Canine trypanosomiasis in an endemic Costa Rican community: Demonstration of the active infection cycle.

A cross-sectional study was conducted to determine the prevalence of canine trypanosomiasis in an endemic community of Costa Rica. The indirect hemagglutination and indirect immunofluorescence assay yielded positive results in 6.4% (20/314) of canine samples analyzed; polymerase chain reaction (PCR) and light microscopy yielded positive results in one dog. Subsequently, a longitudinal study was carried out with 55 negative T. cruzi canines in the cross-sectional study. These dogs were divided into two groups: Group 1, which consisted of 25 individuals that lived in dwellings where triatomines were found in their homes; and Group 2, which consisted of 30 dogs that lived in dwellings where triatomines were not found during the previous study in their homes. Seroconversion occurred in six dogs (10.9%) in Group 1 in the first months of the year (dry season); these dogs remained seropositive until the end of the study. Only one of the six seropositive canines was also found positive once in T. cruzi PCR. The analysis of the amplified T. cruzi sequences of dogs and triatomines showed that all of them belonged to the TcI lineage. It is recommended that residents be made aware of the need to eliminate vectors in their homes and their surroundings.

Serological detection of antibodies to Anaplasma spp., Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen in dogs from Costa Rica.

In a study in Costa Rica 314 serum samples from dogs throughout all seven provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato and Ehrlichia canis, and of circulating antigen of Dirofilaria immitis. A total of 6.4% (20/314) and 38.2% (120/314) were positive for Anaplasma spp. (An) and E. canis (Ec) antibodies. Overall, 8.0% (25/314) were positive for D. immitis (Di) antigen. One single dog reacted positive with B. burgdorferi s.l. (Bb) antigen (0.3%, 1/314). E. canis positive dogs were detected in all provinces (highest percentages in Guanacaste, Puntarenas [both significantly different compared to the overall] and Limón). Guanacaste and Puntarenas also showed the highest prevalences of Anaplasma spp. (both significantly different compared to the overall). The highest prevalence of D. immitis was detected in Puntarenas (significantly different compared to the overall). Double pathogen exposure (Ec plus An; Ec plus Di; Ec plus Bb) were recorded in 8.9% (28/314). Two dogs showed a triple pathogen exposure (0.6%, 2/314; An, Ec and Di). There was a significant difference between male (11.5%, 18/156) and female (4.4%, 7/158) animals for D. immitis positive results. There was also a significant difference between breed and no breed dogs regarding the characteristics of a general positive test, as well as seropositivity to the single pathogens of Anaplasma spp., E. canis and D. immitis. Finally there was a significant difference in the presence of clinical signs again regarding the characteristics of a general positive test, as well as seropositivity to Anaplasma spp., E. canis and D. immitis. Practitioners in Costa Rica should be aware of the canine vector-borne diseases mentioned as dogs are at risk of becoming infected. Concerning the positive B. burgdorferi s.l. dog, an autochthonous occurrence cannot be confirmed due to a history of adoption and an unusual tattoo number. Veterinary advice to protect dogs and limit transmission of vector-borne pathogens, also to humans, by using prophylactic measures is strongly recommended.

Characterization of Anaplasma spp. infection in dogs from Costa Rica.

A cross-sectional study combining serological and molecular techniques for detecting selected Anaplasma species was conducted between 2011 and 2012 in dogs and ticks sampled in all provinces of Costa Rica. Global Anaplasma spp. seroprevalence was 2.7% (11/408) by indirect immunofluorescence assay. The 16S rRNA PCR confirmed active A. phagocytophilum infection only in one dog (0.3%, 1/374); however, the same sample was negative to groEL PCR. Out of 122 Rhipicephalus sanguineus s.l. ticks analyzed, one (0.8%) was found positive to A. phagocytophilum 16S rRNA PCR but negative when tested by groEL PCR; this tick was collected from a seronegative and PCR negative dog. Both 16S rRNA sequences were 100% (510/510bp) identical to A. phagocytophilum strains isolated in different countries from different hosts. The presence of A. platys was established in four dogs (1%, 4/374) by both 16SrRNA and groEL PCR. Ticks collected from the same dogs tested negative by PCR. The 16S rRNA sequences were 100% identical to the corresponding sequences of A. platys strains isolated from dogs in Croatia and Brazil, however groEL sequences showed variable similarity levels (99-100%) with different strains of A. platys isolated in Chile, Japan and Thailand, pointing out the possible presence of different variants in Central America. Collectively data indicate low prevalence of A. phagocytophilum and A. platys in dogs from Costa Rica. Furthermore, infections seem to occur without clinical signs but with some hematological changes, and seem to resolve without treatment.

Germination and soil seed bank traits of Podocarpus angustifolius (Podocarpaceae): an endemic tree species from Cuban rain forests.

Podocarpus angustifolius is an endangered recalcitrant-seeded small tree, endemic to mountain rain forests in the central and Pinar del Río regions in Cuba. In this study, the germination patterns of P. angustifolius seeds were evaluated and the nature of the soil seed bank was determined. Using a weighted two-factor design, we analyzed the combined germination response to seed source (i.e. freshly matured seeds directly collected from trees versus seeds extracted from soil samples) and pretreatment (i.e. seed water-immersion for 48h at room temperature). Germination was delayed for four weeks (= 30 days) in all cases, regardless of both factors analyzed. Moreover, nine additional days were necessary to achieve high germination values (in the case of fresh, pretreated seeds). These results overall may indicate the existence of a non-deep simple morphophysiological dormancy in P. angustifolius seeds. The water-immersion significantly enhanced seed germination, probably as a result of the hydration of recalcitrant seeds. Although germination of seeds extracted from soil samples was low, probably due to aging and pathogen effects throughout the time of burial, the study revealed the existence of a persistent soil seed bank (at least short-termed) of approximately 42 viable seeds per m2 in the upper 10cm of soil. Such a record is noteworthy since references to persistent soil seed banks in recalcitrant-seeded species are scarce in the literature. The population consequences derived from the formation of persistent soil seed banks in this endangered species are discussed.

Germination and soil seed bank traits of Podocarpus angustifolius (Podocarpaceae): an endemic tree species from Cuban rain forests

Podocarpus angustifolius is an endangered recalcitrant-seeded small tree, endemic to mountain rain forests in the central and Pinar del Río regions in Cuba. In this study, the germination patterns of P. angustifolius seeds were evaluated and the nature of the soil seed bank was determined. Using a weighted two-factor design, we analyzed the combined germination response to seed source (i.e. freshly matured seeds directly collected from trees versus seeds extracted from soil samples) and pretreatment (i.e. seed water-immersion for 48h at room temperature). Germination was delayed for four weeks (≈30 days) in all cases, regardless of both factors analyzed. Moreover, nine additional days were necessary to achieve high germination values (in the case of fresh, pretreated seeds). These results overall may indicate the existence of a non-deep simple morphophysiological dormancy in P. angustifolius seeds. The water-immersion significantly enhanced seed germination, probably as a result of the hydration of recalcitrant seeds. Although germination of seeds extracted from soil samples was low, probably due to aging and pathogen effects throughout the time of burial, the study revealed the existence of a persistent soil seed bank (at least short-termed) of ≈42 viable seeds per m² in the upper 10cm of soil. Such a record is noteworthy since references to persistent soil seed banks in recalcitrant-seeded species are scarce in the literature. The population consequences derived from the formation of persistent soil seed banks in this endangered species are discussed. Rev. Biol. Trop. 59 (3): 1061-1069. Epub 2011 September 01.

Podocarpus angustifolius es un árbol endémico de los bosques lluviosos de la región de Pinar del Río y la parte central de Cuba, que se encuentra en peligro de extinción. En este estudio se evaluó la germinación de sus semillas y la naturaleza del banco de semillas del suelo. Específicamente, se analizó la respuesta germinativa de las semillas a dos factores: su procedencia (recolectadas directamente de los árboles versus extraídas de muestras de suelo) y el pretratamiento (inmersión en agua durante 48h a temperatura ambiente). La germinación no comenzó hasta las cuatro semanas (≈30 días) en todos los casos. Además, fueron necesarios 9 días más para alcanzar un valor elevado de germinación (en el caso de semillas recolectadas del árbol con pretratamiento), lo que parece indicar la existencia de un letargo de tipo morfofisiológico simple no profundo. El pretratamiento incrementó significativamente la germinación, posiblemente por hidratación de las semillas recalcitrantes. El estudio demostró la existencia de un banco de semillas persistente en el suelo, al menos de corta duración de ≈42 semillas viables por m² en los 10 primeros cm del suelo. Dicho hallazgo es destacable, ya que los casos descritos en la literatura sobre bancos persistentes en especies con semillas recalcitrantes son escasos. Además, se discute la importancia que tiene para esta especie amenazada la existencia de un banco de semillas persistente.