Characterization and Purification of Glutathione S-Transferase from the Liver and Gill Tissues of Ağrı Balık Lake Trout Salmo trutta labrax and the Effects of Heavy Metal Ions on Its Activity

Glutathione S-transferase (GST) from the liver and gill tissues of A?r? Bal?k Lake Trout (also known as Black Sea Trout) Salmo trutta labrax was characterized and purified, and the toxic effects of some heavy metal ions on the enzyme's activity were analyzed. Liver GST was purified 930 times, resulting in 56% yield using glutathione–agarose affinity chromatography and a specific activity of 60.87 endotoxin units (EU)/mg protein. Using the same procedure, gill GST was purified 576 times, resulting in a 60% yield and specific activity of 46.8 EU/mg protein. The purity check of the purified enzymes was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Optimal pH, ionic strength, and stable pH were found for each tissue, and separate K_M and V_max values were determined for reduced glutathione and 2,4-dinitrochlorobenzene substrates. Heavy metal ions that have toxic effects on living organisms and are known to contribute to environmental pollution were selected, and their in vitro effects on enzyme activity were analyzed. The IC50 values and K_i constants of those metal ions showing an inhibitory effect on GST activity were determined.

Received November 24, 2014; accepted March 11, 2015     

Anatomical Structure of the Brachial Plexus in the Merlin (Falco columbarius)

This study aimed to document the detailed features of the morphological structure and the innervation areas of the brachial plexus in Merlin (Falco columbarius). The skin and muscles of five adult male Merlins were dissected under the stereo microscope. The Merlin had two plexus trunks. The accessory brachial plexus consisted of ventral rami C10 and C11. C11 was divided into two branches: the cranial and caudal. The brachial plexus was composed of a rather complex network involving the ventral rami of C11‐C13, T1 and T2. In addition, a thin branch from the last two cervical sympathetic nerves participated in the plexus formation. C12, C13 and T1 had rather thick trunk. C12, C13 and T1 were also involved in the formation of the brachial plexus emerging after 1 cm from the foramen inter‐vertebrale as three trunk roots.     

Comparison of Cryoprotective Effects of Lycopene and Cysteamine in Different Cryoprotectants on Bull Semen and Fertility Results

The objectives of this study were to compare glycerol and ethylene glycol at different concentrations as cryoprotectants and lycopene or cysteamine (with/without) as antioxidants in Tris extender for bull semen. Twenty‐four ejaculates were obtained from three bulls. Each ejaculate was split into four equal aliquots and diluted using both of the Tris extenders with glycerol (5% or 7%) or ethylene glycol (3% or 5%). After that, each extenders were split into three equal aliquots and added using both of the cysteamine 5 mm or lycopene 500 μg/ml, and control (without additives). The addition of 7% glycerol with cysteamine, 5% ethylene glycol with cysteamine and 3% ethylene glycol with cysteamine groups gave the lowest CASA motility than the other groups. However, 7% glycerol and 7% glycerol with lycopene resulted in a better rate of CASA progressive motility compared with that of other groups. Generally, all the lycopene groups signed better protective effects on acrosome and total morphology than the other groups. Glycerol 7% and 3% ethylene glycol with lycopene groups yielded to slight higher percentages of membrane integrity assessed by HOST than that of the other groups, but 7% glycerol with cysteamine and 3% ethylene glycol with cysteamine showed the worst percentages of membrane integrity. Glycerol 7% and 5% glycerol with lycopene gave rise to a higher value of VAP, VSL and VCL compared with that of the other groups. On the contrary, adding to 5% glycerol with cysteamine showed negative effect for VAP, VSL, VCL and ALH values. All cryoprotectant groups with lycopene decreased chromatin damage than the other groups. Ethylene glycol 3% led to lower non‐return rates of inseminated cows. However, this result was not considered to be statistically important.     

Treatment Method Affects Color,Chemical, and Mineral Composition of Seabream (Sparus aurata) Fish Bone Powder from by-Products of Fish Fillet

ABSTRACT

Seabream fish bone powder was produced using different chemical methods and tap water. The effect of different chemical procedures (sodium hydroxide (NaOH); NaOH + citric acid; NaOH + sodium hypochlorite (NaOCl); NaOH + ethanol (EtOH); and NaOH + hydrogen chloride (HCl)) and tap water on proximate composition, yield, mineral composition, color, and sensorial odor was investigated. Despite its high bone powder yield (59.39%), tap water treatment was not an efficient method due to low calcium (Ca²⁺, 232.13 g/kg) and phosphorus (P, 111.63 g/kg) concentration and heavy fish odor. Sensorial odor analysis of seabream fish bone powder showed that tap water received the lowest scores (1.71/5), while chemically treated samples received sensorial odor scores higher than 4.00, with an average of 4.61, indicating that they had a very slight odor. The best fish bone powder yield was obtained using NaOH treatment (21.46%), where Ca²⁺ (276.73 g/kg) and P (147.23 g/kg) content was also high. The utilization of chemicals in combination with NaOH did not increase the sensorial odor score of seabream fish powders but resulted in a decrease in powder yield. Moreover, the mineral composition of all chemical processing techniques was comparable. Processing by-products of seabream fillet production with 8% NaOH will contribute to daily Ca²⁺ and P intake of individuals.     

Molecular Characterization of Some Apple Accessions Using DNA Markers Associated with Fruit Skin and Flesh Colour

Apple (Malus domestica Borkh.) is one of the most widely grown and economically important fruit crops in the world. Skin colour is one of the most important criteria for marketing of apple fruits. Thus, improving fruit colouring is one of the main objectives of apple breeding programs. Even though environmental conditions may affect the colouring of apple, understanding the genetic basis of colouring is important to accelerate the breeding process. In recent years, molecular biology and genetic studies were conducted to explore the molecular basis of colouring in apple. Many genes responsible for the anthocyanin synthesis were identified and their associations with colouring of apple fruit flesh and/or skin were demonstrated. In addition, some DNA markers associated with fruit flesh and skin colour have been developed for screening apple cultivars and hybrids. In this study, 90 apple accessions of international, national, and local decents were selected from the genetic resources collection. Genomic DNA was isolated from leaf tissue of all acession and screened with four different DNA markers associated with fruit flesh and skin colour. Apple accessions in the collection were characterized and their genotypes and genetic potential for fruit colouration were determined by different DNA markers. Additionally, the use of these DNA markers in different apple accessions, well-characterized cultivars and uncharacterized local and national types were investigated.

     

Clavibacter michiganensis subsp. michiganesis: Tracking strains using their genetic differentiations by ISSR markers in Southern Turkey

There is no available data published related to the dissemination of bacterial canker caused by Clavibacter michiganensis subsp. michiganensis (Cmm) and its genetic diversity in Turkey. It is important to control new introduced inoculum sources by commercial seeds and seedlings. Pathogens were identified by morphological features and the identity was confirmed by PCR amplification using a specific primer PSA-4, PSA-R in addition to microbiological tests. ISSR markers showing high polymorphism ratios were selected and used to characterize Cmm strains. The collected strains were classified into different groups on the basis of ISSR-PCR fingerprints, which showed remarkable genetic specificity and diversity not previously identified in Cmm, suggesting that genetic differences are related to dissemination of the pathogen in the region. This is the first ever study carried out on the characterization of Cmm using ISSR. The selected ISSR primers to characterize Cmm can be used to determine genetic differences in further studies.     

Development and validation of glycoprotein‐based native‐subunit vaccine for fish against Aeromonas hydrophila

Aeromonas hydrophila is known to be causative agent of an infection named as Bacterial haemorrhagic septicaemia or red pest in freshwater fish. The aim of this study was to develop and validate the glycoprotein‐based fish vaccine against Aeromonas hydrophila. For this aim, after identification and characterization of A. hydrophila isolates from fish farms, one A. hydrophila isolate was selected as vaccine strain. Antigenic glycoproteins of this vaccine strain were determined by Western blotting and glycan detection kit. The connection types of these glycoproteins were examined by glycoprotein differentiation kit. Two glycoproteins, molecular weights of 19 and 38 kDa, with SNA connection type were selected for use in vaccination trials. After their purification by SNA‐specific lectin and size‐exclusion chromatography, protection studies with purified proteins were performed. For challenge trials, four experimental fish groups were designated: Group I (with montanide), Group II (with montanide and ginseng), Group III [with Al(OH)₃] and Group IV [with Al(OH)₃ and ginseng]. The survival ratings of fish were determined, and protection was calculated as 21.56%, 29.41%, 69.83% and 78.88% in groups I, II, III and IV, respectively. In conclusion, A. hydrophila glycoproteins with Al(OH)3 and ginseng could be used as a safe and effective vaccine for fish.     

New records of endoparasitoidBassus fabricius, 1804 (Hymenoptera: Braconidae: Agathidinae) species from Turkey

SixBassus species were recorded for the first time in Turkey:B. calculator (Fabricius, 1798),B. cingulipes (Nees, 1814),B. conspicuus (Wesmael, 1837),B. graecus Achterberg, 1992,B. rugulosus (Needs, 1834) andB. zaykovi (Nixon, 1986). By adding these new records, the number ofBassus species in Turkey has been raısed from seven to 13. Distribution and known hosts are given.