جستجوی مقالات مرتبط با کلیدواژه « In vitro » در نشریات گروه « علوم دام »

Heat treatment of a wide variety of feeds has been traditionally used to improve the utilization of their protein by ruminants. Heat treatment of legume seeds can inactivate anti-nutritional factors, it is also effective in increasing the digestibility (Seifdavati 2012). Chemical composition and heat processing can affects fermentability of Vicia Ervilia grain. Particle size and moisture during processing also influence the possible effects of processing. Digestibility may be directly determined in vitro or estimated by using in vitro procedures, which are cheaper and more convenient. In vitro gas production technique was widely used to evaluate the nutritive value of legume forages (Evitayani et al 2004) and tannin containing tree leaves (Rubanza et al 2003). This experiment was conducted to investigate of the nutritive value of Vicia Ervilia using the in vitro disappearance and gas production techniques.

The experimental treatments were grinding Vicia Ervilia grain, steam flak, roasting and microwave. Processing steam flak, treated steam at 100°C for 30 min and then immediately passed through rollers were allowed to air for 48 h and then sealed in plastic bags. Roasting treatments at 120°C for 10 min and microwave irradiation (Nasional at a power of 800w) for 3 min. The treatments were milled through a 2.0 mm screen. Samples of treatments were dried in oven at 65°C for 24h. The gas production of treatment were recorded at 2, 4, 6, 8, 12, 16, 20, 24, 36 and 48 h of incubation time using a water displacement apparatus. For this purpose two ruminal fistulated sheep were used. Rumen liquor samples were obtained from the two weathers that were fed a diet content 60% roughage: 40% concentrate. Rumen fluid was pumped with a manually operated vacuum pump and transferred into prewarmed thermos flask, combined, filtered through four layers of cheesecloth and flushed with CO2. McDougall (1948) buffer solution was prepared and placed in a water bath at 39°C. Samples (300 mg) were weighed into 50 ml serum vial. Each feed sample was incubated in triplicate with 20 ml of rumen liquor and buffer solution (1:2). The vials were sealed immediately after loading and were affixed to a rotary shaker platform set as (120 rpm) housed on a incubator. Vials for each time point, as well as blanks, were prepared in quadruplicate. The metabolizable energy, net energy for lactation content of feeds, short chain fatty acid and digestible organic matter were calculated using equations of Menke & Steingass (1988) and Getachew et al. (2002). For in vitro disappearance method, the incubation time were 2, 8, 12, 24 and 48 h. The rumen liquor was obtained from the same sheep used in the gas production technique and receiving the same diet to similar conditions for both techniques. Mc Dougall (1948) buffer prewarmed to 39 ºC. The inoculum was dispensed 20 ml per vial into 50 ml serum vial (containing of 300 mg sample per vial) which had been warmed to 39 ºC and flushed with oxygen free CO2. The vials were sealed immediately after loading and were affixed to a rotary shaker platform set as (120 rpm) housed in incubator. Vials for each time point, as well as blank (containing no substrate), were prepared in triplicate. Triplicate vials were removed after 2, 8, 12, 24 and 48h of incubation. The residues were washed three times in phosphate buffer followed by centrifugation (2500 rpm, 10 min) and dried at 105 ºC and determined DM disappearance. The data was analyzed by completely randomized design.

Dry matter disappearance and gas product increased with increasing incubation time. Heat processing of steam flak increasing of DM disappearance and gas produced at 48 h and higher than the other treatments (P<0.05). The gas production volume at 48 h incubation for treatments of control, microwave, roasting and steam flaking were 141, 140, 134.57 and 147 (ml/gDM), respectively. The gas production for sum of soluble and insoluble (a+b) fractions (ml/gDM) and gas production rate (ml/h) in control treatment were 142.9 and 0.106, respectively. Dry matter disappearance from in vitro increased with increasing incubation time. The disappearance percentage of DM in control treatment for 2 and 48 h of incubation were 39.88 and 83.48 and degradabilities coefficients (a, b and c) were 31.08, 48.03 and 0.088, respectively. For roasting treatments DM disappearance and gas produced at 48 h were lower than the other treatments. Barley grain treated by the steam flak, flame rusting and exploring can be decreased ruminal crude protein (McNiven 1994). Ljokjel et al (2003) reported that heat treatment decreased ruminal degradation of starch in both barley grain and pea grain. Parnian et al (2010) showed that, cumulative gas production of sorghum grain was linearly increased by altering period of microwave irradiation. Parand et al (2010) reported that the gas production of barley grain for steam flak, microwave and roasting at 48 h were obtained 239.4, 275.9 and 239.7 (ml/gDM), respectively, these values does not match with the results of this study. Similar to the latter, previous in vitro and in vivo studies (Hironaka et al 1992; Huntington 1997) have shown that steam flaking increased ruminal starch digestibility of barley in feedlot cattle. Seifdavati et al (2012) showed the gas production of Vicia Ervilia at 24 h was obtained about 246.29 ml/gDM and rate of gas product was achieved 0.063 (%h) that decreased with autoclave processing. Hadji Panayioton and Economids (2001), reported degradability characteristics of lupin (a, b and c) about 12.45%, 84.41% and 0.1 h-1, respectively. There was strong positive correlation between gas production, in vitro and in situ dry matter disappearance so that the gas production technique can be suitable replacement for in situ and in vitro DM disappearances. The steam flaking treatment showed high potential for producing of gas and high degradability of DM compared to the other treatments (P<0.05). Regarding to high gas production and in vitro degradability in steam flaking treatment, this treatment can be used to improve of digestion process in animal nutrition.

Investigation of fermentation parameters by gas production method is one of the common methods for evaluating feeds and can be a reflection of the fermentation pattern in the rumen. The gas production method in a relatively short time can evaluate the fermentation process of feeds and a large number of samples, and also used to determine the quantity and quality of value and digestion rate of a feed. In recent years, the flaxseed and its oil, due to high concentration of α-linolenic acid extremely considered and used as a fat supplementation in diets of ruminant, but one of the most important concerns in the nutrition of oilseeds is the negative effects of their unsaturated fatty acids on ruminal fermentation. The aim of this study was to evaluate the digestibility, gas and microbial protein production of diets with different forms of flaxseed (whole, grounded and extruded) and the high (40%) and low (20%) levels of rumen undegradable protein (RUP).

Two hundred mg of each feed sample was incubated with ruminal fluid with buffer and gas production was measured at 2, 4, 6, 8, 10, 12, 24, 36, 48, 72, 96 and 120 h. The experimental diets were: 1) diet no flaxseed + 20% RUP; 2) diet no flaxseed + 40% RUP; 3) diet containing 10% whole flaxseed + 20% RUP; 4)diet containing 10% whole flaxseed + 40% RUP; 5) diet containing10% grounded flaxseed + 20% RUP; 6) diet containing10% grounded flaxseed + 40% RUP; 7) diet containing10% extruded flaxseed + 20% RUP, and 8)diet containing 10% extruded flaxseed + 40% RUP.

The results showed that the amount of gas production during 96 hours incubation was higher with control diets compared to other diets (255.8 vs 217.8, 198.1 and 200.4 ml/g DM, P=0.001) and also at 20% RUP level (255.5 vs 210.1 ml/g DM, P=0.049). True digestibility of control diets (684 vs 568, 619 and 538 mg/g DM) and NH3-Nof diets containing grounded flaxseed compared to other diets (1.12 vs 0.41, 0.49 and 0.21 mg), was higher (P>0.05), but microbial nitrogen production by diets containing grounded flaxseed compared to other diets (2.72 vs 3.29, 3.36 and 3.45 mg), was lower (P>0.01). The disappeared DM (161.6 vs 149.9 mg), neutral detergent insoluble nitrogen (1.63 vs 1.35 mg) and NH3-N (0.76 vs 0.35 mg) with 40% RUP, and microbial nitrogen production (3.48 vs 2.93 mg) in 20%RUP (P>0.05), was higher.

Since one of the main objectives of the present experiment was to examine the production of microbial protein in experimental diets, diets containing whole and extruded flaxseed despite their low digestibility, but showed higher microbial protein production. On the other hand, 20% RUP level also showed the highest amount of gas and microbial protein production. So, it seems from an economic point of view as well as the performance of the diets, using a diet containing 10% extruded flaxseed along with 20% RUP can be beneficial and effective.

Plants produce an enormous array of phytochemicals arising from various biosynthetic pathways. More than 200,000 defined structures of phytochemicals have been recognized. The growing concerns over bacterial resistance to antibiotics and chemical residues in animal derived foods have led to use phytochemicals as alternatives to antibiotics, other chemotherapeutic agents, and chemical and growth promoting antibiotic feed additives. Phytochemicals exhibit antibacterial, antiviral and antifungal activities against a wide range of pathogenic and non-pathogenic microorganisms. The antimicrobial properties of phytochemicals are being explored to utilize as feed additives in livestock production system (Patra 2012). One of the plant secondary metabolites which researchers recently focus on beneficial effects is saponins. Saponin-containing plants and their extracts by suppressing the bacteriolytic activity of rumen ciliate protozoa could enhance total microbial protein flow from the rumen. Saponins also have selective antibacterial effects which may prove useful in, for example, controlling starch digestion (Wallace et al 2002). Extract of licorice root can be used to manipulate rumen fermentation due to its saponin content. To date, there are limited research on the effect of plant phytochemicals on ruminal microbial fermentation at low pH. Most research studied the effects of plant phytochemicals on high-forage diets at pH > 6.2. The effects of plant extracts on ruminal microbial fermentation are pH-dependent.Microbial populations and ruminal fermentation conditions in cattle fed high-concentrate diets may be very different from cattle fed high-forage diets, because of the type of substrate being fermented or the resulting pH. Therefore, the search for additives that will help to replace ionophores in high concentrate diets needs to be tested in a high-concentrate and low-pH environment. It was hypothesized that extract of licorice root could exert some antimicrobial activities in low pH. Hence, the objective of this experiment was to study the effect of Licorice extract on in vitro gas production parameters, pH, volatile fatty acids (VFA), N- ammonia concentration and protozoa populations in diets containing different levels of concentrate to forage ratio.

Dried licorice extract obtained from Zagros Company (Kermanshah). Two experimental diets with different forage to concentrate ratio (40 to 60% or 60 to 40 %) were formulated. Licorice extract was added to diets at three levels, 1 or 2 and or 3 milligram /liter of incubation media. The saponin content of Licorice extract was 134.75 mg/g of dired licorice extract. For in vitro gas production, rumen fluid was taken from two rumen fistulated Kordish rams. For measuring gas production, 200 mg of experimental diets with four levels of licorice extract (0, 1, 2 and 3 mg/L incubation medium) were incubated with 40 ml of buffered-rumen fluid for 120 hours. The cumulative produced gas was recorded at different times of incubation and gas production parameters were fitted with Blummel et al equation (2003). Organic matter digestibility (OMD) was estimated after 24 hours of incubation (Menke and Steingass 1988). N-ammonia concentration was measured based Broderickand Kang (1980). Rumen protozoa were identified according to the method of Dehority (2003). After 24 incubation, 5 ml of buffered rumen fluid was pipetted into a screw-capped test tube containing 5 ml of formalin. Thereafter, two drops of brilliant green dye (2 g brilliant green and 2 ml glacial acetic diluted to 100 ml with distilled water) were added to the test tube, mixed thoroughly and allowed to stand overnight at room temperature. Total and differential counts of protozoa were made with five replications. In vitro rumen concentration of volatile fatty acids (VFA) was measured by gas chromatography (Ottenstein and Bartley 1971). All in vitro gas production trials were carried out in three runs. Rumen fermentation parameters, protozoa population and organic matter digestibility date were analyzed in a factorial arrangement based on a completely randomized design and gas production data were analyzed in a factorial arrangement based on a complete randomized block design using Proc GLM of SAS software. The differences among treatments were evaluated using Tukey adjustment when the overall F-test was P ≤ 0.05. Trends were declared when 0.05 < P ≤ 0.10.

The results showed that the interaction of diets and licorice extract levels were not significant on in vitro gas production, rate of gas production, estimated organic matter digestibility N-ammonia concentration, pH, total protozoa population, Entodinium, Epidinium, Diplodinium, Eudiplodinium and Isotricha population and in vitro ruminal concentrations of total VFA, acetate, propionate, butyrate, isobutyrate, valerate and isovalerate. The effect of diets on gas production (P≤0.05) and estimated OMD (p < 0.01) was significant and high concentrate diets in compare to low concentrate diets had greater gas production and greater estimated OMD. Inconsistent to these results, adding saponin to diet containing low level of concentrate decreased gas production (Yogianto et al 2014) and adding saponin to diet containing high level of concentrate increased gas production (Aazami et al 2013). Furthermore, high concentrate diet in compare to low concentrate diet had lower organic matter diegestability (Yogianto et al 2014). Diets containing high concentrate had lower acetate and greater isovalerate concentrations (p < 0.01). Addition of licorice extract reduced estimated OMD and total protozoa population and Entodinium population (p < 0.01) and tended to increase total VFA concentration (P=0.09) and to decrease isovalrate concentration (P=0.06).

Based on this study, it is concluded that different levels of licorice extract had no effect on fermentation parameters of diets containing different concentrate: forage ratios. Further studies are necessary to determine the effectiveness of saponin-containing plants extracts on rumen microbial fermentation and digestion kinetics.

When aflatoxin contaminated food is given to lactating animals, the metabolite of aflatoxin, called M, is secreted in milk. Since pasteurization, sterilization, and milk processing have little effect on the survival and reduction of AFM1 toxicity, this poison ultimately transports to various dairy products and endangers consumer health. Along with the negative effects of mycotoxins on health and livestock products, these compounds can to be effective in digestion, metabolism and ruminal microbial populations. In ruminants (especially lactating cows) fed with AFB1 contaminated feeds, health problems such as liver cancer, reduce immunity, reproductive disorders, malformation, decreased feed intake and milk production have also been reported. An increase in liver enzymes can be attributed to the signs of an abnormal body. In addition, previous studies aluminosilicate has been show to tightly bind aflatoxins in vitro. This significantly reduce mortality and morbidity in animals, decrease molecular biomarkers of aflatoxin exposure in humans and animals.

Two experiments were conducted to investigate the effect of aflatoxin B1 and aluminosilicate toxin adsorbents on the parameters of gas production contains gas production potential (b) and gas production rate (c), in vitro fermentation parameters includes pH, ammonia nitrogen concentration, volatile fatty acids and ruminal digestion. In the first experiment, the effects of different levels of AFB1, including 0, 300, 600 and 900 ng/ml, were investigated on the parameters of gas production, fermentation and digestion using batch culture method. In the second experiment the effectiveness of three aluminosilicate adsorbents on the AFB1 detoxification was investigated. MegaBond and MycoBond as native adsorbents and MilBond as a commercial adsorbent were used in 6% of DM. The gas produced was recorded at 2, 4, 6, 8, 12, 16, 24, 48, 72 and 96 h of the incubation. The data obtained were fitted to the non-linear equation to calculate parameters of gas production. Also, at the end of 24 h incubation, four bottles were transferred to refrigerator to stop fermentation. Then, pH, ammonia nitrogen concentration and volatile fatty acid (VFA) of batch culture medium was measured, as well as the dry matter digestibility.

Results of first experiment indicated that with increasing the AFB1 from 0.0 to 900 ng/ml, the gas production rate (c) decreased from 0.134 to 0.092 ml/h and the gas production potential (b) decreased from 160.7 to 131.3, but there was no significant difference between the treatments 0 and 300 ng/ml AFB1. In addition, the gas production lag phase increased significantly with increasing level of AFB1 (P<0.05). Addition of AFB1 to the batch culture did not affect its pH, but the dry matter digestibility significantly decreased (P<0.05) with increasing AFB1. Ammonia nitrogen concentration decreased significantly (P<0.05) with AFB1 addition, so that the lowest concentration of ammonia nitrogen was observed at 600 and 900 ng/ml AFB1 (15.2 and 15.3 mg/dL, respectively). In this experiment the total VFA concentration decreased significantly with AFB1 (P<0.05), but the molar ratio of acetate, propionate, butyrate, valerate and isovalerate was not affected (P>0.05). In the second addition of different aluminosilicate adsorbents significantly reduced the rate and potential of gas production. Likewise, dry matter digestibility and ammonia nitrogen concentration reduced significantly (P<0.05). Significant increase in pH of the culture medium by addition of aluminosilicate adsorbents can be attributed to the fact that aluminosilicate acts as a modifier of hydrogen ion in the environment due to the replacement of cations with hydrogen ion and prevents a significant decrease in rumen pH. Probably lowering the ammonia nitrogen concentration is due to the fact that the protozoan population is affected by aluminosilicate adsorbent and decreases; consequently, the population of the ruminal bacteria increases, which results in the removal of more ammonia nitrogen by microorganisms, and ultimately the concentration ammonia nitrogen decreases in the rumen.

The results of this study showed that AFB1 reduced gas production rate (c), the gas production potential (b), the concentration of ammonia and digestibility, but the pH is not affected in vitro. Also, none of the adsorbents was able to neutralize or reduce the negative effects of AFB1 on the parameters of gas production, fermentation and rumen digestion, which could be due to the absorption mechanism of aluminosilicate adsorbents for AFB1 absorption. The results of this study indicate that adsorbents cannot reduce the negative effects of AFB1 on digestion and rumen fermentation, therefore only proposed strategy is to prevent the contamination animal feed with mycotoxins.

The effect of addition of different concentrations of folic acid (vitamin B9) to the culture medium on in vitro maturation of bovine oocytes was investigated in two experiments. Optimization of bovine oocyte maturation medium is one of the strategies affecting quality of maturity and fertility, ultimately resulting in increasing the quality of produced bovine embryos. In the 1st experiment, to specify the concentrations of folic acid and homocysteine, ovarian follicles were divided into two categories including follicles with more or less than eight mm of diameter. Then, the concentrations were measured in both follicle groups as well as a commercial culture medium. In the second experiment, bovine ovaries were transferred to the laboratory immediately after slaughter and the cumulus-oocyte complexes were collected by aspiration from 2-8 mm diameter follicles. Oocytes were randomly divided into four groups and different levels of folic acid (0, 10, 100 and 1000 ng/ml) were added to maturation medium based on the results of the first experiment. Following incubation of oocytes, the percentage of mature oocytes was determined by Hoechst staining. The concentrations of folic acid and homocysteine in follicles with diameter of less than eight mm was higher than those with diameter of greater than eight mm, as well as the culture medium Supplementation of maturation medium with 100 ng/ml folic acid increased the percentage of metaphase-II oocytes compared to the control (P <0.05). Considering the concentrations of folic acid and homocysteine in the medium, results showed that the addition of folic acid improves in vitro maturation of bovine oocytes.

A study was conducted to evaluate the effect of adding different levels of ethanol, methanol and their 50:50 (v/v) binary mixture of propolis extract on in vitro gas production parameters, dry matter and organic matter digestibility, protozoa counts and methane production in a completely randomized design. Crude propolis sample was collected from Ramian region and its extraction was taken by ethanol, methanol and their mixture. Experimental treatments were including: 1) control containing basal diet with forage: concentrate ratio of 50:50 (without any extract), treatments 2 till 5 (control treatment + 25, 50, 100, 200 µl levels of ethanol extract, respectively), treatments 6 to 9 (control treatment + levels 25, 50, 100, 200 µl of methanolic extract, respectively), treatments 10 to 13 (control treatment + 25, 50, 100, 200 µl levels of binary solvents mixture extract, respectively). Results showed that the highest and lowest potential gas productions were related to treatment containing 200 µl and 25 µl of methanolic extract, respectively (P<0.05). Treatment containing 200 µl of binary solvents mixture extract had the highest metabolizable energy, organic matter digestibility and short chain fatty acid concentration (P<0.05). There was significant difference between treatments on dry matter and organic matter digestibility (P<0.05). Hence, treatments containing 25 and 100 µl of methanolic extract had the highest and control treatment had the lowest dry matter and organic matter digestibility, respectively (P<0.05). Using 25 µl and 200 µl levels of binary solvents mixture extract reduced Halotrish more than other levels and other extract. However, treatments containing propolis extract reduced methaneproduction remarkably. Generally, it can be concluded that propolis methanolic extract has potential to improve ruminal fermentation efficiency. However, more research is warranted to manipulate rumen microbial fermentation.

Bentonite is a combination of aluminosilicate which has a high capacity to adsorb toxins such as aflatoxins and other substances (Magnoli et al. 2010). Many researchers have used this substance as a mineral to control and balance pH of rumen (Bringe and Schultz 1969; Rinsig et al. 1969; Britton et al. 1978). It is identified with a clay composition that 90% of which contains montmorillonite (Aghashahi et al. 2005). It has a high absorption property due to the presence of negative charges on the surface of clay materials (Aghashahi et al. 2005). Bentonite is used for different purposes such as performance improvement of male calves (Aghashahi et al. 2005), reduce radiocaesium contamination of soil (Vandenhove et al. 2005), organic modification for the adsorption of organic contamination (Sreedharan and Sivapullaiah 2012) and improvement the strength properties of dry pellets (Timofeeva and Nikitchenko 2014). The physical and chemical structure of bentonite, allows it to superficially absorb proteins and amino acids (Fenn and Leng 1989). This property of bentonite is hypothesized to protect the proteins and amino acids from microbial fermentation occurring in the rumen. On the other hand, ion exchange attribute makes ammonium ions and cations exchanged (Fenn and Leng 1989) and then, it prompts a more optimized use of rumen microorganisms for microbial protein synthesis, by gradually discharging ammonium ions (Nikkhah et al. 2001). Many commercial bentonites produce in Iran, but their benefits for ruminants is unknown; so, this experiment was conducted to evaluate the effect of sodium, calcium, processed and natural bentonites on ammonia nitrogen concentration using in vitro methods.
Material and At the first experiment, ammonia nitrogen concentration was compared in a medium containing natural and processed (mass percent of sulfuric acid to bentonite was 10, 15 and 20) bentonites after 4 and 24 h incubation. The culture medium was prepared according to Menke and Steingass (1988) and Theodorou et al (2001). Ammonia nitrogen concentration was determined according to kjeldahl (Mojtahedi and Danesh Mesgaran 2011). Rumen fluid was collected via fistula of three Baluchi male sheep. Bentonite samples were processed and acivated with sulfuric acid and temperature. Applied feed in this culture medium was composed of ground barley (50%), urea (40%) and processed bentonite (10%). At the second experiment, the effect of adding sodium or calcium bentonite on ammonia nitrogen concentration at different incubation time in a culture containing feed rations with different rumen degradability was investigated. The crude protein was similar between treatments with different ruminal degradability (12.02 vs. 15.15). The culture medium was prepared just as experiment 1 with different feed ingredient (table 1). Bentonite samples were taken from Vivan Company. Sodium bentonite was composed of 69%SiO2, 11.71% Al2O3, 0.04% BaO, 1.56% CaO, 1.99% Fe2O3, 1.13% K2O, 1.82 MgO, 0.04 MnO, 3.08 Na2O, 0.08% P2O5, 1.17 SO3, 0.16 TiO2, 0.01> Cr2O3, 8.22% LOI and calcium bentonite was composed of 64.63%SiO2, 10.02% Al2O3, 0.06% BaO, 4.68% CaO, 2.21% Fe2O3, 0.4% K2O, 1.5 MgO, 0.05 MnO, 2.1 Na2O, 0.08% P2O5, 2.31 SO3, 0.3 TiO2, 0.01> Cr2O3, 11.66% LOI.
At the first experiment, the ammonia nitrogen concentration was affected by the treatments after 4 h incubation (p Generally, the results indicated that the ammonia nitrogen concentration was affected by the processing procedure of bentonite and sodium or calcium bentonites and the highest ammonia nitrogen absorption was observed at the first incubation times (2 and 4 h). It seems that sodium bentonite compared to calcium bentonite can be more effective in ammonia nitrogen adsorption.

Introduction Aflatoxins (AF) as secondary metabolites are produced by Aspergillus flavus and Aspergillus parasiticus. The most abundant aflatoxin B1 (AFB1) is toxic and carcinogenic to humans and animals. Utilization of mycotoxin adsorbents are noted as the most practical methods for protection of feed ingredients. The purpose of this study was to investigate the effect of organic and inorganic adsorbents for the adsorption of AFB1 and its effects on dry matter digestibility and rumen fermentation characteristics.
Materials and Methods The experimental diet was a mixture of alfalfa silage and concentrate. Procedure of in vitro batch culture was performed according to the Menke and Steingass procedure. In an anaerobic condition, 30 ml of buffered rumen fluid was dispensed with pipetor pump into a 120-ml serum bottle containing 0.5 g DM of the experimental diet. The content of each bottle was contaminated with 0.5 ppm AFB1. Experimental treatments were: the control diet, commercial bentonite, activated bentonite, activated charcoal, the cell wall of Saccharomyces serevisia with 75% purity, bentonite activated charcoal yeast, (0.5 0.4 0.1 percent), bentonite activated charcoal yeast, (0.4 0.45 0.15 percent) and , bentonite activated charcoal yeast(0.3 0.5 0.2 percent) The amount of absorbents for all treatments was 1% of the experimental diets. All bottles were purged with anaerobic CO2, sealed with rubber stoppers and placed in a shaking water bath for 72 h at 38.6 degree centigrade. The amount of produced gas was recorded at 2, 4, 6, 8, 12, 16, 24, 48 and 72 h of the incubation. At the end of incubation, all the bottles were transferred to refrigerator to stop fermentation, and then opened. After pH measurements 2-ml sample of each filtrate bottle was taken and frozen at 20 degree centigrade after acidification with 2-ml of 0.2 N HCl. The biomass residues were centrifuged at 1000×g for 10 min at 4 degree centigrade. The supernatant in each bottle was decanted and the pellet was dried at 65 degree centigrade to a constant weight for the determination of the residues.
Results and Discussion Absorbents addition increased the amount of produced gas in all treatments in comparison with the control treatment significantly (P

A study was conducted to evaluate the effect of different additives on chemical composition, fermentation characteristic, gas production and digestibility parameters of triticale silage and its comparison with barley silage in a complete randomized design. Whole crop triticale and barley were harvested and chopped with a conventional forage harvester under farm condition to length of 2-3 cm. Representative of triticale and barley forage samples were packed manually, in triplicate into plastic bags. The filled silos were stored at ambient temperature and allowed to ensile for 3, 7, 21 and 45 days. The following treatments were applied to the forage samples: 1) triticale silage without any additives (control), 2) control bacterial inoculants, 3) control organic acid, 4) control urea and 5) barley silage. Results showed that triticale silages had higher DM content than barley silage, although there were no significant differences among triticale silages. However, barley silage had lowest pH than other treatment in all times after ensilage. Triticale silage treated with organic had highest aerobic stability (p

Introduction Since the legislation of European Union has prohibited the use of growth-promoting antibiotics such as: monensin, there is an interest in alternatives to manipulate the rumen fermentation. The use of growth-promoting antibiotics in animal feeds is banned in Europe due to having potential risks such as the spread of antibiotic resistance genes or the contamination of milk or meat with antibiotic residues. Recently, probiotics have been increasingly evaluated to replace or facilitate reductions in the use of antibiotics. Thus, the aim of this study was investigating the comparison of antibiotic (sodium monensin) and probiotic (protexin) on the gas production parameters and organic matter digestibility of feedstuffs (alfalfa hay, barley grain, and alfalfa barley mixture)
Materials and Methods Experimental treatments were included control (basal feeds without additive), basal feeds supplemented with sodium monensin or protexin probiotic at levels of 500 or 1000 mg per kg of DM in a rumen fungi culture. Ruminal fluid was collected from two fistulated sheep (49.5±2.5 kg) and all samples were withdrawn 2 h after the morning ration had been consumed. Collected ruminal contents were strained through four layers of cheesecloth and brought immediately to the laboratory. To have a pure ruminal fungi culture, whole ruminal fluid was centrifuged at 1000 g for 10 min and added 0.100 mg/ml antibacterial agent (streptomycin sulfate, penicillin G, and chloramphenicol (14, 35). Gas production technique was used to detect the fermentation parameters of the treatments (16).Three parallel syringes of each treatment were prepared in this experiment. To measure the total gas production (A) and the rate of gas production (c), cumulative gas production, organic digestibility and metabolizable energy of treatments until 120 h. Gas production was measured directly from the volume of the syringes at 0, 3, 6, 16, 24, 48, 72, 96, and 120 h. Statistical analysis of data were statistically analyzed in a completely randomized design was performed by SAS (9.1 version) and the least square of means.
Results and Discussion Results showed the higher level of sodium monensin (1000 mg/kg) decreased fermentable fraction (b), organic digestibility and metabolizable energy of both alfalfa hay and barley grain compared with other treatments significantly. The total gas production (A) and the rate of gas production (c) of gas production, cumulative gas production, organic digestibility and metabolizable energy of alfalfa were highest for control treatment (alfalfa without additives). But the level of 1000 mg/kg of protexin supplemented with barley increased fermentable fraction (b), cumulative gas production, organic digestibility and metabolizable energy than other treatments by ruminal fungi (p

Using of agro-industrial by-products in diet of livestock not only reduces the production costs but also can dwindle the use of human foods in animal nutrition and competition between human and livestock. Thus, proper use of these by-products in animal nutrition and identification of new and low cost feed resources may be one of the priorities in livestock husbandry of our country. Pomegranate seed pulp (PSP) is a by-product of the industrial decoction of pomegranate. Iran with annually production of more than 900 tons of pomegranates is one of the most important regions of pomegranate cultivation and PSP produced from pomegranate processing factories may be extensively used in animal nutrition. Digestibility determination of feeds is one of the most effective ways to evaluate their nutritional value. In addition, there is a strong relationship between feeds digestibility and performance of animal. There are in vivo, in vitro and in situ methods to determine the digestibility of feeds. Although in vivo methods are reference for digestibility values of feeds and are of high precision but they are usually expensive and time consuming. In addition, these methods do not provide any information related to ruminal degradability kinetic of nutrients. The aim of this study was comparison of two methods of ruminal degradability determination methods namely, in situ (nylon bag) and in vitro (Daisy incubator), using dried and ensiled pomegranate seed pulp (PSP).
The PSP used in this study was prepared from Anaryan Co. in Ferdows, Iran. Decocted pomegranate was a mixture of Yazd varieties which were harvested at early autumn of 1389. Two types of PSP as dried and ensiled were used to compare the methods of ruminal digestibility determination, pervasively. The PSP silage was prepared by ensiling of wet PSP (containing 475 g/kg DM) in 3 kg bins (4 replicates)and with density of 650 kg wet PSP per cubic meter for 60 days and dried PSP was prepared by drying of wet PSP in oven at 60ºC. The digestibility of each feed was determined by both in situ and in vitro methods using two Holstein fistulated heifers and Daisy incubator, respectively. Animals were fed a total mixed ration containing 1.8 kg/d of alfalfa hay, 1.8 kg/d of concentrate, 0.5 kg/d of corn silage and 1.8 kg/d of wheat straw (DM basis) at two meals. The ingredients of concentrate were barley grain (35%), corn grain (18%), soybeanmeal (10%), canola meal (15%), wheat bran (11.5%), molasses (7%), vitamin-mineral supplement (1%), oyster shell (2%) and salt (0.5%) (DM basis). Comparison between treatments for each of digestibility methods at 24 and 48 h of incubation was done separately based on completely randomized design using SAS software. Correlation between digestibility determination methods was estimated using Corr proc of SAS and regression coefficient and also regression equation of in situ digestibility method on in vitro digestibility method was determined using Reg proc of SAS.
The results showed that DM digestibility of PSP (both ensiled and dried) was estimated higher when measured by in vitro than in situ method at both incubation times. The correlation between two methods of digestibility determination for dried and ensiled PSP after 24h incubation was 0.81 and 0.96, respectively and after 48h incubation was 0.99 and 0.75, respectively. The regression equations of DM in situ digestibility estimation from DM in vitro digestibility data were of high accuracy. Tagliapietra et al. (27) compared the in vitro and in situ DM digestibility of 11 different feeds and found a high correlation coefficient of 0.90 (P It was concluded that two studied methods of digestibility determination were highly correlated and regression equation may be used to estimate the in situ DM digestibility of experimental feed samples from in vitro digestibility data precisely.

A study was conducted to evaluate the effect of adding essence and leaves of two species of Oregano (Mentaaquatica and Mentalongifolia) on gas production parameters, digestibility of dry matter and organic matter and microbial biomass production in two experiments separately. Collected two species of Oregano was taken essence by macro wave set at 110 0C and 450 nm for 45 minutes. The experimental treatments were: 1) control containing basal diet with forage: concentrate ratio 50:50 (without additive), 2) basal diet + Mentaaquatica lamiaceae essence, 3) basal diet+ Mentalongifolia lamiaceae essence, 4) basal diet+ Mentaaquatica lamiaceae leave and 5) basal diet+ Mentalongifolia lamiaceae leave. 8 macro liter of essence along 8 macro liter of ethanol in treatment 2 and 3 and 20 mg leaves of two kind of Oregano in treatment 4 and 5 were added to glass vial. The results showed that treatments containing essence and leaves had lower gas production than control treatment. Hence, treatment control and treatmenl) gas production, respectively. In second experiment, there were significantly differences aents that containing leaves of two kind of Oregano had lowest DM and OM digestibility compared with other treatments. Treatments containing essence had higher microbial biomass production and microbial biomass production efficiency. Generally, it can be concluded that essence of two kind of Oregano (even their leaves) has the potential to affect ruminal fermentation efficiency.

This study was conducted to investigating the effects of adding thymus plant on digestibility and fermentable gas production parameters in some feedstuffs on in vitro status, by 3×4 factorial experiment in complete randomize design. The treatments were wheat straw, alfalfa and barley grain with 0, 1.5, 3, and 6% levels of thymus plant mash. Test gas syringes volume to 100 ml and the incubation time was 24 hours. The result showed that, gas production from the fermentation process rapid part degradation (a) the grain was affected by treatments. Also the gas production rate of slow degradable fraction (b) and potential gas production (a+b) in the wheat straw treatment with increasing of thymus level was decreased. Degradation rate in alfalfa hay treatment with increasing levels of thymus decreased, but in wheat straw were increase. Metabolizable energy and organic matter digestibility of alfalfa hay treated with thymus 6% was significantly lower than other treatments, but the adding most level of thymus increased of metabolizable energy and organic matter digestibility of wheat straw. The addition of different levels of thymus plants to barley grain and wheat straw treatments decreased total gas production compared to the control group. Also addition of thymus plants in rate of 6% significantly decreased the CO2 production compared to the other levels. The amount of methane production from fermentation of alfalfa hay was decreased significantly with increasing of thymus levels compared with control group. The results of this study it can be concluded, the thymus plant additive for ruminant diets can be used as a suitable to optimize rumen fermentation.

This study was conducted to evaluate the starchy feed effect (the wheat flour and the wheat starches grade1 and 2) on alfalfa hay digestibility using in vivo and in vitro methods. The in vivo experiment was achieved with 4 ruminally fistulated Makui sheep (BW, 43.20±1.2 kg) in a completely randomized design with a change over arrangement and the in vitro Telly and Terry experiment was conducted in a completely randomized design. In either experiment the DM and OM digestibility for mixed (starchy) rations was significantly higher than those of basal (alfalfa hay) ration, but the NDF digestibility of basal ration was higher than that of mixed ration. The rumen pH for alfalfa was the highest, and the lowest pH was obtained with the starch grade1 containing diet. The highest and the lowest correlation of regression equations were related to in vivo DM digestibility and the experimental mixed rations NDF digestibility, respectively. Results of this study showed that the starch grade 1 containing diet had a highest negative effect on the alfalfa hay fibre digestibility. Moreover, the regression equations may encompass better prediction of DM digestibility of mixed rations than that of the fibre digestibility.

The chemical and fermentative parameters of three fresh forages and silages of sorghum including Sweet, Pegah and Speedfeed varieties were compared with corn using in vitro method, also degradability coefficients of forages and silages were determined by in situ method. Forages were planted in the same condition and harvested in soft dough stage, then ensilaged in four replicates for each time of 30, 60 and 90 days of preservation in mini silos. Buffering capacity in green Sweet sorghum was lower than corn and Speedfeed, and acid detergent fiber and water soluble carbohydrates respectively were significantly highest and lowest in fresh forage of Speedfeed sorghum. In time of 60 days, percent of acid detergent lignin of corn silage was lower than Sweet and Speedfeed sorghum silages; similarly, residual water soluble carbohydrate was lowest in corn silage. The lactate Concentration in corn and Pegah sorghums was higher than Sweet and Speedfeed silages. In corn and Sweet sorghum silages, Contents of acetic acid and ammonium nitrogen were highest and lowest, respectively. In nylon bag experiment, Degradation rate of corn and Pegah sorghum forages were significantly higher than Sweet and Speedfeed sorghums that cause to more effective degradability with passage rate of 0.08 in this forages. Also, the slowly degradation coefficient of corn silage was higher than sorghums silages. In conclusion, Speedfeed sorghum forage is not suitable for making silage in comparison others, and corn silage had more potential of degradability.