Fish photobacteriosis—The importance of rapid and accurate identification of Photobacterium damselae subsp. piscicida

MALDI‐TOF MS was tested for the identification of Photobacterium damselae subsp. piscicida on isolates grown on two media, cultured at three incubation times and applied on the target plate by the direct sample spotting (DS), by the on‐target extraction (OTE) and by the full extraction (FE) method, in triplicates. The identification of samples grown on blood agar (BA) outperformed identification on tryptic soya agar (TSA) by 0.64% for DS and OTE. The OTE gave the highest scores in both culture media, all incubation times and replicates. Reliable 24‐hr species identification was 61.54%, 84.61% and 53.85% for samples grown on TSA and identified by DS, OTE and FE, respectively. For isolates grown on BA, they were 76.92%, 96.15% and 30.77%, respectively. When identified by OTE, the 48‐hr identification was 93.58%, but for 72 hr declined to 71.79%. The reliable identification with the highest score from the first measurement was 100% only for OTE from BA (24 hr), whereas OTE from TSA gave 84.61% (24 hr), 76.92% (48 hr) and 84.61% (72 hr). The reliable MALDI‐TOF MS identification of Ph. damselae subsp. piscicida is incubation time, media, target plate preparation and replicate‐dependent.     

The effects of diet supplemented with Lactobacillus rhamnosus on tissue parameters of rainbow trout,Oncorhynchus mykiss (Walbaum)

This study was carried out to establish the effects of a 6 week treatment with the diet supplemented with L. rhamnosus in concentrations of 10⁷ CFU g^?1 (G1 group) and 10⁸ CFU g^?1 (G2 group) on the condition expressed by condition factors (Fulton's, Clark's and B), intestinal microbiology, haematological, histological and selected antioxidative parameters of rainbow trout. A significantly higher condition factors were found in G1 group indicating that higher concentration of probiotic (10⁸ CFU g^?1) did not result in the better condition. Cholesterol and urea levels were significantly higher in both G1 and G2 groups, albumin in G1 and creatinine in G2 group with respect to control. A significantly higher liver TBARS level was observed in G2 group. The feeding with supplemented probiont apparently changed the resident microbiota. Three weeks after withdrawal of the supplemented feed, the microflora mostly reverted to the control composition, although L. rhamnosus in faecal matter of fish remained inherent. The epithelial structure of the proximal and distal intestine revealed the increased absorptive area in both treated groups, as well as the increase in the mucin‐secreting goblet cells. The L. rhamnosus‐treated groups demonstrated the capacity for the augmentation of the innate host defence.     

Fatty acid and proximate composition of bluefin tuna (Thunnus thynnus) muscle with regard to plasma lipids

The composition of tail muscle fatty acids from wild and cultured bluefin tuna reared on a diet based on herring and sardine, along with the plasma lipid profile of the farmed individuals, was determined. The total lipid content of farmed bluefin in this study was 0.922 g/100 g or 3.49 g of saturated fatty acids (SAFA), 4.48 g of monounsaturated fatty acids (MUFA), 2.58 g polyunsaturated fatty acids (PUFA) n‐3 and 0.37 g of PUFA n‐6 fatty acids; for wild specimens, it was 0.920 g/100 g, or 2.85 g of SAFA, 4.82 g of MUFA, 2.78 g PUFA n‐3 and 0.27 g of PUFA n‐6 fatty acids. The major fatty acids in this study were 16:0; 16:1, n‐7; 18:1, n‐9 and DHA 22:6, n‐3 acids. The sum of these major components accounted for more than 57% and 80% of the total fatty acids in all the samples of farmed and wild tuna respectively. No significant differences in the proximate composition were demonstrated between farmed and wild samples, except for the energy value, in favour of the farmed tuna. Statistically, glucose tends to increase together with cholesterol (CHOL) and plasma triglyceride, as for these pairs, it showed positive correlation coefficients and P>0.05. Some measured tuna metabolites demonstrated strong mutual correlations, especially GLU, CHOL and TRIG, which are crucial factors in the lipid profile of animals.     

Biochemical parameters in the blood of gilthead sea bream (Sparus aurata Linnaeus, 1758) supplemented with commercially available β‐glucan‐based product (IMUNO‐2865®)

The aim of this study was to evaluate the effects of a promising new immune stimulant in aquaculture (IMUNO‐2865^®) on biochemical parameters in sea bream during the winter stress period. A total of 640 sea bream were fed throughout 90 days with diets containing 0 (Group 1), 1 (Group 2), 10 (Group 3) and 25 g (Group 4) of IMUNO‐2865^® kg^?1 of feed. Samples were taken each month and 90 days after supplementation. No statistical differences among treatment groups were noticed for the following biochemical parameters: glucose (GLU), aspartate aminotransferase (AST), plasma cholesterol (CHOL), lactate dehydrogenase (LDH), urea (URE) and creatinine (CREA). At the final sampling, total ammonia (NH₃) was higher in Groups 3 and 4 compared to the control and the low supplementation group (p < .05), while total proteins (TP) was significantly higher in Group 4 compared to all other groups, and in Groups 2 and 3 compared to the control group (p < .05). Blood Ca⁺⁺ levels were significantly higher after 60 days of feeding in all treatment groups compared to the control, and remained elevated in Group 4 even after 90 days following cessation of supplementation (p < .05). The results of this study described the increase of biochemical parameters in the blood of sea bream after use of IMUNO‐2865^® but future research is needed to evaluate its potentially immunostimulative effect on fish in commercial aquaculture.     

Bioprospecting of Less-Polar Constituents from Endemic Brown Macroalga Fucus virsoides J. Agardh from the Adriatic Sea and Targeted Antioxidant Effects In Vitro and In Vivo (Zebrafish Model)

The endemic brown macroalga Fucus virsoides J. Agardh from the Adriatic Sea was in the focus of the present research. The volatiles of fresh (FrFv) and air-dried (DrFv) samples of F. virsoides obtained by headspace solid-phase microextraction (HS-SPME) and hydrodistillation (HD) were analyzed by gas chromatography equipped with flame ionization detector and mass spectrometry (GC-FID/MS). The major HS-FrFv compound was pentadecane (61.90–71.55%) followed by pentadec-1-ene (11.00–7.98%). In HS-DrFv, pentadec-1-ene was not present, and few lower aliphatic compounds appeared, as well as benzaldehyde and benzyl alcohol. In HD-FrFv, particularly abundant were alkenes (such as pentadec-1-ene (19.32%), or (E)-pentadec-7-ene (8.35%)). In HD-DrFv, more oxidation products were present (e.g., carbonyl compounds such as tridecanal (18.51%)). The fatty acids profile of freeze-dried sample (FdFv) after conversion to methyl esters was determined by GC-FID, and oleic acid was dominant (42.28%), followed by arachidonic acid (15.00%). High-performance liquid chromatography-high-resolution mass spectrometry with electrospray ionization (HPLC-ESI-HRMS) was used for the screening of less polar fractions (F3 and F4) of F. virsoides. Mono- and diglycerides of stearic, palmitic, oleic, and arachidonic acids were found. Terpenoids and steroids comprised the compounds C₂₀H₃₀₍₃₂₎O₂ and C₂₉H₄₈O₍₂₎. Among carotenoids, fucoxanthin was identified. Chlorophyll derivatives were also found (C₅₅H₇₄₍₇₂₎N₄O_(5-7)), dominated by pheophytin a. The antioxidant activity of the fractions was investigated by in vitro assays (oxygen radical absorbance capacity (ORAC), reduction of radical cation (ABTS•⁺), 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, and ferric reducing antioxidant power (FRAP)) and by in vivo zebrafish model (along with fish embryotoxicity). In vitro experiments proved good radical scavenging abilities of F3 and F4 fractions, which were additionally supported by the protective effect against hydrogen peroxide-induced oxidative stress in zebrafish embryos.     

Piscine cytochromes P450 (CYP) and their response to antimicrobial drugs

Most research on the P450 (CYP) system in teleosts has been done on environmental factors that influence their induction. Less is known about CYP metabolism of aquaculture antibiotics. This article outlines the impact of aquaculture antimicrobials on the piscine CYP system, with particular regard to interactions with tetracyclines, fluoroquinolones, sulphonamides, berberine and chloramphenicol, as paradigms for current, potential and discontinued piscine antimicrobial drugs. It gives an overview of literature reports and advances in the field of biological significance of the CYP in fish. Emphasis has been placed on highlighting the most significant isoforms for biotransformation of drugs, and their drug response mechanisms. The challenge is to elucidate the differences in responses of CYP enzymes in different species to antimicrobial treatment as they may have relevance for the use of antimicrobials in aquaculture, especially as drug interactions with the fish CYP may alter their distribution, metabolism and elimination. They can impact the metabolism of other drugs metabolized by the same system with an effect on the physiology of fish administered these antimicrobials. Also, they can affect the persistence of residues and the length of the withdrawal period. For food animals such as farmed fish, this knowledge is a fundamental biomedical goal.     

Seasonality of nuclear abnormalities in gilthead sea bream <Emphasis Type="Italic">Sparus aurata</Emphasis> (L.) erythrocytes

The frequencies of erythrocyte nuclear abnormalities, such as irregularly shaped (ISN), vacuolated nuclei (VN), binucleated cells (BN), including micronuclei (MN), were monitored in the peripheral blood of cultured gilthead sea bream (Sparus aurata L.) from the southern Adriatic Sea (Croatia) and assessed for a relation to seasonality. Fish were sampled in February, April, June, and September. The lowest frequencies of ISN and MN were measured in April, and the lowest frequencies of VN and BN were found in February. The highest values of ISN and BN were detected in September. The highest values of MN and the most frequent occurrence of VN occurred in June and April, respectively. The Spearman rank order correlation test revealed a significant positive correlation between the frequencies of ISN, BN and MN; VN did not correlate with any of the measured parameters. The ISN had significantly higher nuclear surface area values than normal nuclei (P < 0.001). Our results demonstrate that the measured parameters displayed a distinct seasonality. Consequently, we conclude that seasonal variations should be taken into account when nuclear abnormalities are used as biomarkers. This variation should be well understood in order to successfully separate true contamination effects from seasonal factors that have the potential to affect the frequency and occurrence of nuclear abnormalities.