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.     

Evaluation of genetic diversity among some Iranian wild asparagus populations using morphological characteristics and RAPD markers

An evaluation of genetic diversity in 39 wild asparagus populations was carried out using morphological and RAPD markers. A combination of morphological traits and random RAPD primers was used to examine the level of genetic variation and polymorphisms among the populations. A factor analysis using Ward's method on mean values of morphological characteristics indicated seven main factors resulting in four groups. Analysis of polymorphic bands using Jaccard's similarity coefficient indicated that genetic similarity ranged between 0.71 and 0.29. At a similarity level of 0.64, the populations were divided in three sub-clusters, containing 34, four and one populations, respectively. Significant regression associations were found between 21 morphological characteristics and 18 RAPD markers, revealing some informative markers associated with some traits. The highest R² was related to 18 RAPD markers associated with gender (53.5%) that among them BA-04₂₀₀₀ had a maximum R². The results showed that Iranian wild asparagus with its high levels of genetic variation could be considered as a valuable gene pool for future asparagus breeding programs. Furthermore, it could be inferred that morphological characteristics and RAPD markers are suitable tools to discriminate asparagus populations for the evaluation of genetic diversity.     

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.     

Multivariate analysis of Prunus subgen. Cerasus germplasm in Iran using morphological variables

Knowledge of relationships among the cultivated and wild species of Cerasus, a subgenus of Prunus L., is important for recognizing gene pools, identifying pitfalls in germplasm collections and developing effective conservation and management strategies. In this study, morphological characterization of wild Cerasus subgenus species naturally growing in Iran, including P. avium L. (mazzard), P. mahaleb L., P. brachypetala Boiss., P. incana Pall., P. yazdiana Mozaff., P. microcarpa Boiss. subsp. microcarpa, P. microcarpa Boiss. subsp. diffusa and P. pseudoprostrata Pojark. and three commercial species, sweet cherry (P. avium L.), sour cherry (P. cerasus L.) and duke cherry (P. × gondouinii Rehd.) was investigated. Twenty-nine variables were recorded in 95 accessions of these species to detect similarities among them. All studied characteristics were showing a high degree of variability, but it was highly pronounced for plant height, leaf characters especially for dimensions and pubescence as well as for petiole and peduncle length. The majority of significant correlation coefficients were found in the characteristics representing plant size and growth habit with leaf size. Principal component analysis was performed for phenotypical diversity determination and grouping of species. Characters with high discriminating values were those related to growth habit, height, compactness, trunk diameter, leaf dimensions, petiole length, leaf tip and serration, flower color, peduncle length, fruit shape and stone shape. Based on the cluster analysis, species included in two main clusters, first those with large structure (sweet, sour and duke cherries, mazzard and mahaleb), second the other species with small structure which might be potentially suitable for rootstock breeding programs. Scatter plot using two first factors also confirmed the grouping by cluster analysis. The conservation of the highly diverse native populations of Iranian wild Cerasus species is recommended.     

Digestive enzyme activity of the red porgy (Pagrus pagrus, L.) during larval development under culture conditions

The ontogenic development of the main digestive enzymes (proteases, amylase and lipase) in the red porgy, Pagrus pagrus, larvae was assayed during the larval development. The green water technique was carried out for larval rearing and whole‐body homogenates were used for enzymatic assays in triplicate. Significant alterations in specific activities of all digestive enzymes measured during the period of this study were mostly related to metamorphosis and weaning. Trypsin‐ and chymotrypsin‐specific activities were first detected on day 3, together with opening of the mouth, and slightly increased until 25 days after hatching (DAH). After this period, the specific activities of these enzymes slightly decreased. Pepsin was first detected on day 28, concurrent with stomach formation, and a sharp increase was observed until 30 DAH. A slight decrease was measured from this date until the end of the experiment. Both amylase and lipase were measured for the first time on days 2 and 4 respectively, and the specific activities of these enzymes showed similar patterns during the first week of the study. Then, slight variations were observed until 30 DAH and while lipase‐specific activity declined, an increase in the specific activity of amylase was found until the end of the experiment. It is concluded that the variations observed in the specific activity of digestive enzymes were related to either metamorphosis, such as the formation of the stomach (28 DAH), or to changes in food composition. The profile of the developmental pattern of the main digestive enzymes detected in P. pagrus is similar to that described for other Sparid species.     

Microsatellite and mitochondrial DNA analyses reveal no genetic difference between two pufferfish species torafugu <Emphasis Type="Italic">Takifugu rubripes</Emphasis> and karasu <Emphasis Type="Italic">T. chinensis</Emphasis>

It has been reported that the nucleotide sequences of the mitochondrial and nuclear genes of Takifugu pufferfish torafugu T. rubripes and karasu T. chinensis show 99–100% sequence identity, indicating a very close relationship between the two species. To further investigate this genetic relationship, we compared genetic variation at four microsatellite loci and at the mitochondrial control region (CR) (561 bp) between groups of T. rubripes caught at two locations [TrG, caught in the Genkai Sea off Tsushima Island in 2003 (n = 50); TrS, caught in the Suwo Sea off Kita-Kyushu in 2008 (n = 50)] and T. chinensis caught at one location (TcK, caught off the east coast of Korea in 2004; n = 50). Analyses using microsatellite loci showed that genetic diversity index values of the TrG, TrS and TcK groups were 0.9505, 0.9350 and 0.9335, respectively, while values of genetic distance and genetic differentiation between TrG and TcK (0.0543 and 0.0189, respectively) were smaller than those between TrG and TrS (0.0857 and 0.0194, respectively). Analyses using CR for the same specimens showed that genetic distances were consistent with those obtained using microsatellite loci. These results, together with our previous observations, suggest that T. rubripes and T. chinensis are very closely related and possibly can be regarded as the same species.     

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.     

Simulating long-term effect of Hyrcanian forest loss on phosphorus loading at the sub-watershed level

Conversion of forest land to farmland in the Hyrcanian forest of northern Iran increases the nutrient input, especially the phosphorus(P) nutrient, thus impacting the water quality. Modeling the effect of forest loss on surface water quality provides valuable information for forest management. This study predicts the future impacts of forest loss between 2010 and 2040 on P loading in the Tajan River watershed at the sub-watershed level. To understand drivers of the land cover, we used Land Change Modeler(LCM) combining with the Soil Water Assessment Tool(SWAT) model to simulate the impacts of land use change on P loading. We characterized priority management areas for locating comprehensive and cost-effective management practices at the sub-watershed level. Results show that agricultural expansion has led to an intense deforestation. During the future period 2010–2040, forest area is expected to decrease by 34,739 hm~2. And the areas of pasture and agriculture are expected to increase by 7668 and 27,071 hm~2, respectively. In most sub-watersheds, P pollution will be intensified with the increase in deforestation by the year 2040. And the P concentration is expected to increase from 0.08 to 2.30 mg/L in all of sub-watersheds by the year 2040. It should be noted that the phosphorous concentration exceeds the American Public Health Association′s water quality standard of 0.2 mg/L for P in drinking water in both current and future scenarios in the Tajan River watershed. Only 30% of sub-watersheds will comply with the water quality standards by the year 2040. The finding of the present study highlights the importance of conserving forest area to maintain a stable water quality.