Pablo Conejeros

Detection and quantification of Chilean strains of infectious pancreatic necrosis virus by real-time RT-PCR assays using segment B as a target

Infectious pancreatic necrosis virus (IPNV) is the causal agent of a highly prevalent disease that affects salmonid fish, mostly during their fresh water life period. Like many other viruses, IPNV produces highly heterogeneous populations. Therefore, diagnostic methods need to be checked constantly so that no variants of the virus escape detection. The IPNV genome is composed of two double-stranded RNA segments: A and B, polymerase chain reaction (PCR) methods normally use segment A as a target. In order to develop an optimized protocol to diagnose IPNV, we present a real-time RT-PCR (reverse transcription) technique, using primers designed to recognize segment B of the virus. To validate the ubiquity of the primers used, the IPNV isolates tested were sequenced and compared with previously published cladograms, which include a wide spectrum of genogroups. These primers made it possible to detect viral isolates belonging to genogroups 1 and 5, which were obtained from different locations linked to fish farming. As expected, we were able to detect the virus from distant Aquabirnavirus genogroups.

Differentiation of Sympatric Arctic Char Morphotypes Using Major Histocompatibility Class II Genes

AbstractArctic Char Salvelinus alpinus have colonized northern postglacial lakes within the last few thousand years. Divergent populations have adapted to thrive in the prevailing oligotrophic environments and thus have developed morphotypes with different ecological behaviors. The morphotypes usually differ in size, morphology, coloration, feeding ecology, and/or habitat occupancy. Although morphotypes that have very divergent spawning seasons should become genetically segregated, genetic differentiation, in most cases, has been weak. Thus, results to date have suggested that Arctic Char morphotype separation has been driven largely by the environmentally mediated phenotypic plasticity of the species, with differentiation between morphotypes having commenced too recently to generate substantial genetic drift. Here we used the major histocompatibility (MH) class II genes in an attempt to isolate sympatric Arctic Char morphotypes known to be ecologically differentiated. These morphotypes are from postglaci...

Global major histocompatibility Class II β (mh-IIβ)-polymorphism in Arctic charr Salvelinus alpinus

This study explored the use of the gene encoding the β subunit of the major histocompatibility (MH) receptor as a population marker in Arctic charr Salvelinus alpinus. The use of this polymorphic marker allowed differentiation of the S. alpinus lineages previously defined using mitochondrial DNA (mtDNA) but also allowed differentiation between the populations studied within those lineages. The majority of the variation observed here occurred prior to the last glaciation event. Nevertheless, all S. alpinus populations were differentiated using both MH Class II β (mh-IIβ) sequences and allelic frequencies. The fact that all the populations studied presented high rates of non-synonymous: synonymous substitutions and high levels of interpopulation variation, suggested mh-IIβ as an ideal marker to assess differentiation among S. alpinus populations in ways that may represent divergence both by genetic drift and natural adaptation to the local environment.

Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line

EJB Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.2 No.2, Issue of August 15, 1999.© 1999 by Universidad Catolica de Valparaiso -- Chile Received March 3 , 1999 / Accepted April 16 , 1999

Coinfection takes its toll: Sea lice override the protective effects of vaccination against a bacterial pathogen in Atlantic salmon

Vaccination is considered crucial for disease prevention and fish health in the global salmon farming industry. Nevertheless, some aspects, such as the efficacy of vaccines, can be largely circumvented during natural coinfections. Sea lice are ectoparasitic copepods that can occur with a high prevalence in the field, are frequently found in co-infection with other pathogens, and are highly detrimental to fish health. The aim of this case-control study was to evaluate the interaction between the detrimental effects of coinfection and the protective effects of vaccination in fish. We used the interaction between the sea louse Caligus rogercresseyi, the bacterial pathogen Piscirickettsia salmonis, and their host, the Atlantic salmon Salmo salar, as a study model. Our results showed that coinfection decreased the accumulated survival (AS) and specific growth rate (SGR) of vaccinated fish (AS = 5.2 ± 0.6%; SGR = −0.05 ± 0.39%) compared to a single infection of P. salmonis (AS = 42.7 ± 1.3%; SGR = 0.21 ± 0.22%). Concomitantly, the bacterial load and clinical signs of disease were significantly increased in coinfected fish. Coinfection may explain the reduced efficacy of vaccines in sea cages and highlights the need to test fish vaccines in more diverse conditions rather than with a single infection.

High immune diversity in farmed Atlantic salmon (Salmo salar L.)

As with any other intensively farmed animal species, the Atlantic salmon has been selectively bred and cross-bred to maximize desirable traits. Selection tends to diminish genetic diversity in target populations, which among other negative effects is hypothesized to decrease their capacity to confront a broad variety of pathogens. We have analyzed mitochondrial (mtDNA) as well as major histocompatibility complex (MHC) DNA sequences from individuals collected from a single aquaculture cage in southern Chile. Interestingly, only two mtDNA haplotypes were obtained; however, several different MH alleles were detected, with divergence values that were compared with those of natural populations of salmonids. Thus, it seems evolutionary processes responsible of keeping MH diversity in the wild managed to retain MH variability in farmed Atlantic salmon, maintaining high immune diversity despite the generally lower levels of observed neutral genetic diversity.