Comparison of genetic variation of ship sturgeon (Acipenser nudiventris) in the southern Caspian Sea and Ural River using PCR-RFLP

Genetic variation of ship sturgeon (Acipenser nudiventris) from the Caspian Sea was investigated using NADF15/6 gene and PCR-RFLP analysis. A total of 80 specimens of the fish were collected from the south Caspian Sea and the Ural River from Kazakhstan. mtDNA ND5i6 gene was amplified by polymerase chain reaction (PCR) digested using 39 Endonucleases Restriction Enzyme. Of the 39 enzymes, five showed polymorphism. Totally, ten composite haplotypes among 80 specimens were detected. Haplotype AAAAA showed maximum frequency (57.5%) whereas haplotypes BBAAA and BABAA showed minimum frequency (12%). Haplotype AAAAB was recognized specifically in Ural River specimens. Average haplotype and nucleotide diversity was 0.8516 and 0.007 respectively. Compared to other sturgeon species living in the Caspian Sea, nucleotide diversity of Ship Sturgeon was much lower (0.007). This may be due to smaller population size of this species. Monte-Carol simulation using 1000 interaction did not show any significant differences between haplotype distribution of the fish sampled in the south Caspian Sea (X^2=35.48 , P=0.74). However, we detected a significant difference between haplotype of Ship Sturgeon from Ural River and the south Caspian Sea. We conclude that Ship Sturgeon from Ural River is different from the fish in the south Caspian Sea and suggest CfrI31 enzyme as a molecular marker for population differentiation in the Caspian Sea

Genetic analysis of wild common carp, Cyprinus carpio L. in the Anzali wetland, the Caspian Sea

The Caspian Sea and its basin (e.g. Anzali wetland) is one of the natural habitats of wild common carp Cyprinus carpio. In this study the genetic structure of this species in the south-west of Caspian Sea (the Anzali wetland) was investigated using PCR-RFLP analysis of D-loop region. Two hundred of mature fish were collected from 5 stations (40 individuals from each station) including Siahkeshim protected area (SK), Selke wild refuge (S), Sorkhankol wild refuge (SO), Abkenar (A) and the Anzali wetland estuary (E) during spawning season. A 420bp fragment of D-loop was amplified and the PCR products were digested with forty endonuclease enzymes. Four out of them: TasI, SmaI, SspI and ApoI showed polymorphism. Seven different composite haplotypes were detected among 5 stations and AAAA was the most frequent. FST ranged from 0.003-0.99. Over all stations, average haplotype and nucleotide diversity were 0.13 and 0.01, respectively. The highest haplotype (0.42) and nucleotide (0.06) diversities were found in (SO) station. AMOVA test showed that the Anzali wetland probably consists of two different populations of wild common carp which are distributed in SK, A-SO-S-E stations. The results of this study will be useful as a guideline for conservation, restocking as well as cultivation purposes of wild common carp in the Caspian Sea

Application of AFLP molecular marker for genetic analysis of black pomfret Parastromateus niger from the Persian Gulf

Black pomfret Parastromateus niger is a commercially important fishery resource in the Persian Gulf but harvesting its stocks lacks genetic identification of populations. AFLP technique was applied to analyze genetic diversity and population structure of 32 fish from coastal waters of Bandar Abbas, Bushehr and Abadan with 7 EcoRI/MseI primer pair combinations. In total, 381 bands were produced of which, 46 were polymorphic (12.07%). Percentage of polymorphic bands was higher in Bushehr samples (91.30%) than in Abadan (84.78%) and Bandar Abbas (73.91%) samples. The highest level of heterozygosity based on Nei’s coefficient and Shannon’s index was observed in Bushehr fish (0.38±0.16 and 0.54±0.21). Observed and effective alleles ranged from 1.73±0.44 and 1.53±0.40 in Bandar Abbas samples to 1.91±0.28 and 1.70±0.34 in Bushehr samples. The average Fst was 0.19 indicating high genetic differentiation among the three locations. Gene flow with mean of 1.93 was the lowest level between Bandar Abbas and Abadan (1.24). Nei's genetic identity revealed the least genetic similarity between the samples of Bandar Abbas and Abadan (0.77). AMOVA analysis demonstrated 81% of the genetic variation within populations and 19% among populations. The UPGMA dendrogram clustered all 32 individuals into 3 groups. In some cases individuals from the same region were grouped together but in most cases, gene exchange was observed to be common among the groups. Analyses provided evidence for genetic differentiation among the three locations, indicating separate populations of black pomfret in the northern Persian Gulf

Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines

Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish

Isolation and proliferation of spermatogonial cells from ghezel sheep

Background: Sheep industry has taken steps toward transforming itself into a more efficient and competitive field. There are many varieties of sheep breeds in the world that each of them serves a useful purpose in the economies of different civilizations. Ghezel sheep is one of the Iranian important breeds that are raised for meat, milk and wool. Field of spermatogonial cell technologies provides tools for genetic improvement of sheep herd and multiple opportunities for research. Spermatogonial cells are the only stem cells capable of transmitting genetic information to future generations. Methods: This study was designed to extend the technique of isolation and in vitro proliferation of spermatogonial cells in Ghezel sheep. Results: Isolated cells were characterized further by using specific markers for type A spermatogonia, including PLZF. Also, sertoli cells were characterized by vimentin which is a specific marker for sertoli cells. After 10 days of co-culture, viability rates of the cells was above 94.7, but after the freezing process the viability rates were 74 percent. Conclusion: In this study, a standard method for isolation and in vitro proliferation of spermatogonial stem cells in Ghezel sheep was developed. © 2018, Avicenna Journal of Medical Biotechnology. All rights reserved

Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and it’s expression in HEK cell lines

Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish

Isolation and proliferation of spermatogonial cells from ghezel sheep

Background: Sheep industry has taken steps toward transforming itself into a more efficient and competitive field. There are many varieties of sheep breeds in the world that each of them serves a useful purpose in the economies of different civilizations. Ghezel sheep is one of the Iranian important breeds that are raised for meat, milk and wool. Field of spermatogonial cell technologies provides tools for genetic improvement of sheep herd and multiple opportunities for research. Spermatogonial cells are the only stem cells capable of transmitting genetic information to future generations. Methods: This study was designed to extend the technique of isolation and in vitro proliferation of spermatogonial cells in Ghezel sheep. Results: Isolated cells were characterized further by using specific markers for type A spermatogonia, including PLZF. Also, sertoli cells were characterized by vimentin which is a specific marker for sertoli cells. After 10 days of co-culture, viability rates of the cells was above 94.7, but after the freezing process the viability rates were 74 percent. Conclusion: In this study, a standard method for isolation and in vitro proliferation of spermatogonial stem cells in Ghezel sheep was developed. © 2018, Avicenna Journal of Medical Biotechnology. All rights reserved

Nestin, a neuroectodermal stem cell marker, is expressed by bovine sertoli cells

Nestin, an intermediate filament protein is expressed by neuroectodermal stem cells and tumors originating from cells of neuroectodermal and mesenchymal lineages. Nestin expression is prominent in embryos and remains upregulated until 3-6 weeks after birth but is downregulated afterward. Sertoli cells are nucleated somatic cells that are spanned in the seminiferous epithelium and play a critical role in supporting and controlling germ-cell development. In this context, we employed immunocytochemical, Western blot, and Flow cytometric analyses to demonstrate nestin expression in bovine sertoli cells. Immunostaining clearly showed that setoli cells express high levels of nestin, a result which was confirmed by Western blot analysis of purified cells. Intracellular staining of sertoli cells by flow cytometry revealed that around 74 of the cells express this marker. Given the high expression of vimentin by sertoli cells, it is proposed that the expression of nestin in these cells might be required for the formation of stable vimentin/nestin intermediate filament network. In light of these findings, it seems that sertoli cells of mature bull have potentiality of proliferation. © 2010 Springer-Verlag London Limited

Study of Fe, Zn, Cu, Cd, Pb concentrations in liver, kidney and muscle tissue of cow and sheep marketed in Hamedan in 2011

Importance of heavy metals in food safety and detrimental effects of their high concentrations in food stuff is well documented. In this study, concentrations of Fe, Zn, Cu, Cd and Pb in kidney, liver and muscle tissues of cow and sheep at Hamedan retails were evaluated. A total number of 180 samples was assessed for the amount of heavy metals as ppb in wet weight. For this, wet-digestion method was used to determine the concentration of given elements by ICP (Varian ES-710). Results showed that the highest concentration of heavy metals was determined in the liver and kidney samples, while the lowest concentration was found in muscle tissue. Among the heavy metals, Fe in cow’s liver had the highest concentration (25507±879 ppb) and Cd in muscle tissue of sheep has the lowest concentration (192±54 ppb). In overall, accumulation of heavy metals in tissues of cows was higher than sheep. Statistical comparison of accumulated metals concentration in various tissues of these two animal groups showed significant difference (

Lead (Pb2+) Removal from Synthetic Aqueous Solution Using Food Waste Ash

Lead (Pb2+) Removal from Synthetic Aqueous Solution Using Food Waste Ash AfsharniaM (Ph.D)1, Shams M (M.Sc)2, Sajjadi SA (Ph.D)1, Qasemi  M (M.Sc)3 1. Assistant Professor, Department of Environmental Health Engineering, ,Gonabad University of Medical Sciences, Gonabad, Iran. 2. Instructor, Department of Environmental Health Engineering, Gonabad University of Medical Sciences, Gonabad, Iran. 3. Corresponding Author: M.Sc Student in Environmental Health Engineering, Gonabad University of Medical Sciences, Gonabad, Iran Abstract Introduction: Lead is a toxic heavy metal in some industrial wastewater that can pose hazards to the environment and human health. Adsorption is a promising technology for decontamination of water. The present study evaluated the efficiency of food waste ash as a cheap, environmental compatible and available adsorbent, in lead removal from synthetic aqueous solution. Methods: In the presentexperimental study, lead removal by  using food waste ash in series of batch experiments were investigated. The influence of different parameters on adsorption such as adsorbent dose, contact time, pH, lead concentrationand adsorption temperature also studied. The experimental data finally were analyzed using excel and the adsorption behavior was fitted  to isotherm and kinetic models. Results: The study showed that lead removal decreased with increasing lead concentration. In addition, it was found that lead adsorption increased by increasing pH and temperature. comparing adsorption capacity of food waste ash with the adsorbents used by scientist also showed the former have a remarkable higher capacity toward lead adsorption. The adsorption process also showed a good conformity to Freundlich model. Conclusion: Current study endorsed the use of food waste ash in real adsorption systems, as a cheap and available adsorbent with high capacity toward lead