Detection of ‘long-haired’ Saprolegnia (S. parasitica) isolates using monoclonal antibodies

P. 726-733The ability of five monoclonal antibodies (Mabs) raised against a pathogenic Saprolegnia parasitica isolate from brown trout to detect and differentiate between isolates with bundles of long hairs (S. parasitica) and other Saprolegnia species was determined by means of an indirect immunofluorescence assay. Four of the Mabs used recognized some of the long-haired S. parasitica isolates but also cross-reacted with other Saprolegnia species without bundles of hairs and with Achlya sp. The other Mab (named 18A6) was able to differentiate between the asexual and most of the sexual isolates in the group of long-haired S. parasitica isolates, but did not recognize Achlya sp. or the Saprolegnia species without bundles of hairs, with the exception of S. hypogyna. These results indicate that isolates with bundles of long hairs are closely related with other members of genus Saprolegnia and share several antigens. However, Mab 18A6 seems to recognize an epitope that is expressed mainly in the asexual isolates in the long-haired S. parasitica isolates.S

Clinical evaluation and antibody responses in sheep after primary and secondary experimental challenges with the mange mite Sarcoptes scabiei var. ovis

P. 109-116In this work the clinical evolution and the specific serum IgG and IgE antibody responses in sheep after primary (n = 10) and secondary (n = 4) experimental challenges with the mange mite Sarcoptes scabiei var. ovis were studied. The primary infection was characterized by the development of mange lesions in all sheep, a detection of live S. scabiei mites in 70% skin scrapings taken in week 10 post-challenge (PC), strongly raised and sustained specific IgG levels and a more moderate but continuous rise in specific IgE levels. Seroconversion was detected for IgG and IgE by ELISA in 90% and 60% of the sheep in week 8 PC, respectively. By Western-blotting (WB), ten IgG-reactive bands (36–120 kDa) and four IgE-reactive bands (90–180 kDa) were observed in week 8 PC. Following the secondary challenge the ewes developed a smaller area of mange lesion than that seen following primary challenge and live S. scabiei mites were not detected in skin scrapings collected in week 8 PC, suggesting that sheep had developed immunity to re-infection. Compared to primary infection, the specific IgG secondary antibody levels were transient, but in contrast there was an anamnestic IgE response, resulting in an elicitation of specific serum IgE levels in week 2 PC significantly higher than those demonstrated after primary infection. WB analysis revealed one additional IgG-reactive band (180 kDa) and no additional IgE-reactive bands. Determining the immunodiagnostic or vaccination value of the IgG-reactive antigens and IgE-reactive allergens detected requires further studies.S

Leer en comunidad: creación y desarrollo de clubes de lectura de literatura escrita por mujeres dentro y fuera de la universidad

Depto. de Lengua Española y Teoría de la LiteraturaFac. de FilologíaFALSEsubmitte

Bóveda, La

Premi 198

Prevalence of serum antibodies against Saprolegnia parasitica in wild and farmed brown trout Salmo trutta

p. 17-22The prevalence of serum antibodies against Saprolegnia parasitica in wild and farmed brown trout Salmo trutta from the province of León (NW Spain) was studied by enzyme-linked immunosorbent assay (ELISA). Blood samples from healthy and Saprolegnia-infected brown trout were collected over 2 yr with a seasonal periodicity (January, April, July and October) from a hatchery and river with frequent presence of saprolegniosis (River Porma) and from a river in which the disease was rarely observed (River Omaña). The individual prevalence was 30.1%, but statistically significant differences were observed between the prevalence in trout from the hatchery (43.0%), from River Porma (31.8%) and from River Omaña (6.4%) and also between the prevalence observed in October (42.9%) and the values obtained in January (24.8%), April (22.7%) and July (27.5%). There was no difference between the seroprevalence in females (34.8%) and males (38.2%), but a positive correlation between raised serum antibody levels and larger (older) fish was found. The low prevalence of antibodies observed in Saprolegnia-infected trout (18.0%) suggests possible immune suppression and the lack of an effective specific immune response in fish with saprolegniosis.S

Adhesion to brown trout skin mucus, antagonism against cyst adhesion and pathogenicity to rainbow trout of some inhibitory bacteria against Saprolegnia parasitica

P. 35-44Biological control of saprolegniosis with bacteria might be an alternative to the use of chemical compounds. Among criteria for the selection of such bacteria are their absence of pathogenicity to fish and their ability to prevent adhesion of the pathogen to the skin mucus. The pathogenicity to rainbow trout of 21 bacterial isolates with in vitro inhibitory activity against Saprolegnia parasitica was studied. Fifteen of the isolates, identified as Aeromonas sobria, Pantoea agglomerans, Pseudomonas fluorescens, Serratia fonticola, Xanthomonas retroflexus and Yersinia kristensenii, were non-pathogenic when injected into rainbow trout. Their capacity to adhere to the skin mucus of male and female brown trout and to reduce the adhesion of S. parasitica cysts under exclusion, competition and displacement conditions was tested. The 15 bacterial isolates showed a low adhesion rate, ranging between 1.7% (for an A. sobria isolate) and 15.3% (a P. fluorescens isolate). This adhesion was greater in the case of mucus from male brown trout than from females. Similarities in the adhesion to male mucus and other substrates and correlation to that observed to polystyrene suggest that adhesion to skin mucus does not depend on the substrate. A high percentage (88.9%) of the S. parasitica cysts adhered to the skin mucus of male brown trout. Almost all of the bacteria reduced this adhesion ratio significantly under exclusion and competition conditions. However, only half of the isolates displaced cysts from skin mucus, and more bacterial cells were necessary for this effect. A novel method to study the adhesion of S. parasitica cysts to skin mucus of trout and their interactions with inhibitory bacteria is described.S

Antibody response of brown trout Salmo trutta injected with pathogenic Saprolegnia parasitica antigenic extracts

P. 107-111Brown trout Salmo trutta injected with antigenic extracts from a pathogenic isolate of Saprolegnia parasitica developed specific antibodies that were detected by enzyme-linked immunosorbent assay (ELISA), immunofluorescence (IF) and Western blotting (WB), but not by immunodiffusion (ID). Three groups of five 2 yr old brown trout were injected intraperitoneally with 3 different antigenic extracts: small hyphal fragments (HF) and soluble extracts from sonicated mycelia grown in medium with or without β-sytosterol (SEB and SE, respectively). In the 2 groups injected with SE and SEB, antibodies were found in 66.7% of the serum samples by ELISA, 54.5% by IF and 48.5% by WB. In the group injected with HF, only 1 trout survived the experiment, and in this fish only 1 sample was positive by ELISA. The results obtained by ELISA and IF were similar and show that there is cross-reaction between the antigens used. By WB, the proteins most frequently recognised were 2 proteins of 25 and 29 kDa. No significant differences were found in the groups injected with SE or SEB.S

Bacterial skin flora variation and in vitro inhibitory activity against Saprolegnia parasitica in brown and rainbow trout

P. 126-135Variations in the number and diversity of bacteria from the skin of brown trout Salmo trutta L. and rainbow trout Oncorhynchus mykiss Walbaum were surveyed from different rivers and fish farms in northern Spain. In addition to determining bacterial populations in skin samples of healthy fish, bacterial populations were determined from skin lesions (of brown trout only) infected with Saprolegnia parasitica, the causal agent of saprolegniosis. Mean bacterial counts from skin lesions of brown trout suffering from saprolegniosis were nearly 1000 times greater than from the skin of uninfected brown and rainbow trout. More than 20 different genera of bacteria were identified, with isolates of Aeromonas and Iodobacter being the predominant genera associated with saprolegniosis lesions. The in vitro inhibitory activity of 72 of these skin isolates was tested against S. parasitica using 3 different assays. These included (1) assessing the inhibition by bacteria of colony growth on agar media, (2) the inhibition of colony growth from colonized hemp seeds in liquid media and (3) the inhibition of cyst germination in liquid media. Finally, the fungicidal effect of the 24 most inhibitory bacterial species, and the inhibitory activity of their culture supernatants, was tested in the same way. Isolates identified as Aeromonas piscicola, A. sobria, Pantoea agglomerans and Pseudomonas fluorescens achieved the highest inhibition against S. parasitica. Many of these inhibitory isolates were obtained primarily from skin lesions of fish with saprolegniosis. It is suggested that some of these isolates might be useful in the biological control of saprolegniosis.S

Dietary protein level during pregnancy. Its influence on maternal-fetal exchange of substrates

[EN]: It is well known that the dietary protein level influences both the pregnancy process and development of the offspring. Therefore, a study was carried out to determine the effect that different protein percentages: 10°/o, 4°/o and 20°/o in diets administered to rats during pregnancy, had on food efficiency and on the substrates partition between rat dams and their neonates. Experimentation was thus carried out over a period of 21 days (pregnancy) and comparisons were made with well-nourished rats receiving 10°/o protein (controls) and between one and other group. Moreover, the effect on pregnancy was observed by comparing pregnant rats with non pregnant rats within each dietary situation. Food intake, weight parameters and food efficiency ratio were recorded in adult rats. Weight parameters were also evaluated in newborns. As results revealed, the highest the protein level in the diet, the highest the food efficiency ratio, both in the pregnant group and in the non pregnant group. Weight changes were determined in rat dams and their neonates, as a consequence of the different protein intakes. These also involved alterations in the substrates partition between the mothers and their offspring. Such findings might lead to the retention o f substrates by the maternal tissues and, hence, to impairment o f neonatal development.[ES]: En vista de la influencia que la concentración proteínica de la dieta materna tiene en el desarrollo del proceso gestacional y sus consecuencias sobre el crecimiento neonatal, se estudiaron, en ratas, las variaciones que el aporte de distintos niveles de proteína dietaria, 10°/o, 4°/o y 2°/o durante la gestación, ejerce en la eficiencia de utilización del alimento y en el reparto matemo-fetal de sustratos. Con este propósito, se hicieron comparaciones entre las tres situaciones dietarias, y el efecto de la gestación se observó comparando las ratas gestantes con las no gestantes, con cada una de las dietas sometidas a estudio. Se determinó la ingesta, parámetros ponderales y eficiencia de conversión alimentaria (ECA) en ratas adultas, así como los parámetros ponderales de los neonatos. Según se pudo apreciar, tanto en los animales gestantes como en los no gestantes la ECA varió en función directa a la cantidad de proteína ingerida, mientras que durante la gestación se elevó en todas las situaciones dietarias. Por otra parte, la variabilidad ponderal de madres y neonatos, consecutiva a los cambios de proteína en la dieta, afectó también el reparto matemo-fetal de sustratos. Este último fue modificado en los animales sometidos a las dietas que contenían 4°/o y 2°/o de proteína, dando lugar a una retención de sustratos en los tejidos maternos, en perjuicio del desarrollo neonatal.Peer reviewe