Domenico Caruso

Investigation of Cyprinid herpesvirus-3 genetic diversity by a multi-locus variable number of tandem repeats analysis

Cyprinid herpesvirus-3 (CyHV-3), or koi herpesvirus (KHV), is responsible for high mortalities in aquaculture of both common carp (Cyprinus carpio carpio) and koi carp (Cyprinus carpio koi) worldwide. The complete genomes of three CyHV-3 isolates showed more than 99% of DNA sequence identity, with the majority of differences located in short tandem repeats, also called VNTR (variable number of tandem repeats). By targeting these variations, eight loci were selected for genotyping CyHV-3 by multiple locus VNTR analysis (MLVA). CyHV-3 strains obtained after sequential in vivo infections exhibited identical MLVA profiles, whereas samples originating from a single isolate passaged 6 and 82 times in vitro exhibited mutations in two of the eight loci, suggesting a relatively slow genetic evolution rate of the VNTRs. The method was subsequently applied on 38 samples collected in Indonesia, France and the Netherlands. Globally, the isolates grouped in two main genetic clusters, each one divided in two subgroups including either CyHV-3-U/I or CyHV3-J. Interestingly, Indonesian strains were rather distant from CyHV-3-J isolate. The results of the present study indicate that these VNTR molecular markers are efficient in estimating the genetic diversity among CyHV-3 isolates and are therefore suitable for further molecular epidemiological studies.

Spatio-temporal analysis of cyprinid herpesvirus 3 genetic diversity at a local scale

Cyprinid herpesvirus 3 (CyHV-3), the causative agent of koi herpesvirus disease, is a major threat for carp populations in many countries worldwide, including Indonesia. It has been shown that many genotypes circulate worldwide, all highly related to one of the two known lineages U/I and J. In this study, we evaluated the spatial and temporal distribution of CyHV-3 strains in a small enzootic area, the lake of Cirata (West Java, Indonesia). Of the 365 samples analysed, from clinical or asymptomatic fish, 244 were found positive for CyHV-3, suggesting a high occurrence of the virus. Genotyping of these viral specimens with a range of molecular markers revealed the presence of numerous haplotypes in the host population, all related to the J lineage. In single individuals, mixed-genotype infections occurred at high frequency. The present results demonstrate that polymorphic molecular markers are suitable to monitor the genetic evolution of a viral population in an enzootic area.

Rapid Detection and Identification of Nontuberculous Mycobacterial Pathogens in Fish by Using High-Resolution Melting Analysis

ABSTRACT Mycobacterial infections in fish are commonly referred to as piscine mycobacteriosis, irrespectively of the specific identity of the causal organism. They usually cause a chronic disease and sometimes may result in high mortalities and severe economic losses. Nearly 20 species of Mycobacterium have been reported to infect fish. Among them, Mycobacterium marinum, M. fortuitum, and M. chelonae are generally considered the major agents responsible for fish mycobacteriosis. As no quick and inexpensive diagnostic test exists, we tested the potential of high-resolution melting analysis (HRMA) to rapidly identify and differentiate several Mycobacterium species involved in fish infections. By analyzing both the melting temperature and melting profile of the 16S-23S rRNA internal transcribed spacer (ITS), we were able to discriminate 12 different species simultaneously. Sensitivity tests conducted on purified M. marinum and M. fortuitum DNA revealed a limit of detection of 10 genome equivalents per reaction. The primers used in this procedure did not lead to any amplification signal with 16 control non-Mycobacterium species, thereby demonstrating their specificity for the genus Mycobacterium.

Evaluation of Aquaculture System Sustainability: A Methodology and Comparative Approaches

Over the last 30 years, aquaculture has experienced an unprecedented development in global animal production with an average yearly growth rate of over 10% between 1980 and 2000 (FAO, 2009). During the same period, capture fisheries saw their progression gradually grind to a standstill and growth stopped from 1995 (total catch fluctuating between 90 and 95 Mt/year according to the year). The growth of aquaculture, despite its benefits and the fact that it is the only way to meet the increase in demand for sea products, evaluated at 270Mt in 2050 (Chevassus au Louis et Lazard, 2009; Wijkstrom, 2003), raises a certain number of issues directly related to its sustainable development. Amongst these are issues related to feed for the farmed organisms, to their biological diversity, to the farms’ economic sustainability, to the impact of aquaculture development on social equity and to the set of arrangements constituting the sector’s governance. Feed, for example, is currently the subject of significant controversy as shown by the emblematic article of Naylor et al. (2000) that exposes the impact on catches of the massive use of fish meal and fish oil in fish and prawn aquaculture and advocates the return to sparser aquaculture systems, directly inspired by traditional Asian systems which use more extensive techniques based on polyculture and fertilisation and where artificial feed is only seen as a potential supplement. This diagnosis, although interesting as it generated much debate, was, however, incomplete and, in fact, inaccurate: by focusing on a single criterion and a single dimension (environmental) of sustainability, the authors were led to make proposals that had no chance of being adopted by the actors. De facto, farming systems have continued to intensify and this has led to a sustained increase in the use of

La pathologie en pisciculture tropicale et l’approche écopathologique : méthodologie et étude de cas sur les élevages de Pangasiidae

The profound changes in breeding and rearing systems along with the intensification of tropical aquaculture production, especially in South-East Asia, tend to increase the occurrence of epizooties and enzootic diseases, of which the socio-economic and environmental impacts amount to hundreds of millions of US

Plasma lysozyme levels in sheatfish Silurus glanis (L.) subjected to stress and experimental infection with Edwardsiella tarda

per year. Because of the variety of cultured species and culture environments, diseases are numerous and their natures are generally multi-factorial, thus complicating health management. Until now, Asian fish-culturists have explored exclusively or essentially the chemotherapeutic register, which is known to result in the accumulation of residues in the environment and/or fish flesh, and to promote antibiotic resistance, including through the transfer of resistance between bacterial species. Alternatives to this chemical strategy are urgently needed to secure the health management of cultured fish, in particular in the field of epidemiology and ecopathology. Ecopathology is a branch of analytical epidemiology that explores and analyzes the different variables of an animal production system in order to individualize the risk factors responsible for creating the setting of a pathogen or disease breakthrough. This holistic approach, which regards pathology as one of the outcomes of a productive system, is very rarely used in aquaculture. This article provides a synthesis on the health issues of fish breeding systems in the tropics, with emphasis on South East Asia, and illustrates the benefits of an ecopathological strategy that was deployed to elucidate then alleviate the mortality of Pangasius spp. reared in floating cages in the Mekong River, Vietnam.