Exposure to Trypanosoma cruzi and Leishmania parasites in dogs from a rural locality of Yucatan, Mexico. A serological survey.

Trypanosoma cruzi, the causal agent of American trypanosomiasis, and Leishmania spp., the causal agents of Leishmaniasis, are prevalent in more than 20 American countries, including Mexico. Dogs have been reported as incidental hosts for both parasites and may be helpful as transmission sentinels. We surveyed the dog population in a rural locality of the Merida municipality in Yucatan, Mexico, to evaluate the seroreactivity against T. cruzi and Leishmania spp. using two antigens, parasite homogenate (H) and iron superoxide dismutase extract (FeSODe), with two serological techniques (ELISA and Western Blot). Our study found that 3.33% of the tested dogs were seroreactive to T. cruzi using ELISA-H, and 29.5% were seroreactive to FeSODe antigen, with a 94.4% consistency between the two tests. Similarly, for L. mexicana, 1.6% were seroreactive using ELISA-H, and 9.8% were seroreactive using ELISA-FeSODe, with an 83.3% consistency between tests. For L. braziliensis, no dogs were seroreactive using ELISA-H, but 16.4% were seroreactive using ELISA-FeSODe, with a 90% consistency between tests. Finally, for L. infantum, 4.9% were seropositive using ELISA-H, and 6.6% were seropositive using ELISA-FeSODe, with a 75% consistency between tests. These results show noticeable evidence of exposure of dogs to trypanosomatid parasites and highlight the potential disease risk for the people and their companion animals in the region.

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.

Infección con Toxoplasma gondii (Eucoccidiorida: Sarcocystidae) en murciélagos de Campeche y Yucatán, México

Toxoplasma gondii es un protozoario parásito reconocido como el agente causal de la toxoplasmosis, enfermedad zoonótica que afecta a humanos y animales domésticos o silvestres. En México, representa un problema de salud pública y veterinaria, sobre todo en regiones con climas tropicales y subtropicales. Los murciélagos han sido identificados como hospederos accidentales en el ciclo de transmisión; no obstante, en México no existe información previa; por lo tanto, el objetivo del presente estudio es reportar la infección con T. gondii en murciélagos capturados en sitios de los estados de Campeche y Yucatán, México. Se capturaron murciélagos en dos sitios de Yucatán y uno de Campeche, ubicados en la Península de Yucatán. Se recolectaron riñones, bazo e hígado y se emplearon en la extracción de ADN total. La infección con T. gondii se detectó a través de la amplificación de un fragmento del gen B1, utilizando PCR anidada. Los productos positivos fueron purificados y enviados a secuenciación para su posterior análisis de alineamiento; adicionalmente, se construyó un árbol filogenético. Se analizaron un total de 69 murciélagos pertenecientes a ocho especies distintas: 41 (59.4 %, 41/69) Artibeus jamaicensis; seis (8.7 %, 6/69) Pteronotus parnellii; seis (8.7 %, 6/69) Noctilio leporinus; seis (8.7 %, 6/69) Chiroderma villosum; cuatro (5.8 %, 4/69) Glossophaga soricina; dos (2.9 %, 2/69) Carollia sowelli; dos (2.89 %, 2/69) Artibeus lituratus y dos (2.9%, 2/69) Rhogeessa aeneus. La PCR anidada identificó ocho (11.6 %, 8/69) murciélagos positivos a la infección: seis (75 %, 6/8) A. jamaicensis, capturados en X'matkuil y Panabá, un (12.5 %, 1/8) G. soricina y un (12.5 %, 1/8) C. villosum, ambos capturados en Panabá. El análisis de alineamiento arrojó 99-100 % para cobertura y 97-99 % para identidad respecto a secuencias de T. gondii. Nuestros resultados aportan al entendimiento del ciclo de transmisión de T. gondii en la región; sin embargo, son necesarias investigaciones futuras para determinar los genotipos circulantes, ya que estudios anteriores han demostrado que estos animales pueden estar infectados con genotipos identificados en otros animales domésticos o silvestres e incluso en humanos.

Toxoplasma gondii is a protozoan parasite recognized as the causative agent of toxoplasmosis, a zoonotic disease that affects humans and domestic or wild animals. In Mexico, it represents a public and animal health problem, especially in regions with tropical and subtropical climates. Bats have been reported as accidental hosts in the transmission cycle; however, there is no preceding information in Mexico. Therefore, the aim of the present study is to report the T. gondii infection in bats captured in sites of Campeche and Yucatan states, Mexico. Bats were captured in two sites in Yucatan (X'matkuil and Panaba) and one in Campeche (Hampolol), located in the Yucatan Peninsula. Kidneys, spleen, and liver were collected and used in the total DNA extraction. Toxoplasma gondii infection was detected through the amplification of a B1 gene fragment, using nested PCR. The positive PCR products were purified and sent to sequencing for a posterior sequence identity analysis. Additionally, a phylogenetic tree was made. A total of 69 bats belonging to eight different species were processed: 41 (59.4 %, 41/69) Artibeus jamaicensis; six (8.7 %, 6/69) Pteronotus parnellii; six (8.7 %, 6/69) Noctilio leporinus; six (8.7 %, 6/69) Chiroderma villosum; four (5.8 %, 4/69) Glossophaga soricina; two (2.9 %, 2/69) Carollia sowelli; two (2.89 %, 2/69) Artibeus lituratus; and two (2.9 %, 2/69) Rhogeessa aeneus. The nested PCR identified eight (11.6 %, 8/69) infected bats: six (75 %, 6/8) A. jamaicensis, captured in X'matkuil and Panaba, one (12.5 %, 1/8) G. soricina, and one (12.5 %, 1/8) C. villosum, both captured in Panaba. The alignment analysis yielded 99-100 % for cover and 97-99 % for identity to T. gondii sequences. Our results contribute to the understanding of the T. gondii transmission cycle in the region; however, future research is needed to determine circulating genotypes, as previous studies have demonstrated that these animals might be infected with identified genotypes in other domestic or wild animals and even in humans.

[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.

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.

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.

Postoperative analgesic effects of either a constant rate infusion of fentanyl, lidocaine, ketamine, dexmedetomidine, or the combination lidocaine-ketamine-dexmedetomidine after ovariohysterectomy in dogs.

OBJECTIVE: To evaluate the postoperative analgesic effects of a constant rate infusion (CRI) of either fentanyl (FENT), lidocaine (LIDO), ketamine (KET), dexmedetomidine (DEX), or the combination lidocaine-ketamine-dexmedetomidine (LKD) in dogs. STUDY DESIGN: Randomized, prospective, blinded, clinical study. ANIMALS: Fifty-four dogs. METHODS: Anesthesia was induced with propofol and maintained with isoflurane. Treatments were intravenous (IV) administration of a bolus at start of anesthesia, followed by an IV CRI until the end of anesthesia, then a CRI at a decreased dose for a further 4 hours: CONTROL/BUT (butorphanol 0.4 mg kg(-1), infusion rate of saline 0.9% 2 mLkg(-1) hour(-1)); FENT (5 µg kg(-1), 10 µg kg(-1) hour(-1), then 2.5 µg kg(-1) hour(-1)); KET (1 mgkg(-1) , 40 µg kg(-1) minute(-1), then 10 µg kg(-1) minute(-1) ; LIDO (2 mg kg(-1), 100 µg kg(-1) minute(-1), then 25 µg kg(-1) minute(-1)); DEX (1 µgkg(-1), 3 µg kg(-1) hour(-1), then 1 µg kg(-1) hour(-1)); or a combination of LKD at the aforementioned doses. Postoperative analgesia was evaluated using the Glasgow composite pain scale, University of Melbourne pain scale, and numerical rating scale. Rescue analgesia was morphine and carprofen. Data were analyzed using Friedman or Kruskal-Wallis test with appropriate post-hoc testing (p < 0.05). RESULTS: Animals requiring rescue analgesia included CONTROL/BUT (n = 8), KET (n = 3), DEX (n = 2), and LIDO (n = 2); significantly higher in CONTROL/BUT than other groups. No dogs in LKD and FENT groups received rescue analgesia. CONTROL/BUT pain scores were significantly higher at 1 hour than FENT, DEX and LKD, but not than KET or LIDO. Fentanyl and LKD sedation scores were higher than CONTROL/BUT at 1 hour. CONCLUSIONS AND CLINICAL RELEVANCE: LKD and FENT resulted in adequate postoperative analgesia. LIDO, CONTROL/BUT, KET and DEX may not be effective for treatment of postoperative pain in dogs undergoing ovariohysterectomy.

Parasitic Zoonoses in Humans and Their Dogs from a Rural Community of Tropical Mexico.

A cross-sectional study was made on 89 inhabitants and their dogs from a rural community of Yucatan, Mexico, to determine the serological prevalence of some zoonotic parasitic agents. Samples were taken to monitor the presence and intensity of infection with gastrointestinal parasites in dogs. In humans, the serological prevalence of T. canis, T. gondii, and T. spiralis was 29.2%, 91.0%, and 6.7%, respectively. No associations were found between positive cases and studied variables. From the total of blood samples taken from dogs, 87 (97.6%) were seropositive to T. gondii; only 52 viable fecal samples were collected from dogs of which 46.2% had the presence of gastrointestinal parasites with low to moderate intensity; from those, 12% had the presence of T. canis. This study demonstrates the presence of the studied zoonotic agents in the area particularly T. gondii which suggest a common source of infection in dogs and humans and a high number of oocyts present in the environment. Preventive measures must be designed towards good prophylactic practices in domestic and backyard animals (T. canis and T. spiralis). Contaminated sources with T. gondii (food and water) should be further investigated in order to design effective control measures.

Evaluation of the isoflurane-sparing effects of fentanyl, lidocaine, ketamine, dexmedetomidine, or the combination lidocaine-ketamine-dexmedetomidine during ovariohysterectomy in dogs.

OBJECTIVE: To evaluate the isoflurane-sparing effects of an intravenous (IV) constant rate infusion (CRI) of fentanyl, lidocaine, ketamine, dexmedetomidine, or lidocaine-ketamine-dexmedetomidine (LKD) in dogs undergoing ovariohysterectomy. STUDY DESIGN: Randomized, prospective, blinded, clinical study. ANIMALS: Fifty four dogs. METHODS: Anesthesia was induced with propofol and maintained with isoflurane with one of the following IV treatments: butorphanol/saline (butorphanol 0.4 mg kg(-1), saline 0.9% CRI, CONTROL/BUT); fentanyl (5 µg kg(-1), 10 µg kg(-1) hour(-1), FENT); ketamine (1 mg kg(-1), 40 µg kg(-1) minute(-1), KET), lidocaine (2 mg kg(-1), 100 µg kg(-1) minute(-1), LIDO); dexmedetomidine (1 µg kg(-1), 3 µg kg(-1) hour(-1), DEX); or a LKD combination. Positive pressure ventilation maintained eucapnia. An anesthetist unaware of treatment and end-tidal isoflurane concentration (Fe'Iso) adjusted vaporizer settings to maintain surgical anesthetic depth. Cardiopulmonary variables and Fe'Iso concentrations were monitored. Data were analyzed using anova (p < 0.05). RESULTS: At most time points, heart rate (HR) was lower in FENT than in other groups, except for DEX and LKD. Mean arterial blood pressure (MAP) was lower in FENT and CONTROL/BUT than in DEX. Overall mean ± SD Fe'Iso and % reduced isoflurane requirements were 1.01 ± 0.31/41.6% (range, 0.75 ± 0.31/56.6% to 1.12 ± 0.80/35.3%, FENT), 1.37 ± 0.19/20.8% (1.23 ± 0.14/28.9% to 1.51 ± 0.22/12.7%, KET), 1.34 ± 0.19/22.5% (1.24 ± 0.19/28.3% to 1.44 ± 0.21/16.8%, LIDO), 1.30 ± 0.28/24.8% (1.16 ± 0.18/32.9% to 1.43 ± 0.32/17.3%, DEX), 0.95 ± 0.19/54.9% (0.7 ± 0.16/59.5% to 1.12 ± 0.16/35.3%, LKD) and 1.73 ± 0.18/0.0% (1.64 ± 0.21 to 1.82 ± 0.14, CONTROL/BUT) during surgery. FENT and LKD significantly reduced Fe'Iso. CONCLUSIONS AND CLINICAL RELEVANCE: At the doses administered, FENT and LKD had greater isoflurane-sparing effect than LIDO, KET or CONTROL/BUT, but not at all times. Low HR during FENT may limit improvement in MAP expected with reduced Fe'Iso.

Preventive and therapeutic DNA vaccination partially protect dogs against an infectious challenge with Trypanosoma cruzi.

American trypanosomiasis, or Chagas disease, is caused by Trypanosoma cruzi, and a vaccine would greatly improve disease control. While some studies in mice suggest that a vaccine is feasible, limited efficacy has been observed in dogs. We evaluated here the safety and efficacy of a DNA vaccine encoding TSA-1 and Tc24 antigens in a dog model of acute T. cruzi infection. Mongrel dogs were immunized with two doses of 500 µg of DNA vaccine, two weeks apart, and infected with T. cruzi (SylvioX10/4 strain) two weeks after the second vaccine dose. Another group of dogs was infected first and treated with the vaccine. Disease progression was monitored for up to 70 days post-infection. The vaccine did not induce any critical change in blood parameters, nor exacerbation of disease in vaccinated animals. On the contrary, it prevented anemia and a decrease in lymphocyte counts following T. cruzi infection in vaccinated dogs. Both preventive and therapeutic vaccination significantly reduced parasitemia, cardiac inflammation and cardiac parasite burden, and tended to reduce the development of cardiac arrhythmias. These results indicate that a preventive or therapeutic DNA vaccine encoding TSA-1 and Tc24 antigens is safe and may reduce both parasite transmission and the clinical progression of Chagas disease in vaccinated dogs. This DNA vaccine may thus be an excellent veterinary vaccine candidate. These data also further strengthen the feasibility of a Chagas disease vaccine for humans.

Evaluation of clinical and immunopathological features of different infective doses of Trypanosoma cruzi in dogs during the acute phase.

Infection with Trypanosoma cruzi is a major risk in Latin America, and dogs are believed to be good models for evaluating Chagas disease. Here, we evaluated the clinical and immunopathological alterations developed by mongrel dogs experimentally infected with different infective doses (2,000, 20,000, and 200,000 metacyclic trypomastigotes of Sylvio X10/4 strain kg(-1) via intraperitoneal). Clinical and electrocardiographic parameters, as well as antibody production and pathologic lesions were evaluated. All three doses of this strain of T. cruzi induced a similar pattern of infection characterized by cardiac arrhythmias and severe and diffuse myocarditis. Specific anti-T. cruzi IgG indicated seroconversion by day 14 after infection, and IgG levels increased during the period of evaluation. Mortality was observed only in dogs infected with the medium or high parasite doses, but not in the group infected with a low dose of 2,000 parasites kg(-1). Infection with a low dose of parasites provides an excellent nonlethal model to evaluate the immunopathology of the acute disease in dogs infected with the Sylvio X10/4 strain of T. cruzi.

Detection of different Leishmania spp. and Trypanosoma cruzi antibodies in cats from the Yucatan Peninsula (Mexico) using an iron superoxide dismutase excreted as antigen.

Although human leishmaniasis has been reported in 20 states in Mexico, no case of leishmaniasis has been reported in cats to date. In the Yucatan Peninsula, it has been found that dogs may act as reservoirs for at least three Leishmania species (Leishmania mexicana, Leishmania braziliensis, and Leishmania panamensis). In this study we identified specific antibodies against these three Leishmania spp. and Trypanosoma cruzi in the sera from 95 cats from two States on the Yucatan Peninsula, namely Quintana Roo and Yucatan, by ELISA and Western blot techniques using whole extract and an iron superoxide dismutase excreted by the parasites as antigens. As well as demonstrating the presence of trypanosomatid antibodies in the feline population on the Yucatan Peninsula, we were also able to confirm the high sensitivity and specificity of the iron superoxide dismutase antigen secreted by them, which may prove to be very useful in epidemiological studies.

Prevalence of antibodies against three species of Leishmania (L. mexicana, L. braziliensis, L. infantum) and possible associated factors in dogs from Mérida, Yucatán, Mexico.

Leishmania spp. has been recorded in humans and in dogs, and numerous studies have demonstrated that dogs act as reservoirs for visceral leishmaniasis. The objective of this study was to determine the prevalence of three species of the Leishmania genus and possible associated factors in sera of 218 dogs from two different populations in Mérida, Yucatán (Mexico). The sera were analyzed to detect antibodies against L. mexicana, L. braziliensis, and L. infantum using the superoxide dismutase- enzyme-linked immunosorbent assay (SOD-ELISA) and Western blot as confirmation. The Fe-SOD excreted was used as the antigenic fraction for the three Leishmania species. The prevalence values found were 30.2% (L. mexicana), 8.2% (L. braziliensis), and 11.9% (L. infantum), with L. mexicana seroprevalence being greater than L. braziliensis and L. infantum (p<0.05). Five percent (11/218) of the dogs showed antibodies against L. mexicana/L. braziliensis, 5.5% (12/218) with L. mexicana/L. infantum and 1.8% (4/218) with L. mexicana/L. braziliensis/L. infantum. No relationship (p>0.05) was found between antibodies against L. mexicana and breed, age, physical condition, or cutaneous lesions in dogs. This study provides evidence of antibodies against L. mexicana, L. braziliensis and L. infantum in dog populations from Mérida, Mexico.

An epidemiological study of intestinal parasites of dogs from Yucatan, Mexico, and their risk to public health.

The prevalence of intestinal parasites in dogs and factors associated were studied in a rural community of Yucatan (southern Mexico), with special attention to those gastrointestinal parasites potentially transmitted to man. One hundred thirty dogs from 91 households were studied. Fecal samples were processed by the centrifugation-flotation and the McMaster techniques. To determine factors associated with zoonotic parasites in dogs, univariate analysis was performed, using sex, age, and body condition as independent variables. Variables with p < 0.2 were analyzed by a logistic regression. Of 130 dogs, 104 were positive for the presence of intestinal parasites, representing a prevalence of 80%. Eggs of four species of helminths (Ancylostoma caninum, Thichuris vulpis, Toxocara canis, and Dipylidium caninum) and coccidian oocysts were detected. A. caninum was the most prevalent parasite (73.8%), followed by T. vulpis (25.4%), T. canis (6.2%), D. caninum (2.3%), and coccidian oocysts (2.3%). The majority of dogs were infected by only one species of parasite (70/130, 53.8%). Mixed infection caused by two or three zoonotic parasites were discovered in 21.3% (30/130) and 3.1% (4/130), respectively. A. caninum showed the highest egg output (42.3% of dogs had ≥ 500 eggs per gram). Factors associated with zoonotic parasites were age (<2 years old; odds ratio = 5.30, p = 0.029) and body condition (poor body condition; odds ratio = 6.69, p = 0.026). In conclusion, young dogs from rural Yucatan, Mexico, with poor body condition had a higher prevalence of intestinal zoonotic parasites as these factors were associated with a higher risk of becoming infected.

An iron-superoxide dismutase antigen-based serological screening of dogs indicates their potential role in the transmission of cutaneous leishmaniasis and trypanosomiasis in Yucatan, Mexico.

An increasing number of studies have reported high infection rates for American cutaneous leishmaniasis in dogs, which have thus been proposed as the reservoir host. Canine leishmaniasis is widespread in different states in Mexico, where a number of Leishmania species have been isolated from dogs. In the present study, the detection of different Leishmania species is described in stray dogs from two localities, namely Tulum and Celestún on the Yucatan Peninsula (Mexico). The use of iron-superoxide dismutase excreted by the parasites as the antigen fraction and enzyme-linked immunosorbent assay and western blot tests allowed us to confirm the presence of at least three species of Leishmania (Le. mexicana, Le. braziliensis, and Le. panamensis), some of which are reported for the first time in this species. In addition to a high prevalence of Le. mexicana and Le. braziliensis, and to a lesser degree, Le. panamensis, there is a significant prevalence of Trypanosoma cruzi, suggesting that the dog may be a source of transmission of trypanosomiasis. However, a more thorough epidemiological study on the dog population, both wild as well as urban, of the Yucatan Peninsula will be required to design a control strategy for these diseases, paying particular attention to the population affected and even broadening the study to other Mexican states as well as neighboring countries. These results again confirm that iron-superoxide dismutase excreted by the different trypanosomatid species constitutes a good source of antigen for serodiagnosis in epidemiological studies.

Prevalence of Dirofilaria immitis infection in dogs from Celestun, Mexico, using polymerase chain reaction test.

The aim of this study was to determine the prevalence of Dirofilaria immitis in dogs and to analyze risk factors associated with infection at Celestun, a coastal locality in southeast Mexico. Blood samples were collected from 279 asymptomatic individuals between August 2007 and March 2008 and analyzed by polymerase chain reaction technique. The association between D. immitis infection and sex, age group, and distance of residence from a wetland of dogs was statistically analyzed. Prevalence of D. immitis infection was of 59.8%. Age of individuals (>2 years) was a risk factor for infection with D. immitis (odds ratio 2.49, confidence interval 1.47-4.23, p=0.001). In conclusion, Celestun can be considered a focus of D. immitis infection with high levels of transmission among the local dog population, as confirmed by the high prevalence reported and the association of age (dogs >2 years) as a risk associated with infection.

Immunopathology of natural infection with Trypanosoma cruzi in dogs.

Chagas disease is caused by Trypanosoma cruzi and dogs are an important reservoir of the parasite as well as a good model for the study of the pathogenesis of the disease. We aimed here at characterizing the immunopathology of naturally infected dogs in Merida, Yucatan, Mexico. Following an initial screening for T. cruzi seropositive stray dogs, we examined 9 seropositive and 10 seronegative animals. High lymphocytes and low monocytes counts were observed in peripheral blood from seropositives dogs. Three of nine seropositive dogs presented electrocardiographic alterations including right bundle branch block, sinusal block and QRS complex alterations and some right ventricle enlargement was noted. Histopathologic analysis of cardiac walls revealed significant inflammation with a clear tropism for the right ventricle, although most walls were affected. Seropositive dogs presented low IgG1 and high IgG2 levels. Higher IgG1 levels were associated with increased cardiac index and myocarditis, suggesting that a Th2 immune response leads to susceptibility and increased disease severity. These observations shed some light on the mechanisms of pathogenesis of Chagas disease in dogs, and provide a good framework for the evaluation of novel drugs and vaccines in this animal model.

Therapeutic DNA vaccine against Trypanosoma cruzi infection in dogs.

Chagas' disease is an important health problem in most Latin American countries, and a concern in dog populations, which act as a reservoir. We showed in previous studies that a therapeutic DNA vaccine could partially control the pathology after Trypanosoma cruzi infection in mice, and this vaccine may represent an alternative treatment for Chagas' disease. Here we evaluated the therapeutic efficacy of this vaccine in experimentally infected dogs for up to 2 months after infection. Our results suggest that DNA vaccine treatment may affect the immune response and delay Chagas' disease progression in T. cruzi-infected dogs, and confirm the potential of this novel treatment.