فرهاد فرهنگ پژوه

Leishmaniasis is one of the intracellular parasitic diseases that is known as the most common infectious disease after AIDS, tuberculosis and malaria. Leishmaniasis is among the 6 most prevalent endemic diseases and is in fact a tropical and subtropical disease caused by intracellular parasites worldwide. In recent years, treatment of leishmaniasis has been associated with risks due to drug resistance and problems related to the side effects of conventional drugs. The disease is found on all continents except Oceania and is endemic to enclosed areas in North Africa, southern Europe, the Middle East, southeastern Mexico, and Central and South America. The aim of this study was to investigate the cytotoxic effect of bioactive peptides obtained from hydrolysis of tiger squid muscle on promastigotes of Leishmania major.

The variables of enzyme type, ratio of enzyme concentration to substrate, time of hydrolysis and drying method were studied on the characteristics of hydrolysis protein. After determining the percentage of white muscle protein of squid caught from Bushehr port, alkalase and papain enzymes were used at 5% levels for 180 minutes for hydrolysis. Production percentage, protein peptide powder, total antioxidant capacity (TAC), radical scavenging activity of DPPH, nitric oxide (NO) and total thiol groups were measured. Also, the cytotoxicity of Leishmania major promastigotes was evaluated using MTT method. Glucantime was used as a positive control and the untreated group was used as a negative control.

Muscle hydrolysis with 5% Alkalase enzyme caused the highest degree of hydrolysis (63%) and in general, the concentration of 5% of enzymes had a higher degree of hydrolysis than the concentration of 2.5%. Analysis of variance showed that the effect of enzyme, drying time and drying method on all measured variables had a statistically significant difference at the level of one percent (p <0.01). The effect of concentration on peptide dry powder, peptide protein, TAC and thiol was significant. However, no statistically significant difference was observed between enzyme concentrations in DPPH and Nitric Oxide. The dual interaction of enzyme × concentration on the variables of dry powder of peptide and Thiol at the level of 1% showed a statistically significant difference (p <0.01). However, the interaction of enzyme × concentration on other variables was not significant. The results showed that the dual interaction of enzyme × time on all studied variables showed a statistically significant difference. (p <0.01). The dual interaction effect of enzyme × drying method was significant only on Nitric Oxide and Thiol variables (p <0.01). Also, statistical results using analysis of variance showed that the dual interaction effect of concentration × time on the variables of dry peptide powder, peptide powder protein and Thiol had a statistically significant difference. (p <0.01). The dual interaction effect of concentration × drying method on the variables of peptide dry powder, peptide powder protein, DPPH and Thiol was also significant (p <0.01). Statistical results showed that the dual interaction of drying time × drying method on all studied variables except TAC showed a statistically significant difference (p <0.01). The results of analysis of variance for the triple interactions of enzyme × concentration × time and enzyme × concentration × drying method showed statistically significant difference only on Thiol at the level of one percent (p <0.01). The triple interaction effect of enzyme × time × drying method on protein peptide powder, DPPH, Nitric Oxide and Thiol showed a statistically significant difference at the level of one percent (p <0.01). The results of analysis of variance showed a statistically significant difference (p <0.01) with the triple interaction of concentration × time × drying method on the protein content of peptide powder, DPPH, TAC, Nitric Oxide and Thiol at the level of one percent. Finally, the results of analysis of variance showed that quadruple interaction effect of enzyme concentration × time × drying method on all studied variables had a statistically significant difference at the level of one percent (p <0.01). The results of comparing the mean of the quadruple interaction effect of enzyme × concentration × time× drying method showed that the highest amount of peptide powder, the lowest protein content and the best antioxidant activity are obtained in TAC, DPPH, Nitric Oxide and Thiol using digestion method with the help of 5% Alkalase enzyme for 180 minutes using spray dryer method. The results showed that the degree of protein hydrolysis in muscle with alkalase enzyme was 5% higher than other enzymes studied. With the help of 5% alkalase enzyme in 180 minutes and using spray drying method, the lowest amount of protein in hydrolyzed powder, the best antioxidant activity were obtained in TAC, DPPH, Nitric Oxide, Thiol and the highest cytotoxicity. The highest amount of muscle hydrolysis powder was obtained in the presence of 5% alkalase enzyme in 180 minutes and cooling drying method.

MTT results showed that with increasing the concentration of peptide powder obtained from enzymatic hydrolysis of white tiger squid muscle, the mortality rate of Leishmania major promastigotes increased in all study groups. The best level of IC50 among the studied groups was observed at a concentration of 10 mg / ml of hydrolyzate from 5% alkalase enzyme by spray dryer compared to the control group. The results of this study showed that the hydrolysis protein of Sepia pharaonis white muscle produced with 5% Alkalase enzyme has cytotoxic effect on Leishmania major promastigotes and this effect has shown a stronger activity in hydrolysis prepared using spray drying method than other methods.

Galvanic replacement reactions are a special method for the synthesis of multi-nucleus nanoparticles of various metals within the bed of clinoptilolite. In this method, first, zinc metal cation (Zn +2) after replacing on the clinoptilolite bed was reduced to zinc metal nanoparticles using sodium borohydride salt. Then, the iron (Fe +2) and copper (Cu +2) salts with zinc metal nanoparticles were located on the clinoptilolite bed participate in the oxidation and reduction reactions by citing the potential of various cations in the electrochemical series. The result of these reactions is the formation of deposits of iron and copper nanoparticles on zinc nanoparticles. The selection of appropriate concentrations of cations and determination of the cause of stopping the growth of nanoparticles in each step was carried out by determining appropriate concentrations. XRD, FT-IR, TEM and SEM techniques were used to prove the various stages of work. The effect of the presence and absence of clinoptilolite on increasing the antimicrobial properties of zinc/ iron/ copper three-nucleus nanocomposites and also stability in aquatic environments using bacterial E-coli and Pasteurella specimens have been investigated.

Aims: The use of immune stimulants to prevent diseases and increase the resistance of aquatic animals to stress has been rising. The aim of the present study was examining the histopathology and immunology of rainbow trout (Oncorhynchus mykiss) Fed with Levamisole.
In this experimental research, 1000 fish were divided in 5 treatments and fed on diet supplemented with levamisole at 0 (control), 100, 250, 500, and 1000mg/kg-1 diet for 45 days. Then, the fish were fed on a diet without levamisole and increased density twice for the following 15 days. Blood samples were collected from all treatments on days 15, 30, 45 and 60 to evaluate the total immunoglobulin and lysozyme activity of serum. At the end of the trial, 15 fish from each treatment (each replicate 5 fish) were randomly selected and tissue samples of gill, kidney, and liver were evaluated. The data were analyzed by SPSS 15 software, using one way ANOVA and Duncan test.
Findings: Treatment fed with 1000mg levamisole on day 30 and 60 had the highest total serum immunoglobulin and had the highest lysozyme activity in comparison to other treatments on 15, 30, and 60 days. The least amount of tissue damage was observed in gills, kidneys, and liver in 1000mg levamisole.
A total of 1000mg/kg-1 levamisole of rainbow trout diet causes the least amount of damage to the gill, kidneys, and liver tissues, as well as the increase of total immunoglobulin and lysozyme activity of serum.

Influenza is one of the most important equine respiratory diseases. Two subtypes of equine influenza virus A/equine-1 (H7N7) and A/equine-2 (H3N8) have been identified. This study was carried out to determine the seroprevalence of equine influenza virus in some provinces of Iran. Blood samples were taken from 290 horses and examined with haemagglutination (HI) test for detection antibodies against to two subtypes: H7N7 and H3N8. According to HI test, antibodies against to H3N8 were detected in 16(5.52%) horses; however, none of them had antibodies against to H7N7. The seroprevalence for H3N8 in the horses in Mazandaran, Hamadan, East Azerbaijan and Kermanshah, was 11.32%, 10.72%, 9.53% and 7.9%, respectively. Statistical analysis showed that there was no significant difference between infection with sex and age of horses. It is concluded that the horses of Iran are infected only with H3N8 and therefore, this should be considered in vaccination to equine influenza virus.

This study was conducted to determine effects of betaine supplementation on growth performance and intestinal morphology of broiler chicks subjected to heat stress condition. Two hundred day-old male broiler chicks (Ross 308) were used in a completely randomized design with four dietary treatments. Each dietary treatment replicated five times with 10 birds per replicate. The dietary treatments consisted of a basal diet and the basal diet supplemented with 0.05, 0.1 and 0.2 % of betaine. The experimental diets were offered to chicks under heat stress condition (32±1ºC from 9 AM to 5 PM) during the final period of rearing (25 to 42 days of age). The results did not show any significant effect for the dietary treatments on feed intake and body weight gain, but betaine addition at the levels of 0.1 (2.001) and 0.2% (1.98) decreased feed conversion ratio compared to the control (2.06) group (P=0.08). Crypt depth of ileum was lower in chicks fed diet with 0.1% betaine (72.40 µm) than the control (85.60 µm) (P

Blood tissue is an important indicator of the physiological condition of the body's organs to diagnose disease and organisms biological control, as aquatic animals. This study aimed to determine the factors and parameters hematological including white blood cells, hemoglobin, hematocrit and percentage differential white blood cell such as neutrophils, lymphocytes and eosinophils with garlic extracts and Echinacea extracts on rainbow trout was impressed. 1050 fishes with an average weight of 50 ± 6.5 g in seven treatments (control, 0.5% garlic, 1% garlic, 0.5% Echinacea, 1% Echinacea, 0.5% garlic + 0.5% Echinacea, 1% garlic + 1% Echinacea) and were divided into three replicates. Rainbow trout in all first stages reared for 30 days whit diets containing plant extracts were fed and in the second stage rearing for 15 days only whit control diet (Without extract) were fed, Until time to remove the effect of additives of the fish's body be determined. In order to compare and evaluate the hematological parameters on days zero, 30 and 45 of the breeding period of per replicate three fishes were randomly selected. From fish's caudal vein using the anticoagulant was bloodletting, then, in order to compare and evaluate the hematological parameters, blood samples were tested by standard methods of Hematology. To determine the effect of plant extracts in the bacterial challenge with pathogen, the bacterium yersinia ruckeri was injected intraperitoneally on 30th day and mortality daily was recorded in two weeks. In the first stage of culture, most of the white blood cells (which was significantly different from control treatments) and most of the lymphocytes, hemoglobin and hematocrit (without significant difference from the control treatments) was observed in garlic extract 1% and echinacea extract 0.5% treatments. In the second phase of culture none of hematologic parameters in the experimental treatments there were no significant difference with control treatments (P > 0.05), but most of the lymphocytes was observed in garlic extract 1% treatments. In experiments bacterial challenge with the bacterium Yersinia ruckeri, Mortality in the treatment garlic extract 1% one day later than other treatments began and the lowest mortality accounted. From the results of this study can be conclude that the use of garlic extract 1 %and echinacea extract 0.5% makes stimulates the immune system and increase resistance in rainbow trout.

This study was conducted to determine the electrophoretic pattern of serum protein in goats infected with Babesia ovis. Serum total protein was calculated and serum electrophoresis from 15 goats naturally infected with B. ovis as well as same number of healthy goats was performed. Microscopic examination of Giemsastained peripheral blood smears revealed B. ovis infection. The parasitological diagnosis was confirmed using polymerase chain reaction (PCR) assay by using a pair of B. ovis-specific primers. Mean values total protein and α and γ globulin fractions in infected goats were significantly higher (P<0.05) than those observed in the healthy group.

This study was conducted to evaluate the effect of phenylbutazone injection on serumic levels of thyroid hormones in the Arabian horse. Twelve Arabian horses were allocated to two groups of control and treatment each consisting of six animals. In the treatment group, 3 mg/kg phenylbutazone was injected intravenously for 6 days while in the control group equal values of 0.9% NaCl solution was used for the injection. Blood samples of all animals were collected from the jugular vein at days 0(before injection), 1, 2, 3, 4, 5 and 6, their sera separated by centrifuging and the levels of T3 and T4 were measured using the ELISA technique. There was a significant difference (P