نشریه تحقیقات دامپزشکی و فرآورده های بیولوژیک
سال سی و ششم شماره 2 (پیاپی 139، تابستان 1402)

The aim of this study was to investigate the effects of two medicinal plants of thyme and rosemary and their essences on gas production, digestibility, ruminal fermentation  and methane production parameters.  The gas production test was conducted with a 96-hour cultivation time in the form of a completely randomized design, along with four repetitions per treatment. The treatments consisted of: 1) basal diet with -  a forage-to-concentrate ratio of 50:50 without addition of the medicinal herbs and essences as the control treatment; 2) control treatment + 15 g/kg of thyme; 3) control treatment + 15 g/kg of rosemary; 4) control treatment + 100 mg/kg essence of thyme and 5) control treatment + 100 mg/kg of rosemary essence.The results of this study indicated that the application of the essences caused a significant increase in the gas production potential from the fermentable segment (b) and the gas production rate constant (c) compared to the control treatment (P <0.05). The treatments containing essences increased dry matter digestibility, organic matter and total volatile fatty acids and propionic acid as compared to the control (P<0.05).  Methane production in milliliters per gram of dry matter and ammonia nitrogen decreased remarkably in all experimental treatments compared to the control treatment(P<0.05). Overall, according to the results of this study, it seems that changes to the ruminal fermentation processes are possible by using the herbs and their essences, although additional –experiments  are - needed to investigate these effects.

Providing a standard diluent for preparing the lyophilized Fowl Pox vaccine not only facilitates the preparation of the vaccine for inoculation and improves the accuracy of vaccination but can also affect the effectiveness of the vaccine. The aim of the present study was to investigate the role of a standard diluent in the preparation of the Fowl Pox vaccine and its effect on virus stability in the ready-to-use vaccine, vaccine efficacy, and effectiveness in field conditions. In this study, a colored diluent specifically prepared for the Razi poultry Fowl pox vaccine was evaluated and compared with conventional diluents recommended by vaccine producers. In the first experiment, the vaccine titer stability was investigated at 0, 12, and 24 hours after vaccine preparation by different diluents including specific diluent, distilled water containing 20% (V/V) of glycerin and PBS (Phosphate-buffered saline), as the positive control. In the second experiment, the efficacy of the vaccine prepared with a specific diluent, distilled water containing 20% (V/V) of glycerin, an imported diluent (containing water for injection), was evaluated following a challenge with the virulent Fowl Pox virus. In the third experiment, the effectiveness of the vaccine prepared with specific diluent or distilled water containing 20% (V/V) of glycerin was studied. The results of the first experiment showed that the use of a specific diluent or distilled water containing 20% glycerin could help to maintain vaccine titer during the post-preparation period compared to PBS (P <0.05). The results of the second experiment revealed that under the controlled laboratory conditions, there was no difference between the experimental diluents in terms of vaccinal take and vaccine efficacy in preventing post-challenge acute disease (P>0.05); however, all the vaccinated groups had higher vaccinal take and protection compared to the positive (inoculation of specific diluents without vaccine) and negative (inoculation of no vaccine and diluent) control groups (P<0.05). In the field trial experiment, the vaccinal take associated with the specific diluent group was 11% higher than the positive control group (P<0.05). In general, the findings of this study showed that the application of a specific diluent for preparing the Fowl Pox vaccine, not only maintains its maximum efficacy but also it has advantages over conventional diluents in terms of applying easiness and improving the vaccine effectiveness.

These days one of the most significant challenges to the control and prevention of tick population in the livestock and agricultural industries is increased drug resistance in ticks and the destructive environmental effects of synthetic chemical compounds. In this study, the effects of ovicidal and larvicidal of Hyaloma anatolicum anatolicum mite in exposure to different extracts of Moringa oleifera plant have been investigated. Aqueous, alcoholic and hydro-alcoholic extracts were prepared from M. Oleifera leaves according to Percolation method. Thence, the main components of the plant extracts were identified and analyzed by the gas chromatography (GC) method. The collected eggs from female ticks were immersed in the proximity of different dilutions of the extract (2.5, 5, 10 and 20%) and kept in the incubator for 10 days and the hatched eggs were counted in the experimental and control groups. Also, 14-dainfant ticks, after immersion in different dilutions of the extract, were counted to check the survival of the infant ticks and the actual mortality rate was calculated. The results showed that hydro-alcoholic and alcoholic extracts of the leaves reduced the hatching rate of tick eggs as well as larval survival compared to aqueous extract. The most important compounds identified by GC were Limonene (32.45%), beta-Bourbonene (49.78%), Terpinolene (25.57), delta-3-Carene (21.99%) and Clovene (58.38%) which were higher in the hydroalcoholic extracts than in alcoholic and aqueous extractsrespectively. Therefore, M. oleifera leaves contain compounds which can be effective in reducing the population of Hyalomma anatolicum ticks.

Hydatidosis is one of the most important parasitic diseases common to human beings and animals. Various  drugs are used to cure this disease.. Due to the side effects of the currently available drug treatments, efforts are being made to find effectual alternatives with fewer side effects. In this current research, curcumin, as an effective ingredient of turmeric, was examined on protoscolices of hydatid cysts in vitro. For this purpose, sheep livers infected with hydatid cysts  were obtained from Mashhad industrial slaughterhouse, the cyst protoscolices  were collected, and the viability of protoscolices  was determined by 0.1% eosin staining. To study scolicidal effects of curcumin, different concentrations of curcumin (40, 80, 160, 320 micromol ) along with positive control (Savlon) and negative control (normal saline) were applied on protoscolices  with 90% viability, 30, 60, 120, 240, 480 minutes after 37̊ C heat exposure . Subsequently, vital staining was done , and pictures were captured with a camera equipped with Tcapture software, finally, the viability of the protoscolices was calculated. All  the experiments were repeated four times. The scolicidal effects of curcumin concentrations of 160 and 320 μmol one hour after exposure were significant in comparison with the negative control group. Therefore, curcumin in the used form showed  protoscolicidal effects only in high doses. However, it may be possible  to increase the anti-protoscolices  effects of curcumin in future studies by  modifications or  combination with substances that increase its penetration and absorption.

In order to evaluate the cholesterol lowering effects of psyllium seeds and also to evaluate the effects of this medicinal plant on growth efficiency, blood parameters, carcass traits and quail egg quality, 160 21-day-old chicks were studied in cages using a 2×2 factorial design with two dietary levels of psyllium seeds (0 and 3%) and two dietary levels of cholesterol (0 and 1%) in a randomized complete block design with 16 experimental units including 4 treatments and 4 replications and 10 birds (male and female) in each replication for 40 days, which were divided into two 20-day periods (as first and second breeding periods). Experimental treatments included a diet without psyllium seed and cholesterol, a diet containing cholesterol, a diet containing psyllium seeds and a diet containing psyllium seed and cholesterol. Serum, egg yolk and muscles (thigh and breast) cholesterol were evaluated in the treatments along with growth efficiency, blood parameters and egg and carcass quality. The results showed that oral administration of cholesterol had no effects on increasing serum lipids and cholesterol (P>0.05). In quails fed diets containing psyllium seeds compared to diets without psyllium seeds, breast and thigh muscles and egg yolk cholesterol, lymphocytes, relative weight of visceral organs, cecum, testis, thigh and liver were decreased and white blood cells, neutrophils, relative weights of carcass, duodenum, breast and pancreas were increased (P<0.01). The use of psyllium seeds throughout the entire study period increased feed intake and weight gain. The feed conversion ratio improved in the first period of rearing due to the use of psyllium seeds in the birds' diets. It was found that while cholesterol is  used solely in the diet, it increases the level of cholesterol in the muscles of the breast and thighs and egg yolk compared to the control group, but while cholesterol is used in combination with psyllium seed in diet, no increase in the cholesterol of those organs will be observed. Therefore, the use of psyllium seeds in diets with high cholesterol levels can play a beneficial role in preventing the increase of muscle and yolk cholesterol.

Reduction of honey bee colonies in recent years has been  a matter of concern in the beekeeping and agricultural industries. One of the main causes of  mortality in colonies is infection with viruses. In the present study, the molecular epidemiology of six honey bee pathogenic viruses, which are deformed wing virus (DWV)و acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), black queen cell virus (BQCV), Sacbrood virus (SBV) and Kashmir bee virus (KBV) and was studied in apiaries of eight provinces of the country (Fars, Sistan and Blouchestan, Kerman, Hormozgan, , Bushehr, Kermanshah, Kohkilouyeh and Boyer Ahmad and Khouzestan). 45 samples of honey bees were collected. Samples were examined by RT-PCR method and the PCR product was electrophoresed on a 1.2% agarose gel. Also, PCR products of positive samples were sent for sequencing. Results were positive for deformed wing virus 17 (37.7%), chronic bee paralysis virus 16 (35.5%), black queen cell virus 6 (13.3%) and acute bee paralysis virus 1 (2.2%) , but no sacbrood virus and keshmir bee virus  were detected. The highest percentage of chronic paralysis virus (100%) was observed in samples of Kermanshah province and the highest percentage of black queen cell virus (62.5%) and deformed wing virus (75%) were observed in samples of Fars province. Acute paralysis virus (14.29%) was observed only in samples of Hormozgan province. The results of this study showed that epidemiological studies of honey bee viruses are vital to know the dynamics of virus prevalence and can provide key information for designing surveillance programs against these beekeeping threats.

Based on the RNA-seq method, a thorough set of information can be obtained from the identification of geneswhich the results  may be beneficial  in the genetics and breeding of livestock and poultry. So far, few studies have been reported regarding the use of this method in the evaluation of native poultry transcriptome. Considering the importance of lncRNAs in biological processes, the present study has investigated the lncRNAs from the transcriptome of native Isfahan  chicken and the commercial Ross 708 strain as well as differences  in their expression. In this study using next generation sequencing, 568 lncRNAs were identified, out of which the expression changes of 116 lncRNAs were meaningful (P-value ≤0.05). The results showed differently-expressed incRNAs which are associated with the biological pathways including histone and lysine methylation, myeloid leukocyte activation, positive regulation of cytokine production ,, , and regulation of lymphocyte activation. The analysis of molecular functions of the genes indicated the activity of non-membrane-spanning tyrosine kinase, protein methyltransferase, S-adenosylmethionine-dependent methyltransferase, and transfer of one-carbon groupsacting on a protein, and signaling receptor binding are enhanced meaningfully. Based on the results, lncRNAs that vary in expression between native and commercial groups are associated with immune and methylation pathways, and can be used as biomarkers.

The aim of this study was to investigate the effect of saturated and unsaturated fat sources in the diet of late pregnancy and early lactation on histological changes in the mammary gland of nulliparous Saanen goats. Thirty does from the second half of pregnancy were divided into three groups based on body weight. For each group, the diet contained either: 1) saturated fat (positive control, C+), 2) roasted soybeans (omega-6, SB), or 3) extruded flaxseeds (omega-3, FS). Milk production was determined weekly and mammary tissue sampling was performed three times after kidding and the histological information of the mammary tissue was obtained using Image J software. The results showed that consumption of flaxseed and soybean increased milk production in the whole period compared to the C+ group (P<0.05).  The percentages of epithelial cells and stroma were the highest and the lowest, respectively in the FS group compared to the other groups(P<0.05). The areas of alveoli, lumen, epithelium and the number of epithelial cells per alveolus were higher in the SB group; however, the number of alveoli per unit area  was lower in SB group than  in the other groups (P<0.05). Based on the results, it is concluded that the consumption of diets rich in unsaturated fatty acids, especially omega-3 fatty acids, in critical periods of mammary gland growth, affects growth and development of mammary tissue, improves lactation performance and increases milk production.

H9N2 subtype of avian influenza virus (AIV) is the most common strain of low pathogenic AIV in the Middle East. The aim of this study was to investigate the humoral and cellular immune responses of commercial layer chickens immunized with gamma-inactivated H9N2 AIV. For this purpose, 120 one-day-old chickens of layer breed were divided into four groups including 3 replicates and 10 birds per replicate. Vaccination of chickens was done on days 11 and 25 of age. The first and second groups received the irradiated vaccine (IV) by intranasal and subcutaneous injection methods respectively, the third group was injected with formalin-inactivated vaccine and the fourth group was not vaccinated as the control. Anti-H9N2 serum antibodies, proliferation rate of splenocytes and the expression levels of interleukins 2, 6 and 10 and interferon gamma in these cells were investigated. The results indicated that in the groups immunized with intranasal IV the level of HI titers at 25 days of age was significantly higher than those in other groups. Additionally, stimulation index of splenocytes in 39 and 85 days of age and the level of gene expression of all the measured cytokines, especially interferon gamma in splenocytes at 39 and 55 days of age was significantly higher compared to formalin-inactivated vaccine. Based on the findings of the present study, the use of H9N2 AIV vaccine inactivated by gamma irradiation could be considered as a good candidate in layer chickens because it could induce antibody responses and cellular immunity at the levels higher than the formalin-inactivated vaccine.

Selenium is an essential element in dairy cattle, particulrarly for reproduction. The present study was conducted in order to determine the amount of selenium in the blood of different groups of cattle in Isfahan province industrial cattle farms and also the effect of season (summer and winter) on its concentration in the form of a randomized complete block design. The present study was investigated the blood selenium level in dairy cattle in Isfahan province. Also, the effect of season (summer and winter) on selenium values was studied. This experiment was carried out in a complete block randomized design for 553 cows in 17 farms from different regions of the Isfahan province. Acordingly, 553 cows were selected from 17 industrial cattle breeding units in different geographical parts of the province. The population of cows in each cattle farm wasere divided into five groups which included high producing cows (n=154), fresh cows (n=118), dry and pregnant heifers (n=121), non-pregnant heifers (n=81) and calves. Blood selenium concentration was indirectly assessed determined through the measurement of glutathione peroxidase activity by Alpha Classic Auto Analyzer. According to the results, the mean whole blood selenium concentration mean for high producing cows, fresh cows, dry and pregnant heifers, non-pregnant heifers and calves were measured at 0.21, 0.18, 0.24, 0.24, 0.21 mg/L, respectively. Dry and pregnant heifers and non-pregnant heifers with higher selenium concentration showed significant differences compared to other groups (P<0.05). The lowest selenium concentration was significantly observed for fresh cows (P<0.05). No significant differences were found among different groups of dairy cows in terms of peroxidase enzyme activity for glutathione peroxidase activity. The mean blood selenium concentrations in summer and winter with no significant differences were at 0.21± 0.08  (n=129) and 0.20±0.06  (n=174) mg/L, respectively which showed no significant effect of season on blood selenium level. Also, the comparison of the same group in different farms showed significant difference in terms of blood selenium levels s between them (P<0.05).Therefore, it is recommended to check the blood selenium level of dairy cattle and then ,use a selenium supplement under supervision of animal nutritionists.

Salmonella Typhimurium is a zoonotic gram-negative bacterium of the Enterobacteriaceae family. One of the newest ways against the pathogen is the use of recombinant vaccines. In this study, a recombinant construct consisting of OmpL antigen from Salmonella Typhimurium and HBHA protein as a molecular adjuvant from Mycobacterium was designed by an in silico method and a peptide linker. For this purpose, sequences of these proteins were extracted from the database and the HBHA-OmpL recombinant construct was designed based on the correct  reading frame. Then, reliable online servers were used to evaluate the immunogenicity score and  physicochemical properties as well as the secondary and tertiary structures of the designed construct. The best refined 3D model was used for the molecular docking process. A codon optimization was done for expression in E. coli, and the possible cloning  of the optimized nucleotide sequence in pET22b(+) expression vector was evaluated. Based on the results obtained in this study, the HBHA-OmpL chimeric construct with a molecular weight of 49.45 kDa had an instability index of 32.75 and an antigenicity index of 0.688, and alpha helices and random coil structures had the largest contribution in its secondary structure. The protein-protein docking results showed that despite being conjugated with OmpL protein, HBHA molecule was able to connect to TLR4/MD2 receptor by 23 hydrogen bonds. Also, cloning results confirmed that nucleotide sequence of the designed vaccine can be successfully inserted in pET22b(+) vector.

Wound healing is a complex process in which a series of events takes place in a coordinated manner, which ultimately leads to tissue regeneration. One of the new treatment methods in medicine and veterinary medicine is the use of platelet-rich plasma (PRP) and plasma medicine.  The present study was conducted in order to compare the effect of cold argon plasma and platelet gel on the amounts of hexosamine, collagen and total protein in the tissue, in the wound healing process of rats; accordingly, 48 Wistar rats were used. After the adaptation period, the mice were classified into one control group and four treatment groups. The treatments included: nitrofurazone ointment treatment, platelet-rich plasma gel treatment, cold argon plasma treatment, and platelet gel treatment with cold argon plasma, and no treatment was performed in the control group. After the treatment and creation of skin wound, samples from the wounds were taken from the entire thickness between the shoulders of the rats, on days three, seven, 14 and 21 and collagen factors (hydroxyproline), hexosamine and total protein were measured. The results of measuring these three factors showed a significant difference between the control group and the plasma and plasma + PRP group (P<0/05). Meanwhile, no significant difference was observed between the control group and the nitrofurazone ointment group (P>0.05). As a result, it seems that the use of plasma and plasma + PRP accelerates the wound healing process significantly compared to the control group.

The aim of this study was to evaluate the sperm quality preservation in sheep using glutathione antioxidant during cooling process. In this experiment, semen samples were collected from 5 Zell rams and added to the extender containing 0, 1, 2, 4 and 8 mM glutathione and stored at 4 ˚C until 48 hours. Sperm quality was evaluated at 0, 24 and 48 hours after cooling and total motility, progressive motility, viability, membrane integrity, mitochondrial activity and lipid peroxidation were assessed. No difference was observed between treatments at time 0 of cooling. At 24 and 48 hours of cooling period, the use of 4 mM glutathione increased total motility, progressive motility, viability, membrane integrity, mitochondrial activity and decreased lipid peroxidation in ram cooled sperm. In conclusion, using 4 mM glutathione antioxidant treatment could be a suitable method to preserve ram sperm quality during cooling process.

In recent years, due to advances in vaccine design, bioinformatic studies on Leptospira have undergone significant changes and advances. The aim of this study was to design a fusion protein based on OmpL1 and LipL41 surface proteins that have a conserved structure over time. Based on the previous field studies, the target proteins were examined and the OmpL1-LipL41 fusion protein was selected. After examining the chemical properties of the target proteins using appropriate servers, the initial structure of the fusion protein was determined and finally this protein was placed in the PET32a+ vector between the two enzymatic regions of HindIII and BamHI. The results revealed that the protein weighs 73 kDa and has 2097 bp and 610 amino acids. The isoelectric pH of the above protein is 7 and its solubility degree in the prokaryotic system is 100%. Finally, according to the above data, it is predicted that the designed construct structure has acceptable immunogenic properties and can be expressed to an acceptable and successful extent in prokaryotic systems and used for immunogenicity studies of Leptospira as a new candidate in the production of leptospirosis vaccine and in the design and manufacture of diagnostic kits.