Evidence of resistance to ivermectin in the gastrointestinal nematodes of horses from Mexican southeast.

Gastrointestinal nematodes (GIN) are a major impediment to breeding and exploitation of horses. Traditional control of GIN has generated resistance to main anthelmintics, including ivermectin. An analysis of five ranches with a history of IVM use was done to determine the efficacy and resistance of GIN to IVM treatment in horses from the Mexican southeast. Predesigned questionnaires were applied to collect information on previous treatment protocols. The fecal egg count reduction test (FECRT) was applied to determine resistance. Before IVM application, a McMaster test was used to diagnose GIN infection in horses, and feces cultures were done to identify L3 larvae for Strongylida eggs. Pre-treatment samples showed that 72.7% (80/110) of horses were GIN positive, with cyathostomins being the most frequent (91.8%), followed by Oxyuris equi (7.0%), Parascaris equorum (1.0%), and Strongylus vulgaris (0.2%). Based on the results, the horses at each ranch were divided in control (CG) and experimental (EG) groups with similar eggs per gram of feces (EPG). The EG (40/80) was dewormed with IVM (0.2 mg/kg orally) and the CG (40/80) remained untreated. After 14 days, EPG were measured, and feces cultures were done again to identify L3 larvae. After treatment of EG, 40% (16/40) of horses were positive, the most frequently identified GIN were cyathostomins (98.6%), followed by P. equorum (1.0%) and S. vulgaris (0.2%). Three of the five ranches were classified as resistant, according to the FECRT, with a percentage of reduction from 53 to 68%, all of which used IVM ≥4 times annually. This is the first evidence of resistance in cyathostomins to IVM treatment in horses from the Mexican southeast, adding to the current problem of anthelmintic resistance in equine GIN.

Distribution and Risk of Cutaneous Leishmaniasis in Khyber Pakhtunkhwa, Pakistan.

Cutaneous leishmaniasis (CL) is a zoonotic infection caused by obligate intracellular protozoa of the genus Leishmania. This study aimed to investigate CL in Khyber Pakhtunkhwa, Pakistan and to estimate the risk of epidemics. Clinico-epidemiological data of 3188 CL patients were collected from health facilities in 2021. Risk factors were analyzed using the chi-square test. ArcGIS V.10.7.1 was applied for spatial analysis. The association between CL occurrence and climatic variables was examined by Bayesian geostatistical analysis. The clinical data revealed males or individuals younger than 20 years old were more affected. Most patients presented with a single lesion, and the face was the most attacked body part. CL was prevalent in the southern region in winter. A proportional symbol map, a choropleth map, and a digital elevation model map were built to show the distribution of CL. Focal transmission was predicted by inverse distance weighting interpolation. Cluster and outlier analysis identified clusters in Bannu, Dir Lower, and Mardan, and hotspot analysis suggested Bannu as a high-risk foci. Bayesian geostatistical analysis indicated that increasing precipitation and temperature as well as low altitudes were associated with CL infection. The study has provided important information for public health sectors to develop intervention strategies for future CL epidemics.

Epidemiological Study of the Occurrence of Typhus Group Rickettsia Natural Infection in Domiciliated Dogs from a Rural Community in South-Eastern Mexico.

The aim is to describe the Typhus group (TG) Rickettsia infection in dogs and to identify factors associated with this infection. We collected blood samples and gathered exposure and clinical data of 142 dogs from a rural community of Yucatan. The Rickettsia group was determined by semi-nested PCR. Generalized linear models with binomial error distribution were used to model the associated factors from the dog sample for risk ratio (RR) estimation. Thirty-four dogs (23.9%) showed molecular evidence of TG Rickettsia DNA. The multivariate model showed that mixed-breed dogs (RR = 0.06) and dogs that had received antiparasitic treatment (RR = 0.049) had a lower risk of getting infected, taking as reference the purebred group and the non-treated dogs, respectively. Looking at variable interactions, adult dogs without outdoor activities had a lower infection risk than puppies (RR = 0.26). Among dogs with antiparasitic treatment, females had a higher infection risk than male dogs (RR = 26.2). The results showed enzootic TG Rickettsia circulation in dogs of a rural community. The factors outdoor activities, age and previous antiparasitic treatment, as well as the clinical variables signs of hemorrhages and epistaxis, were associated with a less chance of natural infection in the studied dogs. Prevention and control of the enzootic transmission risk of TG Rickettsia should help to reduce the potential zoonotic transmission of this pathogen.

Frequency of sheep farms with anthelmintic resistant gastrointestinal nematodes in the Mexican Yucatán peninsula.

The present study explored the frequency of hair-sheep farms with gastrointestinal nematodes (GIN) resistant to albendazole sulfoxide (AS), ivermectin (IVM) and levamisole (LEV) in the Yucatán peninsula, México, using the faecal egg count reduction test (FECRT), and compared the frequency of farms diagnosed with resistance using three different formulae. The survey included farms from the states of Campeche (9) and Yucatán (14) (2016-2019). Collaborating farms had >100 grazing ewes. Animals in the FECRT were > 12 months old, received no anthelmintic for >8 weeks and had ≥150 GIN eggs per gramme of faeces (EPG). Animals were distributed to respective groups: untreated controls, AS (5 mg/kg BW), IVM (0.2 mg/kg BW), and LEV (7.5 mg/kg BW). Due to low EPG, some farms only included one or two AH groups. Second faecal samples were obtained on day 14 post-treatment to estimate the percentage reduction (%R) and 95% confidence interval (95%CI). Criteria to declare resistance were those proposed by the World Association for the Advancement of Veterinary Parasitology. Three formulae were used to estimate resistance frequency: The RESO© and eggCounts-T:C, which considered treated and control EPG means post-treatment, but differed in their 95%CI estimation, while the eggCounts-T:T only considered pre- and post-treatment EPG means with 95%CI. The RESO© and eggCounts-T:C formulae resulted in the same frequency of IVM resistant farms for Campeche (100%; 9/9) and Yucatán (92.9%; 13/14), while, the eggCount-T:T formula resulted in a frequency of 85.7% (12/14) IVM resistance in Yucatán. The three formulae estimated the same frequency of AS resistant farms in Campeche (100%; 9/9) and Yucatán (87.5%; 7/8). The RESO© and the eggCounts-T:C formulae resulted in the same frequency of LEV resistant farms for Campeche (44.4%; 4/9), and Yucatán (60.0%; 6/10), but the eggCounts-T:T formula resulted in a frequency of 40.0% (4/10) LEV resistance in Yucatán. The FECRT using RESO© or eggCounts-T:C formulae are stricter than the eggCounts-T:T as the latter cannot identify what proportion of the %R cannot be attributed to the AH. The untreated control group helped adjusting the %R calculation and seemed more adequate considering the propensity of hair-sheep to reduce their EPG on their own. .

Anthelmintic Activity of Extracts and Active Compounds From Diospyros anisandra on Ancylostoma caninum, Haemonchus placei and Cyathostomins.

The present study aimed to evaluate the anthelmintic activity of leaf and bark extracts of Diospyros anisandra collected during different seasons and their major constituents on eggs of Ancylostoma caninum, Haemonchus placei, and cyathostomins. Specifically, the eclosion inhibition of the methanolic extracts of the leaves and bark of D. anisandra collected during the dry and rainy seasons (600-37.5 µg/ml) were evaluated in addition to the fractions, sub-fractions (300-37.5 µg/ml) and active major constituents (150-2.3 µg/ml). The rainy season bark extract had the highest percentage of eclosion inhibition (PEI) against the evaluated nematodes (≥ 90% at 75 µg/ml) along with high ovicidal activity (90.0 to 93.4% at 75 µg/ml). The purification of the rainy season bark extract showed that its biological activity came from the non-polar n-hexane fraction (≥ 93% at 75 µg/ml). The bioguided fractionation pointed to sub-fraction 5 as having the highest anthelmintic activity against the three evaluated genera of nematodes (PEI ≥ 93% at 37.5 µg/ml). Gas chromatography and mass spectrometry revealed that the major constituent in sub-fraction 5 was plumbagin. Upon evaluation, plumbagin was confirmed to be responsible for the anthelmintic activity of D. anisandra, with a PEI ≥ 90% at 2.3 µg/ml on the three evaluated nematodes. Additionally, the compounds betulin and lupeol in the bark of D. anisandra were evaluated but presented low anthelmintic activity (PEI ≤ 5.3% at 2.3 µg/ml). In conclusion, the rainy season bark extract of D. anisandra exerts a high ovicidal activity against the eggs of the three studied nematodes. Plumbagin is the active compound responsible for this activity and represents a potential alternative for the control of different genera of gastrointestinal nematodes given the current scenario of anthelmintic resistance.

Canine babesiosis: A literature review of prevalence, distribution, and diagnosis in Latin America and the Caribbean.

Canine babesiosis is a tick-borne disease with worldwide distribution and global significance. Traditionally, canine babesiosis was caused by B. canis (large Babesia) and B. gibsoni (small Babesia) based on cytological examination of stained blood smears. Currently, molecular techniques have demonstrated that several Babesia species infect dogs: B. canis, B. vogeli, and B. rossi (large forms) and B. gibsoni, B. conradae, and B. vulpes (small forms). In this study, we compiled and reviewed currently available data on Babesia infections in dogs in Latin America and the Caribbean, as well as on distribution of Babesia species with respect to prevalence, geographic location, and methods of detection. Forty-three studies on canine babesiosis published from 2005 to 2019 were included. The publications retrieved reported three species of Babesia (B. vogeli, B. gibsoni, and B. caballi) based on molecular confirmation of the species. Babesia vogeli was reported in Mexico, Costa Rica, Granada, Haiti, Nicaragua, Saint Kitts and Nevis, Trinidad and Tobago, Argentina, Brazil, Chile, Colombia, Paraguay, Peru, and Venezuela. In contrast, B. gibsoni was recorded in Costa Rica, Nicaragua, Saint Kitts and Nevis, and Brazil. Babesia caballi was found in a dog from Brazil. Babesia prevalence in dogs varied considerably based on parasite species and geographic location, with values close zero to 26.2%. Besides molecular techniques such as PCR, studies included examination of blood smears by microscopy and/or serologic tests. Few countries in the region, e.g., Brazil and Costa Rica, possess profound data availability, whereas the majority of them have scarce information or no data. A deeper understanding of the ecology and epidemiology of Babesia spp. in dogs is needed for the region.

Personal and household factors involved in recent Rickettsia exposure in a rural population from Yucatán, Mexico.

The aim of the study was to describe the epidemiological factors associated with the risks of recent Rickettsia exposure in inhabitants of a rural population from Yucatán, Mexico. The study included 130 inhabitants from Maxcanú, Yucatán. Blood samples were collected to detect IgM and IgG antibodies against Rickettsia typhi and Rickettsia rickettsii by an indirect immunofluorescence antibody test. Additionally, nested polymerase chain reaction was performed to amplify fragments of the 17kDa and sca5 genes. Previously, an epidemiological questionnaire was applied focused on collecting information on personal and housing exposure variables related to the recent infection with Rickettsia to determine epidemiological associations. Results that exhibited a p-value < .25 were included in a generalized multinomial logistic linear model to determine the variables involved with the risk of contact or Rickettsia infection. In all, 76% (99/130) of the participants presented with immunoglobulin titres against the Rickettsia species evaluated, while rickettsial DNA was detected in 35.4% (46/130) of the participants. The association analysis with the personal exposure variables showed that the productive age group (OR = 0.32; 95% CI = 0.10-1.03) and the elders group (OR = 0.12; 95% CI = 0.01-0.83) were protective factors for recent infection with R. typhi, taking as reference the school group. The presence of a family orchard in the home (OR = 7.56; 95% CI = 1.62-35.29) was a risk factor for recent infection with R. rickettsii. Presumably, the presence of ectoparasites (OR = 2.71; 95% CI = 0.90-8.09) at home was a risk factor for recent infection with both Rickettsia species. Recent infection was demonstrated in inhabitants from Maxcanú, Yucatán. A high seropositive frequency was obtained. The results highlight the importance of the family garden and the presence of ectoparasites in the home as risk factors associated with recent infection with Rickettsia in inhabitants from Maxcanú.

Worldwide overview of human infections with Hymenolepis diminuta.

Hymenolepis diminuta is a zoonotic cestode parasitizing the small intestine of rodents (definitive hosts). Humans can accidentally enter into the life cycle of this tapeworm via the ingestion of infected insects (intermediate hosts) containing cestode cysticercoids in their body cavity. More than two centuries after the first record in humans, there are no accurate estimates of the number of human cases around the world. In order to have a more precise idea about the number of human cases with H. diminuta and the current status of the disease (hymenolepiasis) worldwide, we conducted a literature review of published records on human infection with H. diminuta. One thousand five hundred and sixty-one published records of infection with H. diminuta from 80 countries were identified. This review presents an estimate of the number of human cases with H. diminuta and a current overview of the prevalence, geographic distribution, symptoms, diagnosis, exposure to infective stages, and therapeutic approaches for this underestimated zoonotic tapeworm.

Onthophagus landolti and Canthon indigaceus chevrolati (Coleoptera: Scarabaeinae) are attracted to the feces of ivermectin-treated cattle in the Mexican tropics / Onthophagus landolti y Canthon indigaceus chevrolati (Coleoptera: Scarabaeinae) son atraídos por las heces de ganado tratado con ivermectina en el trópico mexicano

Abstract Ivermectin (IVM) is widely used for parasite control in livestock in the tropics. Residual IVM in feces conserves its insecticide activity for weeks and can harm dung beetle (DB) species. Attraction to the feces of IVM-treated cattle was tested using the DB species Onthophagus landolti (Harold) and Canthon indigaceus chevrolati (Harold) as models. Experiments were done under controlled laboratory conditions, semi-controlled field conditions and uncontrolled field conditions. Olfactometers were used in the controlled and semi-controlled trials. The control treatment was baited IVM-free feces, and the experimental treatments were the feces of cattle treated with 1 % IVM (subcutaneous administration; single, 0.2 mg/kg bw dosage) and collected at 5, 14, 21 and 28 days post-treatment. The uncontrolled field trial involved pitfall traps baited with IVM-free feces or feces from IVM-treated cattle collected five days post-treatment. Under controlled and semi-controlled conditions, the feces of IVM-treated cattle (at 5, 14, 21 or 28 days post-treatment) attracted more O. landolti and C. i. chevrolati individuals than IVM-free feces (P < 0.05). The same response occurred under uncontrolled conditions. This clear attraction for IVM-containing cattle feces by the studied DB species highlights that incorrect IVM use may pose a risk to DB communities in cattle production systems.(AU)

Resumen La ivermectina (IVM) es ampliamente utilizada para el control de parásitos en el ganado en los trópicos. La IVM residual en las heces conserva su actividad insecticida durante semanas y puede dañar diversas especies de escarabajos estercoleros. La atracción a las heces del ganado tratado con IVM se probó usando a las especies Onthophagus landolti (Harold) y Canthon indigaceus chevrolati (Harold) como modelos de estudio. Los experimentos se realizaron bajo condiciones de laboratorio controladas, condiciones de campo semicontroladas y condiciones de campo no controladas. Se utilizaron olfatómetros en los ensayos controlados y semicontrolados. El tratamiento de control consistió en heces exentas de IVM, y los tratamientos experimentales fueron heces de ganado tratado con IVM al 1 % (administración subcutánea, dosis única, 0.2 mg / kg pv) las cuales se recogieron a los 5, 14, 21 y 28 días después del tratamiento al ganado. El ensayo de campo no controlado incluyó trampas de caída libre o pitfall cebadas con heces libres de IVM y con heces de ganado tratado con IVM recogido cinco días después del tratamiento. En condiciones controladas y semicontroladas, las heces del ganado tratado con IVM (a los 5, 14, 21 y 28 días después del tratamiento) atrajeron más individuos O. landolti yC. i. chevrolati que las heces sin IVM (P < 0.05). La misma respuesta ocurrió bajo condiciones no controladas. Esta clara atracción a las heces de ganado que contienen IVM por las especies estudiadas pone de relieve que el uso incorrecto de IVM puede plantear un riesgo para las comunidades de escarabajos estercoleros en los sistemas de producción ganadera.(AU)

Molecular detection of rickettsial tick-borne agents in white-tailed deer (Odocoileus virginianus yucatanensis), mazama deer (Mazama temama), and the ticks they host in Yucatan, Mexico.

Twenty-five white-tailed deer (Odocoileus virginianus yucatanensis) (WTD), 4 mazama deer (Mazama temama) (MD), and the ticks they host in Yucatan, Mexico were sampled to run a molecular survey for tick-borne rickettsial agents. The prevalence of rickettsial agents was 20% in WTD (5/25) and 50% in MD (2/4). When sequencing the nested PCR products, E. chaffeensis, A. phagocytophilum and A. odocoilei, were identified as single infection or coinfecting cervids. None of the cervid samples were positive for E. ewingii, E. canis, nor Rickettsia spp. Overall, 355 individual ticks were collected. Species identified based on adult stages infesting cervids included Amblyomma mixtum, A. parvum, A. cf. oblongoguttatum, Ixodes affinis, Rhipicephalus microplus, R. sanguineus sensu lato, and Haemaphysalis juxtakochi. Rhipicephalus microplus was the tick species most commonly found infesting cervids with a frequency of 28.4%, and intensity of 25.2 ticks per animal. A pool of Amblyomma cf. oblongoguttatum adults and one of Amblyomma spp. nymphs were positive for E. canis and E. chaffeensis, respectively. None of the studied tick pools were positive for E. ewingii, A. phagocytophilum, nor R. rickettsii. To the best of our knowledge, this study is the first to report the prevalence of rickettsial agents in WTD and MD in Mexico. Our molecular study is the first to report the detection of E. chaffeensis, A. phagocytophilum, and A. odocoilei in MD in Mexico. The molecular detection of E. chaffeensis, A. phagocytophilum, and A. odocoilei in deer, and E. chaffeensis in Amblyomma spp. nymphs reported here raises the concern for the risk of human exposure to tick-borne rickettsial pathogens. Our findings highlight the need to apply the "One Health" approach to study ticks and tick-borne diseases. This science-based information could be used by state public-health programs to assess the risk for exposure to tick-borne Anaplasmataceae in Yucatan, Mexico.

Ticks infesting dogs in rural communities of Yucatan, Mexico and molecular diagnosis of rickettsial infection.

Rickettsial infection in dog-associated ticks in three rural communities of Yucatan, Mexico was investigated using qPCR and nested PCR assays. A total of 319 dogs were studied and ticks samples were collected. A total of 170 dogs were infested with ticks (frequency of 53.4%). Overall, 1,380 ticks representing seven species were collected: Amblyomma mixtum, A. ovale, A. parvum, A. cf. oblongoguttatum, Ixodes affinis, Rhipicephalus microplus, and R. sanguineus sensu lato. The most abundant species was R. sanguineus s.l. with a mean intensity of 7.4 ticks/host. Dogs in the communities of Chan San Antonio and Yaxcheku were 2.84 and 2.41 times more likely to be infected with R. sanguineus compared with Sucopo (p < 0.05). Adult pools of A. mixtum, A. parvum, I. affinis, R. microplus, and A. c.f. oblongoguttatum were negative to E. chaffeensis, E. ewingii, A. phagocytophilum, and R. rickettsii. However, pools of R. sanguineus s.l. adults and A. ovale adults, as well as nymphs of Amblyomma spp. were positive to E. canis. Sequencing analysis of the nested PCR products amplifying the 16S rRNA gene fragment of E. canis confirmed the results and revealed 100% identity with sequences of E. canis. This is the first report worldwide of E. canis infection in A. ovale by PCR. This finding does not necessarily indicate that A. ovale is a competent vector of E. canis because pathogen transmission of this specific tick to a naïve dog remains to be documented. This study documented that different tick species parasitize dogs in Yucatan, Mexico, where R. sanguineus s.l., A. ovale, and nymphs of Amblyomma spp. were shown to be infected with E. canis. These findings highlight the need for control strategies against tick infestations in dogs to prevent the risk of tick-borne disease transmission among companion animal and probably human populations.

[Zoonotic intestinal nematodes in dogs from public parks in Yucatán, México].

INTRODUCTION: Dogs represent a potential public health risk because of the natural transmission of zoonotic parasitic infections. OBJECTIVE: To estimate the frequency and to determine factors associated with the presence of intestinal nematode eggs in dog feces collected in public parks of Mérida,Yucatán, México. MATERIALS AND METHODS: A total of 100 dog fecal samples collected from 20 public parks in two areas of Mérida were analyzed. Samples were processed by the centrifugation-flotation and the McMaster techniques to confirm the presence and to quantify the excretion of intestinal nematode eggs per gram of feces. The factors associated with the presence of nematode eggs were identified using the chi square univariate analysis. RESULTS: We found an 11% frequency of fecal samples positive for intestinal nematode eggs. Eggs of three species of parasites were identified: Ancylostoma caninum was the most common (10%), followed by Toxocara canis (10%), and Trichuris vulpis (1%). Most positive samples were infected with only one intestinal nematode (10%), and only 1 % was positive for a mixed infection by A. caninum and T. vulpis. The presence of stray dogs in public parks was an associated factor (p=0.046) with a higher number of fecal samples positive for intestinal nematode eggs. CONCLUSIONS: The frequency of intestinal nematodes in dog feces with zoonotic potential was high in parks of Mérida, Yucatán, México; samples from parks where there were stray dogs had a higher possibility of being positive.

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.

Nematodos intestinales de perros en parques públicos de Yucatán, México / Zoonotic intestinal nematodes in dogs from public parks in Yucatán, México

Resumen Introducción. Los perros representan un potencial riesgo para la salud pública debido a que transmiten infecciones parasitarias al hombre. Objetivo. Estimar la frecuencia y determinar los factores asociados a la presencia de huevos de nematodos intestinales en heces de perros recolectadas en parques públicos de Mérida, Yucatán, México. Materiales y métodos. Se analizaron 100 muestras de heces de perros recolectadas en 20 parques públicos de dos zonas de la ciudad. Las muestras se procesaron mediante las técnicas de flotación centrifugada y de McMaster para confirmar la presencia de huevos de nematodos intestinales y cuantificarlos por gramo de heces. Se determinaron los factores asociados a la presencia de los huevos mediante un análisis univariado de χ2. Resultados. Se encontró una frecuencia de 11 %. Se identificaron huevos de tres especies de parásitos y Ancylostoma caninum fue el más frecuente (10 %), seguido por Toxocara canis (1 %) y Trichuris vulpis (1 %). La mayoría de las muestras positivas presentaba infección con un nematodo intestinal únicamente (10 %) y solo el 1 % resultó positivo para infección mixta por A. caninum y T. vulpis. La presencia de perros sin dueño en los parques públicos fue el factor asociado (p=0,046) con un mayor número de heces positivas para huevos de nematodos intestinales. Conclusiones. En los parques de la ciudad se encontraron heces de perros con huevos de nematodos intestinales con potencial zoonótico; la probabilidad de que las muestras fueran positivas fue mayor en los parques con presencia de perros sin dueño.

Abstract Introduction: Dogs represent a potential public health risk because of the natural transmission of zoonotic parasitic infections. Objective: To estimate the frequency and to determine factors associated with the presence of intestinal nematode eggs in dog feces collected in public parks of Mérida,Yucatán, México. Materials and methods: A total of 100 dog fecal samples collected from 20 public parks in two areas of Mérida were analyzed. Samples were processed by the centrifugation-flotation and the McMaster techniques to confirm the presence and to quantify the excretion of intestinal nematode eggs per gram of feces. The factors associated with the presence of nematode eggs were identified using the chi square univariate analysis. Results: We found an 11% frequency of fecal samples positive for intestinal nematode eggs. Eggs of three species of parasites were identified: Ancylostoma caninum was the most common (10%), followed by Toxocara canis (10%), and Trichuris vulpis (1%). Most positive samples were infected with only one intestinal nematode (10%), and only 1 % was positive for a mixed infection by A. caninum and T. vulpis. The presence of stray dogs in public parks was an associated factor (p=0.046) with a higher number of fecal samples positive for intestinal nematode eggs. Conclusions: The frequency of intestinal nematodes in dog feces with zoonotic potential was high in parks of Mérida, Yucatán, México; samples from parks where there were stray dogs had a higher possibility of being positive.

Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium.

An attenuated live vaccine containing Babesia bovis and B. bigemina cultured in vitro with a serum-free medium was assessed for its clinical protection conferred of naïve cattle, under natural tick-challenge in a high endemicity zone to Babesia spp. Three groups of six animals were treated as follows: group I (GI) received a vaccine derived from parasites cultured with a free-serum medium; group II (GII) were immunized with the standard vaccine, with parasites cultured in a medium supplemented with 40% (v/v) bovine serum; and a control group (GIII) inoculated with non-infected bovine erythrocytes. Inocula were administered by IM route. Experimental animals were kept during 23days after vaccination in a cattle farm free of ticks and Babesia spp. Thereafter, cattle were moved to a high endemicity farm for natural exposure to Babesia spp. transmitted by Rhipicephalus microplus ticks. Protection against clinical babesiosis was observed in bovines belonging to GI (100%) and GII (83.33%), while the control animals (GIII) were not protected, and showed severe clinical signs, closely related to babesiosis, were observed for at least three consecutive days during the challenge. These were fever, anemia, which were measured simultaneously, and circulating parasites were detected by optic light microscopy. All cattle showed B. bovis and B. bigemina in stained blood films during the challenge; B. bovis antibody titers were higher than those to B. bigemina in GI and GII, and lower titers were determined in GIII. The protective capacity of the vaccine derived from B. bovis and B. bigemina cultured in vitro in a serum-free medium was demonstrated.

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.

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.