ABSTRACT Global advances in reproductive biotechnology have allowed for the transfer of embryos from donor females with high genetic merit to recipients using the cryopreservation technique, which preserves an embryo of excellent quality and viability, thereby achieving a feasible pregnancy rate. The objective of this study was to evaluate the quality and viability of Holstein embryos that have been cryopreserved for more than 40 years under glycerol freezing. The embryos were transferred to the recipient heifers using a non-surgical method. Two 17-month-old Holstein heifers (360 kg live weights) which were clinically healthy and reproductively active were used as the recipients. Two bovine embryos of Grade 1 quality were thawed and evaluated for their morphology. Of the two embryo transfers, one pregnancy was achieved, resulting in the birth of a calf. Therefore, embryos frozen in liquid nitrogen and glycerol as a cryopreservative for more than 40 years maintained their quality and viability to produce a live calf.

RESUMO Os avanços globais em biotecnologia reprodutiva permitiram a transferência de embriões de fêmeas doadoras com alto mérito genético para receptoras, usando-se a técnica de criopreservação, que preserva um embrião de excelente qualidade e viabilidade, alcançando, assim, uma taxa de gravidez viável. O objetivo deste estudo foi avaliar a qualidade e a viabilidade de embriões Holstein, criopreservados por mais de 40 anos, sob congelamento de glicerol. Os embriões foram transferidos para as novilhas receptoras, usando-se um método não cirúrgico. Duas novilhas Holstein de 17 meses de idade (360kg de peso vivo), que eram clinicamente saudáveis e reprodutivamente ativas, foram utilizadas como receptoras. Dois embriões bovinos de qualidade Grau 1 foram descongelados e avaliados quanto à sua morfologia. Das duas transferências embrionárias, uma gravidez foi obtida, resultando no nascimento de um bezerro. Portanto, os embriões congelados em nitrogênio líquido e glicerol como criopreservante por mais de 40 anos mantiveram sua qualidade e viabilidade para produzir um bezerro vivo.

ABSTRACT Background: Since 2011, the coasts of North America, Mexico, and the Caribbean have experienced an unusual increase in the arrival of Sargassum. As a consequence of this massive accumulations of Sargassum holopelagic species in Caribbean Sea have been generated and that threaten local biodiversity and trigger economic losses associated with beach deterioration and impact on fisheries and tourism. Goals: This document quantifies the occurrence of an excessive drift of Sargassum in two sites (Puerto Viejo and Punta Uva) of the Caribbean coast of Costa Rica, Limón province, in March-April of 2019. A macroalgae species checklist is presented from six sites (Punta Cahuita, Puerto Vargas, Puerto Viejo, Playa Chiquita, Punta Uva and Manzanillo) in the Caribbean. In addition, the Sargassum morphotypes are quantified for the study area. Methods: Transects (50 m) were located parallel to the shoreline and, in points previously estimated, a quadrat of 25 x 25 cm covered the entire Sargassum band width on the beach until the water line, by fliping the quadrats and collecting the biomass for as many quadrats were necessary for the band. Then, pelagic Sargassum biomass was weighted in every point, and the dry weight biomass was estimated. Three quadrats were collected by the transect method. Associated species were identified as well as Sargassum morphotypes in all sites. Results: Descriptions that illustrate the morphology of the three morphotypes of pelagic species. The putatively rare S. natans VIII dominated the dritf (65 % dry weight), followed by S. natans I (25% dry weight), and S. fluitans (10 % dry). Twenty eight species were identified from Chlorophyta (9), Ochrophyta (15), and Rhodophyta (4) Divisions. Conclusions: There were no significant differences between months (March-April) and sites (Puerto Viejo and Punta Uva) for the dritf biomass.

RESUMEN Antecedentes: Desde 2011, las costas de América del Norte, México y el Caribe han experimentado un aumento inusual en la llegada de Sargassum de arribazón. Como consecuencia, se han generado acumulaciones masivas de especies holopelágicas del género Sargassum en áreas costeras del Caribe que amenazan la biodiversidad local y desencadenan pérdidas económicas asociadas con el deterioro de las playas y el impacto en la pesca y el turismo. Objetivo: Se cuantifica la presencia de una deriva excesiva de Sargassum en dos sitios (Puerto Viejo y Punta Uva) de la costa caribeña de Costa Rica, provincia de Limón, en los meses de marzo y abril de 2019. Se presenta la lista de especies de macroalgas conspicuas asociadas al fenómeno en seis sitios (Punta Cahuita, Puerto Vargas, Puerto Viejo, Playa Chiquita, Punta Uva y Manzanillo) en el Caribe. Además, se cuantifican los morfotipos de Sargassum para el área de estudio. Métodos: Se ubicaron transectos (50 m) paralelos a la línea de costa y, en puntos previamente estimados, un cuadrante de 25 x 25 cm cubrió todo el ancho de banda de Sargassum en la playa hasta la línea de flotación, volteando los cuadrantes y recolectando la biomasa para tantos cuadrantes como fueron necesarios para la cubrir la zona arribada. Luego, se pesó la biomasa pelágica de Sargassum en cada punto, y se estimó la biomasa en peso seco. Se recolectaron tres cuadrantes mediante el método de transectos. Se identificaron especies asociadas, así como morfotipos de Sargassum en todos los sitios. Resultados: se presenta la morfología de los tres morfotipos de especies pelágicas. El supuestamente raro S. natans VIII dominó el arribo de algas (65% de peso seco), seguido por S. natans I (25% de peso seco), S. fluitans (10% de peso seco). Se identificaron 28 especies de las Divisiones: Chlorophyta (9), Ochrophyta (15) y Rhodophyta (4). Conclusiones: No existen diferencias significativas entre meses (marzo-abril) y sitios (Puerto Viejo y Punta Uva) para la biomasa arribada.

ABSTRACT Background: Bacteria of the genus Brucella yield the disease known as brucellosis (the most critical zoonosis in the world). Currently, 14 species are recognized, reported in marine mammals, amphibians, and commercial fish. In recent years, in the Eastern Basin, Brucella melitensis was detected in agricultural soil and the Crater Lakes’ water, so it was speculated whether B. melitensis could infect native ichthyofauna. Objective: Isolate and identify Brucella melitensis in the charal of La Preciosa Crater Lake (Poblana letholepis). Methods: The first isolation was carried out on agar-plate from soft tissues and identifying the microorganism through standard microbiological tests. Which was corroborated by PCR-PF, using reference strains, by amplifying the bp26 gene, specific for the genus Brucella. Results: B. melitensis was isolated in 66.66% of the samples studied n=60, the microbiological profiles were comparatively identical with the reference strains. The PCR test amplified the genus-specific gene bp26 with 1029 bp in 40% of the samples analyzed. Conclusions: The study generated the first isolation and identification report for Brucella melitensis in a native fish, which supports future studies to better understand this zoonotic disease and recognize the importance of native fauna in the transmission and permanence of brucellosis in natural aquatic environments.

RESUMEN Antecedentes: Las bacterias del género Brucella producen la enfermedad conocida como brucelosis (la zoonosis más importante del orbe). Actualmente se reconocen 14 especies, reportadas en mamíferos marinos, anfibios y peces comerciales, En los últimos años, en la Cuenca Oriental se detectó a Brucella melitensis en suelo agrícola y en el agua de los Lagos Cráter, por lo que se especulaba si B. melitensis tenía la capacidad de infectar a la ictiofauna nativa. Objetivo: Aislar e identificar a Brucella melitensis en el charal Poblana letholepis del Lago Cráter La Preciosa. Métodos: El primoaislamiento se realizó en placa-Agar a partir de tejidos blandos y la identificación del microorganismo a través de pruebas microbiológicas estándar. La identificación se corroboró por PCR-PF, utilizando cepas de referencia, mediante la amplificación el gen bp26, específico para el género Brucella. Resultados: Se aisló a B. melitensis en 66.66% de las muestras estudiadas n=60, los perfiles microbiológicos resultaron comparativamente idénticos con las cepas de referencia. La prueba por PCR amplificó el gen específico del género bp26 con 1029 pb en 40% de las muestras analizadas. Conclusiones: El estudio generó el primer reporte de aislamiento e identificación para B. melitensis en un pez nativo, el cual da soporte a futuros estudios para comprender mejor esta enfermedad zoonotica y reconocer la importancia de la fauna nativa en la transmisión y permanencia de la brucelosis en ambientes acuáticos naturales.