The benzothiadiazole compound acibenzolar-S -methyl (ASM) was assessed as an inducer of resistance against Crinipellis perniciosa , agent of witches' broom, and Verticillium dahliae , agent of vascular wilt, both on cocoa. ASM induced a reduction in incidence of witches' broom of up to 84·5% when sprayed 30 days before inoculation on cocoa seedlings of cv. Catongo. ASM also induced a reduction in severity of Verticillium wilt to 55·4% on cv. Theobahia. For both pathosystems, effects of dose on disease were not clearly observed. The efficacy of the inducer increased with the interval between sprayings and the respective inoculations with the pathogens. In another experiment, the effect of ASM on the control of witches' broom on cocoa seedlings was compared with that of cuprous oxide and tebuconazole, all sprayed 15 days before inoculation. ASM reduced disease incidence by 60·1% compared with the inoculated control. ASM was superior to tebuconazole, and there was also a tendency for ASM to be better than cuprous oxide. To understand the mechanism of action of ASM as an inducer of resistance, alterations in the levels of total phenolics, polyphenol oxidases and peroxidases were evaluated 3, 15 and 30 days after spraying of seedlings of cv. Catongo. Enzyme activities from seedlings of cv. Theobahia were evaluated 30 days after spraying. On cv. Catongo, no significant differences in total phenolic content and polyphenol oxidase activity were detected after spraying. However, an increase in peroxidase activity was detected at all times of evaluation. On cv. Theobahia, significant increases in activities of peroxidase and polyphenol oxidase were detected, indicating that defence responses due to ASM were dependent on host genotype.

One hundred and fifty-two blood samples of non-human primates of thirteen rescue centers in Costa Rica were analyzed to determine the presence of species of Plasmodium using thick blood smears, semi-nested multiplex polymerase chain reaction (SnM-PCR) for species differentiation, cloning and sequencing for confirmation. Using thick blood smears, two samples were determined to contain the Plasmodium malariae parasite, with SnM-PCR, a total of five (3.3%) samples were positive to P. malariae, cloning and sequencing confirmed both smear samples as P. malariae. One sample amplified a larger and conserved region of 18S rDNA for the genus Plasmodium and sequencing confirmed the results obtained microscopically and through SnM-PCR tests. Sequencing and construction of a phylogenetic tree of this sample revealed that the P. malariae/P. brasilianum parasite (GenBank KU999995) found in a howler monkey (Alouatta palliata) is identical to that recently reported in humans in Costa Rica. The SnM-PCR detected P. malariae/P. brasilianum parasite in different non-human primate species in captivity and in various regions of the southern Atlantic and Pacific coast of Costa Rica. The similarity of the sequences of parasites found in humans and a monkey suggests that monkeys may be acting as reservoirs of P.malariae/P. brasilianum, for which reason it is important, to include them in control and eradication programs.

The trematode Paragonimus mexicanus is the etiological agent of paragonimiasis, a food-borne zoonotic disease in Latin America. This species, as well as Paragonimus caliensis, have been reported from Costa Rica, but it is not known if the two are synonymous. Two types of Paragonimus metacercariae from freshwater pseudothelphusid crabs from several localities in Costa Rica were recognized by light microscopy. Morphologically, these corresponded to descriptions of P. mexicanus and P. caliensis. Metacercariae of the former species lacked a membrane or cyst and their bodies were yellow in color. Those of P. caliensis were contained in a transparent thin cyst and were pink in color. Morphotypes of metacercariae were determined using scanning electron microscopy (SEM). Based on the number and distribution of papillae in the ventral sucker, three morphotypes were found for P. mexicanus and two for P. caliensis. Analysis of DNA sequences (nuclear ribosomal 28S and ITS2 genes, and partial mitochondrial cox1 gene) confirmed the presence of P. mexicanus and provided the first molecular data for P. caliensis. The two species are phylogenetically distinct from each other and distant from the Asian species. The confirmation of P. caliensis as a separate species from P. mexicanus raises several questions about the ecology, biological diversity, and epidemiology of the genus Paragonimus in Costa Rica.

In early 2007, severe yellowing and chlorosis symptoms were observed in field-grown and greenhouse tomato (Solanum lycopersicum L.) plants in Costa Rica. Symptoms resembled those of the genus Crinivirus (family Closteroviridae), and large populations of whiteflies, including the greenhouse whitefly Trialeurodes vaporariorum (Westwood), were observed in the fields and on symptomatic plants. Total RNA was extracted from silica gel-dried tomato leaf tissue of 47 representative samples (all were from symptomatic plants) using TRI Reagent (Molecular Research Inc., Cincinnati, OH). Reverse transcription (RT)-PCR reactions were performed separately with each of the four primer sets with the Titan One-Tube RT-PCR Kit (Roche Diagnostics Corp., Chicago IL). Specific primers used for the detection of the criniviruses, Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV), were primer pair ToCV-p22-F (5′-ATGGATCTCACTGGTTGCTTGC-3′) and ToCV-p22-R (5′-TTATATATCACTCCCAAAGAAA-3′) specific for the p22 gene of ToCV RNA1 (1), primer pair ToCVCPmF (5′-TCTGGCAGTACCCGTTCGTGA-3′) and ToCVCPmR (5′-TACCGGCAGTCGTCCCATACC-3′) designed to be specific for the ToCV CPm gene of ToCV RNA2 (GenBank Accession No. AY903448) (2), primer pair ToCVHSP70F (5′-GGCGGTACTTTCGACACTTCTT-3′) and ToCVHSP70R (5′-ATTAACGCGCAAAACCATCTG-3′) designed to be specific for the Hsp70 gene of RNA2 of ToCV (GenBank Accession No. EU284744) (1), and primer pair TICV-CP-F and TICV-CP-R specific for the coat protein gene of TICV (1). Amplified DNA fragments (582 bp) were obtained from nine samples, four from the greenhouse and five from the open field, with the ToCV-p22 specific primers and were cloned into the pCRII TOPO cloning vector (Invitrogen, Carlsbad, CA). Nucleotide sequence analysis of all purified RT-PCR products verified their identity as ToCV, sharing 99.5 to 100% sequence identity among themselves and 96% to 98% sequence identity with previously reported ToCV p22 sequences from Florida (Accession No. AY903447), Spain (Accession No. DQ983480), and Greece (Accession No. EU284745). The presence of ToCV in the samples was confirmed by additional amplification and sequence analysis of the CPm (449-bp fragment) and Hsp70 (420-bp fragment) genes of ToCV RNA2 and sharing 98 to 99% sequence homology to Accession Nos. AY903448 and EU284774, respectively. One representative sequence of the p22 gene of the Costa Rican isolate was deposited at GenBank (Accession No. FJ809714). No PCR products were obtained using either the TICV-specific primers nor from healthy tomato tissue. The ToCV-positive samples were collected from a region in the Central Valley around Cartago, Costa Rica. To our knowledge, this is the first report of ToCV in Costa Rica. The economic impact on tomato has not yet been determined. Studies are underway to determine the incidence of ToCV in Costa Rica field-grown and greenhouse tomatoes. References: (1) A. R. A. Kataya et al. Plant Pathol. 57:819, 2008. (2) W. M. Wintermantel et al. Arch. Virol. 150:2287, 2005.

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.

Leaf samples of Solanum lycopersicum, Capsicum annuum, Cucurbita moschata, Cucurbita pepo, Sechium edule and Erythrina spp. were collected. All samples were positive for begomoviruses using polymerase chain reaction and degenerate primers. A sequence of ∼1,100 bp was obtained from the genomic component DNA-A of 14 samples. In addition, one sequence of ∼580 bp corresponding to the coat protein (AV1) was obtained from a chayote (S. edule) leaf sample. The presence of Squash yellow mild mottle virus (SYMMoV) and Pepper golden mosaic virus (PepGMV) were confirmed. The host range reported for SYMMoV includes species of the Cucurbitaceae, Caricaceae and Fabaceae families. This report extends the host range of SYMMoV to include the Solanaceae family, and extends the host range of PepGMV to include C. moschata, C. pepo and the Fabaceae Erythrina spp. This is the first report of a begomovirus (PepGMV) infecting chayote in the Western Hemisphere.

A total of six Amblyomma tapirellum ticks were found for the first time in rainforest from the lowland to the southern Pacific of Costa Rica. Tick identification was carried out by morphology and afterward confirmed by molecular analysis, using polymerase chain reaction (PCR) and DNA barcoding. Further studies are required to determine the distribution of A. tapirellum and this species' potential as a vector of bacterial agents to humans and wild hosts.

La acuicultura es uno de los sistemas de producción de alimentos de más rápido crecimiento en elmundo, debido a que contribuye con proteína animal para una población que aumenta día con día, loque refuerza la seguridad alimentaria. Además, aporta al mejoramiento de las condiciones de vida através de la generación de fuentes de empleo y divisas. Desde el entorno productivo, estos productosno contaminan y no alteran los ecosistemas, sino más bien aportan a la flora y fauna de los sistemas.Sin embargo, los problemas en este tipo de actividad productiva son muy frecuentes, sobre todo, por elmanejo inadecuado de las explotaciones y por la presencia de agentes infecciosos. La investigación tuvocomo propósito determinar la presencia de agentes infecciosos en camarones de cultivo en Costa Rica.Entre junio del 2016 y mayo del 2017 se recolectaron muestras pos-larvas y agua del primer bombeo enel momento de la cosecha, y camarones juveniles de seis a ocho semanas pos cosecha en cuatro fincaslocalizadas en el Golfo de Nicoya y el Pacífico Central. Se recolectaron diez camarones juveniles al inicio,a la mitad y al final del estanque, de los cuales se extrajo el hepatopáncreas y el estómago y se sometieron aextracción de ADN. Seguidamente, se analizaron mediante técnica molecular (PCR y secuenciación) parala detección de la toxina binaria (PirA-PirB), de Vibrio parahaemolyticus (causante de la necrosis aguda delHepatopáncreas AHPND), del virus de la necrosis infecciosa hipodérmica y hematopoyética (IHHNV), delvirus del síndrome de la mancha blanca (WSSV) y de Enterocytozoon hepatopenaei (EHP).Se determinó la presencia de IHHNV en todas las fincas analizadas, el virus se encontró en todas lasmuestras de pos-larvas de las cuatro fincas, como también en los estómagos y hepatopáncreas de loscamarones juveniles al inicio, mitad y final del estanque de las fincas 1 y 3, y solamente en los estómagosde los camarones juveniles al final del estanque de las fincas 2 y 4. Las secuencias de IHHNV obtenidasen las cuatro fincas fueron 100 % idénticas con una cepa de IHHNV reportada en China. El virus sedeterminó como del Linaje 3 (cepa patógena). En el agua del primer bombeo no se encontró IHHNV.Además, en dos fincas (Finca 1 y Finca 4) se determinó la presencia de las toxinas PirA y PirB, en loshepatopáncreas de los camarones juveniles, con una identidad del 100 % con una cepa de Vietnam.Estas toxinas se establecieron en la Finca 1, asociadas a V. parahemolyticus. No se encontró WSSV yEHP en las cuatro fincas estudiadas.