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Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of MESO is the property of Zadruzna Stampa D.D. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of MESO is the property of Zadruzna Stampa D.D. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Members of the Cobitidae family are not economically important, thus few studies have been made on their pathology. A survey of parasitic fauna of some representatives of the loaches, such as Cobitis elongata, C. elongatioides, and the recently described C. bilineata, was carried out during the spring and summer of 2006. Fish samples were collected from four Croatian rivers: the Sava, Drava and Kupa draining into the Danubian system, and the Zrmanja draining into the Adriatic system. Specimens were of varying size, weight and age. Ectoparasitological examination revealed infections by the ciliate parasites Trichodina nigra (61; 29%), Epistylis sp. (12; 9%), Chilodonella cyprinid (22; 58%), Ichthyophthyrius multifiliis (81; 81%), dinoflagellate Piscioodinium pillulare (6; 45%) and monogenean Gyrodactylus cobitis (22; 22%). Histopathological examination of haematoxylin-eosin stained tissues showed the presence of Sarcocystis sp. in the muscle of one specimen. Dissections revealed an isolated finding of plerocercoid of Ligula colymbi in a Drava river specimen. A more comprehensive survey is necessary to determine the actual threat of the parasitoses to endangered species of the Cobitidae family. [ABSTRACT FROM AUTHOR]

Nutritive composition, fatty acid profile and health-related lipid indices of natural-born European flat oyster (Ostrea edulis), variegated scallop (Chlamys varia) and smooth scallop (Flexopecten glaber) in 108 samples originating from the Adriatic sea, recovered on a monthly basis were investigated. Out of three shellfish species, the lowest share of saturated fatty acids, the most favourable ratio of polyunsaturated over saturated fatty acids, the most favourable atherogenic and thrombogenic index, and the most favourable ratio of hypocholesterolaemic over hypercholesterolaemic fatty acids were seen in oysters, sampled during springtime. No statistically significant inter-seasonal differences between basic chemical parameters and fat quality indices were established. [ABSTRACT FROM AUTHOR]

We investigated the nutritional quality of commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax), dentex (Dentex dentex), and turbot (Scophthalmus maximus). The omega-3 fatty acid content of dentex was twice as high as that of any other fish species, and its eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid contents were 2-4 times higher. The recommended n-3/n-6 ratio was present in all the fish species, but the recommended polyunsaturated/saturated fatty acid ratio was not present in the turbot. All the fish species, except turbot, met the recommended atherogenic index, thrombogenic index, and ratio of hypocholesterolaemic to hypercholesterolaemic fatty acids, whereas the highest flesh lipid quality value was observed in the dentex. [ABSTRACT FROM AUTHOR]

The European eel ( Anguilla anguilla ) is a catadromous species that inhabits the rivers of the Adriatic watershed in Croatia. It is a critically endangered fish species, according to the IUCN, due to its declining abundance in European rivers caused by overfishing and trafficking and by diseases caused by nematodes, pathogenic bacteria and viruses. An illegal parcel of glass eels was confiscated at the Zagreb Airport and was intended to re-populate Croatian rivers. Barcoding was employed to determine species affiliation, and a thorough health check was carried out. This study reports the evaluation of gross lesions, histological findings, and EVEX virus isolation and identification. Since the confiscated glass eels were of unknown origin and given the serological and genetic similarities of EVA and EVEX, we designed primers and probes for almost whole genome sequencing to elucidate the origin of glass eels based on viral phylogeny. Bayesian phylogeny showed that the isolated strain had the most common ancestor with a Danish isolate and likely evolved from the French isolate of EVEX. These findings are discussed in light of the divergence of recently isolated strains and their possible contribution to the decrease of the abundance of the European eel in European waters., Competing Interests: The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Vibrio harveyi is recognized as one of the major causes of vibriosis, a disease that threatens the long-term sustainability of aquaculture. Current research shows that the Mediterranean strains of V. harveyi are serologically heterogeneous, though research comparing the traits of different strains is scarce. This study aims to describe the biochemical, physiological and genetic characteristics of three serologically different strains of V. harveyi isolated from farmed European Sea bass (Dicentrarchus labrax) from the Adriatic Sea. A total of 32 morphological and biochemical markers were examined and, the susceptibility to 13 antimicrobials tested, and then compared the results of high-throughput sequencing and in silico analyses. This study also presents the first whole genome sequences of V. harveyi isolated from European sea bass. A large number of nonsynonymous variations were detected among sequences of the three strains. The prediction analysis of resistance genes did not correspond with the in vitro antimicrobial susceptibility tests. Six virulence genes previously unrelated to virulence of vibrios were detected in all three studied strains. The results show that differences were detected at every level of comparison among the three studied strains isolated from the same fish species originating from a small geographic area., (© 2022. The Author(s).)

Due to the insufficient capacity of Croatian hatcheries, marine aquaculture depends on the importation of fry from different countries in the Mediterranean basin. Importation enables a risk of spreading pathogenic agents. Viral nervous necrosis (VNN), caused by betanodavirus is devastating for the farming of European sea bass. We described a VNN outbreak that occurred in Croatia in 2014. After the diagnosis of VNN in sea bass fry introduced from the same hatchery to five unconnected marine farms at the Adriatic Coast, we performed surveillance within one of the affected farms. It resulted in proven horizontal spreading of the virus within the farm and to feral fish around farm cages. Real-time RT-PCR tested samples showed the dependence of the virus' proliferation to the water temperature and the fish age. The highest mortality rates were noted during higher sea temperatures. Phylogenetic analysis of partial sequences of RNA1 and RNA2 supported the hypothesis that the virus was introduced to all studied farms from the same hatchery. Moreover, phylogenetic analysis of the whole genome sequences of infected farmed sea bass and thicklip mullet showed high similarity and it is unlikely that infection in Croatian sea bass farms has originated from wild reservoirs, as the first positive record in wild mullet was recorded after the disease outbreak.

During the winter of 2013 and 2016, several Croatian fish farms experienced mortalities in the fry of European sea bass, Dicentrarchus labrax. Affected fish showed abnormal swimming behaviour and reduced appetite, and death ensued several days after the onset of clinical signs of disease. Necropsy revealed pale liver, empty digestive tract, distended gall bladder, and hyperaemia and congestion of the meninges. Routine bacteriological examination tested negative, and virological examination ruled out nodavirus infection. Histological examination revealed multifocal necrosis and extensive inflammation in the brain with abundant cellular debris in the ventricles. Inflammatory cells displayed intra-cytoplasmic basophilic vacuoles leading to suspicion of Piscirickettsia salmonis infection. Fluorescent in situ hybridization using an oligonucleotide probe targeting Domain Bacterium applied to tissue sections tested positive. The pathogen was identified by 16S rRNA gene sequencing of brain material, and the sequence showed 99% similarity with P. salmonis. This result enabled the design of an oligonucleotide probe specifically targeting P. salmonis. In 2016, P. salmonis was successfully isolated on CHAB from the brain of an affected specimen and identified using 16S rRNA gene sequencing and MALDI-TOF. This study describes the first outbreak of disease caused by P. salmonis in sea bass in Croatia, while new diagnostic tools will enable further research on its epidemiology and pathogenicity., (© 2021 John Wiley & Sons Ltd.)

Noble pen shell (Pinna nobilis) is an endemic species and the largest known bivalve in the Mediterranean Sea. By filtering large amounts of water, they maintain a high percentage of organic matter, hence playing an important role in the marine ecosystem. The ecological community of pen shells is impressive, and there are numerous microorganisms present in its soft tissues. Since this species is highly endangered due to recently described mass mortalities throughout the Mediterranean, this study was aimed at finding out more about its microbiome. In this study, we identified the predominant bacterial populations of specimens collected at three separate locations along the Eastern Adriatic coast. The predominant bacteria were isolated and 16S rRNA sequencing was performed to identify eight different bacterial genera: Aestuariibacter sp., Aliivibrio sp., Alteromonas sp., Marinobacter sp., Pseudoalteromonas sp., Rubritalea sp., Thalassospira sp. and the Vibrio splendidus clade. The identified genera are ubiquitous in the marine environment and have previously been described as both beneficial symbionts and potential pathogens in other molluscs. There was a clear difference in the predominant bacterial populations between northern and southern sampling sites, which could be linked to water temperature. These findings indicate the need for expanded sampling over a longer time period, since more exhaustive research would provide information vital to the conservation of this critically endangered species.

Common carp (Cyprinus carpio) is a very important fish species for warm-water aquaculture in Croatia. All Croatian carp farms are subjected to a surveillance programme for the presence of koi herpesvirus (KHV), causing a deadly disease called koi herpesvirus disease (KHVD). However, there is no surveillance for other viral pathogens of importance like carp edema virus (CEV), a causative agent of koi sleepy disease (KSD). During regular testing within the KHVD surveillance programme, we tested samples for CEV simultaneously. The screening indicated possible outbreaks of KHVD and KSD. During 2016, KHVD broke out in an isolated area and soon thereafter a KHV eradication programme was successfully performed. However, during 2018 and 2019, two additional mortality events occurred in lakes in the southern part of Croatia during the spring. Samples from both events tested positive for CEV. An epidemiological investigation confirmed the introduction of infected carps from an infected farm to one of the lakes. To prevent the spreading of CEV into open waters, it is of utmost importance to introduce CEV testing before fish movement or to perform regular testing of all carp farms in the country to determine CEV prevalence for the purpose of implementation of control measures., (© 2020 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

The Croatian part of the Danube River extends over 188 km and comprises 58 % of the country's overall area used for commercial freshwater fishing. To date, the heavy metal contamination of fish in the Croatian part of the Danube has not been studied. The main purpose of this study was to determine heavy metal levels in muscle tissue of sampled fish species and to analyze the measured values according to feeding habits of particular groups. Lead ranged from 0.015 μg(-1) dry weight in planktivorous to 0.039 μg(-1) dry weight in herbivorous fish, cadmium from 0.013 μg(-1) dry weight in herbivorous to 0.018 μg(-1) dry weight in piscivorous fish, mercury from 0.191 μg(-1) dry weight in omnivorous to 0.441 μg(-1) dry weight in planktivorous fish and arsenic from 0.018 μg(-1) dry weight in planktivorous to 0.039 μg(-1) dry weight in omnivorous fish. Among the analyzed metals in muscle tissue of sampled fish, only mercury exceeded the maximal level (0.5 mg kg(-1)) permitted according to the national and EU regulations determining maximum levels for certain contaminants in foodstuffs, indicating a hazard for consumers of fish from the Danube River.

During the spring of 1996 and autumn of 1997 unusual mortality outbreaks among rainbow trout fry and yearlings occurred at two different trout farms, resulting in mortality of 20 and 10 per cent, respectively. Generally, the affected fish, swimming at the water surface, were reluctant to eat and were dark pigmented with visible haemorrhages around and within the oral cavity. Bacterial isolates from moribund fish from both cases were identified as Yersinia ruckeri by standard biochemical tests and API 20E. The isolated strains were found to be sensitive to tetracycline, chloramphenicol, co-trimoxazole, nalidixic acid, flumequine, enrofloxacin, carbenicillin and gentamicin. Microplate agglutination assay confirmed that both isolates belonged to serotype O1. The pathogenicity of the isolated bacteria was confirmed by challenge experiment. Titres of specific antibodies were determined in the sera of survivors. The titre was highest on the 21st day postchallenge and was detectable until the 81st day.

The aim of this study was to assess the suitability of the perirenal fatty tissue for the determination of an organochlorine pesticide. Fatty tissue samples were prepared by the matrix solid phase dispersion (MSPD) method, and pesticide levels were determined by gas chromatography on capillary column using an electron capture detector. Results were confirmed by gas chromatography/mass spectrometry (GC/MS) system. The results showed that the perirenal fatty tissue contained significantly higher levels of hexachlorobenzene (HCB) than the dorsal fatty tissue (P < 0.01). All the levels were below the criteria for maximum residue limits established by Croatia and the EU.

Marteiliosis is a disease of molluscs caused by Marteilia refringens in Europe and M. sydneyi in Australia. During routine examination of cultured mussels Mytilus galloprovinciallis in the northern Adriatic, the occurrence of Marteilia sp. was recorded with a prevalence of 5%. This parasite was not detected in flat oysters reared in the same area. The affiliation of the detected parasite in M. galloprovinciallis was confirmed by in situ hybridization using a M. refringens probe, specific at the genus level. DNA of these infected mussels originating from the same area will be used to clarify the taxonomic position of this species within the genus Marteilia using a molecular approach.