MEDICAL-SURGICAL MANAGEMENT OF INTESTINAL INFECTION BY PROSTHENORCHIS ELEGANS IN NONHUMAN PRIMATES FROM COSTA RICA.

Parasitism is one of the most important diseases in nonhuman primates (NHP). Parasitism by Prosthenorchis elegans can be a threat to health and conservation of NHP in Costa Rica. Surgical management of intestinal acanthocephalan infection in two squirrel monkeys (Saimiri oerstedii) and one white-faced monkey (Cebus imitator) is described as an alternative to the lack of pharmacologic control options when there is a high burden of parasites present. A complete physical evaluation, including medical ultrasound techniques, allow for diagnosis of the parasite and its lesions. When animals present with a high burden of parasites, surgical management has shown to promote good health outcomes and increase the probability of survival.

Exposure of dogs to Rickettsia spp. in Costa Rica: Risk factors for PCR-positive ectoparasites and seropositivity.

Infection of dogs with Rickettsia spp. can result in inapparent, mild, or severe disease. Moreover, common dog ticks and fleas are able to transmit rickettsiae to nearby humans. In this study, the seroprevalence of spotted fever group (SFG) rickettsiae was determined in dogs of Costa Rica, as well as possible risk factors associated with exposure. An interview of owners and clinical examinations were performed in a country-wide sample of 441 dogs. IgG antibodies were determined in 399 dogs by indirect immunofluorescence assay (IFA) using antigens of Rickettsia rickettsii, R. amblyommatis, and R. felis. The presence of Rickettsia spp. gltA gene was evaluated by PCR in ticks and fleas. Poisson regression was performed to assess possible risk factors associated with seropositivity, as well as with having PCR-positive ticks and fleas. The overall seroprevalence to SFG rickettsiae was 10.0% (end titers 64 to 256). Rhipicephalus sanguineus s.l. (116/441; 26.3%) and Ctenocephalides felis (153/441; 34.7%) were the most common ectoparasites. Rickettsia DNA was detected in 30% (39/130) and 32.3% (56/173) of tick and flea pools, respectively. Seropositivity was significantly associated with mean age of 2 to 7 years, scrotal edema, walking problems, large size, and tick and flea infestation. Being a purebred dog was a possible protective factor. The presence of Rickettsia PCR-positive ticks was associated with being a purebred dog, while flea treatment was protective. Having PCR-positive fleas was associated with being purebred and the number of people in the dog's environment; protective factors were free roaming and being an outdoor dog. Results confirm that dogs in Costa Rica are exposed to different species of SFG rickettsiae. This may represent a risk to human health and underscores the need for accurate diagnosis in dogs and humans. Surveillance of rickettsial infection in canines may provide useful indicators to understand the epidemiology of these zoonoses.

Strain diversity of Rickettsia amblyommatis in ticks infesting birds in the North Huetar conservation area of Costa Rica.

Although the presence of rickettsial agents in ticks infesting wild birds in Costa Rica has been recently reported, information on strain diversity is limited to selected rickettsial species. In order to mine deeper into rickettsial agents of ticks infesting Costa Rica wild birds a total of 399 birds from the North Huetar Conservation Area of Costa Rica were captured, and 134 immature ticks (76 larvae and 58 nymphs) were recovered from 61 birds. Ticks were tested for the presence of Rickettsia spp. by conventional PCR and sequencing of the gltA, ompA, ompB, 17 kDa, and groEL genes. Six (11.3%) Amblyomma longirostre and Amblyomma geayi ticks collected from passeriform birds, yielded amplicons of the expected size. Amplicons were sequenced, and BLAST results collectively showed that all sequences had 99-100% nucleotide identity with Rickettsia amblyommatis (formerly, 'Candidatus Rickettsia amblyommii'). Three different R. amblyommatis strains were identified. Four new tick species-host associations and the first detection of R. amblyommatis in A. geayi in Costa Rica are also reported.

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.

Molecular detection and identification of Rickettsiales pathogens in dog ticks from Costa Rica.

Although vector-borne diseases are globally widespread with considerable impact on animal production and on public health, few reports document their presence in Central America. This study focuses on the detection and molecular identification of species belonging to selected bacterial genera (Ehrlichia, Anaplasma and Rickettsia) in ticks sampled from dogs in Costa Rica by targeting several genes: 16S rRNA/dsb genes for Ehrlichia; 16S rRNA/groEL genes for Anaplasma, and ompA/gltA/groEL genes for Rickettsia. PCR and sequence analyses provides evidences of Ehrlichia canis, Anaplasma platys, and Anaplasma phagocytophilum infection in Rhipicephalus sanguineus s.l ticks, and allow establishing the presence of Rickettsia monacensis in Ixodes boliviensis. Furthermore, the presence of recently discovered Mediterranean A. platys-like strains is reported for the first time in Central America. Results provide new background on geographical distribution of selected tick-transmitted bacterial pathogens in Costa Rica and on their molecular epidemiology, and are pivotal to the development of effective and reliable diagnostic tools in Central America.

Serology, molecular detection and risk factors of Ehrlichia canis infection in dogs in Costa Rica.

A cross-sectional study combining different serological and molecular techniques for the detection of Ehrlichia species in dogs and their ticks was carried out with data from all regions of Costa Rica. A seroprevalence of 32.1% (131/408), and infection with E. canis of 3.2% (13/407) was found, whereas 6.9% (9/130) of ticks attached to the dogs were PCR positive to E. canis. Higher prevalences were found outside the Greater Metropolitan Area (GMA). Risk factors associated with E. canis seropositivity were age, between 2 and 7 years (RR: 1.6, 95% CI: 1.2-2.2) and 8-15 years (RR: 1.8, 95% CI: 1.2-3.0), number of dogs/total of households [Dogs per Household Ratio (DHR) ≥3.1 (RR: 2.0; 95% CI: 1.4-3.0)], number of dogs infested with at least one tick/total of dogs sampled [Tick Infestation Prevalence (TIP)≥31% (RR: 2.1; 95% CI:1.3-3.3)] and living outside the GMA (RR: 1.7; 95% CI: 1.2-2.4) and being a mixed-breed dog (RR: 1.5; 95% CI: 1.1-2.1). Risk factors for E. canis PCR positive dogs were a depressive attitude (OR: 11.2; 95% CI: 1.1-115.9), fever (OR:4.8; 95% CI:1.2-19.3), DHR≥3.1 (OR: 5.7; 95% CI:1.7-19.2)], number of ticks/total of dogs sampled [Tick Distribution Ratio (TDR) ≥2.1 (OR: 6.5; 95% CI: 1.3-31.8)], and TIP≥40% (OR: 5.7; 95% CI: 1.7-19.2). This paper describes E. canis seroprevalence, PCR prevalence and tick analysis in dogs from Costa Rica, with associated clinical signs and owner perceptions. In summary, most of the E. canis infections in dogs in our country seemed to pass unnoticed by owners. Since most of the seropositive dogs (97.7%, 131/134) were negative for E. canis DNA in their blood, it is important to determine in future studies if these dogs recovered from the E. canis infection without any medication, or are persistently infected, and will develop chronic disease.

Centrocestus formosanus (Opisthorchiida: Heterophyidae) como causa de muerte de alevines de tilapia gris Oreochromis niloticus (Perciforme: Cichlidae) en el Pacífico seco de Costa Rica

Centrocestus formosanus (Opisthorchiida: Heterophyidae) as a cause of death in gray tilapia fry Oreochromis niloticus (Perciforme: Cichlidae) in the dry Pacific of Costa Rica. Centrocestus formosanus is a zoonotic trematode from Asia and has been mainly associated as cause of death of cultured fish. To identify pathogen trematode species in tilapia fry (Oreochromis niloticus) and to determine mollusks hosting these parasites, freshwater mollusks were collected from tilapia cultured ponds and experimental infections were carried out with tilapia fries and different mollusk species. A total of 907 freshwater mollusks were obtained from tilapia ponds and were identified to species level, four gastropods and one bivalve were determined: Melania tuberculata, Melanoides turricula, Pomacea flagellata, Haitia cubensis and Anodontiles luteola. For the first time, the presence of M. turricula and H. cubensis are reported in Costa Rica. Seven morphotypes of cercariae (Xifiodiocercaria, Equinostoma, Oftalmocercaria, Parapleurolofocercus, Cistocerca, Furcocercaria and Leptocercaria) parasitizing all five species of mollusks were found, all of distome type. Experimental exposure of tilapia fry to M. tuberculata demonstrated that the parapleurolofocercus morphotype found in the mollusk is in accordance with the finding of C. formosanus in tilapia fry. An abundance and mean intensity of 1018-1027 digeneans per gill in each exposed fish was determined. Centrocestus formosanus is reported for the first time in Costa Rica, for which the primary and secondary intermediate hosts were also determined. Rev. Biol. Trop. 58 (4): 1453-1465. Epub 2010 December 01.

Centrocestus formosanus es un parásito trematodo zoonótico originario de Asia asociado con muertes de peces principalmente de cultivo. 907 moluscos provenientes de estanques sembrados con tilapias, seleccionados uno por provincia fueron identificados al nivel taxonómico especifico. Se identificaron cuatro gastrópodos y un bivalvo: M. tuberculata, M. turricula, P. flagellata, H. cubensis y A. luteola. Se reporta, por primera vez, la presencia de dos especies de moluscos en Costa Rica. Se identificaron siete morfotipos de cercarias parasitando las cinco especies de moluscos encontradas. En la segunda exposición experimental se demostró que el morfotipo parapleurolofocercus encontrado en M. tuberculata concuerda con el hallazgo de C. formosanus en alevines de tilapia, después del examen clínico, anatomopatológico y parasitológico realizado a los alevines expuestos. Las metacercarias fueron extraídas del quiste utilizando microagujas y micropinzas lavadas en solución salina fisiológica (0.65%), fijadas en formol caliente al 4% y después esquematizadas con una cámara clara adaptada a un microscopio fotónico, estimándose una abundancia e intensidad media de 1018-1027 digeneos por branquia en cada pez parasitado, determinándose así el hospedador intermediario primario y secundario del parásito. En el presente trabajo se reporta por primera vez Centrocestus formosanus en Costa Rica.

[Centrocestus formosanus (Opisthorchiida: Heterophyidae) as a cause of death in gray tilapia fry Oreochromis niloticus (Perciforme: Cichlidae) in the dry Pacific of Costa Rica]. / Centrocestusformosanus (Opisthorchiida: Heterophyidae) como causa de muerte de alevines de tilapia gris Oreochromis niloticus (Perciforme: Cichlidae) en el Pacifico seco de Costa Rica.

Centrocestusformosanus is a zoonotic trematode from Asia and has been mainly associated as cause of death of cultured fish. To identify pathogen trematode species in tilapia fry (Oreochromis niloticus) and to determine mollusks hosting these parasites, freshwater mollusks were collected from tilapia cultured ponds and experimental infections were carried out with tilapia fries and different mollusk species. A total of 907 freshwater mollusks were obtained from tilapia ponds and were identified to species level, four gastropods and one bivalve were determined: Melania tuberculata, Melanoides turricula, Pomacea flagellata, Haitia cubensis and Anodontiles luteola. For the first time, the presence of M. turricula and H. cubensis are reported in Costa Rica. Seven morphotypes of cercariae (Xifiodiocercaria, Equinostoma, Oftalmocercaria, Parapleurolofocercus, Cistocerca, Furcocercaria and Leptocercaria) parasitizing all five species of mollusks were found, all of distome type. Experimental exposure of tilapia fry to M. tuberculata demonstrated that the parapleurolofocercus morphotype found in the mollusk is in accordance with the finding of C. formosanus in tilapia fry. An abundance and mean intensity of 1018-1027 digeneans per gill in each exposed fish was determined. Centrocestus formosanus is reported for the first time in Costa Rica, for which the primary and secondary intermediate hosts were also determined.

Gastrointestinal parasites and ectoparasites of Bradypus variegatus and Choloepus hoffmanni sloths in captivity from Costa Rica.

Sloths may serve as host to a wide range of parasites. However, there is little information available on the types of parasites that affect Costa Rica's sloth population. During a 1-yr period, 65 specimens of Costa Rican sloth species (Choloepus hoffmanni; n = 56) and Bradypus variegates; n = 9) from a local zoo were sampled. Fecal samples were evaluated using two different diagnostic techniques, Sheather's flotation and sedimentation. Concurrently, these sloths were examined for ectoparasites. Gastrointestinal parasites were found in 14 sloths (21.5%), from which 13 animals were C. hoffmanni and one was B. variegatus. Gastrointestinal parasites were recognized as Coccidia 71.4% (10/14), Cestoda 21.4% (3/14), and Spiruroidea 7.1% (1/14). Coccidia and cestodes were seen in C. hoffmanni, and spirurids were identified in B. variegatus. Among 27 sloths examined, only six had dermal problems (five C. hoffmanni and two B. variegatus). Ectoparasites recovered were Sarcoptes scabiei (Acari, Sarcoptidae) mites and Amblyomma varium (Acari, Ixodidae) ticks. This is the first time that cestode strobilae and nematode eggs are reported in sloth feces and that Monezia benedeni and L. leptocephalus were found in captive sloths.