Inroduction & Objective: Ibuprofen, as a drug contaminant, has deleterious and in some cases irreversible effects on aquatic organisms and in general on living things, which in many cases can result in genetic damage and damage to the DNA structure. The effects of different concentrations of ibuprofen on DNA damage and damage to zebrafish are investigated. In this study, genetic changes of p53 gene and its expression in zebrafish after two weeks of exposure to 0. 1, 1 and 10 mg / l ibuprofen in vitro were investigated. Material and Method: In this study zebrafish were exposed to the above mentioned concentrations for two weeks and then RNA extracted from them. The results were analyzed using qPCR method. Results: The low drug, 0. 1 mg / l had no effect on p53 gene expression, while the other two concentrations, which were 1 and 10 mg / l ibuprofen, respectively, showed a significant difference between the control and the drug containing sample. As a result, the p53 gene was highly expressed. Also for the internal control gene that was GAPDH, there was no significant difference at the lowest drug concentration. Ray did not show a significant difference between the treatment and control samples, while in the subsequent treatments there was a significant difference between control and treatment samples. The final result of this study was that p53 gene expression increased 1. 09, 0. 57 and 2. 2 fold in sample with lowest drug concentration, mean concentration and highest drug concentration, respectively.

Cryopreservation has led to many improvements in domestic animal breeding industry, especially in terms of spreading of superior genes. The improvement of extenders and cryoprotectants can result in a significant increase in transporting and inseminating of frozen stallion semen. In this study, we analysed the genes that would be expressed in stallion cryopreserved sperm and by providing an adequate amplification, they would be selected as reference genes in the future studies. Live sperm with above 70% motility were selected from six different stallions. Sperm samples were divided in to two parts for total mRNA extraction. The selective amplification of three candidate genes (L32, β-actin and Ubiquitin) were transcribed by real-time polymerase chain reaction (RT-qPCR). Gene stabilities were calculated by GeNorm and BestKeeper softwares. Results indicated that the expression of L32 gene is less stable than that of β-actin and Ubiquitin. The expression stability of β-actin and Ubiquitin were similar. These genes can be used for normalizing of RT-qPCR reaction data. The GeNorm software introduced the best combination of genes as, β-actin and Ubiquitin with the stability value (M) of 0. 083. The results of BestKeeper software showed that the most stable house-keeping gene was β-actin.

The present study was conducted to evaluate the effect of Tribulus Tresstrise (TT) herb on sex ratio of semen in Arabic Khouzestan ram using real time-qPCR technique using 18 rams in a completely randomized design with 3 treatments. The SRY and PLP genes were amplified to isolate the specific fragments of Y-and X-chromosome sequences. The treatments included: i) the control group (0% TT), ii) Diet containing 15 g/kg TT, iii) Diet containing 30 g/kg TT. Sperm sampling was taken from all rams at 10 month of age and blood sampling was performed at 8 and 10 month of age. The results showed that expression rate of SRY gene increased with increasing TT level and rams that received 30 g/kg TT diet had the highest SRY gene expression and PLP gene expression decreased with increasing TT level (p-value =0. 004). There was positive correlation between Testosterone concentration and SRY gene expression at 8 (0. 65) and 10 (0. 59) month of age, and the relationship between PLP gene expression and Testosterone concentration was negative and-0. 61 and-0. 66 at 8 and 10 month of age, respectively (p-value= 0. 006). The results indicated that adding Tribulus Tresstrise herb to the ram diet increases the SRY gene expression and also sperm containing Y chromosome. In other words, it increases the sex ratio toward male gens in Arabic Khozestan ram by increasing the androgen hormones.

MicroRNAs (miRNAs) are short, endogenous non-coding RNAs that function as guide molecules to regulate transcription of their target messenger RNAs. Several methods including low-density qPCR arrays are being increasingly used to profile the expression of these molecules in a variety of different biological conditions. Reliable analysis of expression profiles demands removal of technical variations in data, which is achieved via applying normalization techniques. Most normalization techniques have been developed for mRNA microarrays and new and modified methods should be used for miRNA studies in general and RT-qPCR miRNA arrays in particular, because of low number of miRNAs. Here, we introduce a new method based on Procrustes superimposition of arrays to be normalized on a reference array. To assess the performance of our normalization method, we compared this method to the common miRNA normalization methods. Removal of technical variation was assessed by robust modeling of mean square error (MSE) in different subsets of real miRNA datasets before and after applying normalization. We show that our method outperforms the other normalization methods in concurrent reduction of technical variation and retention of biological variability.

Small ruminants theileriosis are widespread in Iraq andacute infections usually with hight mortality. However, the survived animals suffer from low production of meat and milk. Coinfection with more than Theileria sp. And/or Anaplasmosis could have an impact on the disease severity. The main finding was identifying T. lestoquardi, T. ovis, T. annulata, blood samples of infected sheep with a history of chronic theileriosis (n=48) and with acute clinical theileriosis sign (n=24) were being collected from fields located in Babylon province (middle of Iraq) after chlinical examination and Polymerase chain reaction and real time PCR were performed for detection. Theileria. lestoquardi was the highest of these species within the acute and chronic cases. As well as, the load of this species in acute cases was significantly higher (P<0. 01) to that in chronic. However, the load of T. ovis and T. annualta were similar in acute and chronic cases. Importantly, all these cases were coinfected with Anaplasma phagocytophylum. This could be due to the infection of leukocytes meanwhile weakening of the animal’, s immune system. Also, these parasites transmitted by the same tick-vector. The impact of this finding could help in disease prevention and diagnosis.

Introduction: Silver nanoparticles (AgNPs) have a broad spectrum of uses, therefore, AgNPs will be released from those products into many different ecosystems. In the last decades, AgNPs have received substantial attention due to their distinctive physical and chemical properties such as high thermal and electrical conductivity, chemical stability, catalytic activity and antimicrobial properties against microbes such as bacteria, fungi, and viruses. There are many parameters for assessment effect of toxicity due to AgNPs but soil microbial community is one of which considered being an important target for assessing the impact of manufactured nano-materials on the terrestrial environment. Toxicity of AgNPs is due to the physical interaction of AgNPs with microorganisms and the production of reactive oxygen species (ROS). Although as we have been known harmful effects of AgNPs on the soil bacterial community, but the most information about antimicrobial properties of AgNPs come from the routine lab instructions such as soil respiration, substrate induced respiration and microbial biomass and colony forming unite. So, the objective of this paper was to study the effects of silver nanoparticles on microbial activity using the routine lab instructions and compare with the obtained data from the molecular genetic techniques. In this paper, the quantitate population of soil bacterial was estimated using Real time qPCR with the MIQE guidelines. Materials and Methods: In order to study the effect of silver nanoparticles on microbial activity and bacterial population in a calcareous soil, an experiment was conducted as a completely randomized design based on factorial arrangement with three replications. Experimental factors included silver slat forms (AgNPs and AgNO3), Ag concentrations (0, 0. 5, 5, 10, 50, and 100 mg Ag kg-1 dry soil) and incubation time (7 and 42 days). Soil samples (Typic Haplicambids) with clay loam texture and seven percent of calcium carbonate was collected from Research Field of Ferdowsi University of Mashhad, Mashhad, Khorasan Razavi, Iran. The soil samples were amended with different concentrations of AgNPs and incubated at 25 o C for 42 days. The water content of soil samples was adjusted at 70% WHC during the incubation time. After 7 and 42 days of incubation, the soil substrate-induced respiration (SIR), heterotrophic plate count (HPC), and soil urease and dehydrogenase activities were measured. Finally, based on the obtained data, the soil biological quality index was estimated using the soil biological parameters. In order to quantify the total bacterial population, DNA was extracted from soil samples and was estimated using the relative concentration of 16S rDNA gene by a quantitative Polymerase Chain Reaction (qPCR), with a minimum information for publication of quantitative real-time PCR experiments (MIQE) guidelines. Results and Discussion: The results showed that with increasing the concentration of both AgNPs and AgNO3, the activity of dehydrogenase and urease in soil samples decreased during the incubation times. Microbial substrate induced respiration (SIR) and the total bacterial population in soil samples considerably declined at the end of experiment. Bacterial population in AgNPs treatments decreased compared to AgNO3 treatments but the reduction was not statistically significant. Over time, soil dehydrogenase activity and soil SIR decreased in both AgNPs and AgNO3 treatments, while soil urease activity and heterotrophic bacterial populations improved but again in heterotrophic bacterial populations was not statistically significant. The soil biological quality index was estimated from the soil biological data. AgNO3 treatments reduced the soil biological quality index compared to AgNPs treatments. In other words, the results showed that AgNO3 was more toxic to soil bacteria activity compared to AgNPs. The lowest soil urease and dehydrogenase enzyme activity and soil biological quality index were observed in the treatment of 100 mg kg-1 dry soil AgNO3 after 7 days of incubation. The application of 0. 5, 5, 10, 50, and 100 mg Ag kg-1 dry soil decreased relative soil bacterial population by 22%, 40%, 59%, 73%, and 82% in AgNO3 treatment and 10%, 30%, 68%, 76%, and 86% in AgNO3 treatment compared to control after 42 days of incubation, respectively. Conclusion: The results of this study showed that silver nanoparticles can negatively affect the enzymes involved in the nitrogen and carbon cycle. The AgNPs had less toxicity effect on the soil microbial activity compared to AgNO3. However, AgNPs was more toxic to soil bacteria populations compared to AgNO3. Different behavior AgNPs and AgNO3 in calcareous soil needs more investigations but there is no doubt that AgNPs is as an emerging contaminant and it has high toxicity potential for soil microbial community.