Effect of an altitudinal gradient on the morphology, molecular identification and distribution of Rhipicephalus linnaei in Veracruz, Mexico.

Studies of morphological and genetic variation in vector populations across environmental gradients can help researchers to estimate species' responses to climate change scenarios and the potential risk of disease-causing pathogen expansion, which impacts negatively on human health. In this study, we analysed the effect of altitudinal gradients on the phenotypic response of the hard tick of medical and veterinary importance, Rhipicephalus sanguineus sensu lato (s.l.). Specimens of R. sanguineus s.l. were collected from host animals in one of Mexico's regions with high climatic heterogeneity (Veracruz), and geometric morphometric theory was employed to assess the response of three morphological characters to the altitudinal gradient. Additionally, genetic similarity data were provided, and ecological niche models were used to project the climatic distribution in the region. Our results demonstrate that the shape and size of ticks respond to altitude. Molecular identification indicate that all analysed samples correspond to the tropical lineage recently named Rhipicephalus linnaei. According to ecological niche models, the mean annual temperature contributes significantly to the spatial distribution of this tick species, with areas of higher suitability in the mountainous region. These changes in morphological structure and the presence of ticks at higher altitudinal gradients suggest that R. linnaei has a high potential for adaptation. Due to the variability of ecosystems in the state of Veracruz, our results could be valuable in assessing the response of this tick in a changing environment, aiding in predicting future scenarios in the distribution and abundance of this species.

Molecular detection of hemotropic Mycoplasma and Bartonella species in lice from sheep and goats of Mexico.

The knowledge of lice associated with small ruminants, especially sheep and goats, is scarce. In Mexico, there are historical reports of six species of chewing and sucking lice associated with Capra hircus and Ovis canadensis. However, the reports did not analyze the ecology of the infestations or the presence of potentially pathogenic bacteria. For this reason, the objectives of this study were i) to identify the species of lice associated with sheep and goats in three states of the Mexican Republic, ii) to characterize the infestations, and iii) to identify the presence of bacterial pathogens. From October 2019 to August 2021, six ranches with sheep and goats were sampled in the states of Hidalgo and Veracruz. Hosts were visually inspected, and lice were retrieved with forceps. The specimens were sexed and identified using morphological taxonomic keys. DNA extraction was performed individually, and a fragment of the cytochrome oxidase subunit 1 gene (COI) was amplified for the molecular identification of the specimens. Subsequently, Anaplasma, Bartonella, Ehrlichia, Mycoplasma, and Rickettsia were molecularly detected. Additionally, the infestations were characterized by calculating the prevalence and mean abundances. We collected 563 specimens of three species, Bovicola caprae, Bovicola ovis, and Linognathus africanus. The highest infestation levels were recorded for B. ovis (66.7%; 4.4) from Veracruz. Additionally, two Bartonella species were detected: Bartonella mellophagi in B. ovis and Bartonella capreoli in L. africanus. In contrast, Mycoplasma ovis was detected exclusively in one pool of B. ovis. This study provides new bacterial-ectoparasite associations and highlights the possible role of these neglected ectoparasites as vectors in the populations of sheep and goats from Mexico.

Confirmation of the presence of Rickettsia felis infecting Ornithodoros puertoricensis in Mexico.

Soft ticks are neglected competent vectors of a wide range of pathogenic microorganisms, among which bacteria of the genera Rickettsia and Borrelia stand out. In Mexico, previous studies have shown the presence of a member of the Ornithodoros talaje complex in the Virginia opossum (Didelphis virginiana Didelphimorphia: Didelphidae Kerr) from southeastern Mexico. However, its specific identification has not been achieved. Two D. virginiana were treated in a private clinic during the period of April-May 2022. Tick larvae were manually removed, DNA extraction was performed, and some genes from various bacterial and parasitic pathogens were amplified and sequenced. A total of 96 larvae were recovered, which were morphologically identified as Ornithodoros puertoricensis (Ixodida: Argasidae Fox); the 16 S sequences showed a similarity of 96.79%-99.51% with sequences of O. puertoricensis from Panama and Colombia. The presence of Rickettsia felis (Rickettsiales: Rickettsiaceae Bouyer et al.) was detected in 15 specimens from one host. The soft tick O. puertoricensis is recorded for the first time as an ectoparasite of the Virginia opossum in America and represents the second report for this soft tick in Mexico since 1963. This represents the most northern record of this tick species in its geographic distribution and brings a new soft tick-Rickettsia association.

Molecular detection of Coxiella-like endosymbionts and absence of Coxiella burnetii in Amblyomma mixtum from Veracruz, Mexico.

Ticks are obligate ectoparasites associated with a wide range of vertebrate hosts, including domestic animals. Moreover, ticks are capable of transmitting many pathogens such as Coxiella. To date, Coxiella burnetii, the etiological agent of coxiellosis or Q fever, is the only valid species of the genera. Nevertheless, a wide range of agents denominated Coxiella-like have been detected in recent studies, mainly associated with ticks. The pathogenicity of these Coxiella-like agents is controversial as some of them can infect both birds and humans. In Mexico, knowledge about Q fever is scarce and limited to historical serological records, and there is an overall lack of molecular proof of any agent of the genus Coxiella circulating in the country. Therefore, the aim of this study was to detect the presence of Coxiella in ticks associated with cattle in all 10 regions of Veracruz, Mexico. To accomplish this objective, first, we identified ticks collected from cattle and horses in Veracruz. Then, for Coxiella detection, DNA extraction from ticks and PCR amplification of the 16S-rDNA of Coxiella was performed. Finally, we performed a phylogenetic reconstruction to determine the Coxiella lineages detected. From the 10 regions sampled we collected 888 ticks grouped in 180 pools, and only five Amblyomma mixtum from the locality of Castán, and one from Los Angeles from Tuxpan were found positive, which represents a frequency of 20% for each locality. This study represents the first attempt at molecular detection of Coxiella in ticks associated with cattle in the state of Veracruz, the major livestock producer in the country. The findings of the present study are relevant as they establish a precedent regarding the circulation of Coxiella-like agents, as well as the absence in three municipalities of the state of Veracruz of C. burnetii, an abortive agent of livestock importance.

Serosurveillance and risk factors for Neospora caninum infection in water buffaloes (B. bubalis) from central and southern Mexico.

The water buffalo (B. bubalis) is an alternative to cattle ranching in several regions of southern Mexico. Here we report seroprevalence and risk factors associated with the protozoan parasite, Neospora caninum, in water buffaloes in six buffalo production units, in municipalities from central and southern Veracruz, Mexico. Antibodies to N. caninum were assessed in serum samples of 543 buffaloes by a commercial ELISA-kit, and 44.8% (243/543; 95% CI 40.5-49.0) were seropositive. Questionnaires were used to collect epidemiological data and to identify risk factors associated with N. caninum infection. Data analysis indicated that older buffaloes (≥7 year) exhibited the highest seroprevalence for neosporosis 62.3% (38/61; 95% CI 49.7-73.4) (P ≤ 0.05). Buffaloes that were in close contact with cattle had higher seroprevalence 47.6% (168/353; 95% CI 42.3-52.9) (P < 0.01) than those that were not in contact 36.8% (70/190; 95% CI 30.0-44.1). Our findings provide important information to implement preventive measures in the buffalo farms.

Rhipicephalus sanguineus Complex in the Americas: Systematic, Genetic Diversity, and Geographic Insights.

The Rhipicephalus sanguineus group encompasses at least 12 validated species of Palearctic and Afrotropical hard ticks, which are relevant in veterinary medicine and public health. The taxonomy of R. sanguineus s.s., has been particularly intensely debated, due to its wide geographic distribution, morphological variants, parasite-host associations, and its capacity and vectorial competence for the transmission of several pathogens. By sequencing mitochondrial markers, it was possible to identify the existence of multiple lineages, among which the Tropical and the Temperate lineages stand out, particularly in America. However, the northern limit between these lineages is not clear due to the lack of extensive sampling across Mexico. For this reason, the aim of the present study was to determine the genetic diversity and structure of the R. sanguineus group in Mexico and to compare it with the populations reported in the Americas, in order to propose the northern limit of the R. sanguineus Tropical lineage and the potential regions of sympatry with R. sanguineus s.s. The findings of this study now confirm the presence of R. sanguineus s.s. in Mexico, showing a subtle genetic structure and high genetic diversity throughout its distribution in the Americas. In contrast, the Tropical lineage seems to be genetically less diverse in its overall distribution in the Americas. The genetic diversity of these two independent lineages could have important epidemiological implications in the transmission of tick pathogens.

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.

Molecular detection of Theileria equi in horses from Veracruz, Mexico.

Equine piroplasmosis is a tropical parasitic disease caused by several intraerythrocytic members of the genera Babesia and Theileria. These pathogens are transmitted by multiple tick species and are considered as important exotic microorganisms in several countries. Equine piroplasmosis causes significant economic losses due to abortions, decreased activity and even death of equines, making surveillance of these infectious disease essential. In the northern and eastern parts of Mexico, few molecular and serological reports have been made on the presence and exposure of horse to these agents. For this reason, the aim of the current work was to perform a molecular detection study of Babesia and Theileria in equines from the state of Veracruz, Mexico. A total of 100 whole blood samples were tested. Chelex-100 resin was used for DNA extraction and a fragment of 459 bp of the 18S rRNA gene of members of the genera Babesia/Theileria were identified. Of the 100 samples analysed, 18 tested positive for Babesia/Theileria, resulting in a prevalence of 18 %. Identity analyses and phylogenetic reconstruction revealed that all samples were infected with Theileria equi. This work represents the first molecular record of Babesia/Theileria in equines from the state of Veracruz, Mexico, and demonstrates the endemicity of T. equi in this region of the country.

Integrative taxonomic description of the chewing louse Tricholipeurus lipeuroides infesting Odocoileus virginianus veraecrucis white-tailed deer in Veracruz, Mexico.

Chewing lice (Phthiraptera: Amblycera, Ischnocera) represent a component of the ectoparasite fauna associated with large sized mammals as deers. However, the diversity of chewing louse species infesting deer remains to be fully characterized in the Neotropics. Little is known about the chewing lice infesting the extant fourteen subspecies of white-tailed deer (Odocoileus virginianus) in Mexico. Known to infest white-tailed deer (WTD) in Canada and the United States (U.S.), Tricholipeurus lipeuroides is a chewing louse species that was originally described in the nineteenth century infesting O. v. mexicanus in Mexico. For the first time, infestation of O. v. veraecrucis, a Neotropical WTD subspecies in Mexico, with T. lipeuroides is reported herein. An integrative taxonomic approach was taken by combining morphological and molecular analyses to describe the T. lipeuroides infestion of O. v. veraecrucis. Ecological parameters of the T. lipeuroides infestations were also calculated. The prevalence was 91.7% of the 56 O. v. veraecrucis (29 females and 27 males) inspected while under chemical restraint that were sampled at 3 sites in the central region of Veracruz state in Mexico. The amplification and sequencing of previously reported T. lipeuroides Cytochrome Oxidase Subunit I gene confirmed the identity of all the chewing louse life stages. These results are discussed in the context of comparative analyses on the emergence of novel chewing lice-deer associations.

First record of natural infection with Anaplasma marginale in sucking lice infesting the water buffalo (Bubalus bubalis) in Mexico.

The water buffalo (Bubalus bubalis) is an Asian species of bovine which was introduced in Mexico in 1992, as an alternative for milk and meat production. However, little is known about its associated ectoparasites, in particular about their lice. As a part of a project to identify the diversity and prevalence of Anaplasma in cattle and water buffaloes in the state of Veracruz, Mexico, we visited a ranch in the downtown area in which 42 buffaloes were visually inspected for the presence of ectoparasites. Lice were manually recovered, and morphologically identified using taxonomic keys. They were also molecularly identified by the amplification of a fragment of the cytochrome oxidase C subunit I gene (COX1). As a result, we recovered 20 lice from eight animals. All lice were identified as Haematopinus tuberculatus, and the recovered sequences exhibited a similarity of 99.7% with those of the same species deposited in GenBank. Additionally, we detected the presence of Anaplasma marginale in 70% (14/20) of the analysed samples. Our results represent the first record of this exotic ectoparasite for the country.

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.

Apparently high Leptospira antibody seropositivity in donkeys for slaughter in three municipalities in Durango, Mexico.

We determined the seropositive percentage and the determinants of Leptospira exposure in domestic donkeys presented for slaughter in the State of Durango, Mexico. We examined 194 donkeys in 4 gathering premises. Sera of donkeys were analyzed for anti- Leptospira IgG antibodies using a microscopic agglutination assay with a panel of 7 Leptospira antigens. The association between Leptospira seropositivity and general characteristics of donkeys was assessed by bivariate and multivariate analyses. Antibodies against Leptospira (for at least 1 of the 7 Leptospira serovars examined) were found in 151 of 194 (77.8%) donkeys. Seropositive donkeys were found in all regions, municipalities, and premises examined. The predominant serovar was L. interrogans Icterohaemorrhagiae ( n = 117; 60.3%), followed by L. interrogans Sejroe ( n = 96; 49.5%), L. interrogans Canicola ( n = 10; 5.2%), L. kirschneri Grippotyphosa ( n = 6; 3.1%), L. borgpetersenii Ballum ( n = 5; 2.6 %), L. biflexa Semaranga ( n = 3; 1.5%), and L. noguchii Panama ( n = 2; 1%). Leptospira seropositivity was associated with gathering premises (OR = 2.64; 95% CI: 1.27-5.46; p = 0.009) and municipalities (OR = 0.11; 95% CI: 0.01-0.78; p = 0.02). Our results demonstrate an apparently high seropositive percentage of Leptospira infection in the donkeys studied in Mexico.

Seroepidemiology of Infection with Neospora Caninum, Leptospira, and Bovine Herpesvirus Type 1 in Water Buffaloes (Bubalus Bubalis) in Veracruz, Mexico.

We aimed to determine the seroprevalence of infection with Neospora caninum, Leptospira, and bovine herpesvirus type 1 and risk factors associated with these infections in water buffaloes in Veracruz State, Mexico. Through a cross-sectional study, 144 water buffaloes (Bubalus bubalis) raised in 5 ranches of Veracruz were examined for anti-N. caninum and anti-bovine herpesvirus type 1 antibodies by enzyme immunoassays, and anti-Leptospira interrogans antibodies by microscopic agglutination test. Of the 144 buffaloes studied, 35 (24.3%) were positive for N. caninum, 50 (34.7%) for Leptospira, and 83 (57.6%) for bovine herpes virus. The frequencies of leptospiral serovars in buffaloes were as follows: 18.7% for Muenchen (n = 27), 10.4% for Hardjo LT (n = 15), 9.0% for Pyrogenes (n = 13), and 4.8% for Icterohaemorrhagiae (n = 7). Seropositive buffaloes were found in all 5 ranches studied. Logistic regression showed that cohabitation of buffaloes with cows was associated with infection with Leptospira (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.04-4.5; P = 0.03) and bovine herpesvirus (OR, 12.0; 95% CI, 4.0-36.2; P < 0.01). This is the first study that provides serological evidence of N. caninum, Leptospira, and bovine herpesvirus type 1 infections in water buffaloes in Mexico. Our findings could be used to enhance preventive measures against these infections.

Prevalence of Cryptosporidium in small ruminants from Veracruz, Mexico.

BACKGROUND: Cryptosporidiosis is a zoonotic disease caused by the protozoan parasite Cryptosporidium spp. that can affect domestic animal and human populations. In newborn ruminants, cryptosporidiosis is characterized by outbreaks of diarrhea, which can result in high morbidity and economic impact. The aim of the present study was to determine the prevalence of Cryptosporidium spp. in small ruminants from the Perote municipality in Veracruz State, Mexico. One hundred and sixty small ruminants (80 sheep and 80 goats) from eight farms located in four towns of the Perote municipality were examined following a cross-sectional study design. Stool samples were analyzed by a modification of the Faust centrifugation method, and the presence of Cryptosporidium spp. oocysts was examined using a modification of the Ziehl-Neelsen staining procedure. Bivariate and multivariate analyses were used to assess the association of Cryptosporidium infection and the general characteristics of the animals studied. RESULTS: Overall, 112 (70%, 95% CI: 62.3-76.9) of the 160 small ruminants sampled were infected with Cryptosporidium spp. The prevalence of Cryptosporidium spp. infection in goats was 72.5% (95% CI: 61.4-81.9) and in sheep 67.5% (95% CI: 56.1-77.6). Small ruminants aged 1 month old had the highest (88.2%; 95% CI: 63.6-98.5) prevalence of infection. Prevalence varied from 60% to 85% among herds. Animal species, age, sex, breed, farm, town or cohabitation with cattle did not influence the prevalence of Cryptosporidium infection. CONCLUSIONS: A high prevalence of infection with Cryptosporidium spp. was observed in small ruminants from the Perote municipality in Veracruz, Mexico. Infection was widely distributed among sheep and goats regardless of their age, breed or farm location. Further research is required to identify risk factors for, and to assess the veterinary public health significance of Cryptosporidium infection among sheep and goats in the Mexican state of Veracruz.

Seroprevalence of Toxoplasma gondii infection in water buffaloes (Bubalus bubalis) in Veracruz State, Mexico and its association with climatic factors.

BACKGROUND: Infection with Toxoplasma gondii in water buffaloes (Bubalus bubalis) is of epidemiological importance because of the risk for transmission to humans. We sought to determine the seroprevalence of T. gondii infection in 339 water buffaloes in Veracruz State, Mexico using the modified agglutination test (MAT, cut off 1:25). Seroprevalence association with general characteristics of buffaloes and their environment was also investigated. RESULTS: Antibodies to T. gondii were found in 165 (48.7%) of the 339 buffaloes with MAT titers of 1:25 in 104, 1:50 in 52, and 1:100 in 9. Bivariate analysis showed that seroprevalence of T. gondii infection was similar in buffaloes regardless of their general characteristics i.e., age, sex, and breed. In contrast, the seroprevalence in buffaloes varied significantly with environmental characteristics including altitude, mean annual temperature, and mean annual rainfall of the municipalities studied. Multivariate analysis showed that T. gondii seropositivity in buffaloes was associated with a mean annual rainfall between 1266-1650 mm (OR = 1.84; 95% CI: 1.15-2.94; P = 0.01). CONCLUSIONS: Results indicate that environmental characteristics may influence the seroprevalence of T. gondii infection in buffaloes. This is the first report on the seroprevalence of T. gondii infection in buffaloes in Mexico. Further research is needed to assess the risk for infection in humans associated with the ingestion of raw or undercooked meat from buffaloes infected with T. gondii.