فهرست مطالب

  • سال دهم شماره 2 (تابستان 1397)
  • تاریخ انتشار: 1397/05/08
  • تعداد عناوین: 12
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  • A. Naserian*, R. Khodaverdi, R. Valizadeh, A. Tahmasbi Pages 153-162
    Introduction
    The processed crop by-products can be used as a suitable feedstuff to meet livestock dietary requirements. Tomato shoots that left on the field surface after harvest, is a major valuable by-product. In Iran, the estimated annual tomato shoots production is about 100 million ton. We evaluated the nutritional value of ensiled tomato shoots processed by beet pulp as a fermentation enhancer.
    Materials and Methods
    Tomato shoots were collected from tomato farms in Semnan, Iran. The shoots were ensiled with 0, 4 or 8% (W/W) air-dried beet pulp. After 60 days, the silos were opened and ensiled materials were evaluated for sensitive characteristics as well as pH. The ensiled samples were also analyzed for chemical composition after 48 h oven drying at 60° C. Rumen degradability and fermentation characteristics were measured by using a nylon bag method and an in vitro gas production technique, respectively. For in vitro gas production technique, 200 mg of dry matter was incubated for 96 h at 39° C. Cumulative gas volumes were recorded 2, 4, 6, 8, 12, 24, 36, 48, 72 and 96 h after incubation. To estimate the gas production parameters, an exponential model was fitted for data. Rumen degradability parameters of dry matter (DM), crud protein (CP) and neutral detergent fiber (NDF) were determined by incubation of 5 g sample for 0, 2, 4, 8, 16, 24, 48, 72 or 96 h in the rumen of a fistulated steer. Similar analysis and measurements were also performed for oven dried tomato shoot samples. All data were subjected to ANOVA using the GLM procedure of SAS and means were compared by using Duncan's multiple range test at 5% probability level.
    Results and Discussion
    At the end of fruit harvest, the measured DM and CP content of tomato shoot samples were 26 and 14% respectively that are in agreement with previous reports. Ensiled tomato shoots had a significant higher content of CP than oven-dried shoots; more likely due to losses of soluble components during the ensiling process. However, the content of CP was decreased with increase in beet pulp level in the tomato shoot silage. Similarly, the tomato shoot silages with 4 or 8% beet pulp showed lower values of NDF and ADF in comparison with tomato shoot silage without beet pulp. Due to lower protein and fiber percentage of beet pulp, the decreases in CP, ADF and ADF contents of omato shoot silages by adding beet pulp were expected.
    Although the potential of gas production was enhanced by increased levels of beet pulp in the silages, no
    significant difference was observed among tomato shoot silages. The silages were also similar for gas production parameters. However, cumulative volumes of gas produced at 96 h incubation for silages with 4 and 8% beet pulp were significantly higher than other treatment. The highest values of rapidly degradable DM, degradation rate constant and degradation potential were observed in tomato shoot silage supplemented with 8% beet pulp. However, there was no significant difference between tomato shoot silages with 4% beet pulp and without beet
    pulp supplementation.
    Conclusions
    Tomato shoots are a suitable source of nutrient for ruminant. Our results indicated that ensiling increases nutritional value of tomato shoots. The in vitro rumen fermentation characteristics as well as rumen degradability of tomato shoot silages were improved by adding 8% beet pulp. However, future studies are needed to confirm these results and elucidate the potential use of tomato shoot silage in ruminant's diet.
    Keywords: Chemical composition, Degradability, Gas production, Tomato shoot silage
  • A. Vakili *, S. Saki, M. Danesh Mesgaran, F. Alipour Pages 163-178
    Introduction
    In the ruminant animal, rumen fermentation provides nutrients, energy and protein needed for livestock, but this process is accompanied by the loss of energy in the form of methane and protein as ammonia nitrogen. Methane released from ruminants as an environmental pollutant with 38% greenhouse gases is one of the most important reasons for global warming. On the other hand, it reduces 2 to 15 percent of the energy consumed by the livestock. Ammonia nitrogen and nitric oxide are excreted through urine and fertilizers and pollute the environment. Therefore, researchers have tried to eliminate these problems from the past to add some supplements to animal feed to improve fermentation performance. Over the past few decades, ionophors and probiotics have been marketed as methane reducers. However, the antibiotic concerns and the level of these compounds in animal products have limited their use. To solve this problem, the use of herbs, essential oils and their extracts in the replacement of antibiotics and improvement of rumen function has been suggested. Essential oils are aromatic, volatile and oily compounds extracted from plants. They have antimicrobial properties against many bacteria, yeasts, and fungi. Due to the fact that herbal essential oils have strong antimicrobial properties, they are used as additives in livestock feeding. The purpose of this study was to evaluate the effect of some essential oils and pH on rumen fermentation parameters in different diets in in vitro.
    Materials and Methods
    In this study, the essential oils of Fennel (Foeniculu vulgare), Eucalyptus (Eucalyptus camaldulensis) and Pinus pumilio (Pinus pumilio) were used. For the extraction of pine and fennel seeds and Eucalyptus leaves were used. Water extraction was used to extract essential oils from Clevenger apparatus. For this purpose, 1Lit glass balloons (pine and fennel seeds were cut and Eucalyptus leaf grinding), 70 grams were used and 750 ml of distilled water was added to each balloon. Samples and distilled water were warmed up at 100°C. Heating was performed for 4 hours to evaporate the contents of the balloons. The essential oils of each sample were extracted on the cooled water from the evaporation procedure. Finally, each of the essential oils was stored in dark glass containers until refrigerated at 4 ° C. The first phase consisted of incubation 4 basal diet: 100% alfalfa hay, 100% concentrate (barley), mixed concentrate with alfalfa with levels of 80 to 20 and 60 to 40% of concentrates to alfalfa hay with zeros, 3 and 30 mg/l of essentiol oils pine, eucalyptus and fennel mixed with 30 ml of rumen fluid for 72 hours at pH different (5 = pH, 6 = pH and 7 = pH). The pH were set at desired levels using NaOH and HCl at the beginning of incubation to investigate the effect of essential oils and different pH on different parts fermentation, the result of test gas production (a and b) .The second experiment tested the same conditions but in the first 24 hours for examining their effects on pH medium, the concentration of ammonia nitrogen and dry matter disappearance.
    Results and Discussion
    The highest average gas production from fermentable partion in treatment of hay with eucalyptus oil and the lowest at 7 = pH in the treatment of 60 to 40 percent with pine essential oil was observed at 6 = pH. At the end of incubation the treatment of 60% concentrate with essential oils of fennel, pine and eucalyptus in 6 = pH and highest for alfalfa with pine oil was at 7 = pH. The lowest concentration of ammonia nitrogen in the treated hay with pine essential oils 7 = pH and the highest 80 percent concentrate treatment with essential oils in fennel was 7 = pH. The disappearance of dry matter in the hay treated with essential oils of pine was 7 = pH. These results suggest that high levels of essential oils effect on rumen microbial fermentation, which confirmed the antimicrobial activity.
    Conclusions
    The results of this study show that of the added essential oils in tested pH, pine essential oil in 3 μl and less in the amount of 30 μl of eucalyptus essential oil, resulted in further reduction of ammonia nitrogen production, increased pH of the culture medium and decreased disappearance dry matter was found in all foods. Therefore, it can be concluded that the essential oils of pine and eucalyptus can control the degradation of the microbial protein in the rumen by reducing the ammonia nitrogen content and improve the nitrogen yield in ruminants. Since the amount of gas produced from an indigenous feed is due to the fermentation of that feed, and hence its energy value, it can be concluded that the use of pine essential oils, because it reduces gas production and consequently reduces feed fermentation, The fermentation process is not suitable.
    Keywords: Eucalyptus, Fennel, PH, Pine, Ruminal fermentation
  • S. Alaei Baher, H. Mohammadzadeh *, A. Taghizadeh, A. Hoseinkhani Pages 179-193
    Introduction: Ensiling is a popular preservative method for forage crops and some byproducts. In this process water soluble carbohydrates in forages are converted into lactic acid with lactic acid bacteria and prevents nutrients losses by other microorganisms. Low dry matter in fresh forage may lead to higher losses of nutrients in efflunet or fermentation process. Moreover, corn silage are susceptible to spoilages by fungi. Deteriorated silages are rich in mycotoxins and some other harmfull components which may reduce dry matter intake, milk production and milk composition or may led to some acute or chronic disease in ruminatns. Whey powder is a product with high potential to water absorbents which may reduce effluent production at ensilinbg high loisture forages. Lactobacillus buchnery is a lactic acid producing bacteria which produce acetic acid while producing latic acid. Acetic acid is an antifungal compound ahich may inhibit fungi development in silage. This study was conducted to determine the effects of whey poewser and a lactic acid bacteria inoculant (Lalsil Fresh, containing Lactobacillus Buchneri) on chemical composition, pH, aeirobic stabilit and in vitro digestibility of corn silage. Materials and Method: The whole-crop corn was harvested at 1/2 milky maturity stage. Treatments were: 1. Control (corn silage without any inoculant), 2. Corn silage treated with whey powder (1% or 10 kg per ton fresh forage) 3. Corn silage treated with Lalsil Fresh at 1.8 × 10 6 colony forming unit per gram fresh forage and 4. Corn silage treated with whey powder (1% or 10 kg per ton fresh forage) and bacterial additive at 1.8 × 10 6 colony forming unit per gram fresh forage. The additives were solved in water and then sparayed over forages and mixed throughtly. The same amount of water was applied to the control treatment. Laboratory PVC silos -70 cm in height, 10 cm in diameter with a sink at the bottom for measurement of seepage were used for ensilingthe whole corn crops. Corn forage was ensiled in triplicate laboratory mini silos for 90 days at room temperature and in the dark.
    Results and Discussion
    Effluent production was lower and concentration of dry matter (DM) was higher in prebiotic and bacterial treated silages when compared to control (P<0.05). Lower efflunet production from corn silag can inhibit environmental pollution and also retain nutrients in sialge. Application of whey powder to corn silage resulted in silages with higher crude protein concentration and lower concentrations of cell wall compositions (P<0.05). The pH value of silages was lower in bacterial inoculated treatments (P<0.05) compared with control. This was maybe due to accelerated fermentation and rapid fall in pH. Treating corn silage with whey powder reduced aerobic stabilities of corn silages when they were exposed to the air (P<0.05) due to higher concentration of water soluble carbohydrates. But, treating corn silage with Lalsil Fresh improved the aerobic stabilities of corn silages in comparition with control group (P<0.05) due to higher ratio of acetate to lactate. Higher resistance of silage to spoilage can inhibit fungi from mycotoxin production and nutrients deterioration which led to better animal performance and health. The most potential of in vitro gas production and metabolisable energy (ME) concentration was easured in whey powder treatment (P<0.05). Also, treating corn silage with the bacterial inoculant resulted an increase in potential of in vitro gas production and ME concentraion of corn sialge (P<0.05). Higher energy content in corn silage may improve milk production and composition in dairy cattle. Conclusion: Results suggest that whey powder can improve nutritive value of corn silage by increasing dry matter and crude protein concentration and decreasing fiber. Also, Whey powder reduces effluent production from corn silage which consequently prevents environmental pollution. Bacterial inoculant improves corn silage resistance to spoilage by fast falling in pH and improvement of aerobic stability. Higher resistance to spoilage in corn silage can increase milk production and prevents mycotoxin related diseases and problems in ruminants. Both whey powder and bacterial inoculant improve metabolisable energy concentration of corn silage which may lead to higher production in ruminants.
    Keywords: Lactic Acid Bacteria, Lactobacillus Buchneri, Microbial Inoculant, Whey Powder, Yeasts
  • M. Dadashi, A.Hosseinkhani *, H. Mohammadzadeh Pages 195-208
    Introduction
    Weeds constantly invade crop fields and pastures. It is frequently assumed that weeds have low nutritive value and livestock will not eat weeds, so expensive and time consuming methods are often used for their control. Some weeds are toxic or poisonous for livestock, and certain weeds are unpalatable – causing a reduction in total intake. Weeds also compete with cultivated crops and forages for moisture, light, and nutrients, but many weeds are nutrient-rich and digestible. There are large numbers of weeds which are consumed by animals as forage. A study with geographic information system (GIS) in the east Azerbaijan province of Iran, showed that Bromus tectorum,Crepis sancta, Alopecurus myosuroides, Dactylis glomerata and Acroptilon repens were found in 77.8, 66.7, 67.7, 33.3 and 22.2 percent of alfalfa fields respectively. Based on this report only five species of the mentioned weeds consist of about 15 percent of total forages production area at the first cut of alfalfa fields which are harvested and used in the farms as livestock feed. Nonetheless, preliminary results suggest that weeds can play a significant role in livestock industry if their chemical composition and nutritional quality is well known. The main goal of present study was to evaluate nutritional value of seven common species of alfalfa field weeds using in vitro techniques.
    Material and
    methods
    Seven species of alfalfa field weeds including: Crepis sancta, Achillea millefolium and Acroptilon repens from family of Asteraceae, Melilotus officinalis (L.) Pall. from family of Fabaceae and Bromus tectorum, Dactylis glomerata and Alopecurus myosuroides from family of poaceae were harvested from alfalfa field at 10 percent blooming. The samples were dried in 60° oven for 48 hours and grounded to pass through a 2-mm screen. Chemical composition of weeds was determined according to prescribed procedures of AOAC (2003). Neutral detergent finer (NDF) was measured by method of Van-Soest et al. 1991. Rumen fluid was obtained from three fistulated ghezel male lambs before morning feeding. The lambs were fed twice daily at maintenance level. Dry matter fermentation of each weed was determined using in vitro gas production technique. Potential of gas production, organic matter digestibility, small chain fatty acid production and net energy of lactation (NEl) were calculated from the results of gas production. In vitro disappearance of forages was measured. Result and discussion: Alfalfa and Alopecurus myosuroides had the highest and lowest crude protein (CP) content respectively (14.3 vs 8.4 %) (P<0.05). Alopecurus myosuroides and Melilotus officinalis (L.) Pall had the highest and lowest NDF content respectively (58.7 vs 33.8 %) (P<0.05) and Crepis sancta and Acroptilon repens had the highest and lowest ADF content respectively (28.8 vs 19.5 %) (P<0.05). Alopecurus myosuroides had the highes amount of ether extract (i.e. 10.99) and Ash (11.83%) and lowest organic matter content (88.17 % of DM) among the studied forages. Fermentation experiment revealed that fabaceae family had the highest potential of gas production (252 ml/g DM), organic matter digestibility (68.3%), short chain fatty acid production (0.82 mmol) and NEl (5.2 MJ Kg-1DM) (P<0.05). Conversley, Asteraceae family had the lowest amount of organic matter digestibility (54.1%), short chain fatty acid production (0.55 mmol) and NEl (3.7 MJ Kg-1DM) (P<0.05).Among the individual forages and weeds after the alfalfa, Melilotus officinalis had the highest potential of gas production (227 ml/g DM), organic matter digestibility (67.8%), short chain fatty acid production (0.77 mmol) and NEl (5.7 MJ Kg-1DM) (P<0.05), however many of weeds had the same fractional rate of gas production with alfalfa. Accordingly based on the in vitro disappearance experiment, the mentioned family had the highest degradability parameters including soluble and slowly degradable fractions (31.8 and 20.8 % respectively), degradation rate (0.06 % / h) and effective degradability (47.4 %) (P<0.05).
    Conclusion
    According to the results of the present research, it seems that the studied plants can be use as part of ration for ruminant animals. But due to low digestibility and higher NDF content, these forages may limit voluntary feed intake. Maximizing dry matter intake is one of the main goals especially in high producing animals. Therefore considering that many alfalfa field have high proportion of weeds in the first cut, it is recommended that first cut of alfalfa not used in the ration of high producing animals.
    Keywords: Alfalfa field weeds, Chemical composition, Degradation, In vitro digestibility
  • S. A. Mirghelenj *, L. Fathollahzadeh, R Kianfar, M Olyaee Pages 209-223
    Introduction
    CLA is a mixture of several geometrical and positional conjugated isomers of linoleic acid (C 18:2 cis-9, cis-12). Ruminant products, such as milk, meat, and cheese, are major CLA sources in human diets, but foods obtained from non-ruminants, contain much less CLA. Some reports showed positive effects of CLA on production performance of layer hens. Egg internal quality traits may be depressed during storage, but incorporation of CLA, may increase the permeability of the yolk membrane due to the cis-trans arrangements of CLA, therefore ion movement between the yolks and albumen through the yolk membrane, restrict some changes in egg internal quality traits, therefore an experiment was conducted to evaluate the effects of different levels of CLA in post-molting phase of laying hens on production performance and egg quality in different storage times and temperatures.
    Material and
    Methods
    In this experiment, sixty W-36 White leghorn laying hens in post-molting phase (78 week) were assigned to 3 treatments with 5 replications and 4 birds each based on completely randomized design. The purity of CLA source used in this study was above 90% with mixture of 50% c9-t11 and 50% c10- t12 isomers. Experimental diets were 1) Control diet (basal diet containing 0% CLA), 2) basal diet+0. 25% pure CLA and 3) basal diet+0.5% pure CLA and after adaptation period, diets were fed for 4 weeks. During the experiment, hens were provided feed and water ad-libitum. Egg production, egg weight, feed consumption, egg mass and feed conversion ratio were recorded weekly and reported as total period. Every two weeks, internal and external quality traits such as albumen percent, yolk percent, albumen pH, yolk pH, Haugh unit, yolk height, yolk Roche scale, yolk index, egg size, egg surface, specific gravity, shell strength, shell percent and thickness were evaluated. The eggs from treatments having best production performance (0.5 %) collected and stored in different temperatures (4 and 27° C) and duration times (1 day, 1 week and 1 month after production) then evaluated their internal quality.
    Results and Discussion
    Results showed that egg production performance and egg mass of birds fed 0.5 % CLA was significantly higher than control or those fed 0.25 % during whole experimental period, but egg weight and feed consumption were not affected significantly by dietary CLA level and feed conversion ratio of birds fed 0.5 % CLA was significantly (P<0.05) lower than control or those fed 0.25 %. In the end of experiment, the internal egg quality traits such as albumen percent, yolk percent, albumen pH, yolk pH, Haugh unit, yolk height, yolk Roche scale, yolk index were not affected by dietary CLA levels (P>0.05). Also the external egg quality traits such as egg size, egg surface, specific gravity, shell strength, shell percent and thickness were not affected significantly. Results of egg quality traits during different times and temperatures showed that main effects of CLA levels, storage times and storage temperatures were significant (P<0.05) on most important egg internal quality traits such as Haugh unit and pH of albumen. Interaction effects of CLA levels with storage times and temperatures were significant (P<0.05) and in eggs kept within higher temperatures and longer times, albumen pH of eggs for birds fed 0.5 % CLA, were lower compared to albumen pH of eggs for birds fed control diet containing 0 % CLA. These results indicate that ion movement between the yolks and albumen through the yolk membrane in eggs from hens fed 0.5 % CLA was greater than that in the control eggs and caused to restrict pH increase during storage. It is speculated that the incorporation of CLA, most of which was in the trans form, could have increased the permeability of yolk membrane because of the cis-trans arrangements of CLA, which would have increased membrane fluidity and sizes of space between the fatty acid chains of phospholipids that constitute the membrane bilayer.
    Conclusion
    It is concluded that use of 0.5 % CLA in post-molting diet of laying hens improved egg production performance of birds and restricted luctuations in pH and Haugh unit changes of eggs keep during high storage temperatures and duration times, therefore it is suggested to adding 0.5 % pure CLA in post-molted laying hen diets.
    Keywords: Conjugated Linoleic Acid, Haugh Unit, Molting, Layers, Storage temperature
  • S. J. Hosseini Vashan *, T. Ghaznavi Pages 225-236
    Introduction
    One of the main sections of charges in poultry production is the cost of the ration. Thus, the producers are continuously looking to find new compounds that using in poultry diets with low prices. Usage the agricultural and food industry by-products are new strategies to reduce the environmental contamination and animal diet costs. Pomegranate granatum is a fruit from the punicaceae family. The pomegranate pulp is a byproduct that has valuable components for poultry nutrition involved protein, fat, Ash and bioactive compounds. Pomegranate pulp has punicallagin, ellagic acid, tannic acid, flavonoid and anthocyanin. The usage of antibiotic in poultry production was banned because they have several negative effects on poultry and human. Therefore, the medicinal herb contained flavonoid and polyphenolic compounds were extremely increased. An experiment was designed to evaluate the effects of pomegranate pulp and Multi- Enzyme on performance parameters, egg quality, eggshell parameters, egg lipid profiles, blood lipid profile, and the blood concentration of malondialdehyde in peak production of laying hens.
    Materials and Methods
    A total 200 of Hy-Line W-36 at peak production with 32 weeks were selected and allotted to 20 experimental units. The dietary treatments were included the levels of 0, 4, 7 and 10 % of pomegranate pulp with 5gr of multi-enzyme WX. The egg production, egg weight, egg mass feed intake, and feed conversion ratio were weekly recorded. The egg quality parameters involved egg shape, yolk index, yolk color index and haugh unit, the relative weight of yolk and white were determined three consecutive periods (each 28 days). As the same periods, the eggshell parameters such as; eggshell weight, eggshell thickness, specific gravity, and eggshell resistance were evaluated. The egg cholesterol was determined. At the end of experiments, the bloods of two birds from each replication were gathered. The blood plasma was prepared. The blood lipid, total protein, and malondialdehyde were determined by auto analyzer instrument. The data were analyzed with the general linear model by SAS software. The mean differences among treatments were studied by Tukey's test (P<0.05).
    Results and Discussion
    The results revealed that pomegranate pulp did not affect feed intake. The diets contained 10 percentage of pomegranate pulp increased the egg production, egg weight, egg mass. The FCR decreased when birds received 10 % pomegranate pulp. Pomegranate pulp had a valuable percentage of protein and fat and also it has bioactive components involved ellagic acid, punicallagin and flavonoid that they could be affect the metabolism of birds and improved the performance traits. The egg quality parameters included egg shape, yolk index, and yolk colure index did not affect by the levels of pomegranate pulp. However, the haugh unit and the relative weight of white increased when birds fed 10% pomegranate pulp. Haugh unit is synthetized in the magnum of the oviduct. The concentration of nutrients in the blood may alter the white quality and haugh unit. The eggshell parameters involve eggshell thickness and resistance increased when the 4 percentage of pomegranate pulp were included to laying hen diets, however; the higher levels of pomegranate pulp has not the same effects. Thus, higher concentration of fiber in upper levels of pomegranate pulp may negative influence on the absorption of minerals. The egg yolk lipids included LDL, triglyceride was decreased, and the HDL was increased when pomegranate pulp was added to layer diets. The pomegranate pulp decreased the blood cholesterol. The previous studies were revealed that fiber and flavonoid or polyphenolic components have hypocholesterolemia effects. The pomegranate pulp suppressed the plasma malondialdehyde. The concentration of malondialdehyde is an index of lipid peroxidation. This antioxidant effects on the blood of birds is due to flavonoid, polphenolic, ellagic acid, punicallagin components in pomegranate pulp.
    Conclusion
    It is concluded that pomegranate pulp may be combined with multi enzyme be included to laying hen diets with no any negative effects on performance and egg quality parameters; it also may improve the yolk cholesterol and antioxidant properties of laying hens.
    Keywords: Egg quality, Layers, Malondialdehyde, Performance, Pomegranate pulp
  • M.R. Sheikhlou *, E. Abdi Ghezeljeh, R. Safari, Z. Nemati Pages 237-247
    Introduction
    The practices that make breeding programs effective in generating genetic gain results the reduction in genetic diversity. The loss of diversity and the resulting increase in homozygosity may result in decreased production and fitness of inbred animals. One of the important parameters in assessing the level of genetic diversity of population is effective population size. Effective population size is a key parameter in population and conservation genetics due to its direct relationship with the rate of inbreeding and the amount of genetic variation lost because of random genetic drift. As a consequence, effective population size is usually considered as a useful criterion for classifying the livestock breeds according to the degree of endangerment. Classically, effective population size can be estimated from the rate of inbreeding between two successive generations. However, in real populations with overlapping generations, the definition of a ‘previous’ generation is quite difficult to establish and assignment of animals to various generations is not simple. A number of methods are available to estimate the effective population size on the basis of pedigree in livestock populations. However, when it comes to monitoring animal genetic resources not all methods are equally well suited and depending on the conditions in the population under consideration, different methods may have to be chosen. The objective of this study was to estimate effective population size in the breeding flock of sangsari sheep using different methods and to select the best estimates among them according to the pedigree structure.
    Material and
    Methods
    In this study the pedigree information of Sangsari sheep collected in 28 years (1988- 2015) at breeding station of Damghan were used to estimate effective population size. The quality of available pedigree information is of great importance when interpreting the results of pedigree analysis. Thus, the degree of completeness of pedigree was assessed before analysis. For the whole file, the proportion of animals with both parents known was computed by simple counting. The pedigree completeness index (PCI) was used to describe the degree of completeness of pedigree. In addition, for each individual, the number of equivalent complete generations (EqG) was computed. Effective population size was estimated using the six different methods. The best method was selected considering some criteria such as time period used, subpopulation stratification, stability of estimates and negative estimate for effective population size.
    Results and Discussion
    The proportion of animals from the whole file with two known parents was 70%. The completeness of pedigree was low in the early years of the foundation of the center, however, pedigree filling improved over time, with the most recent cohort of lambs having pedigree completeness index of 0.61 and equivalent complete generations of 4.1. Accordingly, the pedigree had an acceptable completeness level for estimation of the effective population size. Estimates for effective population size were different according to the methods used for estimation. Estimated effective population size from and Ne-Coan resulted negative values for some years in the time period of last generation interval, which is clearly meaningless and leads to the rejection of these estimates. Another criteria for choosing the best method is the variability of estimates that should be as small as possible. Here, we consider the square root of the residual after fitting a linear regression to the yearly Ne estimates that should not be greater than 20. In all methods this criteria was greater than the critical value (20) except for the methods Ne-Cens and Ne-Ecg, and Ne-Ecg has the smaller value between the methods. Considering the necessary criteria, estimates from the method of individual increase in inbreeding (Ne-Ecg) were chosen as the best estimates for effective population size in this pedigree. Average estimated effective population size using this method was 131 animals.
    Conclusion
    Estimated realized effective population size was greater than the critical levels reported by FAO (50) and Meuwissen (100). Also, the estimated effective population size in this study was in the range of estimates for some foreign and Iranian sheep breeds. Considering the closed nucleus with no entry of animals from other herds, implementation of methods for preventing the losses of effective population size in the future such as optimum contribution of parents is suggested.
    Keywords: Effective population size, Inbreeding, Pedigree analysis, Sangsari sheep
  • V. Taghizadeh, M. R. Nassiri *, M. Tahmoorespur, M. R. Bakhtiarizadeh, A. Javadmanesh Pages 249-262
    Introduction
    Ascites syndrome is a metabolic disorder in late ages of meat-type chickens. The only commercial broiler strain of Iran, Arian, has been incurred by this syndrome during a couple of decades ago. Due to susceptibility to ascites and some poor performance issues this strain are wiping out from indigenous market. Prevalence of ascites syndrome in commercial broiler of Arian is nearly 10% in average and in paternal lines it gets more fatality. Role of genetic factors in arising ascites has been proven by former researchers. Fighting against this syndrome through old methods are expensive and time consuming. Since presence of hypertrophy in a tissue needs to make facilities such as blood vessels to providing oxygen and nutrients so studying on angiogenesis pathway and its key important genes was directed. Whereas numerous organs engage the disease and among them heart has key role for initiating and developing the disease, so genomic study dealing with next generation sequencing (NGS) technology has been conducted to identify genes relating to angiogenesis and hypertrophy in right ventricle of ascites susceptible birds.
    Materials and methods
    464 one-day-old chicks from B-line pedigree of Arian strain have been reared up to 21 days at the same rearing and nutritional conditions. At the age of 21 the rearing house was divided to 3 sections. Normal and cold sections at the ends of the house and buffer section at the middle. 464 chickens were transferred from normal to the cold section and kept the ambient temperature lower than 16 degrees for inducing ascites. 120 birds from cold section were selected randomly and slaughtered at the age of 42. Those of birds having ascites symptoms classified to ascites susceptible (As) and the other assigned to healthy (He). Samples of right ventricle was picked up and stored with RNAase-Later in liquid N tank. Then 12 As samples and 12 He samples were chosen for extracting total RNA by Biozol kit. After that every 6 samples in each groups pooled together and finally 2 As and 2 He samples prepared to make cDNA libraries for high throughput sequencer machine, Ilumina Hiseq 2000. RNA integrity score (RIN) were more than 7 for all samples. All small RNAs such as microRNAs, rRNAs and tRNAs were eliminated by oligo dt beads and finally all mRNAs was used for preparing library. cDNAs as long as 200 bp were selected for library. Sequencing has been done by BGI Company. For mapping, aligning, and DE analyzing several softwares were used such as: Tophat, cufflinks, cuffmerge and cuffdiff. Then significant DEGs imported to String for creating gene expression network and use DAVID 6.8 for investigation gene annotation and pathway analysis and finally Cytoscape v. 3.5.1 was used for network and cluster analysis.
    Results and Discussion
    Nearly 90% of reads mapped on genome reference. In ascites group among 125 DEGs 79 genes was upregulated and 46 loci was down-regulated. Among 125 significant differentially expressed genes (FDR<20%) between As and He birds 59 genes were identified in a pathway in KEGG database. Gene ontology has been shown that the most significant biological process term containing 9 genes in term of gga04512: ECM-receptors interaction. The gene expression network analysis shows that average node neighboring was 3.763, network heterogeneity was 0.9 and network centralization was 0.236. Cluster analysis depicted 3 significant clusters involving key rule in angiogenesis including Cyclin- ependent Kinase1 (CDK1), ECM-receptors interaction and Interleukin 7 (IL7) (p-value<0.05). Up regulating of CDK1 with CCNB1 and CCNB2 may activated maturation promoting factor (MPF) that eventually induce cell division and proliferation in endothelial cells (ECs). Up regulating COL6A2 shows its roles in remodeling of ECM-receptors that in turn involving in proangiogenesis process in right ventricle hypertrophy in As birds. IL7 up regulated significantly (P-value<0.05) in As birds. This factor can induce IL8 and Ang-1 secretion from ECs. These factors have key important role for vascular remodeling and vascular tone in proangiogenesis and angiogenesis process. Studying on gene networks in susceptible bird’s heart cells has been shown that over expression in angiogenesis related genes was seen significantly in susceptible birds especially in ECM-receptor interaction, Cell division and IL-7 pathways as well. All these recent pathways rule actively in angiogenesis process and vascular network remodeling. So after finding these networks and related clusters it can be possible to find more extended studies on down regulated genes in apart or together for using breeding improvement objectives.
    Keywords: Angiogenesis, Ascites, Gene Network, Gene Ontology, Pathway Analysis, RNA-Seq
  • M. Soltani * Pages 263-273
    Introduction: Mycobacterium avium subsp. aratuberculosis (Map) is a very slow growing bacterium that solely infect the digestive tract and leads to Johne’s disease (JD) that is characterized by immedicable diarrhea. Considering innate resistance and different transmission modes of Map and its possible role in development or progression of some human diseases (mainly Crohn’s disease), strong zoonotic potential has been proposed for Map. Diagnosis of JD depends on isolation of viable Map (culture), tracking of its consequences on host immune system (ELISA) or amplification of Map genome (PCR). Fecal culture considered as gold standard for Map detection but obstacles like very slow growth rate of Map, decontamination issues and its high expenses led to development of alternatives like PCR based approaches to gain PCR advantages like high sensitivity, specificity and rapidity for diagnosis of Map. Since no published report was found on Map infection status in Kerman, the current study aimed to investigate the presence of Map in bovine feces samples using 3 different detection methods including culture, IS900 PCR and IS900 nested PCR. Also IS1311 PCR-REA was used for strain typing of Map isolates. Materials and Methods: 212 fecal samples were taken from 4 dairy cattle farms located in Kerman province, southeast Iran. The fecal fractions for culture and PCR ere prepared separately to inhibit potential crosscontaminations. Per each sample, 300 μl of prepared inoculum was cultured on slope of Herrold’s egg yolk medium (HEYM) alone, and two HEYM + 2.0 mg/L of mycobactin J slopes. Inoculated slopes were incubated at 37 °C for 4 months and monitored at biweekly intervals. After extraction of DNA from fecal samples, PCR reaction performed in 25 μl volume and products were analyzed by electrophoresis on 2% agarose gels. The samples were considered as positive if 413 bp amplified band was present. To avoid false positive results, a REA approach using AlwI restriction enzyme was adopted. For IS900 nested PCR reaction, 3 ml from IS900 PCR products were used as template. Resultant products were screened as before. To evaluate the limit of detection of PCR reactions, the DNA extracted from confirmed MAP field strain culture was 10 fold serially diluted (1 μg – 1 fg) and the minimum detection level of PCR and nested PCR were observed. Strain typing of Map isolates was based on IS1311 PCR-REA. Results and Discussion: It takes 3-4 months of incubation for appearance of MAP positive colonies. Map was detected in 32/212 (15.1%) of cultured samples. All colonies were verified as MAP using IS900 PCR and REA with AlwI. 44 positive samples were detected by IS900 PCR (20.7%) while nested PCR was able to find 52 infected samples (24.5%). All isolates detected by PCR based methods were also verified as MAP by IS900 PCR-REA. According to expectations, nPCR offered higher sensitivity than conventional PCR. However, comparing the methods used in the present study showed that as a simple and cheap assay, nested PCR is inherently successful in amplification even with rare amount of starting template and has the better ability to detect the Map infection in fecal samples and can be used as a routine method in diagnostic processes. Although this study was the first attempt to access the Map infection status in Kerman province, as summarized in table 3, many studies were conducted all around Iran that can be compared with. Altogether, Map incidence in other central Iran provinces (Isfahan, Fars and Chaharmahal and Bakhtiari) were between 14.1 to 31.8 percent of tested populations based on examination of fecal samples. Considering that most of reported values came from clinically infected or suspected animals, the estimates obtained in this study showed the possibility that the Map incidence in Kerman province may be higher than other parts of central Iran, but it more investigations needed to create a realistic scheme of Map infection status in Kerman province. Remembering that a copy of genome was considered as equivalent to 5 fg of DNA, sensitivity analysis using serially diluted DNA preparations revealed that the minimum detection level were 1 pg (200 genomes) and 10 fg (2 genomes) for PCR and nested PCR, respectively. According to results, compared to nested PCR, conventional PCR is not sensitive enough for diagnostic tests on field samples. Analysis of fragments produced after IS1311 REA showed the presence of explicit patterns of four bands for all obtained isolates that is the indication of MAP C type. Obtained DNA sequences from the amplified IS1311 locus was exactly consistent with previously published sequence for MAP C type. S type was not detected in current study that was consistent with results of researches performed in Razavi Khorasan province. Failure to find S Map strain was not surprising, since its global prevalence in cow population is very low and hard to find. Also, there are few papers dealing with Map strain typing in Iran, so it is conceivable to find S Map type in Iran’s cattle population if more research conducted in this respect.
    Conclusion
    Although paratuberculosis is a global neglected disease, the situation is worse in developing countries like Iran. Contrary to bovine tuberculosis, due to absence of a national program to control paratuberculosis infection in Iran’s dairy herds, no reliable data is available for arrangement of tools for efficient restriction of disease which leads to inadequate attention to JD and eventually receive the least priority to control. Unfortunately, paratuberculosis as an overlooked disease, is a severe hazard for cows’ health and also for economic activities related to dairy cattle industry. On the other hand, Map is still considered as an important suspicious zoonotic agent which cause serious health problems for humans. Meanwhile, considering aforementioned warnings, enactment of crucial health standards is vital to minimize the chance of infection transmission to non-infected populations and alleviating health problems and economic losses due to Map infection.
    Keywords: Culture, Detection, Johne’s disease, Mycobacterium avium subsp. Paratuberculosis, nested PCR, PCR
  • M. Piri, Gh. R. Dashab *, B. Asghari Pages 275-285
    Introduction
    Leptin is a pleiotropic protein best known for regulation of appetite and fat storage in mammals. While many leptin orthologs have been identified among vertebrates, an authentic leptin in birds hasremained elusive and controversial. Leptin, the ob gene product, is a 167 amino acid polypeptide known to play a major role in regulating the fat stores of the body and is found in all eukaryotes, including mammals and in invertebrates. In mammals, leptin functions as an adiposity signal: circulating leptin fluctuates in proportion to fat mass, and it acts on the hypothalamus to suppress food intake. However, little is known about the molecular evolution of the leptin sequence gene. Therefore, the aim of this study, we conducted an analysis of the evolutionary and phylogenetic of the mammalian’s Leptin nucleotide sequences in native camel of Sistan and Baluchistan province and compare with other species in NCBI gene bank.
    Materials and methods
    In this study, blood samples were collected from 50 camels, randomly from two stock of Sistan and Baluchistan province. DNA is extracted from whole blood with phenol-chloroform method. PCR amplification of 2000 bp from of partial ob gene including intron2 and xon3 of Leptin gene was performed using one pairs of special primers. PCR product was digested with endonuclease Asuhpi enzyme and purified on the agarose gel. Then, the sequencing of the digestion products was performed by the Sanger method. Data sequence for other species was achieved and aligned by searching its genome database (NCBI). The nucleotide substitution rate of the sequences (transition and transversion substitution rate) and molecular evolution (including polymorphism site, conservation site and gene conversion) of the Leptin were calculated by maximum likelihood and neighbor-joining (NJ) method respectively and phylogenetic tree was based on nucleotide sequences constructed. Evolutionary and phylogenetic tree analysis was performed by using MEGA6 and Dnasp v5 software's. Finally, a fundamental measure of the elative importance of selection and genetic drift in causing amino-acid substitutions is the dN/dS ratio. In this study was calculated with online package HIV_SNAP v2.1.1.
    Results and Discussion
    Results of alignment of DNA sequencing of two populations of camel in Sistan and Baluchistan showed 99% similarity, but diversity showed distinct from other mammalian species. The results showed that the transitional substitution was more than transversional substitution and ratio these was 1.58. Totally, there were 591 mutations including insert, deletion and polymorphism sites at the DNA level of leptin gene (ob gene) in different species but sequence alignment of the Leptin gene fragment revealed only 309 polymorphic sites and conservation area of ob gene was very small. Evolutionary pressures on proteins are often quantified by the ratio of substitution rates at non-synonymous and synonymous sites. The dN/dS ratio was originally developed for application to distantly diverged sequences, the differences among which represent substitutions that have fixed along independent lineages. Nevertheless, the dN/dS measure is often applied to sequences sampled from a single population, the differences among which represent segregating polymorphisms. The dN/dS ratio of the Leptin sequences in this study (0.76) indicated that negative selection was accrued during evolution. Phylogenetic tree for the leptin gene in different organisms show that seven categories in the mammals such as camel, cows and buffalo, sheep and goat, pork's, bats, cats and marine. Phylogenetic analysis of leptin gene using Neighbor-Joining method showed that Baluchi camel population in this study has the highest similarity with Lama among the Camelidae family.
    Conclusion
    Phylogenetic analysis of DNA sequence has played an important role in the study on evolution of life. However, recent researches suggest in some cases phylogenetic trees based on the analyses of DNA sequences may be misleading and that based on trees protein-based trees from amino acid sequences may be more reliable. Similarities of between the two populations of Baluchi camel in this study showed that common ancestry and genetic similarity. Mutations and natural selection resulted in the development of new varieties, new proteins and also stabilizes their performance during the evolution and advance progress toward their performance has been purified. According to this study, protected areas make up a small part of leptin gene sequence insures that this reflects the polymorphism of this gene as well as being susceptible to variations and mutations, respectively. The results of dN/dS suggested that evolution in camel is different of other species.
    Keywords: Evolution, Leptin, Native camel, Natural selection, Phylogeny
  • S. Mohamadi *, M. Mamouei, S. Tabatabaei Vakily, J. Fayazi Pages 287-296
    Introduction
    Semen collection evaluation and addition of preservatives to increase storage period of sperm are essential for successful artificial insemination. Cryopreservation of spermatozoa is becoming more important because of new clinical requirements and current clinical practice. Although frozen-thawed semen has great practical benefits for reproduction, it is widely reported that the cryopreservation process involving cooling, freezing, and thawing induces serious detrimental changes in sperm functions. The viability, motility and membrane integrity of mammalian spermatozoa decrease during the cryopreservation process. Many studies exist as regards the effects of antioxidants on the cryopreservation aimed at improving the quality of post-thaw semen. Antioxidant molecules could decrease the impact of oxidative stress and therefore improve Sperm quality following the freeze–thawing process. Melatonin (N-acetyl-5-methoxytryptamine), a derivative of tryptophan, is mainly synthesized and secreted by the pineal gland during the night in reaction to changes in light levels. Melatonin can stimulate the activity of antioxidant enzymes such as SOD and GSH-Px. melatonin scavenges a variety of reactive oxygen and nitrogen species in vivo and in vitro, signifying it has a powerful non-enzymatic antioxidant property. It has been shown that the spermatozoa undergo a freeze–thawing process produced high concentrations of reactive oxygen species. The aim of this study was to investigate the effects different levels of melatonin supplementation (0, 0.5, 1, 2 and 4 mM/ml) in extender on semen characteristics the Arabic ram after freezing-thawing.
    Materials and Methods
    This research was performed at Ramin Agriculture and Natural Resources University of Khuzestan in the fall in 2015. Semen samples were collected from 6 Arabic ram with an average weight 73 ± 3 kg by electro ejaculator twice a week. Sperm samples motility were assessed by Computer Aided
    Sperm Analysis (CASA) after freezing and thawing. The pH, viability, abnormal sperm percentage, membrane integrity and total antioxidant capacity were also assessed after freezing-thawing process. Pearson correlation test was used to assess correlation of melatonin with routine sperm parameters (pH, viability, abnormal sperm percentage, membrane integrity and total antioxidant apacity of plasma). Data analysis was performed using SAS software. P<0.05 was considered significant. Results and Discussion the Results of this experiment showed that the diluent of ram semen containing 0.5 mM/ml of melatonin improved the motility, membrane integrity and viability of Arabic ram spermatozoa compared to the control. The total antioxidant capacity was highest in diluent contained whit 1 mM/ml of melatonin. The effect of melatonin on the pH of semen at all levels was not significant. Cryopreservation causes an irreversible damage to enzymatic activity and sperm organelles leading to a reduction in the sperm kinetic parameters. Composition of extender may also affect the freeze ability of spermatozoa and their fertilizing ability. Many studies reported that melatonin has beneficial effects on preservation of mammalian sperm function and improves the microscopic parameters of spermatozoa. Melatonin supplementation to ram semen freezing extender protected spermatozoa from the cryopreservation injuries, as proved by post-thaw viability, motility, intracellular ATP concentrations, DNA integrity, and fertilizing ability. It is suggested that melatonin stimulates the activities of antioxidant enzymes. In consequence, melatonin reduces the number of free radicals, ROS, and also may increases the production of molecules protecting sperm cells against oxidative stress. As a conclusion, supplementation of melatonin in the freezing medium can counteract the adverse effects of the freeze–thawing process on the motility, viability, normal morphology and plasma membrane integrity in ram spermatozoa. The results were suggested that the protective effects of melatonin on spermatozoa were associated with a reduction in LPO as a consequence of increasing the TAC and antioxidant enzymes activity.
    Conclusion
    Cryopreservation of sperm is an applicable technique in infertility management but it may influence the post-thaw qualities of sperm, including morphology, motility, viability and DNA integrity. In this research, we show that supplementation of cryopreservation extenders with melatonin provide a cryoprotective effect on pH, viability, abnormal sperm percentage, membrane integrity and total antioxidant capacity. Overall addition of 0.5 mM/ml of melatonin to the extender improved the most of spermatozoa quality characteristics as well as total antioxidant capacity of semen after freezing-thawing process in Arabic ram. In the current study, we observed no correlation between melatonin and pH, viability, abnormal sperm percentage, membrane integrity and total antioxidant capacity of plasma.
    Keywords: Sperm, Motility, Membrane Integrity, Survival, Total Antioxidant Capacity
  • M. Barkhordar, O. Safari, M. R. Ahmadi, A. A. Naserian, R. Valizadeh * Pages 297-310
    Introduction
    The aquaculture industry in the world, with an 8-million-dollar transaction rate, has 8% growth annually. It also has the most value-added rate among different fishery activities based on the food and Agriculture Organization (FAO). The angelfish belonging to Cichlid family, is known as a famous species in this industry. This fish feeds on various live feedstuffs. Regarding the significance of suitable feedstuffs in aqua feed industry, the live feeds used to provide nutritional requirements, immunity tuning and stimulation will be of important. Tubifex worm, containing considerable crude protein, diverse sorts of amino acids and essential fatty acids, has a deserving role and checking the nutritional value of Tubifex worm with aim of improving the growth performance, boosting immune responses and survival rate, reducing expenditure cost and increasing productivity is so crucial. The objective of the current experiment was to compare the effect of the cultured and commercial Tubifex worm powder in the diet of juvenile angelfish on growth performance and immune responses.
    Materials and methods
    The experimental rations included 3 rations: 1) control ration as pellet 2) commercial Tubifex powder as freeze-dried food and 3) the rations containing mass-produced Tubifex as freezedried food. One-hundred thirty five Angel fishes with an average initial weight of 0.2± 0.5 gr were tested for 63 days. The fishes were placed randomly into nine 920 liter-capacity of 15 fishes. The average temperature was (27±1˚C), pH (7.5±0.1) and dissolved oxygen (7.68±0.35) was regulated according to required conditions. The length of light-dark cycles was considered 14 and10 hours, respectively.The fishes were fed 3% body weight in three times a day (7, 13 and 20). Biometry was conducted at 1st, 31st and 63rd days and the criteria related to the growth performance, nutrient utilization efficiency and survival rate were computed. At the end of the experimental period, five samples were taken from each aquarium, exposed to the airchallenge for 3 min and bleed 3 h after challenge. After that, biochemical analyses including trypsin, amylase and lipase were measured by using benzoyl-arginine p-nitroanilide substrate, starch and olive oil as substrate, respectively through titration method. At the end of the experimental period, total immunoglobulin, lysosome and complement, liver enzymes including ALT and AST were measured by kinetic enzymatic method and the microbial parameters including (plate count agar, and lactobacilus conut were measured. Chemical analysis of experimental diets including crude protein by Kjeldahl method, crude fat by Soxhelt extractor and ash by electrical furnace were measured. Normality Test of was done by Shapior-wilk test. Analysis of alterations in growth performance, nutritional and biochemical factors were done by one-way analysis of variance (ANOVA) and comparing means based on Duncan multiple test using SPSS software, version19.0.
    Results and Discussion
    The highest rate of final weight (about 30%), initial length (about 14%) and the rate of special growth (about 27%) were belonged to the angel fishes fed on the diet containing cultured Tubifex. The activity of the immunity parameters including total immunoglobulin, lysosome as well as complement were also significantly higher in the treatment with the diets containing Tubifex after air exposure challenge. The activities of the lipase, protease and amylase increased in the culturing Tubifex treatment in compared with two other ones. The number of the intestinal lactobacillus indicated that the most colonies forming unit of lactobacillus was significantly higher in angel fish fed the diet containing cultured Tubifex treatment. The results indicated that the angel fishes fed on the cultured Tubifex showed higher weight increment, special growth rate, survival rate as well as better nutrient utilization efficiency which may be attributed to the existence of higher rate of protein, unsaturated fatty acids Improvement of the immunity in fish fed the diet containing cultured Tubifex can be related to the role of lysosome as an important factor in natural perseverance for fishes, immunoglobulin as an important factor in antibody secretion as well as complement which has a basic role in acquired natural immunity for the fishes. The enzymes ALT and ALP which have an important role in using amino acids in oxidation process The increase of intestinal LAB count in angel fish fed the cultured Tubifex can be enhanced absorbing nutrients, disease resistance and finally, higher growth performance Considering the higher mortality rate among the larvae in the first 20-30 days and the initial time of the active feeding because of changing the nutritional modes, use of Tubifex worm powder as a live food may be effective in better growth function, external form as well as promotion of immunity system for the fishes. Conclusion: Feeding the angel fishes on cultured Tubifex worm powder especially in the initial steps of the active feeding may result in improvement in growth performance and some specific and nonspecific immune responses.
    Keywords: Tubifex Worm, Growth performance, Angel Fish (pterophylum scalare)