Potential distribution of Amblyomma mixtum (Koch, 1844) in climate change scenarios in the Americas.

Amblyomma mixtum is a Neotropical generalist tick of medical and veterinary importance which is widely distributed from United States of America to Ecuador. The aim of this study was to evaluate changes in the geographic projections of the ecological niche models of A. mixtum in climate change scenarios in America. We constructed a database of published scientific publications, personal collections, personal communications, and online databases. Ecological niche modelling was performed with 15 Bioclimatic variables using kuenm in R and was projected to three time periods (Last Glacial Maximum, Current and 2050) for America. Our model indicated a wide distribution for A. mixtum, with higher probability of occurrence along the Gulf of Mexico and occurring in a lesser proportion in the Pacific states, Central America, and the northern part of South America. The areas of new invasion are located mainly on the border of Mexico with Guatemala and Belize, some regions of Central America and Colombia. We conclude that the ecological niche modelling are effective tools to infer the potential distribution of A. mixtum in America, in addition to helping to propose future measures of epidemiological control and surveillance in the new potential areas of invasion.

Morphometrics of Amblyomma mixtum in the State of Veracruz, Mexico.

The tick Amblyomma mixtum is an ectoparasite of veterinary and public health importance because of its role as a vector of zoonotic pathogens. However, little is known about A. mixtum intraspecific variability and if morphological differentiation exists between populations across its geographic range. This study aimed to determine by electron microscopy the morphological variability of A. mixtum populations in the state of Veracruz, which has a large livestock population among states in Mexico. Forty male and 40 female A. mixtum collected from the 10 natural regions of Veracruz state were analyzed microscopically to accomplish main component analysis for each sex. Clusters were calculated with the out-tree method and a dendrogram produced to group the specimens according to their morphometric characteristics. Using 10 main components, 77% of the morphological variation of the ticks was explained. This is a reflection of scarce intraspecific variation between females. The dendogram for females grouped six clusters of specimens with similar characteristics. Morphometric variability in males was described using eight main components. Limited intraspecific variation was also observed between males. In males, the dendogram yielded six groups with similar morphometric characteristics. Morphometric analyses confirmed that the only species from the Amblyomma cajennense complex that are parasites to livestock in Veracruz state is A. mixtum. The eryxenous nature of A. mixtum combined with the frequent movement of livestock hosts may contribute to the apparent homogeneous phenotype of this tick species in Mexico.

Morphological and molecular characterization supporting Amblyomma mixtum presence in Cuba.

Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) is a widely distributed tick taxon. Recent studies have reassessed this taxon as a complex of six species. Amblyomma mixtum Koch, 1844 has been suggested by some authors as the only species of this complex that is present in Cuba. Other authors have pointed a niche overlapping for A. mixtum and A. cajennense s.s. in the country. Detailed taxonomic studies on the Cuban species belonging to this complex are needed in order to evaluate their current distribution according to the recent classification. This study aimed to characterize Cuban populations from the A. cajennense complex by using tick samples obtained from 3 occidental provinces and 1 central province of the country. Morphological identification and measurements of the main relevant taxonomic structures were conducted by using Scanning Electron Microscopy. Phylogenetic analyzes were carried out with 16S ribosomal RNA, internal transcribed spacer 2 and the subunit I of mitochondrial cytochrome c oxidase gene sequences. The results of these studies demonstrated that all samples belonged to the species A. mixtum (Koch, 1844). This study constitutes the first molecular characterization of this Amblyomma species in Cuba. Further studies will be necessary in order to corroborate if A. cajennense s.s. is also present in the island.

Molecular detection and identification of spotted fever group rickettsiae in ticks collected from horses in Cuba.

Spotted fever group (SFG) rickettsiae are obligatory intracellular bacteria that cause disease in humans and other animals. Ixodid ticks are the principal vectors of SFG rickettsiae. The present study aimed to determine the prevalence and species identity of SFG rickettsiae in ticks and horses from urban and rural areas of western Cuba using PCR assays. Tick samples, collected from 79 horses, consisted of 14 Amblyomma mixtum adults, 111 Dermacentor nitens adults and 19 pools of D. nitens nymphs (2-5 individuals/pool). The PCR results revealed the presence of Rickettsia spp. in 64% of the A. mixtum adults, 16% of the D. nitens adults, and 11% of the pooled samples of D. nitens nymphs. In contrast, Rickettsia spp. was not detected in any of the 200 horse blood samples included in this study. DNA sequence data of the rickettsial 17 kDa antigen gene showed that Rickettsia amblyommatis was present in A. mixtum; and Rickettsia felis in D. nitens. This is the first report of R. felis in D. nitens in Cuba. The present study extends our knowledge of the potential vector spectrum and distribution of SFG rickettsiae pathogens in western Cuba.

Detection of Amblyomma mixtum (Acari: Ixodidae) in Germany on a Human Traveler Returning From Cuba.

The importation of exotic ticks to Germany on infested humans is rarely reported. Here we describe the detection of an Amblyomma mixtum nymph harboring Rickettsia amblyommatis by a tourist returning from a holiday trip to Cuba. Tick infestation may be neglected by tourists. Therefore, careful anamnesis and diagnosis should be carried out when tourists return from tropical and subtropical countries suffering from nonspecific symptoms, such as fever and malaise.

Rickettsia species in ticks that parasitize amphibians and reptiles: Novel report from Mexico and review of the worldwide record.

Ticks are obligate haematophagous ectoparasites that are associated with a wide range of vertebrate hosts, among them also reptiles and amphibians. They have dynamic ecological interactions with multiple microorganisms, ranging from endosymbionts to pathogens, such as the members of the genus Rickettsia. The aim of this work was to detect Rickettsia in ticks from amphibians and reptiles from southern Mexico by the amplification, sequencing and phylogenetic reconstruction of the gltA and ompA genes, and also to compile all the published records worldwide of Rickettsia associated with ticks attached to reptiles and amphibians, in order to elucidate the host-parasite relationships, and to identify the geographical distribution of each bacterial species. We record for the first time the presence of Rickettsia sp. strain Colombianensi and Rickettsia amblyommatis in ticks from several reptiles and amphibians collected in three new localities from the states of Guerrero and Veracruz, Mexico. Additionally, we here report 23 Rickettsia taxa associated with 18 tick species attached to 42 host taxa of amphibians and reptiles in 36 countries. Our findings increase the inventory of rickettsia reported in Mexico and summarizes the knowledge of these bacteria associated with ticks of this particular group of vertebrate host worldwide.

Genetic structure analysis of Amblyomma mixtum populations in Veracruz State, Mexico.

Amblyomma mixtum Koch, 1844 parasitizes livestock, humans, and wildlife in Mexico. However, information on population genetics for this tick species in the country is missing. Tick samples were collected from livestock in ten regions across the state of Veracruz (22°28'N, 17°09'S, 93°36'E, 98°39'W) to analyze the genetic structure of A. mixtum populations. Ticks were morphologically identified using taxonomic keys. In order to test the intra-specific variability of A. mixtum fragments of the mitochondrial gene 16S-rRNA and cytochrome oxidase subunit 1 (COI) were amplified. Ninety-six sequences were amplified from the 50 specimens' analyzed (96% amplification success). Eleven haplotypes were detected in 16S-rRNA gene and 10 more for COI. Neutrality tests showed negative results in most of the locations analyzed, which is indicative of an excess of recently derived haplotypes. However, these results were not statistically significant. Minimal union network analysis revealed that there is no separation of populations by geography, and that there is an overlap of several haplotypes among diverse populations. Significant genetic differentiation was not detected in the A. mixtum populations sampled in the state of Veracruz, Mexico, this may be due to the frequent movement of livestock hosts. This is the first report on the genetic structure of A. mixtum populations in Mexico.

Occurrence of Amblyomma mixtum on the water buffalo (Bubalus bubalis) in Mexico.

The water buffalo (Bubalus bubalis) is an Asian species of bovine that was introduced in Mexico in 1992 as an alternative for milk and meat production. To date, no surveys have been conducted to identify ticks acquired by water buffaloes since their arrival in the country. Here we report, for the first time, the presence of Amblyomma mixtum, a neotropical tick in the Amblyomma cajennense complex, found on water buffaloes from Mexico and discuss its possible implications in veterinary public health for the region.

Detección de Borrelia burgdorferi sensu lato en perros y sus garrapatas en comunidades rurales de Yucatán, México / Detection of Borrelia burgdorferi sensu lato in dogs and its ticks in rural communities of Yucatán, Mexico

Resumen Actualmente, el estudio de la borreliosis canina adquiere mayor relevancia, ya que el perro es considerado como un importante centinela del complejo Borrelia burgdorferi sensu lato, el cual podría desempeñar un papel clave en la dispersión de garrapatas de las áreas selváticas al ambiente doméstico. En México, la distribución y presencia de genoespecies patógenas de B. burgdorferi en perros y sus garrapatas aún no ha sido investigada. Por tal motivo, la presente investigación tiene como objetivo detectar y estimar la prevalencia de B burgdorferi s.l. en perros y sus garrapatas en dos comunidades rurales de Yucatán, México. En cada comunidad se visitaron 50 viviendas donde se estudiaron 144 muestras de sangre de perros por punción de la vena safena, así como la colecta de sus garrapatas. Se colectaron un total de 846 garrapatas de las especies Ixodes affinis (33 / 846), Rhipicephalus sanguineus sensu lato (786 / 846) y Amblyomma mixtum (27 / 846). Para detectar la presencia de B. burgdorferi s.l. en dichas muestras, se amplificó el gen conservado flaB y las lipoproteínas de membrana externa, ospC y p66, mediante el uso de la reacción en cadena de la polimerasa. La prevalencia obtenida en sangre de perros fue de 17.3 % (25 / 144) para flaB, 12.50 % (18 / 144) para el gen p66 y 1.38 % (2 / 144) para el gen ospC. De las garrapatas analizadas, R. sanguineus s.l. tuvo una prevalencia de infección de 0.89 %, A. mixtum de 5.88 % e I. affinis de 15.15 %, siendo esta última especie la que presentó mayor prevalencia. Dos perros y sus garrapatas I. affinis fueron positivos al gen flaB. Solamente una garrapata R. sanguineus s.l. fue positiva al gen p66 y ninguna especie de garrapata fue positiva al gen ospC. Este estudio confirma la existencia de B. burgdorferi s.l. en perros y sus garrapatas en comunidades rurales de Yucatán, México. La detección de Borrelia en perros podría ser un criterio importante para la evaluación del riesgo de borreliosis en humanos, ya que el perro puede emplearse como indicador epidemiológico para la identificación de nuevos focos de esta enfermedad.

Abstract In Mexico, the distribution and the presence of pathogenic genospecies of B. burgdorferi in dogs and their ticks has not been extensively investigated. The study of canine borreliosis is acquiring greater relevance, since the dog is considered to be an important sentinel for pathogens pertaining to the complex Borrelia burgdorferi sensu lato; in addition, dogs could be playing a key role in the spread of ticks from forested areas into the domestic environment. This study aimed to detect and estimate the prevalence of B. burgdorferi s.l. in dogs and their ticks in two rural communities of Yucatán, Mexico. In each community, 50 houses were visited, where 144 blood samples from dogs were studied by puncture of the saphenous vein, as well as the collection of their ticks. To detect the presence of B. burgdorferi s.l. in these samples, the conserved gene flaB, p66 and ospC were PCR amplified. A total of 144 dog blood samples, and 846 of ticks were obtained from the examined animals. Considering tick species, Rhipicephalus sanguineus sensu lato (786 / 846) was common, while Ixodes affinis (33 / 846), and Amblyomma mixtum (27 / 846) resulted less frequent. As per gene conservation, the prevalence of B. burgdorferi in canine blood was 17.3 % (25 / 144) to flaB, 12.50 % (18 / 144) for p66 and 1.38 % (2 / 144) for the ospC gene. Within the analyzed ticks, R. sanguineus s.l. had a prevalence of 0.89 %, A. mixtum 5.88 % and I. affinis 15.15 %, being this last species the one that presented higher prevalence. Two dogs and their ticks I. affinis were positive to the flaB gene. Only a tick R. sanguineus s.l. was positive to the gene p66 and no tick species was positive the ospC gene. This study confirmed the existence of B. burgdorferi s.l. in dogs and their ticks in rural communities of Yucatán, Mexico. The detection of Borrelia in dogs may be an important criterion for the evaluation of the risk of borreliosis in humans, since the dog can be used as an epidemiological indicator for the identification of new outbreaks of this disease. Rev. Biol. Trop. 66(1): 428-437. Epub 2018 March 01.

Contributions to the knowledge of hard ticks (Acari: Ixodidae) in Colombia.

The known tick fauna of Colombia includes 58 species (15 Argasidae and 43 Ixodidae). To add to the knowledge of the biology of ticks in Colombia, hard ticks (Ixodidae) were collected from domestic animals or vegetation during 2014-2016 in 10 of Colombia's Departments. Ticks were identified to species through morphological examinations. Taxonomic identification was confirmed for some specimens by molecular methods, including phylogenetic analyses inferred from three tick genes (cytochrome c oxidase, 16S rDNA, second internal transcribed spacer). A total of 1745 tick specimens encompassing 8 species were collected. Overall, 5 tick species were recorded on cattle [Amblyomma dissimile, Amblyomma mixtum, Dermacentor nitens, Rhipicephalus microplus, Rhipicephalus sanguineus sensu lato (s.l.)], 5 on dogs (Amblyomma maculatum, Amblyomma ovale, Amblyomma varium, R. microplus, R. sanguineus s.l.), 3 on horses (A. mixtum, D. nitens, R. sanguineus s.l.), 3 on donkeys (A. mixtum, D. nitens, R. microplus), 1 on pig (D. nitens), and 2 from vegetation (A. mixtum, A. dissimile). This included the first records of A. mixtum from two Colombian Departments, indicating that the distribution of this tick in Colombia may be broader than currently known. Phylogenetic analyses confirmed that R. sanguineus s.l. specimens from 8 Departments belong to the "tropical species". Moreover, Colombian specimens of A. maculatum formed a large clade with GenBank sequences of A. maculatum and A. triste, although some Colombian specimens grouped with A. maculatum from the United States while others grouped with A. triste from Brazil. Significant polymorphisms were observed between specimens of A. ovale or D. nitens; for the former species, it is noteworthy that two distinct clades were observed. Our study provides new records for 8 tick species parasitizing domestic animals in Colombia, including species with veterinary and medical importance in the Neotropical region, such as R. microplus, R. sanguineus, D. nitens, A. mixtum, and A. maculatum. Noteworthy, we provide the first record of A. varium infesting a domestic mammal.

Prevalence, intensity and population dynamics of hard ticks (Acari: Ixodidae) on sheep in the humid tropics of Mexico.

Hard ticks are a perennial problem in livestock systems throughout Mexico. No data are currently available on tick prevalence, infestation intensity and population dynamics for sheep in the humid tropics of Mexico. Blackbelly sheep (n = 30) in Tabasco state, Mexico, were examined every 3 weeks for 1 year, and all Ixodidae ticks were counted and removed for analysis. Tick species were identified, infestation prevalence and intensity calculated per animal, and infestation per body zone determined. Overall infestation on the studied animals was 51.9%. Four tick species were identified (Amblyomma mixtum, A. imitator, Rhipicephalus microplus and R. annulatus), the most abundant being A. mixtum (94.9%). Prevalence was highest during the northwinds season (61.9%), followed by the dry (48.5%) and rainy (47.3%) seasons; however, the intensity (2.01-2.07 ticks/sheep) did not differ between seasons. Infestation was most frequent in certain zones including the axilla, crotch, udder, abdomen, thorax and rib area. Sheep in the humid tropics of Tabasco, Mexico, are parasitized by four hard tick species and prevalence is highest in the northwinds season. The present data constitute an important baseline for developing sustainable tick control programs for sheep in the humid tropics of Mexico.

First report of "Candidatus Rickettsia amblyommii" in west coast of Mexico.

We report the first case of "Candidatus Rickettsia amblyommii" detected in Amblyomma mixtum ticks on humans on the west coast of Mexico. This is the most western record of "Ca. R. amblyommii" in the Western Hemisphere, representing the first record for the western coast of the Americas. Even if the record is far from the previously known locations for the species it does not represent a new record regarding temperature, precipitation and topographic parameters. Since "Ca. R. amblyommii" antibodies have been detected in patients suspected of Rocky Mountain spotted fever, and the tick A. mixtum has been associated with humans, it is important to consider "Ca. R. amblyommii" as a potential risk for the human population that has not been considered at risk before.