Baculovirus mediated high-level expression of luciferase in silkworm cells and larvae

Bombyx mori nuclear polyhedrosis virus (BmNPV)-based baculovirus expression system exploits silkworm larvae as an economical alternative to large-scale cell cultures for production of biomolecules. To generate recombinant BmNPV at high efficiency, we have achieved high efficiency transfection of B. mori cells, BmN, through lipofection. Optimal conditions for lipofection were standardized by quantification of the transient expression level of firefly luciferase (luc) reporter gene under control of an immediate early gene promoter of BmNPV Lipofection was 50-fold and 100-fold more efficient than the calcium phosphate method for transfecting BmN and Sf9 cells, respectively. Lipofection enabled us to generate a recombinant BmNPV (vBmluc), harboring luc under control of the strong polyhedrin promoter On infection with vBmluc, luciferase was expressed at very high levels, 170 mu g/10(6) BmN cells or 13 mg/larva. Expression of luciferase in vBmluc-infected larvae was visualized by luminescence emission instantaneously following luciferin injection generating ''glowing silkworms''.

A new genus and species of arboreal toad with phytotelmonous larvae, from the Andaman Islands, India (Lissamphibia, Anura, Bufonidae)

A new bufonid amphibian, belonging to a new monotypic genus, is described from the Andaman Islands, in the Bay of Bengal, Republic of India, based on unique external morphological and skeletal characters which are compared with those of known Oriental and other relevant bufonid genera. Blythophryne gen. n. is distinguished from other bufonid genera by its small adult size (mean SVL 24.02 mm), the presence of six presacral vertebrae, an absence of coccygeal expansions, presence of an elongated pair of parotoid glands, expanded discs at digit tips and phytotelmonous tadpoles that lack oral denticles. The taxonomic and phylogenetic position of the new taxon (that we named as Blythophryne beryet gen. et sp. n.) was ascertained by comparing its 12S and 16S partial genes with those of Oriental and other relevant bufonid lineages. Resulting molecular phylogeny supports the erection of a novel monotypic genus for this lineage from the Andaman Islands of India.

Caracterizacion del comportamiento de la estomatitis vesicular en animales domesticos de pezuña hendida en la sexta region (Jinotega y Matagalpa) en el periodo 2008

El presente trabajo se realizó con el fin de contribuir al conocimiento dando a conocer la Caracterización del comportamiento de la estomatitis Vesicular en animales domésticos de pezuña hendida en la región VI (Jinotega y Matagalpa) en el periodo 2008, debido a su gran poder de difusión considerando que en Nicaragua no existe un programa de erradicación o control de la estomatitis vesicular, aunque los humanos también pueden contraer estomatitis vesicular. Se hizo necesario realizar un monitoreo de la enfermedad en esta región, determinar el tipo de cepa que es mas prevalente en la especie de pezuña hendida y el lugar anatómico que más afecta , se tomaron muestras en conjunto con técnicos del MAGFOR y fueron enviadas al Laboratorio de Diagnóstico de Enfermedades Vesiculares (LADIVES) ,localizado en Panamá, se llevó a cabo un análisis estadístico descriptivo, para este análisis se utilizó la información que se colectó en los casos atendidos entre el mes de Julio a Diciembre 2008 y la información retrospectiva de Enero a Junio, con apoyo de la base de datos de Enfermedades Vesiculares que lleva la oficina del Convenio Bilateral Antiaftosa (CAB) del MAGFOR Se elaboraron distribuciones de frecuencia, para las variables; diagnóstico, prevalencia, especie animal y región anatómica, lo que nos accedió informarnos sobre los valores concretos que adoptaron las variables a analizar y sobre el número(o porcentaje) de veces que se repite cada uno de esos valores y nos permitió construir los diagramas con los resultados, concluyendo que de un total de 132 muestras enviadas al laboratorio 82 resultaron positivas a Estomatitis Vesicular, 44 de ellas resultaron al serotipo New Jersey en la zona de Matagalpa y 33 en Jinotega, 5 resultaron positivas al serotipo Indian a en Matagalpa y en la región de Jinotega no se presentó ninguna positiva a este serotipo, la especie más afectada fue el bovino, de las 132 muestras enviadas 129 fueron tomadas en esta especie, siendo las pezuñas el lugar anatómico de mayor predilección para el virus, del total de muestras 91 se tomaran esta región, manifestándose con mayor frecuencia en épocas de lluvia sobre todo en los meses de mayo a octubre.

Evaluación de seis fungicidas in vitro para el manejo de tres patógenos del cafeto (Coffea arabica L.) Región sexta

Con el propósito de evaluar la efectividad "in vitro'' de algunos fungicidas usados para el manejo de patógenos causantes de enfermedades del cafeto y reducir sus dosis, se condujo el presente trabajo que se llevó a cabo en el Laboratorio de Sanidad Vegetal del Centro Experimental de la Comisión Nacional del Café (CONCAFE) ubicado en el Km. 7, empalme San Francisco Carretera a San Ramón-Matagalpa y en el Laboratorio de Micología del Centro Nacional de Protección Vegetal (CENAPROVE) que está ubicado en el Km. 12.5 Carretera Sur, San José de la Cañada, Managua, a partir de Mayo a Diciembre de 1991. Se probaron seis fungicidas: Benomyl, Fermate, Propiconazol, Cupravit, Propineb y Captafol, evaluándose en cada uno cuatro dosis que se escogieron partiendo de la dosis comercial recomendada por las casas distribuidoras, la que tomamos como dosis alta, de la cual se derivaron las subsiguientes dosis, para él manejo de: Colletotrichum coffeanum Noack., Rhizoctonia solani Kuhn y Fusarium oxysporum f. sp. Se usó un Diseño Completo al Azar (DCA) con cuatro repeticiones, 24 tratamientos y 1 testigo absoluto sin tratamiento. Se midió el crecimiento del hongo en medio de cultivo PDA, (Papa-Dextrosa-Agar} con los fungicidas, encontrándose que el Propiconazol fue el que mayor efectividad tuvo aún en la dosis más baja, en cualquiera de los patógenos. El Fermate en su dosis media a baja y el Captafol en su dosis media son los productos que lograron en los 3 patógenos ejercer un buen control. Cupravit no mostró ninguna efectividad sobre R. solani y F oxysporum, pero si sobre C.coffeanum. Benomyl ejercio un buen efecto para R. solani y f. oxysporum pero no para C. coffeanum. Propineb mostro su efectividad en R. solani y C. coffeanum pero no ejerció ningun efecto en F. oxysporum.

The mechanical effects of water flow on fish eggs and larvae

The impact of mechanical stresses upon ichthyoplankton entrained in power plant cooling systems has long been considered negligible. Arguments and evidence exist, however, to show that such a supposition is not universally true, especially in nuclear power plants. The mechanisms of mechanical damage can be detailed in terms of pressure change, acceleration, and shear stress with in the fluid flow field. Laboratory efforts to quantify the effects of mechanical stress have been very sparse. A well-planned bioassay is urgently needed. (PDF has 11 pages.)

Effects of shear on eggs and larvae of striped bass, morone saxatilis, and white perch, M. americana

Shear stress, generated by water movement, can kill fish eggs and larvae by causing rotation or deformation. Through the use of an experimental apparatus, a series of shear (as dynes/cm2)-mortality equations for fixed time exposures were generated for striped bass and white perch eggs and larvae. Exposure of striped bass eggs to a shear level of 350 dynes/cm2 kills 36% of the eggs in 1 min; 69% in 2 min, and 88% in 4 min; exposure of larvae to 350 dynes/cm2 kills 9.3% in 1 min, 30.0% in 2 min, and 68.1% in 4 min. A shear level of 350 dynes/cm2 kills 38% of the white perch eggs in 1 min, 41% in 2 min, 89% in 5 min, 96% in 10 min, and 98% in 20 min. A shear level of 350 dynes/cm2 applied to white perch larvae destroys 38% of the larvae in 1 min, 52% in 2 min, and 75% in 4 min. Results are experimentally used in conjunction with the determination of shear levels in the Chesapeake and Delaware Canal and ship movement for the estimation of fish egg and larval mortalities in the field.

The food of the larvae of the northern anchovy Engraulis mordax

ENGLISH: Hitherto the only investigation dealing with the food and feeding of the larvae of the northern anchovy, Engraulis mordax Girard, was that of Arthur (1956). His main consideration was, however, with the Pacific sardine, Sardinops caerulea (Girard), and his work on the anchovy can only be considered preliminary. The present investigation is a continuation of Arthur's work on the food of the larval northern anchovy. SPANISH:El único trabajo publicado hasta ahora que trata sobre el alimento y nutrición de las larvas de la anchoa norteña, Engraulis mordax Girard, es el de Arthur (1956); pero su objeto principal fué la sardina del Pacifico, Sardinops caendea (Girard), y el estudio dedicado a la anchoa solo puede considerarse como preliminar. La presente investigación es una continuación del estudio de Arthur sobre el alimento de las larvas de la anchoa norteña.

Artificial fertilization of the eggs, and rearing and identification of the larvae of the anchoveta, Cetengraulis mysticetus

ENGLISH: The anchoveta, Cetengraulis mysticetus (Günther), is an important bait fish used to capture tunas in the Eastern Tropical Pacific Ocean. Contributions to the early life history of this species in the Gulf of Panama were made by Simpson (1959), who was able to identify deductively the planktonic egg of the anchoveta from 10 other anchovy eggs concurrently present. He also reared these planktonic eggs in the laboratory and described the resultant larvae to the age of 48 hours after hatching. Because of the lack of differences among the anchovy larvae, this description does not permit the identification of anchoveta larvae from those of other engraulid species. Furthermore, while adult specimens are easily recognized, up to the present it has not been possible to extend the identification of the juvenile anchoveta to specimens smaller than about 25 mm. The purpose of this study, therefore, was to identify anchoveta from the time of hatching to about 25 mm. SPANISH: La anchoveta, Cetengraulis mysticetus (Günther), es un importante pez de carnada que se emplea en la captura de los atunes en el Océano Pacífico Oriental Tropical. Simpson (1959) logró identificar deductivamente el huevo planctónico de la anchoveta al separarlo de otros diez huevos de anchoas que se encuentran al mismo tiempo, contribuyendo de esta manera a conocer los primeros estados de la historia natural de esta especie en el Golfo de Panamá. El también estableció un criadero en el laboratorio con estos huevos planctónicos y describió las larvas resultantes hasta la edad de 48 horas después de la eclosión. Debido a que no hay diferencias entre las larvas de las anchoas, esta descripción no permite identificar las larvas de la anchoveta de las otras especies de engráulidos. Más aun, a pesar de que los especímenes adultos son fácilmente reconocibles, hasta ahora no ha sido posible identificar la anchoveta juvenil de menos de unos 25 mm. Consecuentemente, el propósito del presente estudio ha sido el de identificar al anchoveta desde el momento de la eclosión hasta que tiene unos 25 mm.

Observations on the spawning of four species on tuna (Neothunnus macropterus, Katsuwonus pelamis, Auxis thazard and Euthynnus lineatus) in the Eastern Pacific Ocean, based on the distribution of their larvae and juveniles

ENGLISH: Knowledge of spawning habits is useful in the elucidation of the life history, ecology and population structure of tropical tunas, and is essential to the sound management of these resources. Until recently, little was known concerning the spawning of tunas, or about the distribution of their larval and juvenile stages, in the Eastern Pacific Ocean. Nichols and Murphy (1944) reported the capture off Colombia of young scombroids ultimately identified as frigate mackerel, Auxis thazard (Schaefer and Marr, 1948a). Fowler (1944) reported the capture off Manzanillo, Mexico of two young tunas, one of which is definitely and the other most likely Neothunnus macropterus (Klawe, 1959). In 1947, young of N. macropterus, K. pelamis, A. thazard and E. lineatus were caught offshore from Central America (Schaefer and Marr, 1948a, 1948b, and Schaefer, 1948). Further collections of young N. macropterus, A. thazard and E. lineatus were made in the same general area in the spring of 1949 (Mead, 1951). In January and February 1955, Clemens (1956) carried Out experiments in rearing young tunas, E. lineatus and A. thazard, in shipboard aquaria, using fish caught off Central America. Matsumoto (1958) reported captures of larval N. macropterus and K. pelamis in the area along the 120th meridian of west longitude. Klawe (1958 and 1961b) reported captures of larval N. macropterus and Auxis from the Revillagigedo Islands. Captures of young Auxis and E. lineatus in the Gulf of Panama in January 1922 during the Dana Expedition have recently been reported by Matsumoto (1959). Capture of juveniles of K. pelamis, E. lineatus and Auxis in the area off tropical Mexico and in the area of outlying islands during the SCOT Expedition has been reported by Klawe (1960a). SPANISH: El conocimiento sobre los hábitos del desove es útil para el esclarecimiento de la historia natural, ecología y estructura de las poblaciones de atunes tropicales, y es esencial para la acertada administración de estos recursos. Hasta hace poco tiempo no se sabía mucho sobre el desove de los atunes o acerca de la distribución de sus larvas y juveniles en el Océano Pacífico Oriental. Nichols y Murphy (1944) informaron sobre la captura frente a Colombia de escómbridos jóvenes últimamente identificados como melva, Auxis thazard (Schaefer y Marr, 1948a). Fowler (1944) también informó sobre la captura de dos atunes jóvenes frente a Manzanillo, México, uno de los cuales era definitivamente Neothunnus macropterus y el otro era lo más probable que también lo fuera (Klawe, 1959). En 1947 se capturaron especímenes juveniles de N. macropterus, K. pelamis, A. thazard y E. lineatus frente a la América Central (Schaefer y Marr, 1948a, 1948b, y Schaefer, 1948). Otras recolecciones de ejemplares jóvenes de N. macropterus, A. thazard y E. lineatus fueron hechas en la misma área general durante la primavera de 1949 (Mead, 1951). En enero y febrero de 1955, Clemens (1956) efectuó experimentos de crianza de atunes jóvenes, E. lineatus y A.. thazard, en acuarios a bordo para lo que empleó peces capturados frente a la América Central. Matsumoto (1958) informó sobre capturas de larvas de N. macropterus y K. pelamis en el área a lo largo del meridiano 120 de longitud oeste. Klawe (1958 y 1961b) ha dado cuenta también de capturas de larvas de N. macropterus y Auxis en las Islas Revillagigedo. Matsumoto (1959) ha informado recientemente acerca de capturas de ejemplares jóvenes de Auxis y E. lineatus en el Golfo de Panamá en enero de 1922 durante la Expedición Dana. Klawe (1960a) informó así mismo que durante la Expedición SCOT se capturaron juveniles de K. pelamis, E. lineatus y Auxis en el área frente a la zona tropical de México y en la región de las islas alejadas del continente.

Description of Early Larvae of Four Northern California Species of Rockfishes (Scorpaenidae: Sebastes) from Rearing Studies

About 72 species of Sebastes (Family Scorpaenidae) are found along the eastern Pacific coast of North America, some of which are heavily exploited by both commercial and sport fisheries. Because of the large number of species, the identification of early life stages has progressed slowly. The objectives of this study were 1) to rear the larvae of four species of rockfish (Sebastes mystinus, S. carnatus, S. atrovirens, and S. rastrelliger); and 2) to describe the larvae using morphometric measurements, pigmentation patterns, and head spination. Pigmentation was the most useful feature for identification purposes. Two general patterns were found: 1) a short row of ventral midline melanophores on the tail, and none or very little postero-dorsal pigmentation (S. mystinus); and 2) complete ventral midline pigmentation on the tail, and anterior and postero-dorsal melanophores (S. carnatus, S. atrovirens, and S. rastrelliger). With the exception of very early stages of S. carnatus and S. atrovirens, these species can be readily identified. Morphometric proportions and head spination did not show major differences among species. Because of the great similarities found among species in this genus, descriptions from field studies are uncertain to some extent. Laboratory rearings, although difficult, can at least provide early larvae from known species which allow precise identification as well as an estimation ofvariability of characters (e.g., pigmentation) within and between broods.(PDF file contains 22 pages.)

Larvae of Nearshore Fishes in Oceanic Waters near Oahu, Hawaii

Larvae of over 50 families of nearshore fishes were taken in oceanic waters about 13 km offshore of the leeward (southwest) coast of Oahu, Hawaii during 1977-78, The five most frequently taken families (Labridae, Parapercidae, Serranidae, Gobiidae, and Carangidae) made up over 50% of the total nearshore larvae. Most other families were taken very infrequently. Comparison of catch data from three types of nets indicated that 1.25-m diameter bongo nets often sampled larvae as well or better than a 3-m Isaacs-Kidd trawl and that smaller, 70-cm diameter bongo nets were often as effective as the larger nets for certain abundant taxa. Only a few taxa showed evidence of seasonal patterns in abundance. Irregular temporal variability in abundance of some taxa may have been related to occasional recent influxes of surface water from closer to shore. Most larvae taken were late preflexion stage or older. Densities of even the most abundant taxa were rarely greater than 0.001 m-3. The nearshore fish larvae were not dominated by taxa with large larvae or with larvae possessing apparent specializations to pelagic existence, Most taxa taken were pelagic spawners as adults, but larvae of demersal spawners were roughly as well represented as demersal spawners are among the nearshore fish fauna. Previous studies of waters closer to shore probably sampled insufficient volumes for any but a few exceptionally abundant taxa. Sampling with volumes filtered of the order of 104-105 m3 will be necessary to determine if the dominant taxa taken by the present study are ever more abundant closer to shore, (PDF file contains 23 pages.)