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.

Prevalencia de Borrelia burgdorferi sensu lato en roedores sinantrópicos de dos comunidades rurales de Yucatán, México / Prevalence of Borrelia burgdorferi sensu lato in synanthropic rodents in two rural communities of Yucatán, México

Introducción. La enfermedad de Lyme es una zoonosis multisistémica causada por Borrelia burgdorferi sensu lato. Esta espiroqueta circula en un ciclo enzoótico entre un reservorio vertebrado primario y las garrapatas. Se ha encontrado que varias especies de roedores son eficientes reservorios naturales de B. burgdorferi s.l. Objetivo. Estimar la prevalencia de B. burgdorferi s.l. en roedores sinantrópicos en dos comunidades rurales de Yucatán, México. Materiales y métodos. Se capturaron 123 roedores (94 Mus musculus y 29 Rattus rattus ) para obtener muestras de tejidos de oreja y vejiga. Para la detección de B. burgdorferi s.l. en las muestras, se amplificaron los genes de la flagelina B ( fla B ) y las lipoproteínas de membrana externa, ospC y p66 , mediante reacción en cadena de la polimerasa, y se secuenciaron los amplicones obtenidos. Resultados. La frecuencia de infección por B. burgdorferi s.l. en roedores fue de 36,5 % para flaB (45/123), de 10,5 % (13/123) para p66 y de 3,2 % (4/123) para ospC . En R. rattus la frecuencia de infección fue de 17,2 % y en M. musculus fue de 42,5 %. La frecuencia de infección de B. burgdorferi s.l. en los tejidos estudiados fue de 11,3 % (14/123) en muestras de tejido de vejiga y de 17,0 % (21/123) en las de oreja. No se encontraron diferencias estadísticas (p>0,05) en la frecuencia de infección entre los dos tipos de muestras de tejido utilizadas para el diagnóstico. El gen ospC presentó 98 % de homología con la especie Borrelia garinii , una de las especies heterogéneas del complejo B. burgdorferi s.l. Conclusiones. Los roedores presentaron una alta prevalencia de infección con B. burgdorferi s.l.; las especies M. musculus y R. rattus podrían jugar un papel importante en la continuidad de la presencia de esta bacteria en comunidades rurales de Yucatán, México.

Introduction: Lyme disease is a multisystemic zoonotic disease caused by Borrelia burgdorferi sensu lato. This spirochete circulates in an enzootic cycle between the primary vertebrate reservoir and its tick vectors. Different species of rodents are known to be efficient natural reservoirs for B. burgdorferi s.l. Objective: To estimate the prevalence of B. burgdorferi s.l. in synanthropic rodents from two rural communities of Yucatán, México. Materials and methods: A total of 123 rodents (94 Mus musculus and 29 Rattus rattus ) were trapped, and ear and bladder samples were collected. Flagelin B ( flaB ) genes and outer membrane lipoproteins ospC y p66 were amplified in order to detect B. burgdorferi s.l. presence in the samples. The obtained amplicons were sequenced. Results: The overall infection rates in rodents were 36.5% for flaB (45/123), 10.5% (13/123) for p66, and 3.2% (4/123) for ospC . Rattus rattus had 17.2% of infection and M. musculus , 42.5%. From all examined tissue, 11.3% (14/123) of bladders, and 17.0% (21/123) of ears were infected with the spirochete Borrelia burgdorferi s.l. No statistical differences (p>0.05) were found between the two tissue samples used for diagnosis. The ospC gen was 98% homologous to Borrelia garinii , one species of the B. burgdorferi s.l. complex. Conclusions: We concluded that rodents have a high prevalence of B. burgdorferi s.l. infection, and both species of rodents, M. musculus and R. rattus, might be playing an important role in the maintenance of this bacterium in rural communities of Yucatán, México.

Prevalence of Borrelia burgdorferi sensu lato in synanthropic rodents in two rural communities of Yucatán, México.

INTRODUCTION: Lyme disease is a multisystemic zoonotic disease caused by Borrelia burgdorferi sensu lato. This spirochete circulates in an enzootic cycle between the primary vertebrate reservoir and its tick vectors. Different species of rodents are known to be efficient natural reservoirs for B. burgdorferi s.l.  OBJECTIVE: To estimate the prevalence of B. burgdorferi s.l. in synanthropic rodents from two rural communities of Yucatán, México.  MATERIALS AND METHODS: A total of 123 rodents (94 Mus musculus and 29 Rattus rattus) were trapped, and ear and bladder samples were collected. Flagelin B (flaB) genes and outer membrane lipoproteins ospC y p66 were amplified in order to detect B. burgdorferi s.l. presence in the samples. The obtained amplicons were sequenced.  RESULTS: The overall infection rates in rodents were 36.5% for flaB (45/123), 10.5% (13/123) for p66, and 3.2% (4/123) for ospC. Rattus rattus had 17.2% of infection and M. musculus, 42.5%. From all examined tissue, 11.3% (14/123) of bladders, and 17.0% (21/123) of ears were infected with the spirochete Borrelia burgdorferi s.l. No statistical differences (p>0.05) were found between the two tissue samples used for diagnosis. The ospC gen was 98% homologous to Borrelia garinii, one species of the B. burgdorferi s.l. complex.  CONCLUSIONS: We concluded that rodents have a high prevalence of B. burgdorferi s.l. infection, and both species of rodents, M. musculus and R. rattus, might be playing an important role in the maintenance of this bacterium in rural communities of Yucatán, México.

Associated Factors to Seroprevalence of Ehrlichia spp. in Dogs of Quintana Roo, Mexico.

The objective of this study was to determine the seroprevalence to Ehrlichia spp. in dogs from Xcalak, Quintana Roo, Mexico, and the associated factors. Serum samples were obtained from 118 dogs and used in an indirect immunofluorescent assay test for the detection of antibodies against Ehrlichia spp. A questionnaire was used to obtain information about possible variables associated with seroprevalence. These variables were analyzed through Chi2 test and logistic regression. Dog seroprevalence of antibodies against Ehrlichia spp. was 64% (75/118). Fifty-two percent (61/118) of dogs had tick infestation which was identified as Rhipicephalus sanguineus sensu lato. Anemia was observed in 36% of dogs. Leucopenia (2.5%), thrombocytopenia (70%), and hemorrhage (14%) were also observed. Thirty-one percent (23/75) of dogs with anemia, 4% (3/75) of dogs with leucopenia, 80% (60/75) of dogs with thrombocytopenia, 17% (13/75) of dogs with hemorrhages, and 59% (44/75) of dogs with ticks were positive for Ehrlichia spp. antibodies. The factors associated with seroprevalence were age (1-3 and >3 years old, OR = 7.77 and OR = 15.39, resp.), tick infestation (OR = 3.13), and thrombocytopenia (OR = 3.36). In conclusion, seroprevalence of Ehrlichia spp. was high in the community of Xcalak and its associated factors were age, tick infestation, and thrombocytopenia.

Molecular Diagnosis of Ehrlichia canis in Dogs and Ticks Rhipicephalus sanguineus (Acari: Ixodidae) in Yucatan, Mexico.

Ehrlichia canis is the etiological agent behind canine monocytic ehrlichiosis, and the tick Rhipicephalus sanguineus (Latreille) is its main vector. Blood smear and nested polymerase chain reaction (PCR) techniques were used to identify E. canis infection in dogs and R. sanguineus, and explore factors possibly associated with infection in dogs in Yucatan, Mexico. Blood samples were taken and ticks R. sanguineus collected from 50 dogs (10 house dogs and 40 in an animal control center). Data were collected on dog age, sex, body condition, and signs associated with platelet deficiencies (epistaxis). Blood smears were analyzed to identify E. canis morulae and generate platelet counts. Nested PCR analysis was done on blood samples and 200 ticks. A χ(2) test was done to identify factors associated with the E. canis infection in the tested dogs. The overall prevalence for infection, as determined by PCR, was 36% (18 out of 50). All positive dogs were from samples collected from the animal shelter, representing prevalence, for this sampling site, of 45% (18 out of 40). Morulae in monocytes were identified in only 4% of samples. Dog origin (i.e. animal control center) was the only variable associated with E. canis infection (P < 0.01). Male ticks had a higher (P < 0.05) infection rate than female ticks (24.5 vs 13.5%). It is concluded that E. canis infection is present in both dogs and the brown dog ticks R. sanguineus in Yucatan, Mexico.