Recent update on the prevalence of Vibrio species among cultured grouper in Peninsular Malaysia

Vibrio infections are common among marine fish and lead to serious problems in the aquaculture sector. This study reports a recent occurrence of Vibrio species (spp.) isolated from cultured groupers in Peninsular Malaysia using the gyrB and pyrH genes. A total of 147 Vibrio strains were successfully isolated from 77 (64%) groupers using culture method and subjected to gyrB and pyrH sequencing for species identification and confirmation. Results showed that 89% of Vibrio strains were identified and clustered to six groups of Vibrio spp., while 11% were not clustered to any Vibrio spp. using the gyrB sequences. Meanwhile, by analysis of the pyrH sequences all the 147 Vibrio strains (100%) were successfully identified and clustered into 11 groups of Vibrio spp., including the gyrB non‐identified strains. The pyrH gene provides a better resolution for identification of Vibrio spp. compared with the gyrB gene. Thus, the pyrH gene was more suitable for a rapid determination of Vibrio spp. distribution in Peninsular Malaysia. Using the pyrH gene, our study found higher prevalence of Vibrio vulnificus (33%), V. alginolyticus (24%) and V. parahaemolyticus (22%), followed by V. rotiferianus (5%), V. harveyi (3%), V. tubiashii (2%), V. campbellii (2%), V. ponticus (1%), V. diabolicus (1%), V. owensii (1%) and others Vibrio sp. (7%). Thus, the results of this study revealed that the occurrence of pathogenic vibrios among grouper fish is still high in Malaysian aquaculture. In addition, the pyrH gene was proved as a suitable marker for rapid identification of Vibrio species compared with the gyrB gene.     

Rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) anesthesia with myrcene: efficacy and physiological responses in comparison with eugenol

The aim of the present study was to investigate anesthetic efficacy of myrcene in rainbow trout (Oncorhynchus mykiss) along with the fish biochemical response to anesthesia in comparison with eugenol. In the first experiment, 240 fish were stocked in 12 tanks and acclimatized to experimental conditions for 2 weeks. Then, the fish of each tank were subjected to one concentration of either eugenol (12, 20, 30, 50, 80, and 130 μL/L) or myrcene (100, 150, 200, 300, 400, and 500 μL/L) concentrations. Induction time of and recovery time from anesthesia were recorded for each fish separately. Using these results, desired concentrations to induce anesthesia within 60, 180, 300, and 600 s were determined, being 81, 30, 19, and 10 μL/L eugenol and 531, 251, 177, and 111 μL/L myrcene. In the second experiment, 96 fish were stocked in 8 tanks. Six fish were netted from each tank and exposed to the calculated eugenol or myrcene concentrations. Blood samples were taken after the fish reached anesthesia. The results showed that there was no significant difference in serum lactate, alanine transaminase, and alkaline phosphatase. Increase in the induction time of anesthesia resulted in increased serum glucose with no significant difference between the anesthetics. Increase in induction time of anesthesia led to increase in serum lactate dehydrogenase activity in the eugenol-anesthetized fish and aspartate transaminase activity in myrcene-anesthetized fish. In conclusion, myrcene is capable to anesthetize rainbow trout, but at higher concentrations compared to eugenol. In addition, biochemical analysis showed that increase in induction time of anesthesia leads to hyperglycemia and increase in AST and LDH activities depending on anesthetic type.     

Non‐specific immune responses and intestinal immunity of common carp (Cyprinus carpio) fed Jujube (Ziziphus jujube) fruit extract

This study investigated the effects of Jujube (Ziziphus jujube) fruit extract (JFE) as an excellent source of nutrients and phytochemicals on the innate immune responses and expression of genes involved in intestinal immunity of common carp (Cyprinus carpio) fingerlings. Three hundred healthy carps (10.78 ± 0.05 g) fingerlings were randomly allocated to 12 fibre glass tanks (300L; 15 fish per tank) and fed for 8 weeks with experimental diets which contained different levels of JFE (0%, 0.25%, 0.5%, or 1%). Thereafter, serum non‐specific immune parameters [total immunoglobulin (Ig) as well as alternative complement (ACH50) and lysozyme activity (SL)] and expression of genes involved in intestinal immunity (lyz, tnf‐alpha, il1b, il8, il10, and tgfb) were measured. The results revealed a significant elevation of Ig level in the JFE‐fed fish (p < 0.05) over the control fish. However, there was no significant difference among treated groups in case of serum total Ig. The highest levels of SL and ACH50 were observed in fish fed 1% of JFE. Nonetheless, no significant difference was noticed regarding SL and ACH50 of control and other supplemented groups (0.25% and 0.5% JFE). Also, expression of lyz, tnf‐alpha, and il1b genes increased significantly (p < 0.05) in JFE‐fed fish, in a dose‐dependent manner. On the other hand, significant (p < 0.05) down‐regulation of il110 and tgfb were noticed in treated groups. The present findings suggested that extraction of Jujube, Z. jujube fruit, possesses beneficial effects on immune responses of common carp.     

Maternal age influence on diapause induction in sexual and asexual Trichogramma brassicae

In this study, the effect of maternal age on prepupae diapause of progeny in sexual (Wolbachia free) and asexual (Wolbachia infected) Trichogramma brassicae was studied at two temperatures. Maternal generation (G₁) of both populations was reared at 10L:14D photoperiod, 14 and 20 °C, separately. Mated females from G₁ were supplied with eggs of Ephestia kuehniella, each 48 h from one day after emergence to death. The tendency to diapause in progeny (G₂) was determined by rearing at 10 °C. According to the results, reproductive modes, maternal temperature and age have significant effect on the mean number of eggs laid by G₁ and the diapause percentage of G₂. Offspring whose maternal generation was reared at 20 °C had higher diapause percentage than those reared at 14 °C. Diapause percentage in sexual population was higher than the asexual, which demonstrates a negative effect of Wolbachia on this process. Wolbachia infection modifies the effect of maternal age on the diapause percentage in the offspring at 20 °C but not at 14 °C. The result can be due to lower Wolbachia density at 14 °C and greater Wolbachia density at 20 °C.     

Dietary nucleotide mixture effects on reproductive and performance,ovary fatty acid profile and biochemical parameters of female Pacific shrimp Litopenaeus vannamei

The effects of different dietary nucleotide mixture (NT) levels (0, 2, 4 and 6 g/kg) were investigated on the reproductive performance, fatty acid profile and biochemical parameters in Litopenaeus vannamei female that were co‐fed with fresh food (two times daily) and experimental diets (two times daily) for 30 days. Sampling was carried out at the first day of the trial, before eyestalk ablation (ESA) (day 21) and after ESA (day 30). Reproductive performance parameters including the hepatopancreatic index, absolute fecundity, egg diameter and latency period were higher in the NT‐supplemented groups than the control group (p < .05). Total n‐3 polyunsaturated fatty acids in the ovaries, especially eicosapentaenoic acid, were higher in the NT‐supplemented groups than in the control (222.6 ± 7.3 versus 204.7 ± 3.0 mg/g extracted lipid) (p < .05). Total hemocyte count, granular cells, plasma glucose (Glu), total protein (TP), calcium (Ca²⁺), total cholesterol (Chol), triglyceride (Tg) and high‐density lipoprotein (HDL) levels were significantly increased in the NT‐supplemented groups in comparison with the control group (p < .05). After ESA, the concentration of plasma Glu, TP, Chol and Tg significantly decreased, but HDL levels increased in all groups (p < .05). These results suggested that the application of dietary NT may present a novel strategy for increasing reproductive performance and health status in L. vannamei.     

Growth,morphology and gas exchange responses of two-year-old Quercus castaneifolia seedlings to flooding stress

Tolerance to flooding is crucial when thinking in promissory species for restoration of ecosystems prone to suffer soil water excess. In this study, we tested the flooding tolerance of two-years-old seedlings of Chestnut-leaved oak (Quercus castaneifolia C.A.Mey.) to determine whether it can be recommended for use in wetland restoration programs. Seedlings of Q. castaneifolia were subjected to three treatments: (1) control (C), (2) flooding for 60 days followed by a 42-day recovery period (F?+?R) and (3) continuous flooding for 102 days (F). Physiological performance, plant morphological changes and biomass accumulation were assessed. Results showed that, although net photosynthetic rates, stomatal conductance and transpiration decreased with prolonged flooding, when flood waters were removed, plants were able to recover their physiological activity (49–80% compared to controls). By contrast, when plants were continually flooded, their physiological activity decreased as well as the leaves experienced precocious senescence and wilting. Biomass responses paralleled physiological responses: leaf and root biomass were 42–49% higher under F?+?R treatment than under continuous flooding, and all plants under the F?+?R survived. Therefore, Q. castaneifolia appears as a promising species to be further studied when thinking re-vegetation of riverine areas and other temporarily flooded wetlands.     

Improvement of Tissue Survival of Skin Flaps by 5α-Reductase Inhibitors: Possible Involvement of Nitric Oxide and Inducible Nitric Oxide Synthase

Background:

Skin flap grafting is a popular approach for reconstruction of critical skin and underlying soft tissue injuries. In a previous study, we demonstrated the beneficial effects of two 5α-reductase inhibitors, azelaic acid and finasteride, on tissue survival in a rat model of skin flap grafting. In the current study, we investigated the involvement of nitric oxide and inducible nitric oxide synthase (iNOS) in graft survival mediated by these agents.

Methods:

A number of 42 male rats were randomly allocated into six groups: 1, normal saline topical application; 2, azelaic acid (100 mg/flap); 3, finasteride (1 mg/flap); 4, injection of L-N^G-nitroarginine methyl ester (L-NAME) (i.p., 20 mg/kg); 5, L-NAME (20 mg/kg, i.p.) + azelaic acid (100 mg/flap, topical); 6, L-NAME (20 mg/kg, i.p.) + finasteride (1 mg/flap, topical). Tissue survival, level of nitric oxide, and iNOS expression in groups were measured.

Results:

Our data revealed that azelaic acid and finasteride significantly increased the expression of iNOS protein and nitric oxide (NO) levels in graft tissue (P < 0.05). These increases in iNOS expression and NO level were associated with higher survival of the graft tissue.

Conclusion:

It appears that alterations of the NO metabolism are implicated in the azelaic acid- and finasteride-mediated survival of the skin flaps.Key Words: Finasteride, Azelaic acid, Surgical flaps, Nitric oxide, Nitric oxide synthase Type II