Key molecules regulating the blood meals of Rhipicephalus sanguineus (Acari: Ixodidae) revealed by transcriptomics.

Rhipicephalus sanguineus, a repulsive obligate blood feeder, is a three-host tick inflicting tremendous damage. Blood-sucking initiates tick-pathogen-host interactions along with alterations in the expression levels of numerous bioactive ingredients. Key molecules regulating blood meals were identified using the transcriptomic approach. A total number of 744 transcripts showed statistically significantly differential expression including 309 significantly upregulated transcripts and 435 significantly downregulated transcripts in semiengorged female ticks compared to unfed ticks, all collected in 2021. The top 10 differentially upregulated transcripts with explicit functional annotations included turripeptide OL55-like protein, valine tRNA ligase-like protein and ice-structuring glycoprotein-like protein. The top 10 differentially down-regulated transcripts were uncharacterized proteins. Gene Ontology (GO) enrichment analysis revealed four associated terms in the cellular component category and 16 in the molecular function category among the top 20 terms. Differentially expressed genes (DEGs) were enriched in GO terms ID 0000323 (lytic vacuole) and ID 0005773 (vacuole). The top 20 enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways included metabolism, cellular processes, organismal systems and human diseases. The DEGs were enriched in the KEGG term ID: ko-04142 (lysosome pathway) associated with intracellular digestion in the tick midgut epithelium. Molecular markers annotated via comparative transcriptomic profiling were expected to be candidate markers for the purpose of tick control.

Description of Rhipicephalus hibericus sp. nov. (Ixodoidea: Ixodidae), a species of the Rhipicephalus sanguineus group in southwestern Europe.

We describe all the life stages of Rhipicephalus hibericus n. sp., provide the types, and present molecular support for a new species of the Rhipicephalus sanguineus sensu lato group, present in southwestern Europe, that has been historically confused with Rhipicephalus turanicus Pomerantzev, 1940. A new name is proposed for this taxon because it was impossible to ascribe to types of already described species in the group, deposited for more than 100 years in natural history institutions. The males have a dorsum showing deep and coarse punctations (absent in Rhipicephalus sanguineus sensu stricto) and adanal plates with large punctations (absent in R. sanguineus s.s.); the tail of the spiracular plate is as wide as the closest festoon (half the width in R. sanguineus s.s.). Females have large punctations in dorsal fields, a wide spiracular plate, and a "V" shaped genital opening; such a combination of characters cannot be found in other species of the group. Immatures are described from specimens collected on hosts (Rodentia and Eulipotyphla). Both larvae and nymphs are markedly smaller than R. sanguineus s.s. Nymphs display long, backward-projected auriculae; larvae are almost half the size of R. sanguineus s.s. The new species can hybridize with R. sanguineus s.s. in laboratory colonies producing an unfertile F2, laying brown and dry eggs that did not hatch. Phylogenetic analysis of partial coxI gene sequences placed R. hibericus in a well-supported clade with other sequences of R. sanguineus s.l. from Portugal, as a sister clade of R. sanguineus s.s. The new species does not belong to the R. turanicus group of species. Both 12S and 16S partial gene sequences were not as precise in the correct phylogenetic placement of R. hibericus, in part probably due to the existence of erroneously identified sequences in GenBank©. This description, together with the previous reinstatement of Rhipicephalus secundus and Rhipicephalus rutilus, and the description of the neotypes of R. sanguineus s.s. should help researchers to adequately identify their collections. Our findings demonstrate that R. turanicus is absent in southwestern Europe. Old collections should be re-examined to provide the actual range of the new species.

A mutation associated with resistance to synthetic pyrethroids is widespread in US populations of the tropical lineage of Rhipicephalus sanguineus s.l.

The brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.), is an important vector for Rickettsia rickettsii, causative agent of Rocky Mountain spotted fever. Current public health prevention and control efforts to protect people involve preventing tick infestations on domestic animals and in and around houses. Primary prevention tools rely on acaricides, often synthetic pyrethroids (SPs); resistance to this chemical class is widespread in ticks and other arthropods. Rhipicephalus sanguineus s.l. is a complex that likely contains multiple unique species and although the distribution of this complex is global, there are differences in morphology, ecology, and perhaps vector competence among these major lineages. Two major lineages within Rh. sanguineus s.l., commonly referred to as temperate and tropical, have been documented from multiple locations in North America, but are thought to occupy different ecological niches. To evaluate potential acaricide resistance and better define the distributions of the tropical and temperate lineages throughout the US and in northern Mexico, we employed a highly multiplexed amplicon sequencing approach to characterize sequence diversity at: 1) three loci within the voltage-gated sodium channel (VGSC) gene, which contains numerous genetic mutations associated with resistance to SPs; 2) a region of the gamma-aminobutyric acid-gated chloride channel gene (GABA-Cl) containing several mutations associated with dieldrin/fipronil resistance in other species; and 3) three mitochondrial genes (COI, 12S, and 16S). We utilized a geographically diverse set of Rh sanguineus s.l. collected from domestic pets in the US in 2013 and a smaller set of ticks collected from canines in Baja California, Mexico in 2021. We determined that a single nucleotide polymorphism (T2134C) in domain III segment 6 of the VGSC, which has previously been associated with SP resistance in Rh. sanguineus s.l., was widespread and abundant in tropical lineage ticks (>50 %) but absent from the temperate lineage, suggesting that resistance to SPs may be common in the tropical lineage. We found evidence of multiple copies of GABA-Cl in ticks from both lineages, with some copies containing mutations associated with fipronil resistance in other species, but the effects of these patterns on fipronil resistance in Rh. sanguineus s.l. are currently unknown. The tropical lineage was abundant and geographically widespread, accounting for 79 % of analyzed ticks and present at 13/14 collection sites. The temperate and tropical lineages co-occurred in four US states, and as far north as New York. None of the ticks we examined were positive for Rickettsia rickettsii or Rickettsia massiliae.

Rhipicephalus sanguineus from Hungarian dogs: Tick identification and detection of tick-borne pathogens.

The brown dog tick, Rhipicephalus sanguineus is a complex of tick species with an unsettled species concept. In Europe, R. sanguineus is considered mainly a Mediterranean tick with sporadic findings in central and northern Europe. R. sanguineus is known as a vector of a range of pathogens of medical and veterinary importance, most of which not yet reported as autochthonous in Hungary. A total of 1839 ticks collected by veterinarians from dogs and cats were obtained in Hungary. The study aims at precise determination of ticks identified as R. sanguineus and detection of pathogens in collected ticks. All ticks were morphologically determined and 169 individuals were identified as R. sanguineus. A subset of 15 ticks was selected for molecular analysis (16S rDNA, 12S rDNA, COI). Phylogenetic analyses invariably placed sequences of all three markers into a single haplotype identified as R. sanguineus sensu stricto. All 169 brown dog ticks were tested for the presence of A. platys, E. canis, R. conorii, B. vogeli and H. canis. None of the investigated ticks was positive for the screened pathogens, though A. phagocytophilum sequence was detected in a single tick.

The emergence of Rocky Mountain spotted fever in the southwestern United States and northern Mexico requires a binational One Health approach.

Rocky Mountain spotted fever (RMSF) is an international and quintessential One Health problem. This paper synthesizes recent knowledge in One Health, binational RMSF concerns, and veterinary and human medical perspectives to this fatal, reemerging problem. RMSF, a life-threatening tick-borne disease caused by the bacterium Rickettsia rickettsii, emerged during the first decade of the 21st century in impoverished communities in the southwestern US and northern Mexico. Lack of an index of suspicion, delay in diagnosis, and delayed initiation of antibiotic treatment contribute to fatality. Campaigns targeting dog neutering, restraint to residents' properties, and on-dog and on-premises treatment with acaricides temporarily reduce prevalence but are often untenable economically. Contemporary Mexican RMSF is hyperendemic in small communities and cities, whereas epidemics occur in the western US primarily in small tribal communities. In in both locations, the epidemics are fueled by free-roaming dogs and massive brown dog tick populations. In the US, RMSF has a case fatality rate of 5% to 7%; among thousands of annual cases in Mexico, case fatality often exceeds 30%.1,2 Numerous case patients in US border states have recent travel histories to northern Mexico. Veterinarians and physicians should alert the public to RMSF risk, methods of prevention, and the importance of urgent treatment with doxycycline if symptomatic. One Health professionals contribute ideas to manage ticks and rickettsial disease and provide broad education for the public and medical professionals. Novel management approaches include vaccine development and deployment, acaricide resistance monitoring, and modeling to guide targeted dog population management and other interventions.

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.

Distribution of Rhipicephalus sanguineus and Heamaphysalis elliptica dog ticks and pathogens they are carrying: A systematic review.

The role of ixodid ticks especially Rhipicephalus sanguineus and Heamaphysalis elliptica in the epidemiology of several diseases of veterinary and public health importance have been documented. This study conducted a systematic review focusing on the distribution of R. sanguineus and H. elliptica, as well as the common tick-borne pathogens they harbour. The Scopus, ScienceDirect, PubMed, and Web of Science databases were used to search for English journal articles published between January 1990 and June 2021. The articles were assessed by following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. This systematic review was registered on PROSPERO [(ID no: CRD42022327372). Of the studies included in the systematic review, 247 and 19 articles had identified R. sanguineus and H. elliptica respectively, whereas 15 articles had identified both tick species. There is a reported worldwide distribution of R. sanguineus from 64 countries, whereas H. elliptica was only reported in the African continent from 6 countries. In total, 120 articles that were included in this systematic review reported detection of tick-borne pathogens from R. sanguineus (n = 118 articles) and/or H. elliptica (n = 2 articles) ticks. According to the studies tick-borne pathogens harboured by R. sanguineus included protozoa such as Babesia spp., Hepatozoon spp., Leishmania spp., and Theileria spp., as well as bacteria such as Acinetobacter spp. Anaplasma spp., Bacillus spp., Borrelia spp., Brucella spp., Coxiella spp., and Staphylococcus spp. The H. elliptica was reported to be harbouring Babesia spp., Ehrlichia spp. and Rickettsia spp. Most of the studies (50%) used the conventional polymerase chain reaction (PCR) technique for the detection of tick-borne pathogens, followed by real-time PCR (qPCR) (n = 26), and nested PCR (n = 22). This systematic review has shed light on the distribution of two common dog ticks as well as the tick-borne pathogens of veterinary and zoonotic importance they are harbouring. This data will enable surveillance studies that can report whether the distribution of these ticks and their associated tick-borne pathogens is expanding or shrinking or is stable.

Cross-species immunoprotective antigens (subolesin, ferritin 2 and P0) provide protection against Rhipicephalus sanguineus sensu lato.

BACKGROUND: Tick control is mostly hampered by the rise of acaricide-resistant tick populations. Significant efforts have focused on developing alternative control methods, including cross-species protective and/or cocktail-based anti-tick vaccines, to achieve protection against various tick species. METHODS: In this study, full-length open reading frames encoding subolesin (SUB) from Rhipicephalus microplus and ferritin 2 (FER2) from Hyalomma anatolicum as well as the partial 60S acidic ribosomal protein (P0) from R. microplus were cloned, expressed in Escherichia coli and used as vaccine antigens against Rhipicephalus sanguineus sensu lato (R. sanguineus s.l.) infestation in rabbits. RESULTS: In silico analyses revealed that the SUB, P0 and FER2 proteins were antigenic and displayed limited similarity to the host's homologous proteins. The proteins shared identities of 97.5%, 100% and 89.5% with their SUB, P0 and FER2 R. sanguineus s.l. orthologous sequences, respectively. Antibodies against each recombinant protein cross-recognized the native proteins in the different tissues and developmental stages of R. sanguineus s.l. Overall efficacy of the SUB, FER2 and cocktail (SUB+FER2+P0) vaccines against R. sanguineus s.l. infestation was 86.3%, 95.9% and 90.9%, respectively. CONCLUSIONS: Both mono-antigen and the cocktail anti-tick vaccines affected the biological parameters of R. sanguineus s.l. infestation in the rabbit model, which could be extrapolated to its infested host under natural conditions. These findings support the possibility of using mono-antigenic and cocktail-based vaccines for large-scale anti-tick vaccine development against multiple tick species.

Ecological factors shaping ectoparasite communities on heteromyid rodents at Médanos de Samalayuca.

Rodent ectoparasites are vectors for important pathogens of wildlife, domestic animals, and even zoonosis. Nevertheless, distribution patterns of ectoparasites are not fully understood; habitat, season, and host species are important predictors of distribution and prevalence. Heteromyid rodents are considered important reservoirs of diseases, given the presence of different ectoparasites and pathogens in them, and they offer the opportunity to learn about the ecology of parasites. The aim of the present work was to survey ectoparasites associated with heteromyid rodents near a National Protected Area in Chihuahua Mexico, south of the USA-Mexico border, and asses the effects of ecological factors (season, vegetation type, host species, and host body condition) on parasite infestation. We sampled five different locations from January 2018 to July 2022; 845 heteromyid rodents were examined and 49 fleas and 33 ticks were collected. Ectoparasites belonged to the Siphonaptera and Ixodida orders, including three families Ixodidae (Riphicephalus sanguineus), Pulicidae (Pulex irritans), and Ctenophthalmidae (Meringins altipecten, M. dipodomys). Five species of host rodents were captured, Dipodomys merriami, D. ordii, Chaetodipus eremicus, C. hispidus, and C. intermedius, but the last two species did not present any ectoparasites. Dipodomys merriami presented the highest flea and tick prevalence followed by D. ordii. We found parasitic partnerships between heteromyids according to ecological factors. The infestation in C. eremicus was related to body condition, vegetation type, and sex; in D. merriami, it was related to vegetation type and season, while D. ordii did not present a clear pattern of infestation. Our results suggest that the infestation patterns of heteromyid rodents in desert habitats are species dependent.

Molecular Evidence of Ehrlichia canis (Rickettsiales: Anaplasmataceae) in Ticks (Ixodida: Ixodidae) Associated with Dogs (Carnivora: Canidae) from Nuevo Leon, Mexico.

Background: Ehrlichia canis is transmitted by ticks causing Canine monocytic ehrlichiosis, which is considered one of the most critical tickborne pathogens. Materials and Methods: This study aimed to identify by PCR technique E. canis in ticks associated with dogs from urban and rural homes in Nuevo Leon, Mexico. The study was conducted at 13 localities in eight municipalities from 2012 to 2021. Results: A total of 1873 ticks of three species were captured: Amblyomma tenellum, Dermacentor variabilis, and Rhipicephalus sanguineus s.l. The overall infection rate of E. canis in ticks was 59.12% (149/252). Of the 15 sequences, three haplotypes were identified. Conclusion: The urban transmission cycle of canine ehrlichiosis is demonstrated, where the potential vector is the tick R. sanguineus s.l.

Optimal control of a tick population with a view to control of Rocky Mountain Spotted Fever.

In some regions of the Americas, domestic dogs are the host for the tick vector Rhipicephalus sanguineus, and spread the tick-borne pathogen Rickettsia rickettsii, which causes Rocky Mountain Spotted Fever (RMSF) in humans. Interventions are carried out against the vector via dog collars and acaricidal wall treatments. This paper investigates the optimal control of acaricidal wall treatments, using a prior model for populations and disease transmission developed for this particular vector, host, and pathogen. It is modified with a death term during questing stages reflecting the cost of control and level of coverage. In the presence of the control, the percentage of dogs and ticks infected with Ri. rickettsii decreases in a short period and remains suppressed for a longer period, including after treatment is discontinued. Risk of RMSF infection declines by 90% during this time. In the absence of re-application, infected tick and dog populations rebound, indicating the eventual need for repeated treatment.

Efficacy of D-Limonene Nanoemulsion Against Rhipicephalus annulatus and Rhipicephalus sanguineus Ticks.

PURPOSE: Ticks infestation has a negative impact against human and animal health through blood sucking, transmission of blood-borne diseases and also caused economic losses. METHODS: In the present study the adulticidal, ovicidal and larvicidal activity of D-limonene nanoemulsion (DLN) were evaluated against two tick species; Rhipicephalus annulatus and Rhipicephalus sanguineus. Nanoemulsion form of D-limonene was prepared, and its characteristics were evaluated using a UV spectrophotometer and zeta droplet size measurement. Acetylcholinesterase activity was determined. RESULTS: The results revealed significant adulticidal effect with low LC50 and LC90 for D-limonene pure form (DL) against both adult tick spp. (R. annulatus and R. sanguineus) ((0.958 and 1.559%) and (2.26 and 3.51%), respectively). DLN LC50 and LC90 values were ((1.277 and 2.396) and (3.97 and 7.28), respectively) against R. annulatus and R. sanguineus, respectively. DL and DLN showed significant ovicidal effect against R. sanguineus at high concentrations (10 and 5%). In larval packet test, LC50 and LC90 values of DL were ((1.53 and 2.22%) and (6.81 and 12.07%), respectively) against R. annulatus and R. sanguineus, respectively, while LC50 and LC90 values of DLN were ((6.48 and 11.26%) and (7.82 and 13.59%), respectively) against R. annulatus and R. sanguineus, respectively. Significant acetylcholinesterase inhibition percentage was detected for both ticks spp. which treated by DL and DLN. CONCLUSION: Pure DL is more effective than DLN form against R. annulatus and R. sanguineus.

Year-round efficacy of a single treatment of fluralaner injectable suspension (Bravecto Quantum™) against repeated infestations with Rhipicephalus sanguineus (sensu lato) and Ctenocephalides felis in dogs.

BACKGROUND: Poor owner compliance with monthly control measures means that dogs in Australia can remain susceptible to infestations with fleas, present throughout the whole year, and brown dog ticks, which thrive in tropical and subtropical areas. A 150 mg/ml injectable fluralaner suspension (Bravecto Quantum™) was developed to help ensure year-round protection against these parasites. A study investigated the persistent efficacy of this formulation against repeated challenges with Rhipicephalus sanguineus (sensu lato) and Ctenocephalides felis throughout 12 months following a single subcutaneous treatment. METHODS: Twenty dogs were blocked by pre-treatment R. sanguineus s.l. counts and randomized to an untreated control group or to a group treated once, on day 0, with the fluralaner injection (15 mg/kg). Infestations of 50 mixed-sex, unfed adult R. sanguineus s.l. and up to 100 C. felis were done on days 7, 14, 35, 63, 91, 126, 154, 182, 210, 245, 273, 301, 336 and 365. Live flea and tick counts were completed 48 h post-infestation. Flea infestations were also done on day -1, with counts on day 2. Infestations were considered adequate if at each evaluation, at least six dogs in the control group retained at least 20% of tick challenges and 25% of flea challenges. RESULTS: The fluralaner injectable suspension was well tolerated. Efficacy against existing flea infestations was > 99% (arithmetic and geometric means) at 48 h post-treatment. At all subsequent assessments throughout the year following treatment, efficacy against fleas remained at 100%. Arithmetic mean tick count reductions relative to the control group ranged from 97.6% to 100% from day 7 through 11 months and was 92.6% at 12 months (geometric means 95.2% to 100% through 12 months). CONCLUSION: The injectable fluralaner suspension was effective against fleas and brown dog ticks for 12 months following a single treatment. Compared with more frequently administered products where compliance may be suboptimal, the year-round efficacy of this veterinarian-administered fluralaner formulation has the potential to improve owner compliance for control of fleas and ticks. In turn, by reducing the detrimental effects of flea and tick infestations and risk of transmission of flea- and tick-borne pathogens, canine health can be enhanced.

History and Current Status of Mediterranean Spotted Fever (MSF) in the Crimean Peninsula and Neighboring Regions along the Black Sea Coast.

Mediterranean spotted fever (MSF) is a tick-borne rickettsiosis caused by Rickettsia conorii subspecies conorii and transmitted to humans by Rhipicephalus sanguineus ticks. The disease was first discovered in Tunisia in 1910 and was subsequently reported from other Mediterranean countries. The first cases of MSF in the former Soviet Union were detected in 1936 on the Crimean Peninsula. This review summarizes the historic information and main features of MSF in that region and contemporary surveillance and control efforts for this rickettsiosis. Current data pertinent to the epidemiology of the disease, circulation of the ticks and distribution of animal hosts are discussed and compared for each of the countries in the Black Sea basin where MSF occurs.

Detection of ivermectin and fipronil resistance in Rhipicephalus sanguineus sensu lato in Maha Sarakham, Thailand.

Background and Aim: Administration is the main strategy for controlling ectoparasites in dogs. Ivermectin and fipronil are most extensively used to prevent and treat Rhipicephalus sanguineus sensu lato infestation in dogs in Thailand. Several researchers have reported resistance to acaricides in R. sanguineus s.l. globally, but documentation of acaricide resistance in the tick population in Thailand is lacking. In this study, we investigated the status of resistance to commonly used acaricides in Thailand in R. sanguineus s.l. Materials and Methods: Engorged brown dog tick females (10 tick populations) were field-collected directly from parasitized dogs in Maha Sarakham, Thailand, for toxicological bioassays with ivermectin and fipronil. Bioassays were performed in three replicates at 25°C-27°C and 80%-85% relative humidity under a 12-h/12-h photoperiod. The 50% of lethal concentration and its confidence intervals and the slope were estimated for each tick population using probit analysis. Resistance ratios (RRs) of field ticks were characterized based on the relative susceptible population of each acaricide. Results: Six tick populations (P1-6) were tested for resistance to ivermectin, three of which (P2-4) exhibited low-level resistance to ivermectin (RR = 2.115-2.176). Of four tick populations (P7-10) treated with fipronil, two exhibited moderate-to-severe resistance (P7 and P9, RR = 21.684 and 4.387, respectively). All tick populations deemed resistant to acaricides had a history of exposure. Conclusion: Based on RR values, four R. sanguineus s.l. tick populations from Maha Sarakham province were resistant to ivermectin and fipronil. To the best of our knowledge, this represents the first documentation of acaricide-resistant populations of R. sanguineus s.l. in Thailand, and recommendations on tick control programs must be formulated with veterinarians and pet owners to prevent the development of further resistance.

First detection and genetic identification of Rickettsia spp. from ticks collected from rodents in north India.

BACKGROUND: Changing climatic conditions and invasion of ticks in urban areas have led to a greater number of cases of tick-borne diseases, thus, becoming a matter of increasing concern. Tick borne rickettsioses are one of the important emerging diseases worldwide. Knowledge of epidemiology of the vector and pathogen in the community is essential in order to understand and prevent the transmission of the disease to healthy population. METHODS: In our present study, we trapped rodents in selected areas of Chandigarh and Punjab in north India. The rodents were screened for the presence of ticks which were further screened for the presence of rickettsial agents. PCRs targeting 17 â€‹kDa and gltA genes were carried out followed by Sanger sequencing of the positive amplicons followed by phylogenetic analysis of the sequences. RESULTS: A total of 17 ticks were collected out of which one (Rhipicephalus sanguineus) was found to be harboring a Rickettsia sp. PCR targeting gltA and 17 â€‹kDa genes of rickettsia were put up and Sanger sequencing was performed. Phylogenetic analysis revealed the sequences to be closely related to Rickettsia rhipicephali. CONCLUSION: The current study establishes the presence of rickettsial agents in the community. Although Rickettsia rhipicephali is a non-pathogenic agent, the study encourages more vigorous community surveillance should be carried out in order to determine the exact burden of rickettsial agents in our community. To our knowledge, this is the first study reporting Rickettsia rhipicephali in India.

Morphological and molecular identification of the brown dog tick in Mexico.

Ticks of the Rhipicephalus sanguineus complex are known as the brown dog ticks. This complex groups at least 12 species of ticks that are distributed worldwide. On the American continents, R. sanguineus sensu stricto (s.s.), is distributed in temperate areas, while Rhipicephalus sanguineus sensu lato (s.l.), also called "tropical lineage" is distributed in tropical regions. Previous analyses of brown dog ticks from Mexico have identified the so-called tropical lineage and the country generally has a climate more favorable for these ticks (> 20o C in average). In addition, some pathogens thought to be transmitted by this lineage (such as Ehrlichia canis, and Rickettsia rickettsii) are prevalent in Mexico. Herein we aim to contribute to the study of brown dog ticks by providing morphological identification and molecular analysis of mt 12S rDNA and 16S rDNA sequences from ticks collected from 12 states in Mexico. Our results indicate that the tropical lineage of R. sanguineus s.l., recently redescribed as R. linnaei is widely distributed in Mexico.

Molecular Detection and Characterization of Ehrlichia canis Isolates from Three Geographic Regions in Mexico: A Retrospective Study.

Canine monocytic ehrlichiosis (CME) is the most common tick-borne disease affecting domestic dogs and other wild canids. It has a worldwide distribution and is associated with the presence of the brown dog tick. Few studies have been conducted in Mexico to identify and characterize Ehrlichia canis genetic variability. In the present study, 111 dogs of different sex, breed, and age from three geographic regions in Mexico were included. All of them had a previous history of tick infestation and/or the presence of one or more clinical signs compatible with CME. All dogs were tested by a commercial ELISA and nested PCR assay for the detection of E. canis. In addition, we analyzed the E. canis genetic diversity from the 16S rRNA gene sequences obtained in this study, along with 15 additional sequences described for E. canis in Mexico and obtained from GeneBank. Serological detection by commercial ELISA results showed overall infection rates of 85.58% (95/111), including 73.1% (30/41) in samples from Guerrero state; 75% (15/20) in Morelos; and 100% (50/50) in Chihuahua. On the other hand, molecular detection (nPCR assay) showed 31.5% (35/111) overall infection rate, with 41.4% (17/41) in Guerrero state; 55% (11/20) in Morelos; and 14% (7/50) in Chihuahua. We observed a high 16S rRNA gene sequence conservancy in most of the E. canis isolates in the three geographical areas from Mexico, including those analyzed in this research, suggesting a common geographic origin among isolates.

First report of multiple Rickettsia sp., Anaplasma sp., and Ehrlichia sp. in the San Miguel Department of El Salvador from zoonotic tick vectors.

Neglected bacterial zoonoses are a group of Neglected Tropical Diseases (NTDs) that are commonly underdiagnosed and underreported due to their undifferentiated febrile illness symptomology. Spotted fever group rickettsioses (SFGR), a subset of tick-borne bacterial zoonoses, belong in this group. There is a dichotomy in the reporting and recognition of these pathogens in Central America: countries with reduced human development scores-like El Salvador-have little to no research or surveillance dedicated to these pathogens and the diseases they cause. This was the third-ever tick survey in El Salvador, highlighting the knowledge gap in this country. A total of 253 ticks were collected from 11 animals at two farm sites and one veterinary office. Standard and quantitative PCR were used to detect presence of SFGR, Ehrlichia, and Anaplasma sp. pathogens in ticks. Ehrlichia sp. were detected in 2.4% of all collected ticks and Anaplasma sp. were detected in 5.5% of all ticks. Rickettsia rickettsii was amplified in 18.2% of ticks, and amplicons similar to R. parkeri, and R. felis were found in 0.8% and 0.4%, of collected ticks, respectively. This is the first report of these pathogenic bacterial species in El Salvador. This study emphasizes the need for further surveillance and research including incorporating additional human seroprevalence and testing to understand the public health burden in this country.

Correlation between Babesia Species Affecting Dogs in Taiwan and the Local Distribution of the Vector Ticks.

The objective of our study was to survey Babesia infection rates by PCR and tick species on stray dogs to correlate the distribution of Babesia with the distribution of ticks infesting dogs in Taiwan. Three hundred eighty-eight blood samples and 3037 ticks were collected from 388 roaming, and free-ranging owned dogs at residential sites in Taiwan between January 2015 and December 2017. The prevalence of B. gibsoni and B. vogeli was 15.7% (61/388) and 9.5% (37/388), respectively. Most positive B. gibsoni dogs were found in the northern part of the country 56/61 (91.8%), whereas a few were found in the middle 5/61 (8.2%). Babesia vogeli infection rates were 10%, 3.6%, and 18.2% in the northern, central, and southern regions, respectively. Five species of ticks were found: Rhipicephalus sanguineus (throughout Taiwan), Rhipicephalus haemaphysaloides (in the north), Haemaphysalis hystricis (in the north and middle of Taiwan), and Amblyomma testidunarium and Ixodes ovatus (both in the north). None of the dogs in the south were infected with B gibsoni, which correlated with the absence of H. hystricis, a tick recently identified as the local vector for B gibsoni. Babesia vogeli was more equally distributed, coinciding with R. sanguineus, a tick that is present throughout Taiwan. Anaemia was detected in 86.9% of infected dogs; among these dogs, approximately 19.7% showed severe anaemia (HCT < 20). These findings provide useful advice for owners regarding outdoor activities with their dogs and local veterinarians with a regional differential diagnosis of babesiosis in Taiwan.