Evaluation of physiological aspects and molecular identification of Saprolegnia isolates from rainbow trout (Oncorhynchus mykiss) and Caspian trout (Salmo trutta caspius) eggs based on RAPD–PCR

The genus of saprolegnia is one of the most important pathogenic aquatic fungi in farmed and wild fish. In the present study, fungal infected egss were collected from rainbow trout (Oncorhyncus mykiss) and Caspian trout (Salmo trutta caspius). After purification, 16 isolates were obtained (8 isolates from rainbow trout and 8 isolates from Caspian salmon, respectively). The isolates were then coded as R2 – R9 (rainbow trout) and S2 – S9 (Caspian trout).The registered DNA for S. parasitica (ACTT # 200048) and S. diclina (ACTT # 4206) were used and coded as R1 and S1, respectively. Based on the RAPD profile obtained all samples were divided to 3 groups and members of each group had more than 90% similarity among themselves. According to matrix of similarity and reference strains, the isolates were classified as three groups. Therefore, all of isolates in group 1 and 3 were S. parasitica and S. diclina, respectively, and the members of group 2 were known as Saprolegnia sp. The results of thermal resistance assessment showed that the isolates of rainbow trout and Caspian salmon eggs had slow growth in the temperature between 18 – 20 °C. Thus, R2 and S8 isolates had the lowest radial growth compared to other isolates. The isolates categorized in S. parasitica (group 1) created secoundry zoospores but not observed in two other groups. Thus, catenulated gamme was found in 78% and 55.55% isolates of rainbow trout and Caspian trout eggs, respectively. This study indicated that molecular methods were the best methods for identification of Saprolegnia spp. and it could be applied as a supplementary confirming method

Quantitative evaluation and identification of fungi in Shahid Rajaeii Dam Lake, Mazandaran Province (Sari)

The present study is carried out to investigate the fungal species present in water of Shahid Rajaeii damlake in Sari, (Mazandaran province). Samples were taken from five stations including, Station 1: Input of Shirinrud river, station 2: Input of Sefidrud river, Station 3: The confluence of the two branches, Station 4: dam crest and stations 5: Output dam from June to February 2012. Every sample was diluted by sterile saline (10-1 and 10-2) and 0.5 mL from each dilution was cultured on SD and incubated at 27-30°C for 3-5 days. Finally, the number of colonies wasrecorded as (colony forming unit = CFU) per 100 mL. Identification of fungal agents were conducted by slide culture preparation and stained in lacto-phenol blue. The results showed that in August and February were significantly highest and lowest rates of fungal colonies were isolated from water in different stations respectively. Moreover, the number of fungal colonies in the crown and the output was significantly higher than other stations. The frequency of identified fungi were: Aspergillus species (31.4%), various types of yeast (mainly Candida) (24.2%), Penicillium sp. (19.3%), Cladosporium sp.(10.3%), Mucor sp. (5.4%), Fusarium sp. (2.9%), sterile hype (2.8%), Alternaria sp. (2.3%) and Paecilomyces sp. (1.4%)

Creation of cryopreservation bank of bony fish

In this study, 11 male of Caspian trout (Salmo truta caspius) (with mean length and weight 37/8 ± 5/3 cm and 523/3 ± 24/7 respectively) and 23 male of Caspian kutum (Rutilus frisii kutum) (with mean length and weight 36/1 ± 7/1 cm and 631/3 ± 21/6 g respectively) were evaluated. All the fish were good at the initial examination of sexual maturity. After sperm sampling, their quality were tested. In this step, the parameters such as motility, duration of mobility, density, pH and osmolality were measured. After this stage, the sperm samples of Caspian trout in the ratio 1: 3 were diluted with the aqueous solution containing compounds (0.3M Glucose, 10% Methanol, 10% egg yolk) and the freezing process was done manually and the sperm was frozen in liquid nitrogen. The sperm samples of Caspian kutums were diluted (ratio of 1: 3) with two soluble diluent containing compounds (350 mM glucose, 30 mM Tris and 4% Polyethylene glycol) and (350 mM glucose, 30 mM Tris and 2% Glycerol) and were frizzed automatically by Planner Kryo instrument and placed in liquid nitrogen. The sperm samples were thawed 1 to 3 months after the date of first freezing and their quality were assessed by measuring percent and timing motility. The results showed that the obtained semen volume of Caspian trout was more than Caspian kutum. Moreover, percentage of motile sperm, timing motility and sperm density of Caspian trout were higher than those of Caspian kutum but osmolality and pH of Caspian trout were lower than those of Caspian kutum. Over time, the percentage of sperm motility and mobility for both species declined compared with fresh samples. After thawing, percentage of motile sperm and timing motility of Caspian kutum were lower than those factors Caspian trout. The results showed that the sample of Caspian kutum sperm that were diluted by ethylene glycol after thawing and were immotile ll of them. However, the samples were diluted by glycerol, after thawing, were alive and motile. According to the results, it seems very important species differences that must be fully considered in the process of freezing sperm. The use of a single protocol would not be successful in cryopreservation because the reaction of sperm against to chemical agents is variable. Therefore, it is essential to get the right information to protect valuable Caspian fish by using cryopreservation. Further studies on the characteristics of each species, as well as the freezing process take appropriate diluent