نشریه پژوهش های علوم دامی
سال سی و یکم شماره 1 (بهار 1400)

Freezing is one branch of cryobiology knowledge that process to protecting and long term keeping of cell in very low temperature. Sperm freezing commonly done in infertility treatment centers and keeping sperm banks and livestock industry. Sperm can survive decrease out of body and environment temperature. freezing is so important one of influential destructive factors in reducing duration of sperm storage is creation of ROS. It needs very low, but it is increasing has destructive effects on sperm (lenzi et al. 1994). Use of antioxidants in sperm extender have better results to deal with these reactive oxygen species. It is believed that ROS induces lipid peroxidation in the sperm membrane and The peroxidation of the resulting fatty acids has toxic effects on the sperm that Leading to a decrease in sperm function (Sanuka and Corps 2004). Antioxidants are compounds that control, stop and neutralize intermediates derived from the energy production process in the electron transport chain of cells (aerobic activity) known as active oxygen species, and in a group of free radicals (Zinee et al. 2000). Adding antioxidants to semen diluent during freezing of sperm improves the quality and improvement of semen parameters such as motility, viability and acrosome membrane integrity (Mironoksuk et al. 2018). Selenium is an essential micronutrient element to human health Which is toxic at high concentrations. Selenium is a constituent of selenoproteins that they have an enzymatic and structural role in biochemistry. Selenium is the main component of selenoenzymes in the center of all of these proteins, there is the amino acid of selenocysteine Which acts as an oxidation-reducing agent. However, one of the limiting factors for selenium consumption is bioavailability (the rate of entry into circulation and tissues) and its toxicity (Tarz et al. 2007). Therefore, the use of other forms of selenium that is less toxic and more beneficial is more appropriate. Selenium nanoparticles are low toxicity compounds and bioavailable and effectively increase the production of selenoproteins in the body (Peng et al. 2007). Selenium is a cofactor or activator of the glutathione peroxidase enzyme Which is one of the strongest antioxidants in the body and catalyzes the decomposition of lipid peroxides and hydrogen peroxide (Hill et al. 1996). The glutathione peroxidase enzyme destroys free radicals within the cytoplasm (Ozball et al. 2008). Typically, the concentration of most of the elements in the liver is higher than the other organs, but in the case of selenium, it has the highest concentration in the testis. High concentration of selenium in testis indicates the protective role of this element and its related enzymes in spermatogenesis (Schumberger et al. 1983).

Semen was collected with an artificial vagina twice a week from 4 selected adult Ghezel rams during non-reproductive season. Ejaculates were immediately evaluated primarily for parameters including volume, concentration, motility, viability and abnormal sperm. And if they had the required standards (high concentration of 3 billion sperm per ml, motility above 70% and volume greater than 0.5 ml), they were selected as suitable samples for dilution. The semen was diluted with extender with no antioxidants (control) and containing 1 µg/ml Nano Selenium and 2 µg/ml Nano Selenium and then immediately aspirated to 0.25 ml freezing straw. After recording, treatments were packaged and refrigerated in 5°C for 1.5 hours. Then, the packaged straw containing semen were placed on 4 cm above the liquid nitrogen, then were immersed in liquid nitrogen immediately after 10 minutes. Qualitative parameters of sperm included viability, total motility (TM), progressive motility (PM), plasma membrane integrity (HOST), percentage of abnormal sperm and acrosome membrane damaged was studied in days 0, 15 and 30 of freezing process.

Percentage of viability, total motility, progressive motility, plasma membrane integrity (HOST) of sperm decreased significantly (p < 0.01). Also the percentage of acrosome membrane damage and abnormal sperm increased during experiment (p < 0.01). However, the addition of different levels of Nano Selenium to semen extender significantly increased the percentage of viability, percentage of sperm with total motility, progressive motility and health plasma membrane (positive HOST) on days 0, 15 and 30 compared with control group (p < 0.01). Also, the addition of different levels of Nano Selenium to extender significantly decrease the percentage of abnormal sperm and acrosome membrane damaged (p < 0.01).

According to the results of this study, the addition of different levels of Nano Selenium, especially Nano Selenium 1 µg/ml to ram sperm extender increases the quality of sperm during storage.

Study field:

 The amount of residues of some pesticides in some animal and poultry feeds in Iran is considered higher than the international standard.

The purpose of this study was to determine the effect of reducing the amount of patulin and diazinon in ensiled apple pomace on some rumen parameters (pH, protozoa, ammonia nitrogen) and the concentration of volatile fatty acids in the mahabadi lactation goats that consuming diets containing high apple pomace (28.88% in dry matter).

In this study, 30 mature newborn Mahabadi lactation goats with mean live weight of 55 ± 5 kg, with 5 experimental treatments and 6 replications were used in a completely randomized design. Experimental treatments consisted of unprocessed apple pomace (control group), microwave apple pomace (treatment 2), processed apple pomace with two commercially active adsorbent and inactive pesticides (50 g / d respectively For every goat head and and one kg / t of apple pomace (3 and 4) and apple pomace processed with an acidifiers commercial compound (treatment 5).

The results showed that the highest amount of patulin and diazinon in different treatments were in the control group (178 Kg / μg and 936.0 mg / kg) and the lowest levels were related to treatments treated with Bio-Tox toxin binder and Mycofix Plus (58 Kg / μg and 279/0 mg /kg). Therefore, only the pH of treatment 5 decreased significantly in comparison with control group and decreased to 6.36 in comparison with 6.42 control group. Also, the results showed that ruminal protozoa population was not affected by apple pomace processing in different treatments, and the ammonia nitrogen concentration of rumen fluid in test goats showed a significant difference between treatment 2, 5 and 3, in relation to, control and other treatments respectively. The lowest amount for control group (without processing) was 21.42 mg /dl and the highest amount for 3 was 12.22 mg /dl by adding Mycofix-Plus to the diet (0.05 mg /dl). P). The results also showed a significant increase in the concentration of total volatile fatty acids, except treatment 4 compared to control (P ˂ 0.05).

Generally reducing the patholin and diazinon toxicity by processing with adsorbent and inactive compounds of toxins in high-level diets of apple pomace can improve some of the animal's rumen parameters, and, in addition to ensuring health, will increase the efficiency and performance of the animal. Key words: Patulin, Diazinon, Apple pomace, Toxic Adsorbent, Mahabadi Lactation Goats Study field: The amount of residues of some pesticides in some animal and poultry feeds in Iran is considered higher than the international standard.

The analysis of blood cells is used for detecting several diseases in human and animals. White blood cells, also called leukocytes, are an important part of the body’s defense system against disease. In response to infection, the white blood cells (WBC) count typically increases (Sparavigna 2017). Therefore, measurement of the WBC count able to give an indication if there is infection or inflammation occurring. Increases in WBC can also occur in some cancer conditions such as leukemia. A bird under stress may also have a high level WBC, but greater elevations are a definite indication of disease. Reduced numbers of WBCs can occur due to bone marrow disease, severe acute disease and other conditions. White blood cell counts can be determined by various methods either through estimation or varied counting techniques. At this time, there is no automated system, such as is used with humans or dogs and cats that has proven effective for determining avian white blood cell counts (Sakas 2002); this is because of avian blood cells are nucleated, therefore, leukocytes cannot be separated from other blood cells. Bare nuclei from lysed cells, nucleated thrombocytes, and lymphocytes are all of similar sizes and difficult to distinguish by impedance or light scatter properties, and the contents of the leukocyte granules of birds differ from those of mammals. In addition, morphologic differences between avian species commonly limits the development of automatic protocols. These qualities limit the use of centrifugal hematologic analyzers such as quantitative buffy coat analysis, impedance, and laser cytometry tools to differentiate all types of cells (Harrison and Lightfoot 2005). Total WBC count is usually performed in laboratory by using manual techniques and hemocytometer. Differential counting has also been done by stained blood smear and light microscopy. Present of nucleated erythrocytes and thrombocytes in avian blood prevents from automated method that have been developed for mammals. Also, manual counting is time-consuming and requires to an experienced technician (Beaufrere and et al. 2013). The evaluation of blood smears for cell differentiation and detection of anomalies is a complex process and many morphological aspects of the cells are assessed and analyzed. Therefore, because human visual perception is a type of computation, it is possible that digital images can be analyzed by using a similar process. Content-based image indexing and retrieval has been an important research area in computer science for the last few decades. Many digital images are being captured and stored such as medical images, architectural, advertising, design and fashion images, etc. As a result large image databases are being created and being used in many applications (Maitra and et al. 2012). Therefore, this project carried out to examine the applicability of image processing system for rapid detection of white blood cell in broiler chickens.

Blood samples were taken from 15 broiler chicks, and then blood smears stained with Giemsa color and finally were photographed by Canon E500. A total number of 80 RGB color image were created in jpg format, 42 images were selected and WBC counted by confirmation hematology expert. After that the images were processed by MATLAB R2017a and CellProfiler (CP 2.0 r10997) softwares. The appearance of WBC such as the color, shape and size were considered by MATLAB software. The images were examined in RGB, HSV and L*a*b color spaces. Visual examination and index recording showed that the color feature in all color spaces is not useful for detecting WBC. Since the cell nucleus was more distinct in a-layer images than other layers, the image of this layer was used for further processing. The a-layer images converted to binary images by a threshold. Then obtained binary image examined by the implementation of different conditional ring based on their size and shape. Two methods were used based on size; the first method is based on remaining four larger objects, and the second method removed the areas less than 5000 pixels in the image. In the shape method the interval 1-1.25 was considered to be form a factor for remaining the round objects and in the combined method, the removal of area less than 5000 pixels with an interval 1-1.25 have been considered. In the image processing by cellProfiler software, different modules and typical diameter 18-52 pixel for objects were used.

Among the different methods, the image processing with MATLAB software, and the use of second method (removed areas less than 5000 pixels in the image) had the highest number of diagnostic cells (84% of WBC were correctly detected). The rate of diagnostic cells of other method (first, third and fourth) were 35, 48 and 39% respectively. In the image processing by cellProfiler software, by using various modules and typical diameter 18-52 pixel for objects, WBC was identified in the rate of 84%. The results of our study show that our methodology was not consistent enough to agree with WBC counts obtained by using manual differential, which invalidates our initial hypothesis. Therefore, it is speculated that variability in the blood smear features, such as smear thickness, cell spread, and, therefore, surface area, cell touching, uneven spatial cell distribution (depending on areas of the blood smears), and the similar appearance of thrombocytes and small avian lymphocytes could contribute to the results. Color error and nucleated red blood cells are the biggest problem in this project and more advance algorithms are required to solve this problem. Based on our results it seems that nothing can replace the expertise of a trained technician or veterinarian, but manual CBC determination is time consuming, partially subjective, and semi quantitative. Despite tremendous advances in the machine learning, recent investigations have shown that humans have better visual categorization abilities than machines. However, machines are better, for instance, at computing complex geometric properties, performing more precise measurements and statistical evaluation, and evaluating complex sets of rules, and they can adapt quickly from training samples.

Although our technique did not provide satisfactory results in its present form, we hope that this approach can still stimulate further research in the application of digital image analysis. Generally, the results showed that more advanced machine-learning algorithms and neural networks are required to be test for counting and differential white blood cell diagnostics.

Meat and dairy products from ruminants are the main source of vaccenic acid (VA) and conjugated linoleic acid (CLA) in human nutrition. In particular, the isomer of cis-9, trans-11 CLA is active in the prevention of cancer and atherosclerosis in human (Belury, 2002). CLA is partially synthesized in the rumen by cellulolytic bacteria, and mainly by Butyrivibrio spp. during the biohydrogenation (BH) of linoleic acid (cis-9, cis-12 C18:2, LA). However, the largest proportion of CLA present in milk is endogenously produced in the mammary gland by the action of the enzyme Δ9-desaturase on vaccenic acid (trans-11 C18:1, VA), which is another product of the ruminal BH (Jenkins et al., 2008; Shingfield et al., 2013). In the last decade several efforts have been made in order to develop efficient enrichment strategies of VA and CLA in ruminant products (Shingfield et al., 2013). The main strategy to enhance these beneficial fatty acids (FAs) content in food is manipulating ruminant feeding and modulating rumen BH. Tannins are phenolic compounds that are chemically classified as hydrolysable (HT) or condensed (CT). Tannins extracted from chestnut and oak are HT and those extracted from quebracho are an example of CT (Waghorn, 2008). Tannic acid (TA) is a commercial HT. Tannins have been reported to modify ruminal fermentation by inhibiting ammonia and methane production (Carulla et al., 2005) partially by their ability to form complexes with dietary protein and fiber (Waghorn, 2008). It is widely known and accepted that tannins are able to bind proteins and inhibit the growth of ruminal bacteria (Min et al., 2003). Concerning lipid metabolism, tannins were shown to inhibit Butyrivibrio fibrisolvens (Jones et al., 1994), one of the bacterial species known to be a major microbial species involved in ruminal BH (Jenkins et al., 2008). In vitro and in vivo studies have suggested that feeding tannins to ruminants can favourably alter ruminal BH of dietary linoleic acid, enhancing accumulation of VA in the rumen and thereby the content of some human health promoting FAs, such as VA and CLA in dairy or meat products. However, reports on impacts of these phenolic compounds are very limited and inconsistent. The main objective of the present study was to verify whether the TA inhibit the ruminal BH of diet containing extruded soybean seeds in vitro and whether there is a dose-dependent effect of TA.

Basal diet formulated to contain high concentrate to forage ratio (58.2:41.8) and extruded soybeans as unsaturated FA source. Experimental treatments consisted of control (C; without additive), 0.1 mg/L of monensin (M), 250 mg/L of tannic acid (TA1), 500 mg/L of tannic acid (TA2) and 750 mg/L of tannic acid (TA3). Sample of basal diet was dried at 60°C in a forced air oven for 48 h, ground to pass through a 1-mm screen using a Wiley mill. The rumen fluid was collected from two Holstein cows fed alfalfa a mixture of hay and wheat straw (700:300 g/kg on a DM basis) ad libitum. Ruminal fluid was filtered through four layers of cheese cloth and transferred quickly to the laboratory in anaerobic condition at 39° C and was continuously purged with oxygen free CO2 to ensure anaerobic conditions. The buffer solution was prepared according to Makkar et al. (1995) procedure and mixed with rumen fluid as 3:1 (v/v). Incubation was carried out in 150 mL bottles containing 200 mg of the basal diet and 40 mL of buffered rumen fluid. Gas production (GP) after 24 h incubation, ruminal volatile FA (VFA), ammonia-N and pH and FA profile were measured.

The results showed that gas production (GP) and ruminal fluid concentration of ammonia-N and volatile fatty acid (VFA) were not affected by additives (P > 0.05). Alipanahi el al. (2019) reported no effect of oak acorn (HT source) on ruminal pH and VFA concentration in lactating does. The capacity and trend of tannins to bind to specific proteins may be dependent on the type of protein (Gonzalez et al., 2002). In our experiment, it was expected to reduce ruminal ammonia-N concentrations by TA. The lack of effect of TA on ruminal ammonia-N may be related to the type of protein source and processing method. In the current experiment, we used extruded soybeans, which may loss its natural structure to bind to TA. Ruminal fluid C10:0 to C14:0 concentrations decreased and C16:0 increased by TA (P < 0.05). The M and TA1 treatments had lower ruminal fluid C18:0 concentration compared to other treatments (P < 0.05). The highest trans-11 C18:1 (VA) and cis-9, trans-11 conjugated linoleic acid (CLA) were observed in M treatment (P < 0.05). Ruminal fluid unsaturated (USFA) and long-chain (LCFA) fatty acid concentrations were increased in TA2 compared to other treatments (P < 0.05). Vasta et al (2009) reported higher VA and lower stearic acid concentrations by different sources of tannin on in vitro study, but concentrations of CLA isomers were not affected. They concluded that tannins prevented BH of USFA by inhibiting the microorganisms. Similar to our results, feeding diet containing oak acorn (as a source of HT) increased and decreased USFA and SFA concentrations in does milk fat, respectively (Alipanahi et al., 2019).

Our results confirmed that addition of TA to a diet containing extruded soybean seeds can modify BH pathways without any negative effect on ruminal fermentation. Consequently, TA has potential to enhance beneficial FA in ruminant products, although more researches need to confirm these results.

Zinc is an essential trace mineral for all forms of life because it plays an important role in ‎DNA and RNA synthesis by increasing replication and cell proliferation. Zn deficiency in grazing and ‎forage feed is widespread in Iran and other countries, so Zn supplements are generally required for ‎ruminants. Several kinds of Zn sources have been used in diet ruminants. Supplemental Zn is usually ‎added to animal diets in the form of inorganic Zn (Zno and Znso4), However,. Many studies reported ‎that organic Zn is absorbed more readily by ruminants than inorganic Zn. Recently, a novel elemental ‎Zn source used in animal diets. The development of nano technology holds unique properties for nano ‎Zn, because nanometer particulates exhibit novel characteristics, such as great specific surface area, high ‎surface activity, high catalytic efficiency and strong absorbing ability. The objectives of this study were to evaluate the effects different forms of zn on feed digestibility, rumen fermentation of diatery dairy ‎cows with high production using gas production and nylon bags techniques.‎

The Zno naoparticles were prepared by chemical co-precipitation method. The ‎basal diet for sheep and dairy cows were provided according to NRC (2007) and NRC (2001) ‎respectivelely, to meet requirements, except of Zn. The four various tretments in the current experiment ‎were as: (1) the basal without Zn supplementation. (2) the basal diet plus 40 mg Zn/kg DM as zinc ‎oxide (Zno), (3) the basal diet plus 40 mg Zn/kg DM as zinc glycine (ZnGly, ZnGly, B-TRA MIX R ‎‎2C, 18.5% Zn) and (4) the basal diet plus 40 mg Zn/kg DM as Zn nano (zn>99%, 15-20 nm). Four ‎ruminal fistuled male sheep were used in a replicated 4×4 Latin square, experiment with four 28-day ‎periods (21 day adaptation and 7 day sampling collection each period). Nylon bags methods 5g of ‎samples were incubated in the rumen of fistulated sheep for 0, 2, 4, 8, 12, 16, 24, 36, 48, 72 and 96 h ‎for determination degradability of dry matter (DM) and crude protein (CP). The gas production was ‎recorded after 2, 4, 6, 8, 12, 16, 24, 36, 48,72 and 96 h of incubation. Ruminal fluid were used for pH, ‎ammomia-N, voliate fatty acids (VFA). ‎

In present study, the highest amount of gas production was obtained in ‎organic, nano and in organic zin supplemented treatments respectively. Zinc supplementation ‎significantly increased the OMD, SCFA and ME concentrations (p < 0.05). The results of this study ‎showed that Zinc supplementation significantly (p < 0.05) decreased the N-NH3 concentration in ‎different experimental groups. There was no effect on molar proportion of acetate and butyrate ‎‎(p < 0.05), whereas that of propionate was linearly (p < 0.05) increased with increasing nano-zn and ‎glycine-zn supplementation. As a result, ratio of acetate to propionate was decreased. Total ruminal ‎VFA concentration was increased (p < 0.05) as Zinc Supplementation. Increasing the concentration of ‎total volatile fatty acids in the rumen can be due to improved microbial fermentation in the rumen. In ‎situ trial, adding zn in forms nano and organic increased the DM disappearance (p < 0.05). Also zn ‎supplementation significantly increased ;b; fraction of DM of diatery cows with high production in ‎different experimental groups. The present results suggested zn functions to maintain the production of ‎proteolytic digestive enzymes and activity of protein decomposing bacteria could be improved by zn. In ‎‎24 to 96 hours incubations crude protein degradability zn-glycine and nano-zn was significantly ‎‎(p < 0.05) lower than zn oxide and control treatments. Also zinc Supplementation in forms nano and ‎organic decreased (p < 0.05) ;b; fraction and ;ED; of crude protein of diatery cows with high production ‎compared to control and zinc oxide treatments. The reason for this can be because of the low solubility ‎of proteins dietary due to bonding with the zinc element in the rumen or the change in the activity of ‎the proteolytic microbes.‎

The results suggested that the addition of zinc in the forms of organic and nano increased ‎the fermentation and degradability of the dietary high production dairy cows compared with inorganic ‎form. Also, there was no difference in digestibility and fermentation between organic and nano forms; ‎hence, in order to select the nanoparticulate or organic form of zinc element in the diet of animals, it ‎was necessary to pay attention to their price and price in the market.‎

Description of the subject. Increasing the concentrations of blood glucose lead to decreased appetite and decreased concentration of stimulant in the hunger center of the bird's hypothalamus and consequently decreased feed intake. High intakes of vitamin C may alter blood glucose concentration, although the evdienec is contradictory. Objectives. The aim of this study was to evaluate the effects of three different levels of vitamin C (0, 200 and 400 mg/kg DM) on performance, carcass and digestive organ characteristics, blood plasma components, liver enzymes, immune system, and cecal microbial flora of broilers. Method. The study had a completely randomized design with 3 treatments and 4 replicates of 10 chickens per pen. The study lasted 42 days and started with 120 one-day-old male chickens of commercial Ross 308 strain. Tmt measn were compared using Duncan's multiple-range test. Results. Chicks fed a diet containing 200 mg/kg vitamin C had the highest feed intake and weight gain as well as the best feed conversion ratio, the lowest cost per kilogram of live body and the best European production factor. The effect of vitamin C was significant on live body weight, featherless weight, full abdomen carcass weight, empty abdomen carcass weight, relative crop weight and relative breast weight (p <0.05) and the highest increase was related to the level of 200 mg/kg vitamin C. On the other hand, the use of different levels of vitamin C on blood parameters and liver enzymes of broilers was not significant (P≥0.05). In addition, the percentage of neutrophils and lymphocytes was significant (p <0.05) and the highest percentage of neutrophils was also for using the level of 200 mg/kg vitamin C. Comparing the mean results of antibody titers against SRBC did not show a significant difference (P≥0.05), except for 35 days of age, which was significant (P <0.05). Numerically, the highest antibody titer was related to the level of 200 mg/kg vitamin C. So, based on the results of the present study, the use of 200 mg/kg vitamin C to supplement the diet of Ross 308 strain broiler chickens is recommended. Conclusions. Although vitamin C was not very effective on blood parameters, it improved the immune system and reduced the fat content of the ventricular area, thereby improving the quality of carcass meat. Therefore, according to the results of this experiment, it is recommended to use 200 mg/kg vitamin C in diet as an antioxidant compound and a cheap promoter of growth.