Jolanta Kempter

Susceptibility of koi × crucian carp and koi × goldfish hybrids to koi herpesvirus (KHV) and the development of KHV disease (KHVD)

Hybrids of koi, Cyprinus carpio x crucian carp, Carassius carassius and koi x goldfish, Carassius auratus, proved to be susceptible to koi herpesvirus (KHV, syn. CyHV-3) and developed KHV disease (KHVD). While hybrids of koi x goldfish were partly resistant to mortality following infection by immersion, most koi x crucian carp hybrids died after bath infection. KHV DNA was detected in dead fish but also in all surviving animals by different polymerase chain reactions (PCRs). According to these results, hybrid crossbreeding does not seem to prevent severe losses associated with KHV in terms of inducing KHVD. The present study showed severe losses after a waterborne KHV infection of between 35% and 100% in koi x goldfish and koi x crucian carp hybrids as well as in SPF carp.

Koi herpes virus: do acipenserid restitution programs pose a threat to carp farms in the disease-free zones?

In 2003 and subsequent years, a substantial number of Polish fish farms experienced mass mortality of carp (Cyprinus carpio)—their major culture species. Clinical signs have led to the assumption that these heavy losses may have been caused by infection with the koi herpes virus (KHV), which was first isolated and diagnosed in carps from the Rybacka Stacja Doświadczalna (the Fisheries Research Station) of the Agricultural University in Szczecin (Sadowski and Kempter 2004, Bergmann et al. 2006). In the wake of this finding, many research centres in Poland have initiated projects focusing on KHV, in its different aspects, and attempted to develop diagnostic tests for carp cultures (Kempter et al. 2008a). According to the State Veterinary Research Institute in Pulawy (Panstwowy Instytut Weterynaryjny – Panstwowy Instytut Badawczy), this problem is not only related to the carp market and the stocking material turnover. The research projects carried out at the Pulawy institute focuses mainly on the other species, suspected of being a vector for KHV. This assumption is based on the fact that the clinical signs have hitherto not been observed in any other species except Cyprinus carpio (common carp and koi). Many research facilities, such as the the ACTA ICHTHYOLOGICA ET PISCATORIA (2009) 39 (2): 119–126 DOI: 10.3750/AIP2009.39.2.06

Genetic identifiability of selected populations of Indian mackerel, Rastrelliger kanagurta (Actinopterygii: Perciformes: Scombridae)—Celfish Project—Part 1

Genetic identification of food ingredients is an important method that enables the traceability and confirmation of authenticity of fish products. With regard to common cases where valuable fish species are replaced by lowvalue equivalents it is necessary to provide reliable methods which will precisely identify fish species that are valuable for economy. Such methods help to eliminate counterfeit products from the market but also provide tools that might be used by governmental units which control food safety, including custom services. Population identification by molecular methods also provides inforACTA ICHTHYOLOGICA ET PISCATORIA (2014) 44 (2): 145–152 DOI: 10.3750/AIP2014.44.2.08

Genetic variability in European populations of Coregonus lavaretus (L.): an assessment based on mitochondrial ND-1 gene haplotypes

Genetic variability in European populations of Coregonus lavaretus (L.): an assessment based on mitochondrial ND-1 gene haplotypes The genetic variability of whitefish, Coregonus lavaretus (L.), was studied based on 114 individuals from nine populations inhabiting Polish lakes, including the Szczecin Lagoon, and from one population each from lakes in Austria and Switzerland. Differences within and among populations were assessed with mitochondrial ND-1 gene sequences that were PCR amplified and digested with ten restriction enzymes. The ten composite haplotypes obtained were sequenced and analyzed with MEGA4 software. The highest intrapopulation variability was noted in the whitefish populations inhabiting lakes Ińsko, Miedwie, Marianowo, Wisola, Śremskie, Morzycko, the Szczecin Lagoon, and Lake Lucerne, which possessed from two to five composite haplotypes. In contrast, populations inhabiting lakes Woświn, Czarne, and Traunsee were fixed for the most common haplotype H2. Zróżnicowanie genetyczne europejskiej populacji Coregonus lavaretus (L.): ocena na podstawie mitochondrialnego genu ND-1 Zróżnicowanie genetyczne 9 populacji siei pospolitej (Coregonus lavaretus, L.) określono w grupie 114 osobników pozyskanych ze zbiorników wodnych Polski, Szwajcarii oraz Austrii. Analizę przeprowadzono w oparciu o technikę PCR-RFLP zastosowaną względem mitochondrialnego genu ND-1, którego sekwencję poddano trawieniu 10 enzymom restrykcyjnym. Na tej podstawie otrzymano dziesięć haplotypów, których sekwencje reprezentatywne zostały poddane sekwencjonowaniu oraz analizie z wykorzystaniem programu MEGA4. Najwyższe zróżnicowanie międzypopulacyjne określono dla populacji siei zamieszkujących jeziora Ińsko, Miedwie, Marianowo, Wisola, Śremskie, Morzycko, Zalew Szczeciński oraz szwajcarskie jezioro Lucerne. Zawierały się ona w obrębie haplotypów od 2 do 5. Dla porównania osobniki z jezior Woświn, Czarne (DPN) oraz Traunsee (Austria) były zaklasyfikowane do pojedynczego haplotypu H2.

Genetic Variability of Koi Herpesvirus In vitro—A Natural Event?

Worldwide koi herpesvirus (KHV) causes high mortalities in Cyprinus carpio L. aquaculture. So far, it is unknown how the different variants of KHV have developed and how they spread in the fish, but also in the environmental water bodies. Therefore, a phylogenetic method based on variable number of tandem repeats (VNTR) was improved to gain deeper insights into the phylogeny of KHV and its possible worldwide distribution. Moreover, a VNTR-3 qPCR was designed which allows fast virus typing. This study presents a useful method for molecular tracing of diverse KHV types, variants, and lineages.

Computational modelling of cormorant swarm

Abstract The great cormorant ( Phalacrocorax carbo ) is a species with a strong impact on the environment in the areas inhabited by breeding colonies or migrating colonies. Depending on the size of colonies, destruction of tree cover, soil sterilization due to the supply of enormous amounts of aggressive faeces and overpreying on fish from nearby water bodies can be observed. In this study, an iterative algorithm for modelling the annual life cycle of a swarm of cormorants was described. The procedure was based on a mathematical model of a herd of birds, taking into account both fixed biological factors, such as food demand, availability of habitats and the reproductive cycle, and random factors occurring in the environment, e.g., flooding, storms or human activity. Additionally, the algorithm included the variable time of inhabitance depending on the duration of the ice cover over water bodies, and the possibility of conducting a killing programme. The proposed procedure was tested and verified positively using data obtained from the literature.

Is There Any Species Specificity in Infections with Aquatic Animal Herpesviruses?-The Koi Herpesvirus (KHV): An Alloherpesvirus Model

Most diseases induced by herpesviruses are host-specific; however, exceptions exist within the family Alloherpesviridae. Most members of the Alloherpesviridae are detected in at least two different species, with and without clinical signs of a disease. In the current study the Koi herpesvirus (KHV) was used as a model member of the Alloherpesviridae and rainbow trout as a model salmonid host, which were infected with KHV by immersion. KHV was detected using direct methods (qPCR and semi-nested PCR) and indirect (enzyme-linked immunosorbant assay; ELISA, serum neutralization test; SNT). The non-koi herpesvirus disease (KHVD)-susceptible salmonid fish were demonstrated to transfer KHV to naive carp at two different temperatures including a temperature most suitable for the salmonid (15°C) and cyprinid (20°C). At 20°C KHVD was induced in carp cohabitated with infected trout. KHV was also detected virologically and serologically at the end of the experiment in both rainbow trout and carp.

Development of the method for identification of selected populations of torpedo scad, Megalaspis cordyla (Linnaeus, 1758), using microsatellite DNA analyses. CELFISH project – Part 4

Catch and consumption of torpedo scad (Megalaspis cordyla) over the western Indian Ocean, but also the western Pacific from Japan to Australia is constantly increasing. Taking into account the degree of exploitation and missing information on the population structure of torpedo scad stocks it is crucial to provide population data. The analysis included individuals obtained in 2012 and 2013 from local markets in Madagascar, Tanzania, Vietnam and Cambodia and after successful DNA extraction fragment of the nuclear rhodopsin gene (RH1) and 9 microsatellite regions (SSRs) were amplified and analysed. Based on the obtained results it was found that there was no 100% overlap between the compared RH1 sequences and those from GenBank. In the case of the studied SSRs, the results allowed the initial characterisation and assessment of the genetic diversity of populations. Moreover, population assignment test distinguished the studied populations into two geographically distant subpopulations.

Microsatellite DNA-based genetic traceability of two populations of splendid alfonsino, Beryx splendens (Actinopterygii: Beryciformes: Berycidae)—Project CELFISH—Part 2

Background. The study is a contribution to Project CELFISH which involves genetic identifi cation of populations of fi sh species presenting a particular economic importance or having a potential to be used in the so-called commercial substitutions. The EU fi sh trade has been showing a distinct trend of more and more fi sh species previously unknown to consumers being placed on the market. Molecular assays have become the only way with which to verify the reliability of exporters. This paper is aimed at pinpointing genetic markers with which to label and differentiate between two populations of splendid alfonsino, Beryx splendens Lowe, 1834, a species highly attractive to consumers in Asia and Oceania due to the meat taste and low fat content. Material and methods. DNA was isolated from fragments of fi ns collected at local markets in Japan (MJ) (n = 10) and New Zealand (MNZ) (n = 18). The rhodopsin gene (RH1) fragment and 16 microsatellite DNA fragments (SSR) were analysed in all the individuals. The sequences obtained were processed using the BioEdit and BLAST software, whereas SSR data were processed with the GeAlEX analysis package. Results. The BioEdit software-aided comparison of MJ and MNZ nucleotide sequences of the rhodopsin gene fragments were identical and showed 100% agreement with the alfonsino sequence deposited under access number DQ197832. The preliminary analysis of SSR markers showed all the loci analysed in both populations to be polymorphic, and when randomly selected specimens were assigned to the original populations. The affi nity test correctly identifi ed the provenance of all those specimens. Conclusion. The results obtained constitute a tool for molecular differentiation between alfonsino populations collected in the FAO 81 (New Zealand) and FAO 71 (Japan) areas for the purpose of catch quota control and for checking the agreement between the label declaration and the actual product.