dengue diagnosis, as well as viral isolation to confirm infection. DENV and HCV were detected by RT-PCR. Thus, 31 (12.9%) samples showed DENV-HCV coinfection, but interestingly the highest frequency of coinfection cases was found in male patients presenting hemorrhagic dengue in 19/31 (61.29%), with a predominance of 12 : 7 in males. Firstly, coinfection of DENV-HCV in Mérida, Mexico, was detected in young dengue patients, between 11 and 20 years old (38.7%), followed by those between 21 and 30 years old (32%); only 16.13% were between 0 and 10 years of age. Diagnosis of HCV infection in patients with dengue is highly recommended in order to establish potential risk in clinical manifestations as well as dictate patients' special care.

Antibodies to West Nile virus in wild and farmed crocodiles in southeastern Mexico.

Surveillance for evidence of West Nile virus (WNV) infection in Morelet's crocodiles (Crocodylus moreletii) was conducted in Campeche State, Mexico, in 2007. Sera from 62 crocodiles (32 free-ranging and 30 captive) were assayed for antibodies to WNV by epitope-blocking enzyme-linked immunosorbent assay. Antibodies to WNV were detected in 13 (41%) wild and nine (30%) captive crocodiles, and the overall antibody prevalence was 35%. Although evidence of WNV infection in captive crocodiles has been reported in Mexico, we provide the first evidence of WNV exposure in wild crocodiles in Mexico.