University of Guilan Aquatic Physiology and Biotechnology 2345-3966 11 1 2023 05 22 Investigation on susceptibility of the Caspian kutum (Rutilus kutum ) primary caudal fin cells to spring viremia of carp virus Investigation on susceptibility of the Caspian kutum (Rutilus kutum ) primary caudal fin cells to spring viremia of carp virus 1 18 6833 10.22124/japb.2022.21587.1456 FA Seyed Abolfazl Alaei M.Sc. Student in Fisheries, Department of Fisheries, Bandar Anzali Branch, Islamic Azad University, Bandar Anzali, Iran Somayeh Haghighikarsidani Assistant Professor in Department of Fisheries, Bandar Anzali Branch, Islamic Azad University, Bandar Anzali, Iran Mohaddes Ghasemi Assistant Professor in Inland Waters Aquaculture Research Center, Iranian Fisheries Science Research Institute (IFSRI), Agriculture Research, Education and Extension Organization (AREEO), Bandar Anzali, Iran Journal Article 2022 01 20 In this in vitro study the susceptibility of primary caudal fin cells of Caspian Whitefish (Rutilus kutum)- as the mast commercial endemic fish in the southern Caspian Sea- was examined to the spring viremia of carp virus (SVCV), the acute infectious, hemorrhagic and contagious agent. For this purpose, the primary cell culture was established from the caudal fin of fingerlings, using tissue explant technique and trypsinization of minced tissues. After the propagation of SVCV’s standard strain, the different dilutions of virus, in six replicates, were inoculated to 96-well plates which was contained primary caudal fin monolayer of cells after 5 passages, and the EPC cell line. In order to evaluation of virus characterization, an indirect immunofluorescent antibody test was hired. The clearly observable cytopathic effects and emergence of SVCV antigens as light fluorescence spots in both primary culture of caudal fin cells and the EPC cell line, highly confirmed the susceptibility of caudal fin primary cells to the SVCV. In this in vitro study the susceptibility of primary caudal fin cells of Caspian Whitefish (Rutilus kutum)- as the mast commercial endemic fish in the southern Caspian Sea- was examined to the spring viremia of carp virus (SVCV), the acute infectious, hemorrhagic and contagious agent. For this purpose, the primary cell culture was established from the caudal fin of fingerlings, using tissue explant technique and trypsinization of minced tissues. After the propagation of SVCV’s standard strain, the different dilutions of virus, in six replicates, were inoculated to 96-well plates which was contained primary caudal fin monolayer of cells after 5 passages, and the EPC cell line. In order to evaluation of virus characterization, an indirect immunofluorescent antibody test was hired. The clearly observable cytopathic effects and emergence of SVCV antigens as light fluorescence spots in both primary culture of caudal fin cells and the EPC cell line, highly confirmed the susceptibility of caudal fin primary cells to the SVCV.

https://japb.guilan.ac.ir/article_6833_2ad772d6bc05f8583918c14d0f1f29f2.pdf

University of Guilan Aquatic Physiology and Biotechnology 2345-3966 11 1 2023 05 22 Investigation of Sistan deep aquifer well water potential for culturing microalga Dunaliella tertiolecta Investigation of Sistan deep aquifer well water potential for culturing microalga Dunaliella tertiolecta 19 40 6834 10.22124/japb.2022.21644.1457 FA Abdolali Rahdari Assistant Professor in Department of Fisheries, Hamoun International Wetland Institute, Research Institute of Zabol, Zabol, Iran Ali Khosravanizadeh Assistant Professor in Department of Fisheries, Hamoun International Wetland Institute, Research Institute of Zabol, Zabol, Iran Sahel Pakzad Toochaei Assistant Professor in Department of Natural Ecosystems, Hamoun International Wetland Institute, Research Institute of Zabol, Zabol, Iran Journal Article 2022 01 28 The purpose of this study was to evaluate the potential of deep aquifer well water located in Sistan region (southeastern of Iran) for growing Dunaliella tertiolecta. For this purpose, 5 treatments including deep aquifer well water with salinities of 20 and 35ppt, deep aquifer well water and salt of Urmia Lake (35 ppt) and tap water with salinities of 20 and 35ppt were considered and D. tertiolecta microalgae was reared for 24 days under standard conditions (temperature 26±1ºC, light 2000 lux, 16 hours light and 8 hours darkness). The results showed that the pH value in all treatments increased slightly until day 6 but then decreased and had a steady trend from day 12 to 24. The highest amount of growth, chlorophyll a and carotenoid production, specific growth rate, the least doubling time and the highest rate of division were observed in deep aquifer well water treatment (salinity 20 ppt), although the lowest amount of biomass production was in this treatment. Based on these results, it can be said that if salt is added to deep aquifer well water, by changing the composition and increasing salinity, the amount of biomass produced will increase. In general, D. tertiolecta can be grown in deep aquifer well water (No. 1) extracted in Sistan region The purpose of this study was to evaluate the potential of deep aquifer well water located in Sistan region (southeastern of Iran) for growing Dunaliella tertiolecta. For this purpose, 5 treatments including deep aquifer well water with salinities of 20 and 35ppt, deep aquifer well water and salt of Urmia Lake (35 ppt) and tap water with salinities of 20 and 35ppt were considered and D. tertiolecta microalgae was reared for 24 days under standard conditions (temperature 26±1ºC, light 2000 lux, 16 hours light and 8 hours darkness). The results showed that the pH value in all treatments increased slightly until day 6 but then decreased and had a steady trend from day 12 to 24. The highest amount of growth, chlorophyll a and carotenoid production, specific growth rate, the least doubling time and the highest rate of division were observed in deep aquifer well water treatment (salinity 20 ppt), although the lowest amount of biomass production was in this treatment. Based on these results, it can be said that if salt is added to deep aquifer well water, by changing the composition and increasing salinity, the amount of biomass produced will increase. In general, D. tertiolecta can be grown in deep aquifer well water (No. 1) extracted in Sistan region

https://japb.guilan.ac.ir/article_6834_cb4f77c9372dc168ad24098c0d7af7f0.pdf

University of Guilan Aquatic Physiology and Biotechnology 2345-3966 11 1 2023 07 13 CYP1A gene expression and antioxidant enzymes in green scat (Scatophagus argus) as PAHs biomarker CYP1A gene expression and antioxidant enzymes in green scat (Scatophagus argus) as PAHs biomarker 41 57 6857 10.22124/japb.2022.21777.1458 FA Seyed Ahmad Qasemi Assistant Professor in Department of Biotechnology, Persian Gulf Institute, Persian Gulf University, Bushehr, Iran Sepideh Hosein Janzade M.Sc. Student in Fisheries, Department of Fisheries, Faculty of Fisheries and Environmental Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran Journal Article 2022 02 14 Monitoring of the pollutant effects in the biological environment requires the identification of a suitable biomarker. In this study, the efficacy of the expression of the CYP1 gene and the antioxidant enzymes in the Scatophagus argus were studied as biomarker of oil pollutants at three stations (Bandargah, Solhabad and Jofreh) and the control samples. CYP1A gene expression in liver was significantly different from control samples in all regions. The highest CYP1 gene expression was measured in Solhabad station. CYP1A gene expression in the gills of green scat fish showed a significant increase compared to control samples. The highest and lowest CYP1A gene expression was observed in the gills in Solhabad and Bandargah samples, respectively. The amount of superoxide dismutase and catalase of liver tissue in Jofrah and Solhabad stations were significantly lower than the control (P<0.05). In contrast, the rate of lipid peroxidation and GST were directly correlated with the amount of polycyclic aromatic hydrocarbons (P<0.05, R=0.96). Due to the relationship between CYP1A gene expression and antioxidant markers and the level of contamination, CYP1A gene expression and antioxidant markers can be used as a suitable biomarker for PAH contaminants in this species. Monitoring of the pollutant effects in the biological environment requires the identification of a suitable biomarker. In this study, the efficacy of the expression of the CYP1 gene and the antioxidant enzymes in the Scatophagus argus were studied as biomarker of oil pollutants at three stations (Bandargah, Solhabad and Jofreh) and the control samples. CYP1A gene expression in liver was significantly different from control samples in all regions. The highest CYP1 gene expression was measured in Solhabad station. CYP1A gene expression in the gills of green scat fish showed a significant increase compared to control samples. The highest and lowest CYP1A gene expression was observed in the gills in Solhabad and Bandargah samples, respectively. The amount of superoxide dismutase and catalase of liver tissue in Jofrah and Solhabad stations were significantly lower than the control (P<0.05). In contrast, the rate of lipid peroxidation and GST were directly correlated with the amount of polycyclic aromatic hydrocarbons (P<0.05, R=0.96). Due to the relationship between CYP1A gene expression and antioxidant markers and the level of contamination, CYP1A gene expression and antioxidant markers can be used as a suitable biomarker for PAH contaminants in this species.

https://japb.guilan.ac.ir/article_6857_f9aa1ff40691891321ca0836aac5249b.pdf

University of Guilan Aquatic Physiology and Biotechnology 2345-3966 11 1 2023 06 21 Cloning, expression and purification of a novel carboxypeptidase from a hypersaline lake halophilic Bacillus persicus Cloning, expression and purification of a novel carboxypeptidase from a hypersaline lake halophilic Bacillus persicus 59 75 6851 10.22124/japb.2022.22750.1479 FA Samira Sepehri M.Sc. Student of Biochemistry, Faculty of Science, University of Guilan, Rasht, Iran محمود رضا آقا معالی Associate Professor in Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran Hossein Ghafoori Associate Professor in Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran Sajjad Sarikhan Scientific Member in Molecular Bank, Iranian Biological Resource Center (IBRC), Academic Center for Education, Culture and Research (ACECR), Tehran, Iran Journal Article 2022 08 25 Microbial proteases have a large portion of the market of industrial enzymes, because of their wide application in detergents, drug enzymes and animal feed processing. Bacillus strains potentially produce a significant quantity of proteases, due to their physiological properties. In this study, first, the gene encoding carboxypeptidase from Bacillus persicus was isolated by PCR and using primers with NdeI and XhoI restriction sites. After digestion of the PCR product, it was cloned into the appropriate site at PET28a+ vector, then recombinant plasmids transformed into E. coli BL21)DE3( strain. Plasmids with positive colony PCR result were sent for sequencing for final confirmation, then, recombinant enzyme expression was optimized at different temperature and time conditions, then purified with nickel embedded agarose affinity chromatography. IPTG concentration of 0.4mM at 32°C and incubation time of 20 hours provided the maximum amount of enzyme expression. Molecular weight of the carboxypeptidase was estimated approximately 58KDa. Considering the high amount of expression of the soluble protein and the easy optimization of its expression and purification in laboratory conditions, if the enzyme activity in the presence of standard substrates is adequate, and also if additional studies show the possibility of optimal and cost-effective production of a high amount of this enzyme in a large volume and pilot conditions, it can be hoped that this enzyme is a suitable option for industrial purposes. Microbial proteases have a large portion of the market of industrial enzymes, because of their wide application in detergents, drug enzymes and animal feed processing. Bacillus strains potentially produce a significant quantity of proteases, due to their physiological properties. In this study, first, the gene encoding carboxypeptidase from Bacillus persicus was isolated by PCR and using primers with NdeI and XhoI restriction sites. After digestion of the PCR product, it was cloned into the appropriate site at PET28a+ vector, then recombinant plasmids transformed into E. coli BL21)DE3( strain. Plasmids with positive colony PCR result were sent for sequencing for final confirmation, then, recombinant enzyme expression was optimized at different temperature and time conditions, then purified with nickel embedded agarose affinity chromatography. IPTG concentration of 0.4mM at 32°C and incubation time of 20 hours provided the maximum amount of enzyme expression. Molecular weight of the carboxypeptidase was estimated approximately 58KDa. Considering the high amount of expression of the soluble protein and the easy optimization of its expression and purification in laboratory conditions, if the enzyme activity in the presence of standard substrates is adequate, and also if additional studies show the possibility of optimal and cost-effective production of a high amount of this enzyme in a large volume and pilot conditions, it can be hoped that this enzyme is a suitable option for industrial purposes.

https://japb.guilan.ac.ir/article_6851_cef97c3e2f7c438c0fd36946fdad4f0b.pdf

University of Guilan Aquatic Physiology and Biotechnology 2345-3966 11 1 2023 06 21 Effects of dietary butyrate monoglyceride and probiotic of Bacillus on growth performance and enzymatic antioxidant activity of Nile tilapia (Oreochromis niloticus) Effects of dietary butyrate monoglyceride and probiotic of Bacillus on growth performance and enzymatic antioxidant activity of Nile tilapia (Oreochromis niloticus) 77 103 6852 10.22124/japb.2022.22091.1461 FA Reza Bagherpoor M.Sc. in Aquatic Breeding, Department of Fisheries, Faculty of Nano and Bioscience and Technology, Persian Gulf University, Bushehr, Iran Gholamreza Badzohreh Assistant Professor in Department of Fisheries, Faculty of Nano and Bioscience and Technology, Persian Gulf University, Bushehr, Iran Reza Davoodi Assistant Professor in Department of Fisheries, Faculty of Nano and Bioscience and Technology, Persian Gulf University, Bushehr, Iran Ahmad Qasemi Assistant Professor in Department of Fisheries and Biology, Persian Gulf Research Institute, Persian Gulf University, Bushehr, Iran Journal Article 2022 04 08 In this study, the effect of dietary butyrate monoglyceride and probiotic (Bacillus subtilis and Bacillus licheniformis) supplementation were evaluated on growth performance and the enzymatic antioxidant activity of Nile tilapia (Oreochromis niloticus). Fish were divided into 4 treatments with 3 replications including control, 10g/kg butyrate monoglyceride (AB1), 5g/kg probiotic Bacillus (AB2), 10g/kg butyrate monoglyceride and 5g/kg probiotic Bacillus (AB3). 144 fish with an average weight of 12±0.58g were fed for 8 weeks. This study showed that applying butyrate monoglyceride and probiotic Bacillus in the feeding of Nile tilapia significantly improved the growth indices of final weight gain (FWG), daily weight gain (DWG), protein efficiency ratio (PER), and FCR compared to control (P<0.05). All experimental diets had higher levels of catalase, glutathione peroxidase and superoxide dismutase activity compared to control (P<0.05). However, the highest levels of superoxide dismutase activity were measured in treatment AB3. The present study indicated that singular and combined dietary administration of butyrate monoglyceride and probiotic Bacillus had a significant impact on growth performance, and liver and serum enzymatic antioxidant activity of Nile tilapia. In this study, the effect of dietary butyrate monoglyceride and probiotic (Bacillus subtilis and Bacillus licheniformis) supplementation were evaluated on growth performance and the enzymatic antioxidant activity of Nile tilapia (Oreochromis niloticus). Fish were divided into 4 treatments with 3 replications including control, 10g/kg butyrate monoglyceride (AB1), 5g/kg probiotic Bacillus (AB2), 10g/kg butyrate monoglyceride and 5g/kg probiotic Bacillus (AB3). 144 fish with an average weight of 12±0.58g were fed for 8 weeks. This study showed that applying butyrate monoglyceride and probiotic Bacillus in the feeding of Nile tilapia significantly improved the growth indices of final weight gain (FWG), daily weight gain (DWG), protein efficiency ratio (PER), and FCR compared to control (P<0.05). All experimental diets had higher levels of catalase, glutathione peroxidase and superoxide dismutase activity compared to control (P<0.05). However, the highest levels of superoxide dismutase activity were measured in treatment AB3. The present study indicated that singular and combined dietary administration of butyrate monoglyceride and probiotic Bacillus had a significant impact on growth performance, and liver and serum enzymatic antioxidant activity of Nile tilapia.

https://japb.guilan.ac.ir/article_6852_064472ec85f60999c7d06b987486cfbf.pdf

University of Guilan Aquatic Physiology and Biotechnology 2345-3966 11 1 2023 06 21 The effect of probiotic bacteria Bacillus subtilis and Lactobacillus plantarum on fungal control and egg hatching performance of rainbow trout (Oncorhynchus mykiss) The effect of probiotic bacteria Bacillus subtilis and Lactobacillus plantarum on fungal control and egg hatching performance of rainbow trout (Oncorhynchus mykiss) 105 125 6853 10.22124/japb.2022.22297.1465 FA Omid Rahmati Associate Professor in Department of Fisheries, Faculty of Natural Resources, Behbahan Khatam Alanbia University of Technology, Behbahan, Iran Saeid Aeyaeinejad Associate Professor in Department of Fisheries, Faculty of Natural Resources, Behbahan Khatam Alanbia University of Technology, Behbahan, Iran Ali Abromand Associate Professor in Department of Fisheries, Faculty of Natural Resources, Behbahan Khatam Alanbia University of Technology, Behbahan, Iran Esmaeil Kazemi Ph.D. in Fisheries, Fisheries Research Institute of Iran, Yasouj Shahid Motahari Clod-water Fish Genetics and Breeding Research Center, Fisheries Research Institute of Iran, Yasouj, Iran Journal Article 2022 05 15 In the fish breeding industry, many disinfectants are used to prevent fungal contamination of eggs during the incubation period, including formalin, hydrogen peroxide, potassium permanganate, etc. The purpose of this research is to evaluate the effect of probiotic bacteria Bacillus subtilis 1 and 2 grams and Lactobacillus plantarum 1 and 2 grams (adding to water) in controlling the fungal contamination of rainbow trout eggs during the incubation stage and comparing its effects with formalin and It was distilled water. To conduct this study, 18 Californian troughs were used in 6 treatments and three repetitions, and 1850 fertilized rainbow trout eggs were transferred to each trough. After preparing the trays and distributing the eggs, a separate serum was used for each treatment to disinfect the eggs. The duration of complete emptying of each serum was 50 minutes. The results showed that in the treatments containing formalin (10±1) followed by Lactobacillus 2 grams (15.33±3.21), the lowest rate of fungal infection of eggs was observed, which showed a significant difference with other treatments. P>0.05). The highest rate of eye opening (84.67±0.58) and egg opening (81±1) was observed in formalin treatment, which was significantly different from other treatments (P<0.05) In the fish breeding industry, many disinfectants are used to prevent fungal contamination of eggs during the incubation period, including formalin, hydrogen peroxide, potassium permanganate, etc. The purpose of this research is to evaluate the effect of probiotic bacteria Bacillus subtilis 1 and 2 grams and Lactobacillus plantarum 1 and 2 grams (adding to water) in controlling the fungal contamination of rainbow trout eggs during the incubation stage and comparing its effects with formalin and It was distilled water. To conduct this study, 18 Californian troughs were used in 6 treatments and three repetitions, and 1850 fertilized rainbow trout eggs were transferred to each trough. After preparing the trays and distributing the eggs, a separate serum was used for each treatment to disinfect the eggs. The duration of complete emptying of each serum was 50 minutes. The results showed that in the treatments containing formalin (10±1) followed by Lactobacillus 2 grams (15.33±3.21), the lowest rate of fungal infection of eggs was observed, which showed a significant difference with other treatments. P>0.05). The highest rate of eye opening (84.67±0.58) and egg opening (81±1) was observed in formalin treatment, which was significantly different from other treatments (P<0.05)

https://japb.guilan.ac.ir/article_6853_bda851462ca909f2feba6923ebcffa2f.pdf