Chemical composition and in vitro antifungal activity of Sambucus ebulus and Actinidia deliciosa on the fish pathogenic fungus,Saprolegnia parasitica

The aim of the present study was to determine the chemical composition of Sambucus ebulus and Actinidia deliciosa ethanolic extracts as well as their in vitro antifungal activity on the mycelial growth of the water mold, Saprolegnia parasitica. The preliminary minimum inhibition concentration (MIC) and minimum lethal concentration (MLC) were determined by the HeMP method and finally, concentrations of each extract ranging from 1 to 10% were prepared by an agar dilution method to assess the in vitro antifungal activity, quantitatively. Both herbal extracts inhibited growth of Saprolegnia hyphae in vitro. Complete in vitro growth inhibition was found at a concentration of ≥5% for S. ebulus, whereas it was not observed even at 10% concentration of A. deliciosa. Based on gas chromatography coupled with mass spectrometry (GC/MS) analysis, the main constituents of S. ebulus include monophthalate (66.17%), fatty acids (26.47%), phytol (4.25%), and acetic acid (2.11%). Using colorimetric assays, A. deliciosa contained phenolic contents at 162 mg gallic acid (GAE)/g DW and flavonoid contents at 2.31 mg quercetin (QE)/g DW. In conclusion, the results of this study indicated that S. ebulus and A. deliciosa showed some antifungal activities against S. parasitica with formerly exhibiting stronger activity (p < 0.05).

     

Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.     

Anaesthetic efficacy of eugenol on Flowerhorn (Amphilophus labiatus × Amphilophus trimaculatus)

Anaesthetic efficacy of eugenol was investigated on Flowerhorn (Amphilophus labiatus × Amphilophus trimaculatus). A total of 104 fish with average weights of 12 ± 2.5, 28 ± 5 and 53 ±5.1 g were subjected to 25–200 mg L^?1 eugenol and behavioural responses as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 49.9–127.3 s after exposure to 50–200 mg L^?1 eugenol, fish reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 57.3–140.4 s post exposure to such concentrations. Recovery time was 91.7–312 s in all weight classes for all eugenol concentrations. Mortality (23%) was only observed in 12‐g fish when were subjected to 200 mg L^?1 eugenol. This study showed the behavioural response of Flowerhorn to anaesthesia and eugenol efficacy as an anaesthetic in this important ornamental species. The general quadratic equation revealed that concentrations of eugenol and fish size along with their interactive effects have significantly contributed to the model, with concentration recording the highest beta value in all models (β = ?0.809, ?0.818 and ?0.909, P = 0.000). According to the results, minimum eugenol concentration to induce anaesthesia in less than 3 min was 50 mg L^?1.     

The effect of maternal size on larval characteristics of Persian sturgeon Acipenser persicus

The objective of this work was to study the relationship between female size (weight) and variables of egg and larval stage of Persian sturgeon Acipenser persicus . In this study, 19 female breeders were captured in Caspian Sea and fertilized by routine methods. Positive significant correlations ( P <0.05) were established between female weight and ovulated eggs per female, time of second mitosis division and volume of yolk-sac at hatching. There was not significant correlation ( r =0.33, P =0.161) between female weight and egg diameter. Female weight was not affected weight of larvae at hatching time ( r =0.37, P =0.119), as well as larval length ( r =−0.14, P =0.558) and larval weight at the end of the experiment (48 hours after first feeding) ( r =0.16, P =0.491). Mortality rate during yolk-sac absorption was higher with increased female weight but their correlation was not significant ( r =0.40, P =0.076). During the first feeding stage, mortality rate was 13.39% and there was no significant correlation between mortality rate in this period and female weight ( r =−0.12, P =0.613). Conclusively, as a result female size influenced fecundity, time of second mitosis division and yolk-sac volume at hatching time without affecting mortality rate during yolk-sac absorption and first feeding stage in Persian sturgeon. Thus, smaller female broods do not cause more mortality than larger ones in larval production and they can be used in reproduction procedure.     

Anaesthetic efficacy of eugenol on iridescent shark,Pangasius hypophthalmus (Sauvage, 1878) in different size classes

Anaesthetic efficacy of eugenol was investigated on iridescent shark, Pangasius hypophthalmus. Fish (2, 5, 10 and 20 g) subjected to 20–200 mg L^?1 eugenol and behavioural response as well as induction and recovery times were recorded. Induction and recovery times were significantly affected by eugenol concentration as well as fish weight (P < 0.05). Generally, 27–300 s after exposure to 20–200 mg L^?1 eugenol, iridescent sharks reached stage 3 anaesthesia (suitable for general handling). Fish entered stage 4 anaesthesia (suitable for surgery and blood sampling) over 54–710 s exposure to such concentrations. Recovery time was 109–600 s in all weight classes as well as eugenol concentrations. Mortality (44–100%) was only observed in 2 g fish when subjected to 110–170 mg L^?1 eugenol. This study, for the first time, showed behavioural response of iridescent shark to anaesthesia as well as effectiveness of eugenol as anaesthetic in this important aquaculture‐ornamental species. According to the models obtained in this study, minimum eugenol concentrations to induce anaesthesia over less than 3 min were 53.8–81.5 mg L^?1 in 2–20 g fish. Likewise, maximum eugenol concentrations in which fish recovered over less than 5 min were 65.9–105.8 mg L^?1 in 2–20 g fish.     

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.     

Effects of emulsified versus saline administration of GnRHa on induction of ovulation in rainbow trout, Oncorhynchus mykiss

The effects of saline-dissolved or Freund's incomplete adjuvant (FIA)-emulsified GnRHa treatment on the induction of ovulation in rainbow trout, Oncorhynchus mykiss, were examined. The FIA-emulsified GnRHa was first diluted in 0.25 ml physiological saline and then mixed with an equal volume of FIA. Fish were selected in the beginning of the spawning season and were allocated into four groups and were treated intraperitoneally with (a) 0.5 ml of emulsified GnRHa (GnRHa–FIA), (b) 0.5 ml saline-dissolved GnRHa in a single injection (GnRHa-1), (c) 0.5 ml saline-dissolved GnRHa in two injections spaced 1 d apart (GnRHa-2) and (d) 0.5 ml of saline (Control). The GnRHa dose in all hormone treatments was 25 µg kg^− 1. All fish in the FIA–GnRHa and GnRHa-2 groups ovulated within 10 and 11 d after treatment, respectively. In contrast, only 75% in the Control fish and 60% of the fish in the GnRHa-1 group ovulated within 36 d after treatment. None of the treatments caused any pre- or post-spawning mortality in the broodstock. Fertilization, eyeing and hatching percentages of the produced progeny were normal in all the treatment groups and did not differ significantly among them. In conclusion, FIA-emulsified GnRHa can be effective in advancing the onset of and synchronizing the ovulation of rainbow trout within a two-week period, thus shortening the egg collection period, without affecting broodstock survival and egg quality.     

Effects of triploidy on the Caspian salmon <Emphasis Type="Italic">Salmo trutta caspius</Emphasis> haematology

The aim of this study was a comparison of key haematological features of diploid (2n) and triploid (3n) Caspian salmon (Salmo trutta caspius). Morphometric indices of erythrocytes were determined on blood smears by light microscopy. Triploidy significantly (P < 0.001) increased all morphometric indices measured in the erythrocytes including cell size, cell surface area, and cell volume. The increase in cell size was larger for the major (27%) axis than for the minor (22%) axis, thus making erythrocytes of 3n Caspian salmon more ellipsoidal. The estimated increase in erythrocyte nuclear volume (87%) was bigger than the theoretical expected 50% increase. Haematological indices were measured manually by hemocytometry. Triploids had lower numbers of red blood cells (RBC: 1,120,000 cells/mL in 2n vs. 700,000 cells/mL in 3n; P < 0.001) but they were larger in size (mean erythrocytic volume [MEV]: 363.1 nm³ in 2n vs. 483.3 nm³ in 3n; P < 0.001). The decrease in RBC number was not compensated by the increase in MEV and, thus, triploidy affected the haematocrit (Hct: 38.8% in 2n vs. 33.06% in 3n; P < 0.05). Total blood hemoglobin concentration was lower in triploid fish (Hb: 9.9 g/dL in 2n vs. 8.9 g/dL in 3n; P < 0.05). In contrast, mean erythrocytic hemoglobin (MEH: 95 μg in 2n vs. 133.2 μg in 3n; P < 0.001) was higher for 3n Caspian salmon as a result of their larger erythrocytes, although MEH concentration (MEHC: 0.26 g/dL in 2n vs. 0.27 g/dL in 3n) did not significantly differ (P > 0.05). White blood cell (WBC) counts (lymphocytes and neutrophiles) were measured and WBC/RBC ratios were calculated. There were no significant differences in WBC (15,710 cells/mL in 2n vs. 12,683 cells/mL in 3n; P > 0.05), lymphocytes, and neutrophils as %WBC as well as WBC/RBC ratios between two ploidy levels (P > 0.05). Triploid Caspian salmon showed higher erythrocyte abnormalities such as ‘twisted’, ‘tailed’, and ‘anucleated’ cells as well as high portions of immature RBC in blood smears in comparison with diploids (P < 0.001).