Iranian Journal of Biotechnology
Volume:12 Issue: 1, Winter 2014

The breeding performance is an important parameter to evaluate the breeding success in captivity conditions. The optimum dose of hormone in combination with latency period is desirable for getting best breeding performance in fish.. The objective of the study was to find the spawning latency period in the hormonal induced reproduction of Snow trout with two inducers (Ovaprim and hCG) separately and in combination. The fish spawners were randomly divided into five groups and treated with Ovaprim, Ovaprim and high dose of hCG, Ovaprim and low dose ofhCG, hCG and saline water as control.. The results suggested that Ovaprim and high dose of hCG treatment lead to shorter latency time (40 hours and 40 minutes), but ovulation percent, percentage of live embryos in the eyed stage and ovulation synchronization were lower than treated groups with Ovaprim alone or Ovaprim and low dose of hCG. Females from the control and hCG groups did not spawn.. The highest hormonal stimulation effectiveness was recorded in the group in which one hormonal substance (Ovaprim) was applied. Therefore the accurate ovulation time of snow trout, Schizothorax zarudnyi was difficult to be predicted..

Genetic polymorphism of milk proteins has been associated with composition, manufacture, and traits of milk. Caseins are the most important milk proteins that its genes are strongly linked and inherited together. κ-casein, which is a quantitatively minor constituent of bovine milk, is thought to play a critical role in organization, fixation and aggregation of casein micelles and firmness of curd during cheese making. In this study, we aimed to investigate the polymorphism of κ-casein gene in Iranian Holstein cows. In this study, κ-casein gene polymorphism among 50 DNA samples of Iranian Holstein cows via polymerase chain reaction sequence analysis (PCR sequencing) was analyzed. For data analysis SPSS 11.5 (ANOVA test) was used. Four polymorphic sites that created four variants and seven different genotypes of κ-casein gene were identified. In this population the frequencies of A, B, C, and E alleles were as 0.391, 0.413, 0.087, and 0.109 respectively. We suggest that the frequency of κ-casein gene B allele should be increased in Iranian Holsteins because it is an essential factor in marker-assisted selection for milk traits.

The influenza virus is globally pathogenic and it is usually associated with zoonotic respiratory diseases. This virus has caused a number of pandemics with high mortality rates. The non-structural protein-1 (NS1) of influenza A viruses is a non-essential virulence factor that has multiple accessory functions during a viral infection. This protein is highly conservative. It has been shown that this protein plays a major role against the immunity responses of host cells. The aim of this study was to produce the recombinant influenza NS1 protein by the use of a bacterial production system, in order to evaluate the immunological and structural features of this protein in future researches. The NS1 gene construct was artificially synthesized; subsequently it was sub-cloned into the pQE30 expression vector. The expression vector was then transformed into the BL21 cells and induced by IPTG. Finally, the expression was evaluated by SDS-PAGE and Western blotting techniques. The NS1 gene was successfully cloned and transformed into expression cells. As a result, a 23 kDa band was observed both on the SDS-PAGE and nitrocellulose paper after Western blotting. Based on the results of this study, it could be concluded that the NS1 gene of influenza A H1N1 virus (A/Shiraz/14/2010 strain) could be cloned and the rNS1 protein (recombinant NS1 protein) could be expressed using a bacterial protein translation system. Since this protein is a conservative protein among influenza A viruses, it could be used as a potent vaccine for the prevention of various types of pandemics caused by influenza A.

Equine chorionic gonadotropin (eCG) is commonly used in association with a progestagen treatment to synchronize estrus of goats and ewes during breeding and non-breeding seasons. Classical purification of the eCG from serum includes pH fractionation with metaphosphoric acid two ethanol precipitation steps as well as dialysis followed by fixed-bed chromatography. The aim of the current study was to develop an accurate and fast method for production and purification of eCG using a polyclonal antibody assay. The blood samples (300 mL) were taken from the jugular vein of 17 mares on days 50, 70 and 90 of pregnancy. Plasma of the samples was siphoned and phenol solution was added to the plasma and stored in the refrigerator until eCG extraction. To prepare the polyclonal antibody against eCG, four male rabbits, about four months old with 2 kg weight, were chosen. A basic immunization was done by injecting 25 IU of eCG to the rabbits. Ouchterlony assay or double immunodiffusion test was used to assess the immunization and the titer of antiserum against eCG. eCG was purified from the plasma via the solid-phase extraction (SPE) method. Results based on agarose-gel double immunodiffusion test showed that rabbits were completely immunized. SDS-PAGE analysis showed purified eCG is extracted without any significant contamination. The extraction of eCG with polyclonal antibody using the SPE method and production of anti- eCG antiserum in rabbits is suitable and may be a cost effective method for large scale production of eCG and anti-eCG antiserum in Iran.

Hydrocarbons degradation is principally achieved by microorganisms in natural environments. The extent of hydrocarbons biodegradation is mainly conditioned by environmental factors and its success depends on the optimal condition for the crude oil degrading isolates. The aims of the current study was to isolate and identify crude oil degrading bacterium from surface sediments of Qeshm Island, Iran and to evaluate the efficiency of a statistically-based experimental design for the optimization of crude oil degradation performed by the isolated strain. Crude oil degrading bacteria were isolated by serial dilutions of bacterial consortium. In order to optimize crude oil biodegradation by isolated strains, Plackett-Burman experimental design was used to evaluate nine factors affecting crude oil biodegradation in twelve experimental trials. To observe the best yield in crude oil biodegradation, factors that had higher effects were considered for the next stage in the biodegradation optimization process using the Taguchi experimental design. A gram-negative bacterium strain named as the KK1- strain (with 98% homology with Marinobacter litoralis) was isolated from enrichment consortium. Among the various variables screened using the Plackett-Burman experimental design, pH, temperature, salinity and NH4Cl were determined as the most significant factors and considered for the next stage of the biodegradation optimization process using the Taguchi experimental design. Theoretically, the optimum degradation conditions were determined to be: pH = 8, temperature = 35˚C, salinity = 30 ppt and NH4Cl = 1 g.L-1. The validity of the predicted optimized condition was tested by conducting experiments considering the predicted criteria. Biodegradation efficiency of 58.32±5.57% was achieved under the suggested conditions, which was significantly higher than that achieved by the primary conditions (35%). Indigenous bacteria from surface sediments of Qeshm Island were found to be able to degrade crude oil. Our results showed that a combination of the Plackett-Burman and the Taguchi experimental designs may be successfully used to find the optimal amounts of factors required for crude oil biodegradation.

Chronic active adrenocorticotropin hormone (ACTH) immunization of ewes (with a previous history of immunization) resulted in elevated titres of antibody against ACTH, which induced a reduction in circulating levels of cortisol. The aim of the present study was to investigate: a) the influence of active immunization against ACTH on milk production of the ewe as assessed by milk intake of the lamb measured with two different methods, b) whether the stimulation of levels of β-end and α-MSH in the circulation can result in a similar increase in the expression of these peptides in the milk of the lactating ewe, thereby producing a milk with high analgesic properties. Ten primiparous Merino ewes (4-5 years old) during their first 5 weeks of lactation were used in the experiment. Milk intake of each lamb can be used to give the ewe’s milk yield per unit time, and this can be used for secretion of peptides over time. This was determined by two different techniques: the deuterium oxide method and the weigh-suck-weigh method. β-end, α-MSH and cortisol were measured in blood samples and β-end, α-MSH in milk samples using RIAs (Radio Immune Assay). Plasma β-end and α-MSH in ACTH-immune animals increased to 3 and 38-fold respectively, although the level of α-MSH is most likely exaggerated by the presence of α-MSH antibodies in the ACTH immune plasma, as α-MSH comprises the N-terminal 13 amino acids of ACTH. Antibodies also were detected in the plasma of lambs of immune ewes after birth, at levels, which were comparable to those measured in the ewes during the period of colostrum production and thereafter decreased to much lower levels by the end of the experiment. Plasma β-end and α-MSH in the lambs of immune ewes were significantly higher than those measured in the lambs of control ewes. The immunization procedure had no effect on feed intake, live weight gain and milk production of ewes nor was the growth rate or milk intake of the lambs altered. The expression of β-end and α-MSH in milk was also unaffected by the high concentrations of these hormones in the circulation of immune ewes.

The excellent biocompatibility, biodegradability and biological properties of the hydrogels, fabricated using natural polymers, especially polysaccharides, are very advantageous for biomedical applications. Gum tragacanth (GT) is a heterogeneous highly branched anionic polysaccharide, which has been used extensively in food and pharmaceutical industries. Despite, its desirable properties, the potential biomedical applications of this natural gum have not been fully explored. In this study, an enzyme catalyzed in situ forming hydrogel, based on Iranian gum tragacanth (exudate of Astragalus fluccosus) was prepared and characterized for biomedical applications. The main objective of the present study was to explore the feasibility of using tragacanth natural gum as a base for in situ-forming hydrogels in biomedical applications. First, tyramine (TA) was conjugated to the water-soluble part of GT (TGA) using aqueous-phase carbodiimide activation chemistry. Next, in situ forming hydrogel was prepared via an enzyme catalyzed coupling reaction in the presence of horseradish peroxidase (HRP) and H2O2. Gelation time, swelling/degradation behavior and mechanical properties of the hydrogel and cell viability of the encapsulated cells within these hydrogels were investigated. The gelation time of the hydrogel was less than 30 seconds, which is very desirable for clinical applications. At concentrations ≤ 0.1% (w/v), both GT and TA-TGA showed no toxicity towards human mesenchymal stem cells (hMSCs) and Caco-2 cells. More than 90% of the encapsulated hMSCs in the hydrogels, which were prepared at H2O2 concentrations of less than 15.0 mM, remained viable after 2 hours of incubation. The TA-TGA conjugate can be gelled enzymatically in the presence of HRP and H2O2. This in situ forming hydrogel might be a desirable candidate for biomedical applications.

ß-carbolines, harmaline and harmine, are major alkaloids present in the seeds of Peganum harmala L. These alkaloids are known as herbal active principals with potential use in pharmaceutical and medicine. To assess the growth inhibitory effect of phyto-alkaloids, harmaline and harmine, on cancer cell lines. P. harmala L.’s alkaloids were extracted by acidic/basic extraction method and identified by two methods, Fourier Transform Infra-Red Spectroscopy (FTIR) and High Performance Liquid Chromatography (HPLC). Two breast cancer cell lines, MDA-MB-231 and Mcf-7, were subjected to different concentrations (1–100 μg.mL-1) of the P. harmala extract at different time courses (24, 48, and 72 hours). Methylthiazol Tetrazolium (MTT) test, half maximal inhibitory concentration (IC50) and morphological changes through optical microscopy were evaluated. In both studied cell lines, the P. harmala extract decreased cell viability in longer time exposure in a dose dependent manner. The more concentrated extract led to higher motility of MDA-MB-231 at 24 hours. Although, Mcf-7 cell line required longer exposure time to reach the same motility. It was observed that 30 μg.mL-1 is the minimum lethal dose that kills approximately 50% of cells at 24 hours for MDA-MB-231 cell line (IC50). IC50 for Mcf-7 was calculated 40 μg.mL-1 and 25 μg.mL-1 48 and 72 hours after being exposed against harmala’s extract, respectively. The morphological observation confirmed the apoptosis nature of P. harmala on cells as their membrane kept intact and no membrane permeabilization was observed. The results revealed that the P. harmala extracts significantly decreased the growth rate and cell survival of cancer cell lines. The extract induced cell death regarding natural cell growth rate. MDA-MB-231 cell line naturally has a higher growth rate than Mcf-7 cell line, thus higher growth inhibition of MDA-MB-231 cell line by the P. harmala extract was confirmed.