mohammad hassanzadeh saber

Fish population segregation through diagnostic morphometric traits has a great importance in fish conservation and fisheries management programs. In the current study, morphometric variations of 31 morphometric characteristics of Alosa braschnikowi was investigated in the Southern Caspian Sea in three provinces including Guilan, Mazandaran and Golestan, during winter 2021. The average standard length (±SD) was 231.73±4.38, 231.52±4.14 and 200.93±32 in Guilan, Mazandaran and Golestan, respectively. The results obtained from the one-way ANOVA showed a significant difference (p<0.05) in 17 out of 31 morphometric traits. The PCA showed that the investigated areas can be distinguished through nine components, so that the main changes in the first component (PC1) are in the head area while variations in the pectoral and dorsal fins were identified as important through the second component (PC2). Investigating the growth pattern of A. braschnikowi illustrated a negative allometric growth for Anzali (b=2.78) and Sari (b=2.61) regions while Miankaleh (b=3.12) had a positive allometric growth pattern. Furthermore, condition factors estimation revealed a better nutritional condition in Anzali (CF=1.10) and Miankaleh (CF=1.09) compared to Sari region (CF=0.71). Morphometric-based clustering showed that there are at least two separate populations of A. braschnikowi in the studied areas of the Southern Caspian Sea. Due to the evenness in sampling season and way of sampling, it seems that the environmental features in each region have caused the ensuing high morphological diversity in A. braschnikowi, however, investigations through molecular methods are needed for this claim. Considering the high rate of morphometric variations in Alosa genus in the Caspian Sea, the usage of machine-learning based analytical methods is highly recommended for future studies.

In this study, the sperm quality indicators of rainbow trout breeders in selected Specific Pathogen Free (SPF) farms of the country were investigated in order to plan the long-term preservation of rainbow trout sperm through cryopreservation and creating a sperm bank in the next phase. Sperm samples from 24 breeders from six selected SPF farms from West-Azerbaijan and Mazandaran provinces were collected and evaluated. The average density of sperm is 7.83 ± 4 billion per milliliter, the average volume of extracted sperm is 9.2 ± 1.6 ml, the average percentage of motility and its duration were 78.7 ± 10.3% and 26.6 ± 4.4 seconds, the average pH was 7.74 ± 0.23 and the fertilization percentage of the samples was as 92.8 ± 1.2%. The results showed that the pH 7.7-7.8 is the most appropriate value for inducing motility and its increasing or decreasing relative to this value causes a decrease in fertilization efficiency. Also, a positive correlation between pH and sperm motility (r = 0.533) and fertility (r = 0.67) was determined. According to the obtained information, the characteristics of the best sperm sample is selected for long-term storage and the creation of a genetically diverse sperm bank and targeted management of rainbow trout reproduction.

Identifying the species of sturgeons, the non-mixing of multi-species caviar in products made from sturgeons' caviar is one of the most important challenges in the legal and international trade of this product. One of the ways to identify the original caviar and correctly label caviar products is the use of barcodes and especially molecular barcode tracking.

In this research, fin tissue samples of Russian and Persian sturgeon, Ship, Sterlet, stellate and Siberian sturgeon, as well as their caviar samples, were used to investigate species identification markers of Caspian Sea sturgeons. DNA extraction was performed from three caviar eggs of each species and pure sturgeons fin tissue, and two types of primers were selected based on mitochondrial genes.

sturgeon's marker amplified a 138 new nucleotide fragment using PCR method in all species based on COI gene, but the species-specific marker amplified different sized fragments based on D-loop gene. These fragments were reproduced in both caviar and fin tissue of sturgeons. The overall results showed that the amplified fragment of the sturgeon's marker and the species-specific marker in pure sturgeons can indicate the pure fish species.

By broodstocking and propagating of specific pathogen-free (SPF) rainbow trout in Iran, the need to create a cryopreserved sperm bank from the breeders was deemed necessary. In this study, after evaluation of four different extenders for rainbow trout sperm cryopreservation, the best extender (0.6M sucrose and 8% DMSO) has been selected. The sperm samples were obtained from 24 breeders, including four breeders from each of the selected specific pathogen-free farms of the country in Mazandaran and West Azerbaijan provinces (Sarshar, Maroufi, Fakhari, Hadidi, Malekitabar, and Qorbani) were cryopreserved after qualitative evaluation. The fertilization test of sperm samples on the day of sampling and comparison of their fertilization percentage with frozen-thawed sperms in three time intervals (4, 24 and 48 hours) did not show any significant differences between the sperm of the control group and the frozen-thawed samples. The results indicated that the method used in this research can be well used in cryopreservation the sperm of SPF rainbow trout breeders and if needed, the sperm obtained from the best breeders can be transferred to other centers with this method.

In sturgeon, female-sex rearing is economical and gynogenesis induction is one of the ways to achieve it. In gynogenesis, an egg develops without the genetic influence of the male, or on the other words, it is a method of reproduction in which the offspring are formed exclusively from the genetic information of the mother. In this technique, the sperm used must be genetically inactivated. Rays such as X-ray, gamma (ionizing rays) or ultraviolet (UV) rays can be used to inactivate the sperm. Activation of embryonic development with genetically inactive sperm has resulted in the production of haploid eggs, which in order to diploidize must use physical shocks during the second meiotic division or the first mitotic division. In this case, all the offspring have maternal inheritance. It depends on the type of sex determination system, that amount of production of female offspring is determined. In sturgeons, the offspring sex determination is the responsibility of the mother, so by inducing gynogenesis, both males (lower ratio) and females (higher ratio) are produced. Accordingly, artificial gynogenesis could increase both the caviar production and prevent the extinction of these valuable species.

Using cryopreservation techniques are so important in conducting brood stock selection programs, species protection, and accessibility to the spermatozoa throughout the year. Normally, in the conventional method in Iran, the formulation based on egg yolk and DMSO has been employed to freeze the sturgeon sperm. In the present study, the MT extender (Modified Tsvetkova’s) which consists of the main components of methanol, sucrose, potassium chloride, and Tris has been used for the preservation of Acipenser stellatus spermatozoa, without using chemical substance DMSO and egg yolk. Our results demonstrated that the frozen sperm had at least 30% motility after one year of storage in nitrogen tank. The results of this research show that this extender could be used to preserve Acipenser stellatus spermatozoa. Since the sperm’s concentration of sturgeon species are similar, the usage of the MT extender can be suggested for other species of the sturgeons.

Siberian sturgeon (Acipenser baerii) grows rapidly in breeding ponds. Gynogenesis is a suitable technique for producing sturgeon female sex. The aim of this study was to establish female sex in Siberian sturgeon using induction of gynogenesis. The sperm of Persian sturgeon (Acipenser persicus) was genetically inactivated by UV radiation at a dose of 8514 J/m2. In this case, sperm with 40% motility were combined with Siberian sturgeon oocytes and in 2°C cold shock for 30 minutes, 10 minutes after activity, gynogenic progeny were created. Fertilization rate was 44.5% (control hybrid 62%) and hatching rate was 19.8% (control hybrid 40.5%). Purely maternal heritability in gynogeneic progeny was confirmed using microsatellite markers. By achieving the biotechnique of sperm production with inactivated DNA of Persian sturgeon, it is possible to prevent the risk of extinction and protect other sturgeon by inducing gynogenesis.

This study aims to compare effects of sources levels of zinc on growth and carcass quality, some haematological and serological indices of juvenile beluga sturgeon, Huso huso (Linnaeus, 1758). A number of 315 juvenile beluga sturgeon (8.4±0.29) were fed in 21 500 lit fiberglass tanks (15 fish per tank) under 7 experimental diets including 1 control diet plus 6 diets containing zinc (mg per kg of diet) in two forms of mineral (zinc sulfate) (ZnSul15, ZnSul30, and ZnSul60 treatments) and organic (chelated with methionine) (ZnMet15, ZnMet30, and ZnMet60 treatments) with three replications per treatment for 12 weeks. The best values of final weight and feed conversion ratio were observed in the ZnMet15 and ZnMet60 treatments. Red blood cells, hemoglobin, and hematocrit showed an incremental trend influenced by increased zinc supplement. Increased carcass protein in the ZnMet30 treatment was significant compared to the control, ZnSul15, and, ZnSul30 treatments (P≤0.05). Reduced carcass lipid in the ZnSul30 and ZnSul60 treatments was significant compared to the other treatments (P≤0.05). Serum triglyceride and cholesterol decreased in the ZnMet treatments compared to the control treatment. Reduced serum glucose in the ZnMet15 and ZnMet60 treatments and also increased serum lysozyme in the ZnMet30 and ZnMet60 treatments were significant compared to the control treatment (P≤0.05). Results demonstrated that most of the indices had better performance in the ZnMet treatments comparted to the other treatments and are proposed as an optimal diet for juvenile beluga sturgeon.

The great sturgeon, Huso huso, is one of the most important cultured sturgeon species in Iran, which effective management of aquaculture production of this species requires knowledge of broodstock structure, mating patterns, and genetic diversity of broodstock. The aim of the present study was the application of microsatellite DNA analysis for genetic diversity assessment in the first generation of cultured great sturgeon farmed at two centers of the live gene bank at the International Sturgeon Research Institute and Shahid Dr. Beheshti center for restoration and conservation of genetic stocks of sturgeon in Guilan province. Fin clips were sampled from 147 spawners and pre-spawners of great sturgeon at the two centers (47 samples from gene bank and 100 samples from Shahid Beheshti) in 1397-1399. Genomic DNA for amplification of microsatellite loci was extracted using ammonium acetate. Four microsatellite loci (LS68, LS57, LS19, and LS39) were amplified for examination of the genetic diversity of the two group studied sturgeons. Within 147 individuals of the great sturgeon, 198 alleles were detected (123 alleles in genebank and 75 alleles in Shahid Beheshti stocks) and all loci were disomic. The genetic diversity in gene bank and Shahid Beheshti populations were 0.44 and 0.68, respectively. Results showed that the Shahid Beheshti Center broodstocks had good allelic richness and genetic diversity and the gene bank broodstocks had good allelic richness but with reduced heterozygosity index. The genetic differentiation index (FST) between the two populations was 0.34, which indicated a low genetic differentiation between the two populations. Also, the genetic distance and similarity of Nei based on the population matrix were 1.491 and 0.225, respectively, while based on Analysis of Molecular Variance (AMOVA) genetic diversity between the two populations of great sturgeon was 92% and within the population was 8%. Genetic study of fish populations in two important fisheries centers of the country was able to provide basic information on the genetic conditions of the stocks of these valuable fish from the point of view of conservation of stocks as well as commercial aquaculture management programs.

In this study, sturgeon species were identified based on mitochondrial genome amplification (D loop control region) and band size differences by agarose gel. For this purpose, a total of 140 Caspian Sea sturgeons and Siberian sturgeons were selected. After DNA extraction, primers were used to amplify the D-loop control region. The right primers were the same in all species but the left primers were different and they were selected such that the size of different species was distinct. Based on the obtained results, the desired bands for all species of Caspian Sea sturgeon and Siberian sturgeon, except Beluga, were amplified which represents the species for that sample. This method is a useful approach for detection of fraudulent mislabeling of the fishery products including sturgeon and caviar due to its ease and rapid execution. However, since the mitochondrial genome has only maternal inheritance, for additional tracking, the whole genome of sturgeon should be studied for each species in order to obtain the desired points for tracking the differences of Caspian Sea sturgeon at the nuclear genome level, thereby a diagnostic evaluation as well as barcoding of samples of fishery, health and cosmetic products from Caspian sturgeon can be done by combining differences at the mitochondrial and nuclear genome levels of the species. of the species.

Induction of gynogenesis requires the spermatozoa with degraded DNA with high motility, simultaneously. In this study, to obtain this capability in Siberian sturgeon spermatozoa, UV radiation with a dose of 4257 j/m2 was applied. First, the seminal fluid was separated from the spermatozoa using a centrifuge, and then 10% of the sperm was prepared and irradiated with UV radiation for 30, 60, 90, 120, and 150 seconds to obtain the best motility and percentage of DNA damage using the comet assay method in comparison with the control. Simultaneously, the fertilization ability of these sperms was tested in sterlet sturgeon eggs and the rate of hatched larvae. Based on the results, the best duration of irradiation of spermatozoa was determined to as 90 seconds. The DNA degradation rate was 32.27±1.96 percent, the motility rate was 50% with forwarding movement, the fertilization rate was 52.1±0.52 percent, and the egg hatching rate was 1.1±0.11 percent. Therefore, genetically inactivated Siberian sturgeon sperm could be used in interspecific gynogenesis studies in sturgeon. Either to producing offspring of all females or to protect their extinction.

Based on IUCN criteria, sterlet sturgeon (Acipenser ruthenus) has been shown as vulnerable species. Its male sex is inadequate or absent in the most of propagation and rehabilitation centers of Iran. The aim of this study is the induction of gynogenesis to produce male sterlet sturgeon using heterologous sperm. DNA of heterologous sperm from Siberian sturgeon (Acipenser baerii) was damaged successfully by UV irradiation with 473 µw/cm2 intensity and 90 seconds duration. The irradiated sperm mixed with sterlet sturgeon eggs to induce gynogenetic progenies of sterlet sturgeon. For this purpose, 15 minutes after activation, the fertilized eggs exposed to 34ºC heat shock for 2 minutes. Gynogenetic progenies were verified by microsatellite markers (AFU68 and AFUG195) as only maternal heritability. Histological observations of preliminary gonads of gynogenetic progenies were carried out by biopsy method at 17 months-age. Results showed that gynogenetic progenies consisted of 76.6% female and 23.3% male. Regarding to female heterogametic sex determination system in sterlet sturgeon, this study showed that both female and male as pure could be induced by inactivated heterologous sperm and gynogenesis.

Among vertebrates, most of the fishes are unique for the lack of acrosome in their spermatozoa. The sperm enters the egg through micropyle of the egg. This has facilitated heterologous fertilization in many fishes and led to hybridization and polyploidization. Hybridization is performed both in the nature and domesticated fishes. Today, some of the hybrids have high growth rate than their parents so artificial hybridization be carried out among some economic species. Some of hybrids have high similarity to their parents, so identification and verification of hybrids is impossible. This identification needs both phenotypic (morphometric and meristic) traits and molecular analysis in compared with parents. In this case, the most confidence and the quickest method are molecular markers based on DNA, especially microsatellite markers. It could play an important role in the precise confirmation of fish hybrids.

Through expanding of aquaculture activities, considering of male gamete quality has been increased. Egg fertilization with better quality sperm samples might increase fertilization outputs among all criteria (Fertilization and hatching rates, survival and growth). The quality of gametes involving fertilization relies on extensive factors which could have specific and different effects among species. So, there are many definitions for sperm quality. According to the scientific and technological improvements, the methods for sperm quality assessment have been continuously renewed and assessed through new vision. Study of spermatozoa, is easier than the eggs because of their specific characteristics and ease of sampling and the views of its quality assessment have been developed. In current article, after mentioning to most important fish spermatozoa cellular and morphometric characteristics, the most valuable quality indices and some factors affect them, have been discussed. Respecting to all studies on these indices and quality improvements of each techniques, spermatozoa motility and fertilization indices, still have been recognized as the indices and most reliable and practical indicators for routine sperm quality assessment in fish.

In this study, a HindIII Satellite DNA (168bp) was isolated from Russian sturgeon, Acipenser gueldenstaedtii and was used as a probe for fluorescent in situ hybridization (FISH) with the chromosomes of Persian sturgeon, Acipenser persicus and Russian sturgeon. After obtaining suitable chromosomal preparations from the fishes through leukocyte culture and labeling the probe with Spectrum orange (Orange-dUTP) through Nick translation method, the probe was hybridized with chromosomes of the fishes on the microscopic slide. In the studied chromosomal preparations, the hybridization colored signals were clearly visible. Counting the produced signals in twelve chromosomal preparations of three fishes from each species showed that the mean number of the signals in the chromosomal preparations of the Persian sturgeon and Russian sturgeon were 66±4 and 68±3 respectively. In this study, accurate kind determining of chromosomes and HindIII SatDNA site on the chromosomes were impossible due to the presence of microchromosomes and heterogeneity of shapes and sizes of colored signals

Primary cell lines were established from the culture of explants of caudal fin of Persian sturgeon, Acipenser persicus. The margin of the caudal fin from a two year old specimen (23 cm in length, and 133 g in weight) was aseptically cut (2 cm), washed three times with PBS containing antibiotics and then minced into 1 mm explants and cultured in L-15 medium complemented with antibiotics and FBS (20% ) at 22 °C. After confluent monolayer formation on the bottom of the flask, the cells were washed with PBS containing antibiotics and dislodged by trypsin-EDTA solution (0.25%) and shaking the flask. After washing the cells with PBS containing the antibiotics, they were cultured in 5 ml of L-15 medium, FBS (20%), antibiotics at 22 °C (first subculture). After a confluent monolayer formation, a similar procedure was followed for the second subculture.The emerging cells from explants exhibited fibroblast-like and epithelial-like cells in primary culture but the fibroblast-like cells were seen to predominate after the first subculture. In the present study the establishment of cell lines from the original culture at two passages was investigated.

sox9 is a protein coding gene that plays a crucial role in the regulation of several cellular activities among vertebrates. According to the importance of the beluga as the main species in aquaculture and the importance of sox9 gene in vertebrate evolution, in this study, the identification of gene sequence and expression pattern in different stages of larval and gonadal development of male beluga was developed. The results showed that the sox9 sequence in the beluga has high homology with the sox9 sequence in other vertebrates, which represents a fundamental role of this gene in different animals. Indeed, the study of sox9 gene expression during larval development (1, 3, 6, 15 and 50 days post hatching- dph) revealed that sox9 mRNA is expressed at different stages of larval and the most expression is about 15 dph. However, in differentiated gonads (developmental stages 1-4), sox9 mRNA expressed only at stage 4. In conclusion, it seems that the sox9 expression is related with cartilage synthesis and organogenesis at early life stages and with testicular development at advanced stages of sexual maturation.

The effects of starvation periods on physiological response of the juvenile Persian sturgeon, Acipencer persicus, was assessed through such stress factors as glucose, cortisol and hepatic enzymes for a period of 8 weeks. For this purpose, in a randomly designed experiment, five groups of fish (108.04 ± 0.28 gr) in 3 replicates were starved for a period of 0 (control), 1, 2, 3 and 4 weeks and fed them to satiation after their starvation lags. Blood plasma glucose and cortisol during starvation periods did not significantly change (p>0.05). This indicates high performance of this specific in maintenance of blood glucose during starvation and recovery of glucose level after feeding. However, plasma hepatic enzymes level in fasting treatments increased (p

Hybrids were produced by crossing female Caspian kutum Rutilus frisii kutum with male grass carp Ctenopharyngodon idella. The genome of eight larvae and parents were studied using microsatellite markers for genetic evaluation and verification. After DNA extraction from parent fish and progeny, hybrid heritability of two loci was assessed using two pairs of microsatellite primers. Hybridizied offspring showed as similar banding pattern to that of their maternal parent, without heritability of the paternal genome.