ALBERT

Description: In this study lusiferse gene extracted from Vibrio fischeri. V. fischeri strain was kindly provided by Iranian Research Organization for Science and Technology (IROST). Genomic DNA extraction was carried out by Phenol-chloroform. A DNA fragment encoding the luxA and LuxB was amplified by PCR using sense and antisense primers, specific restriction sites for BamH1 and Kpn1 were introduced into 5´ end of forward and reverse primer, respectively. The PCR product was purified from agarose gel and ligated into cleaved PT257R cloning vector. Following the confirmation of the cloned Luciferase transformed into E. coli. Recombinant clones were confirmed by specific PCR and restriction enzyme digestion analysis. The luxA and LuxB fragment was released subcloned in to the PcDNA3.1\hug and PcDNA3.1\neo expression vectors, respectively. recombinant plasmid was confirmed through restriction digestion using BamH1 and Kpn1 enzymes and subsequently, transformation procedure continued into NIH3T3 eukaryote cells by specific kit. Luminescence ability of recombinant clones was tested by NIH3T3 cells and dechanal (substrate) and Neomysin and hygromysin. The results showed that luminescence start after 2 hours and then increase after 6 hours. Inaddition, the protein identity was verified by western blot analysis, the protein bands 76 kD were detected. , which indicates protein expression of luxB, luxA.

Description: Iranian Fisheries Science Research Institute

Description: Published

Description: Limnologicalstudyof SabalanrunoffDamfor aquaculturecold water fish was conducted in sixtimesat fourstationsfrom JulytoDecember2009.Averageminimumandmaximumparametersmeasuredindifferentstationswere included water temperature, dissolved Oxygen, total hardness, electrical conductivity, pH, BOD5, COD,TDS, NH4, NO2, NO3, PO4, TN, TP, SiO2, respectively (10/3 ± 9/5-14 ± 8/7° C), (9 ± 1/2-11/4 ±2/1 mg/liter), (367 ± 19/8-381 ± 17/8mg/liter), (0/95 ± 0/07-1/05 ± 0/07 mz/cm), (8/3 ± 0/1-8/4 ± 0/1), (3 / 3 ± 1/14-3/88 ± 0/71mg/ liter), (15/5 ± 3/1-16/3 ± 3 mg/liter), (0/48 ± 0/04-0/54 ± 0/06mg/ liter), (0/08 ± 0/02-0/13 ± 0/07 mg/liter), (0/004 ± 0/003- 0/05 ± 0/1 mg/ liter), (0/6 ± 0/2-0/64 ± 0/29 mg/ liter), (0/19 ± 0/07-0/22 ± 0/07 mg/ liter), (10/48 ± 0/6-10/98 ± 0/54mg/liter), (0/39 ± 0/11-0/44 ± 0/18 mg/liter), and(9/9 ±0/6-11/1 ± 1/4 mg/liter). Averageminimumandmaximumof water heavy metalswas included of Cd, Pb, Zn, Fe, Cuand Ni respectively was measured as ND(No Deliver), ND, ND- 0/242, ND- 1/075, ND- 0/018andND- 0/014inmg/ liter. Alsoamong the16organochlorinecompoundsmeasuredthehighestamount of was belonged totoxinHexachlorine Benzene (1/5 mg/ liter). The results of theanalysis ofwater quality parametersshowed that parametersexcept ofCOD, TDS, PO4, TP, Zn, Fe andwater temperature, otherparametersdo not haveany restrictions foraquaculturecold waterfishin therunoffDam. Based on data results it was noticed that there was temperature restriction up to November at downstream and station 2 showed less temperature fluctuation and more suitable condition compared to other stations. BOD5 value was within the allowable range for growing trout, in which station 2 somewhat with lower volatility of value had the most suitable BOD5 condition.In the present study, the amount of TDS in the study area was somewhat higher than the limitation and since TDS values increased with distance from the dam, therefore station 2 had somewhat better conditions for fish farming.In addition the amount of EC in the study area, indicating the inappropriateness of it for various activities in the fisheries.Although none of the stations in terms of the amount of phosphorus didn't show favorable conditions for rainbow trout fish propagation, it appears between the researches stations, station 2 had the most favorable terms.The results of this study (regardless of the unseen), it seems that in August and September; Zn had overestimated value for cold fish propagation. Recent study showed that among heavy metals, Fe in water, sediments and fish was over limitation value. Also, Fe value was measured of 1.08 at station 4, which showed over normal value for cold fish propagation. Therefore, station 2 at downstream seemed to have suitable cold fish propagation due to lower Fe value. On the based on the recent study, planktonic biological index did not endorse water quality for fish propagation. In this study, aquatic plant such as Potamogetonpectinatus and Pheragmitiesaustraliswas found at water of higher and somehow polluted organic matter. Probably, based on the results it was noticed that downstream water of dam was polluted to organic matter. At the recent study, resisted maroinvertebrates to organic matter (Chironomidae) was consisted noticeable frequency compared to other sensitive invertebrates to organic matter (EPT).Statistically, it can be expressed that probably organic pollution resulted to increase of resistance invertebrates and decreased of sensitive groups. In recent study, EPT/CHIR index was measured 0.6 in station 2 and 1.74 in station 4. This ration was less to many other mountain rivers. As ETP sensitive invertebrates has higher sensitive to other groups with the regards of environmental condition and pollution, unpredicted increase of Chironomidae compare to sensitive groups resulted of EPT/CHIR value which showed environmental stress. In this study, HFBI index was measured as 7.49 at station 2 and 7 in station4. Based on HFBI index, station 2 was classified as a high organic pollution and station 4 with noticeable organic pollution. At the recent study, station 2 was catch more fish to station4. For instance, Squaliuscephalus and Alburnoides bpinctatuswere catch in station 2 and were not found in station 4. As a whole, Biologicalindicators whichwas includedPlanktonBioindicator, EPT / CHIR. ,HilsenhoffBiologicalIndicator, diversityanddistribution ofaquaticplantsandfish, showed water situation inrunoffDamrich inorganic matter. Finally, thestudy areawasapprovedfor fish farmabout 50tons and needs managementarrangementswiththeincreasedproduction rate.

Description: Iranian Fisheries Science Research Institute

Description: Published

Description: Mazandaran and Gorgan provinces have temperate climate, thus they have more potential for aquatic animal culture. There are thousands hectare of salt and useless lands in adjacent to Caspian Sea. As these areas have provided a favorable back ground for aquatic animal culture. As a result, the successful results obtained from imported gray mullet (Mugil cephalus) culture project in north climate, it has demonstrated that the gray mullet has a good biocharacteristic for culturing in pond enclosure environment and in different aquatic conditions (fresh water, brackish water and salt water). From 1998 until 2001, the broodstock yield and gray mullet artificial propagation projects were performed by fisheries research center of Mazandaran in Ghomishan prawn culture station in adjacent to Caspian Sea (East north of Mazandaran Province). This investigation executed during two stages (phases). At first stage, the goal of this project included the survey of possibility available about matured fish as well as induction of final maturation and artificial propagation for producing of broodstock and larva. In addition, in this way, we will obtain new information about gray mullet propagation and culture as marine species. We introduce mass production in aquaculture. At present research, the possibility of broodstock yield and artificial propagation of gray mullet have investigated by gray mullet fingerlings imported from Hong kong and then they have cultured in earth ponds of Ghomishan areas during five years (1994-1998). In order to broodstock production in spring 1998, two earth ponds (0.5 hectare) were prepared. 100 specimens of fish stock (1-2.5 kg weight average and 5 years of age) placed in each pond. For providing of suitable water and nutrition, fish were fed by food containing rich protein (40%) with 3-5% body weight and maintained in water with 30-35 ppt salinity. The survey of sexual maturation was performed by sampling of sexual glands through year. There were four stages in dominant female broodstocks. This survey indicated that oocytes have emerged stage 1 (immature) from March to June, stage 2 (yolk vesicle) in September and stage 4 (yolk globule) in October. Three stages (first, second and third) of yolk formation in oocytes will occur but these stages take place in October, December and January respectively. It's obvious that oocytes will progress into the end of third stage (yolk formation) and then their growth was arrested. Ovum with 600 m diameter was observed when the water temperature declined less than 18 c and day time was short (from middle autumn to middle winter), on that time, fish were induced by hormone because lack of final maturation and ovulation, there fore, natural spawning was not occurred in pond condition. Furthermore, artificial propagation of mugil cephalus was occurred by hormonal induction. Hormonal induction was utilized by inject of many hormones (LHRH, Cph, HCG). Using different components of hormones and also their different doses obtained the best results from broodstock that the average ovum diameter was about 600 m. Several types hormones which were injected into gray mullet with two or several intermittent (24 h intervals) along with 20-25 c temperature and 30-35 ppt salinity. This condition can provide stage 4 maturity for fertilization. Totally (as whole), three intermittent fertilization was necessary for exiting of larva, larva production in first, second and third intermittents that were 2000, 2500 and 300 specimens respectively. Larva fed on chlorella algae and rotifera and they have maintained for 14-15 days. The results of obtainable research indicated the possibility of gray mullet broodstock production in cultural condition, artificial propagation and larva production. This research took place for the first time in north climate of Iran.

Description: Iranian Fisheries Science Research Institute

Description: Published

Description: In a research project during 1999-2000, transferrin polymorphism in blood serum as well as several morphometric and meristic characteristics were determined in 7 groups of carps of different fish farms. Transferrin polymorphism in carp is expressed by the occurrence of one or two band phenotypes determined by a series of codominant alleles. In this study, there was significant difference especially between sea and cultured carp (P〈0.01). Sampling of sea carp in general have been from race of carp with BB dominant genotype, but fish sample from shahid rajaii fish farm have shown variety genotype including AA, AB, BB, AC, BC and CC. In morphological investigation, the C.V. of meristic characteristics, which is under genetic bases such as number of fin rays and number of scales on lateral line was higher significantly in those fishes caught from the sea than cultured fishes (P〈0.00). Relative comparative of morphometric characteristics such as head length to standard length and width of body showed also significant difference between those two groups (P〈0.05).

Description: Published

Description: In this study population genetic structure of Mnemiopsis leidyi were examined by 200 samples from the Caspian Sea. Samples were collected from Guilan province (Astara region), Mazandaran province (Amir Abad region), Golestan province (Gomishan region) and north part of the Caspian Sea. 50 samples were colected from each region. DNA was extracted by phenol - chlorophorm method and its concentration was 50 to 100 nanogram . PCR was performed using 18s rRNA gene. The PCR products of samples were digested by 12 restriction endonuclease enzymes. The digested products accompanied with standard marker (50 bp lader ). In order to measure fragment size samples were run on a 6% vertical polyacrylamide gel. The fragments were visualized by silver staining of the polyacrylamide gel. In this study, Dra I and Alu I enzymes showed different digestion pattern, as each of these enzymes had two genotypes. Other enzymes had similar digestion pattern. RAPD method used by 19 random primers. Ten of nineteen primers showed polymorphism patterns. Statistical analysis of data was performed by Popgene software. In this study, the maximum of genetic variation was in north of the Caspian Sea. Also, the maximum of genetic distance was between north area and Golestan coasts (0.089) and the minimum of genetic distance was between Mazandaran and Guilan coasts (0.001). The UOGMA dendogram showed two clusters. The samples of Mazandaran , Guilan and Golestan coasts placed in one cluster and samples of north area in other cluster The genetic diversity was significantly different between samples of north area and Golestan coasts (p≤0.05). As result, there is a significant genetic divergence between some of samples. Therefore, two genetic group of Mnemiopsis leidyi were identified in the Caspian Sea.

Description: Iranian Fisheries Science Research Institute

Description: Published

Description: The project investigates the relationship between the biological parameters (phytoplankton, zooplankton, Macrobenthic and the comb jelly Mnemiopsis leidyi) and environmental parameters, nutrients and environmental pollutant (oil, pesticides, heavy metals, and detergents) in water and sediment, at the southern part of Caspian Sea in 2009-2010. Sampling was performed in four seasons (spring, summer, autumn and winter) and in eight transects perpendicular to the coast in Astara, Anzali, Sefidroud, Tonekabon, Noshahr, Babolsar, Amir Abad and Turkmen. Samples were collected from depths of 5, 10, 20, 50 and 100 meters and the different layers. The relationship between biological and environmental parameters surveyed through parametric and multivariate methods. Based on the result, the annual mean of environmental parameters and nutrients concentration at euphotic layer such as water temperature, salinity, pH, transparency, DO%, ammonium, nitrate, inorganic nitrogen (DIN), organic nitrogen (DON), inorganic phosphorus (DIP), organic phosphorus (DOP) and soluble silicon (DSi) were 17.82±0.32 (ºC), 11.35±0.10 (g/l), 8.43±0.01 (m), 126±1 (%), 3.76±0.19 (µM), 1.55±0.07 (µM), 4.81±0.13 (µM), 29.88±0.66 (µM), 0.24±0.01 (µM), 0.62±0.02 (µM), 8.22±0.22 (µM), respectively. Meanwhile, annual mean of environmental pollutant such as TPH, OCPs and LAS in water were 12.33±1.76 (µg/l), 2.85.0.90 (µg/l), 0.048±0.003 (mg/l), respectively and for TPH and OCPS in sediment were recorded 33.07±9.36 (µg/g) and 2.64±0.34 (µg/g), respectively. In addition, annual mean of heavy metals such as Ni, Hg, Pb and Cd at water were 4.01±0.01 (µg/l), 0.64±0.01 (µg/l), 15.0±0.1 (µg/l) and ND respectively, and in sediment were obtained 43.77±3.55 (µg/g), 0.13±0.29 (µg/g), 14.14±1.07 (µg/g) and 0.07±0.06 (µg/g), respectively. Abundance annual mean of biological parameters such as phytoplankton, zooplankton and M. leidyi were 143±12 (million cells/m3), 6548±700 (individuals/m3) and 86±10 (individuals /m3) respectively, and for biomass were 548±41 (mg/m3), 60±9 (mg/m3), 5.06±0.65 (g/m3). Abundance and biomass annual mean of macrobenthic were 5970±460 (individuals /m2) and 44±10 (g/m2), respectively. The stratification of water column was strongly based on gradient of water temperature and the rule of salinity was low in this area. According to mono and multivariate statistical analysis, the southern of Caspian Sea experienced four seasons without any overlapping based on environmental parameters and nutrients. In 2009-10, the nitrogen concentration was higher than previous years and showed a decrease in the amount of inorganic phosphorus in the study area. Therefore, the system has been lead to phosphorus limitation. No limitations of the DSi in the Caspian ecosystem had caused the dominance of bacillriophyta phylum during spring, autumn and winter. However in summer (highest seasonal mean temperature) which held the most DIP, the cyanophyta was dominant phylum. During spring and winter, the abundance of zooplankton was maximized and conversely the minimum values observed during summer and autumn with growth increasing of M. leidyi. By now, M.leidyi has been preferred the time with highest temperature or salinity during year. However with considering PCA analysis result and high ability of the organism to adaptation, the M.leidyi will probably to growth and reproduce at lower temperature and salinity than now in this area. Result showed that during summer and autumn only abundance of two of six main zooplankton groups namely Copepoda and Cirripedia were noticeable. The other zooplankton groups were low due to high consumption by different predators or were influenced by environmental factors. The dynamics population of M.leidiy is used to consider as first explanation of the Caspian ecosystem happening, however the interaction of different factors such as temperature, salinity, nutrients and predators determine the dynamic of biological parameters. Based on statistical analysis, feeding behavior of zooplankton species was not same in different seasons and was strongly dependent on the phytoplankton structure (species diversity and population). In other words, the classical boundary of phytoplankton species to, edible and suppressed, non-edible and unaffected and finally non-edible, enhanced species was not possible. Copepoda can act as filter-feeder and raptorial-feeder. So Copepoda compare to Cladocera (generally filter- feeder and sometimes raptorial feeder) and Rotifera (mostly suspension feeding and sometimes raptorial feeder) had more chances and opportunities for the achievement food and it support them for dominanane in all seasons. Anthropogenic and eutrophication affect on the structure and function of plankton community and it reflected on ratio of phyto/zoo biomass. Seasonal increasing of different groups of zooplankton were done through their life cycles in 2009-10, but mostly phytoplankton abundance increased several folds than zooplankton populations. This trend made the ratio of phyto/zoo biomass at 10, 4, and14 during spring, summer and autumn, respectively. In summer, with stratification of water column and dominant of Cyanophyta, grazing pressure by herbivores zooplankton was low. However in winter due to the zooplankton population growth, the ratios reached to 11, and declined with 1.3 folds compare to autumn. Because of severe decreasing of zooplankton biomass in 2009-10 compare to years before introduction of M.leidiy in to the Caspian Sea, the ratio of phyto/zoo biomass increased from "less than" 5 "to more than 10" Top-down control of zooplankton grazing on phytoplankton populations was clear in the years before the introduction of M.leidyi in to the Caspian Sea. But in 2009-10 zooplankton Shannon index reduced and phytoplankton Shannon index increased. In other word higher functional diversity at the lower trophic level (phytoplankton) increased the probability of successful defense against top-down control from zooplankton and phytoplankton abundance controled mainly by nutrients materials (down-top control). Seasonal macrobenthos abundance showed the maximum values in summer and minimum in the cold season (winter). The polychaeta had the highest proportion of abundance compare to others main orders (oligochaeta and crustacea). The dominance of deposit feeder species of polychaeta indicates to high level of organic matter in sediment and trophic status of ecosystem. Result of CCA analysis showed that three dominant orders had different behavior respect to environmental and sediments characteristics. The oligochaeta order was directly related to sediments characteristics (TOM) and inverse relationship with environmental parameters at most seasons. In contrast, polychaeta order was linked with environmental parameters in most seasons.Crustacea order was related to the both of them. Two orders of oligochaeta and crustacea were also related directly to OCPs and inverse relationship with TPH and heavy metals. In the other words, the two orders were more affected by TPH and heavy metals in this area. As a last point, increasing of trophic level from oligotrophic to the meso–eutrophic status, an increase of DO% from 105 percent to 120 percent, phytoplankton Shannon index increasing, zooplankton Shannon index decreasing, entrance of the potentially invasive species to the list of dominant phytoplankton species, increasing of phyto/zooplankton biomass ratio from less than 5 to more than 10 and also increasing of deposit feeder species abundance of macrobenthos are some evidences that indicate to disturbance and stress condition of the Caspian Sea.

Description: Iranian Fisheries Science Research Institute

Description: Published

Description: In this study population genetic structure of Mnemiopsis leidyi were examined by 200 samples from the Caspian Sea. Samples were collected from Guilan province (Astara region), Mazandaran province (Amir Abad region), Golestan province (Gomishan region) and north part of the Caspian Sea. 50 samples were collected from each region. DNA was extracted by phenol - chlorophorm method and its concentration was 50 to 100 nanogram. PCR was performed using 18s rRNA gene. The PCR products of samples were digested by 12 restriction endonuclease enzymes. The digested products accompanied with standard marker (50 bp lader ). In order to measure fragment size samples were run on a 6% vertical polyacrylamide gel. The fragments were visualized by silver staining of the polyacrylamide gel. In this study, Dra I and Alu I enzymes showed different digestion pattern, as each of these enzymes had two genotypes. Other enzymes had similar digestion pattern. RAPD method used by 19 random primers. Ten of nineteen primers showed polymorphism patterns. Statistical analysis of data was performed by Popgene software. In this study, the maximum of genetic variation was in north of the Caspian Sea. Also, the maximum of genetic distance was between north area and Golestan coasts (0.089) and the minimum of genetic distance was between Mazandaran and Guilan coasts (0.001). The UOGMA dendogram showed two clusters. The samples of Mazandaran , Guilan and Golestan coasts placed in one cluster and samples of north area in other cluster The genetic diversity was significantly different between samples of north area and Golestan coasts (p≤0.05). As result, there is a significant genetic divergence between some of samples. Therefore, two genetic group of Mnemiopsis leidyi were identified in the Caspian Sea.

Description: In this study lusiferse gene extracted from Vibrio fischeri. V. fischeri strain was kindly provided by Iranian Research Organization for Science and Technology (IROST). Genomic DNA extraction was carried out by Phenol-chloroform. A DNA fragment encoding the luxA and LuxB was amplified by PCR using sense and antisense primers, specific restriction sites for BamH1 and Kpn1 were introduced into 5´ end of forward and reverse primer, respectively. The PCR product was purified from agarose gel and ligated into cleaved PT257R cloning vector. Following the confirmation of the cloned Luciferase transformed into E. coli. Recombinant clones were confirmed by specific PCR and restriction enzyme digestion analysis. The luxA and LuxB fragment was released subcloned in to the PcDNA3.1\hug and PcDNA3.1\neo expression vectors, respectively. recombinant plasmid was confirmed through restriction digestion using BamH1 and Kpn1 enzymes and subsequently, transformation procedure continued into NIH3T3 eukaryote cells by specific kit. Luminescence ability of recombinant clones was tested by NIH3T3 cells and dechanal (substrate) and Neomysin and hygromysin. The results showed that luminescence start after 2 hours and then increase after 6 hours. Inaddition, the protein identity was verified by western blot analysis, the protein bands 76 kD were detected which indicates protein expression of luxB, luxA.

Description: In a research project during 1999-2000, transferrin polymorphism in blood serum as well as several morphometric and meristic characteristics were determined in 7 groups of carps of different fish farms. Transferrin polymorphism in carp is expressed by the occurrence of one or two band phenotypes determined by a series of codominant alleles. In this study, there was significant difference especially between sea and cultured carp (P〈0.01). Sampling of sea carp in general have been from race of carp with BB dominant genotype, but fish sample from shahid rajaii fish farm have shown variety genotype including AA, AB, BB, AC, BC and CC. In morphological investigation, the C.V. of meristic characteristics, which is under genetic bases such as number of fin rays and number of scales on lateral line was higher significantly in those fishes caught from the sea than cultured fishes (P〈0.00). Relative comparative of morphometric characteristics such as head length to standard length and width of body showed also significant difference between those two groups (P〈0.05).

Description: Mazandaran and Gorgan provinces have temperate climate, thus they have more potential for aquatic animal culture. There are thousands hectare of salt and useless lands in adjacent to Caspian Sea. As these areas have provided a favorable back ground for aquatic animal culture. As a result, the successful results obtained from imported gray mullet (Mugil cephalus) culture project in north climate, it has demonstrated that the gray mullet has a good biocharacteristic for culturing in pond enclosure environment and in different aquatic conditions (fresh water, brackish water and salt water). From 1998 until 2001, the broodstock yield and gray mullet artificial propagation projects were performed by fisheries research center of Mazandaran in Ghomishan prawn culture station in adjacent to Caspian Sea (East north of Mazandaran Province). This investigation executed during two stages (phases). At first stage, the goal of this project included the survey of possibility available about matured fish as well as induction of final maturation and artificial propagation for producing of broodstock and larva. In addition, in this way, we will obtain new information about gray mullet propagation and culture as marine species. We introduce mass production in aquaculture. At present research, the possibility of broodstock yield and artificial propagation of gray mullet have investigated by gray mullet fingerlings imported from Hong kong and then they have cultured in earth ponds of Ghomishan areas during five years (1994-1998). In order to broodstock production in spring 1998, two earth ponds (0.5 hectare) were prepared. 100 specimens of fish stock (1-2.5 kg weight average and 5 years of age) placed in each pond. For providing of suitable water and nutrition, fish were fed by food containing rich protein (40%) with 3-5% body weight and maintained in water with 30-35 ppt salinity. The survey of sexual maturation was performed by sampling of sexual glands through year. There were four stages in dominant female broodstocks. This survey indicated that oocytes have emerged stage 1 (immature) from March to June, stage 2 (yolk vesicle) in September and stage 4 (yolk globule) in October. Three stages (first, second and third) of yolk formation in oocytes will occur but these stages take place in October, December and January respectively. It's obvious that oocytes will progress into the end of third stage (yolk formation) and then their growth was arrested. Ovum with 600 m diameter was observed when the water temperature declined less than 18 c and day time was short (from middle autumn to middle winter), on that time, fish were induced by hormone because lack of final maturation and ovulation, therefore, natural spawning was not occurred in pond condition. Furthermore, artificial propagation of mugil cephalus was occurred by hormonal induction. Hormonal induction was utilized by inject of many hormones (LHRH, Cph, HCG). Using different components of hormones and also their different doses obtained the best results from broodstock that the average ovum diameter was about 600 m. Several types hormones which were injected into gray mullet with two or several intermittent (24 h intervals) along with 20-25 c temperature and 30-35 ppt salinity. This condition can provide stage 4 maturity for fertilization. Totally (as whole), three intermittent fertilization was necessary for exiting of larva, larva production in first, second and third intermittents that were 2000, 2500 and 300 specimens respectively. Larva fed on chlorella algae and rotifera and they have maintained for 14-15 days. The results of obtainable research indicated the possibility of gray mullet broodstock production in cultural condition, artificial propagation and larva production. This research took place for the first time in north climate of Iran.