شکمبه

So far, changes in the vaginal microbial population during the estrous cycle have been evaluated in several livestock species such as sheep. However, changes in the rumen microbial population have not been investigated during the estrous period in sheep. In a study, the results showed that the differences in the rumen microbial population in two reproductive seasons were due to the differences caused by the reaction to day length, and nutrition did not have much effect on it. It is possible that during the reproductive and non-reproductive seasons, the animal has a different microbial population in the digestive tract. The available evidence from clinical research in animals shows that microbes in the digestive system can alter brain activity and behavior through hormonal and neural pathways. Therefore, this study mainly aims to investigate changes in the rumen microbial population during the reproductive season in the estrous period in ewes with estrus and those without estrus for any reason or are so-called anestrus. The changes in the rumen microbial population during the breeding season in the estrous period were measured for the animals showing the occurrence of estrus and those not showing this trait or were anestrous.

This research was done at the same time as the non-breeding season (late spring-beginning of July). The ewes were of Grey Shirazi breed (2-3 years old). To investigate changes in the rumen fluid microbial population in the estrus cycle in estrus and non-estrus animals, two groups of 10 animals were formed from estrus and anestrus animals with the use of teaser rams, and rumen fluid samples were collected for microbial culture and other investigations in both groups once per four days (5 times) during 17 days (like estrus cycle days). The colonies were cultured and separated using general culture media with different growth factors for anaerobic bacteria colonies, such as plate count agar, MRS agar (de Man-Rogosa - Sharpe), and starch agar. The colonies were subtracted by subtractive cultures or different diagnostic tests to identify colonies. Data were analyzed using SAS software version 1/9. Means were compared with the least square procedure and adjusted for Tukey's test.

The population difference between the two groups at 5 different times showed that the population of lactic acid bacteria (LAB) on the day of estrus showed a higher value and a significant difference with other times than the anestrus group. Lipolytic bacteria were higher on the day of estrus than on all days in both groups, with a significant difference. The comparison of different groups showed that the estrous group from times 1 to 5 in the breeding season had the highest number compared to the other groups at different times of the study (p < 0.05). Anestrous groups also had the lowest number at all times (p < 0.05). The comparison of the two estrus and anestrus groups showed that heat and ninth days had higher values than the other times. In comparing the average number of lipolytic colonies in the estrus group, the highest and the lowest numbers belonged to the 1st and the 4th times, respectively, which were significantly different from the other times (p < 0.05). The population of proteolytic bacteria had the highest number of bacteria at time 1 and showed the next values at times 2, 5, 3, and 4, respectively, compared to the estrus group at time 5 (p < 0.05). Time 5 was not significantly different from times 2 and 3 (p < 0.05). In this season, the estrus group was the highest and the lowest at times 1 and 4, respectively (p < 0.05). Times 3-5 were no different significantly (p < 0.05). In the anestrus group, times 3 and 1 had the highest number vs. the lowest numbers in times 2 and 5  (p < 0.05). Based on the comparison of the average population resulting from the counting of total bacterial colonies in the examination of different groups, the estrous group had the highest number of the average microbial population in all 5 times (p < 0.05). The lowest number belonged to two anestrus groups (p < 0.05). The population difference between the two groups at 5 different times showed that the population of LAB had a higher value only on the day of estrus, with a significant difference compared to the anestrous group. The population of lipolytic bacteria in two groups of estrus and anestrus in the 5 studied times showed that the estrus group had a higher and significant value than the anestrus group in the 4th time, i.e. the day of estrus and days 5, 6, and 17. The results showed that the total bacterial population was higher in the estrus group at all times, with a significant difference. Furthermore, anaerobic bacteria increased significantly in certain days of estrus (days 1-9) compared to the anestrous group. The population difference between the two groups at 5 different times indicated that the population of LAB had a higher value only on the day of estrus, with a significant difference compared to the anestrous group. The population of lipolytic bacteria in the two estrus and anestrus groups in the 5 studied times revealed that the estrus group had a higher and significant value than the anestrus group in 4 times, i.e. days of estrus, 5, 6, and 17. The total bacterial population in the estrous group was higher and significant at all times.

In general, the colony population of amylolytic and lipolytic bacteria and the total colonies of anaerobic bacteria are significantly different on days of the estrus cycle, which may affect reproduction. Accordingly, it is better to provide more understandable results of rumen microbial effects on estrus in ewes by increasing analyses such as hormonal changes and molecular investigations, along with microbial culture in estrus crops.

Probiotics accelerate and improve rumen development, stability, and balance of beneficial microbes in the digestive system and decrease microflora disruption, which will increase the activity of desirable enzymes. This action of probiotics increases digestibility, feed efficiency, livestock performance, and general defense including antioxidant power. However, the response of different animals to the specific probiotics is not uniform and similar. Therefore, it is necessary to provide factors that improve the conditions for the establishment and functioning of microbial additives in the rumen or to use compounds that have a synergistic effect with probiotics to receive a uniform and reassuring response from the herd. One of the potentially useful materials to achieve this goal is biochar. In the present study, it was hypothesized that if biochar (as a favorable habitat for microorganisms) is added to probiotic-containing diets, the conditions for establishing these microbial products in the fermentation environment may be improved and probiotics can act more efficiently. Moreover, a probable synergy between probiotics and biochar may increase the efficiency of these additives compared to their separate usage. However, there is no special information available in this regard, especially about the effect of including biochar in probiotic-containing diets on different microbial populations and rumen hydrolytic enzymes. Therefore, this study aimed to investigate the potential of biochar in enhancing the effectiveness of the probiotic sources (Bacilli and/or Lactobacilli) on microbial populations, hydrolytic enzyme activity, digestibility, antioxidant capacity, and fermentation products in the sheep rumen, in vitro.
The experimental treatments were: 1. A basal diet without probiotics and biochar (control), 2. Basal diet containing probiotic Bacilli (B. coagulans, B. subtilis, and B. licheniformis, at the ratio of 2×1011, 5×109, and 5×109 CFU/g, respectively), 3. A basal diet containing probiotic Lactobacilli (L. plantarum, L. rhamnosus, and Enterococcus faecium, at the ratio of 2×109, 2×1010, and 2×1010 CFU/g, respectively), 4. A basal diet containing biochar, 5. A basal diet containing Bacilli-biochar, 6. A basal diet containing Lactobacilli-biochar, and 7. A basal diet containing Bacilli-Lactobacilli-biochar. Diet digestibility was determined by the two-stage Tilley and Terry method. In addition, the 24 and 72-h in vitro gas production techniques were conducted. At the end of each incubation, cellulolytic and proteolytic bacteria, as well as protozoa population, enzymatic activity (carboxymethyl cellulose, microcrystalline cellulose, filter paper degrading, and α-amylase), truly digestible substrate (TDS), partitioning factor (PF), microbial biomass production (MBP), methane release, antioxidant capacity, pH, ammonia-N (NH3-N), and volatile fatty acids (VFA) were determined by the standard methods. All in vitro tests were done in three replicates and two batches (runs) in different weeks. Data were analyzed in a completely randomized design using the GLM procedure of SAS.
Separate inclusion of probiotic Bacilli and biochar in the diet increased the cellulolytic bacteria population and activity of fibrolytic enzymes compared to the control (P<0.05), but these variables were not affected by Lactobacilli. The highest values of these variables were observed with the inclusion of biochar in probiotic Bacilli-containing diets (i.e., Bacilli-biochar and Bacilli-Lactobacilli-biochar treatments). These results were due to probiotics' ability to provide superior growth conditions for useful ruminal microbes, as well as the positive influence of the biochar structure, as a desirable habitat, on establishing, attaching, and developing rumen microorganisms. The protozoa population was not affected by biochar, but it was decreased (P<0.05) in the probiotic-containing diets (Bacilli and Lactobacilli treatments without or with biochar) in comparison to the control. The number of proteolytic bacteria and protease activity were not affected by the experimental treatments. Alpha-amylase activity in 24-h incubation in Basilli and Lactobacilli treatments was higher than the control (P<0.05) but was not affected by biochar. The activity of this enzyme in 72-incubation was lower in the control than in probiotic or biochar treatments (P<0.05). The alpha-amylase activity was the highest in the Lactobacilli-biochar and Bacilli-Lactobacilli-biochar groups. Separate inclusion of probiotics and biochar in the diet increased the diet digestibility, degraded substrate, and microbial biomass production (P<0.05). The maximum values of these parameters were detected in the probiotics-biochar diets. The reason for these increases can be related to the improvement of the cellulolytic bacteria population, alpha-amylase, and fibrolytic enzyme activity in the probiotic or biochar groups. The 24-h total antioxidant activity was not affected by the treatments. In 72-h incubation, the probiotics did not affect this variable, but a tendency to increase in antioxidant capacity was observed in diets containing biochar (without or with probiotics). The improving effect of biochar on the antioxidant power could be owing to its activity in trapping pollutants, poisons, and adverse factors in the incubation medium. Methane release, NH3-N, and acetate to propionate ratio were decreased, but total VFA was increased by separate use of the probiotics or biochar in the diet, compared to the control (P<0.05). More importantly, the inclusion of biochar in the diets containing the probiotic (i.e., Bacilli-biochar, Lactobacilli-biochar, and Bacilli-Lactobacilli-biochar) resulted in the greatest total VFA and the lowest NH3-N and methane release (P<0.05). One reason for the increased methane and ammonia was the lower protozoa population in the additives groups. The methane decline may also be related to the decreasing effect of the additives on methanogens, and their increasing effect on methanotrophs. Moreover, the improved bacterial biomass production could be considered as another reason for the decreased ammonia; i.e., more ammonia was assimilated into the bacterial protein. The increased VFA production was due to the higher digestibility and degraded substrate in the probiotic and/or biochar groups. Regarding the valuable characteristics of probiotics and biochar and their probable synergistic effects, their simultaneous application resulted in the highest improvement of fermentation variables.
Separate use of probiotics Bacilli and Lactobacilli or biochar in the diet improved in vitro ruminal microbial and enzymatic activity and reduced energy (as methane) and nitrogen (as ammonia) losses. More importantly, the addition of biochar to the probiotics-containing diets was a suitable strategy for enhancing the effectiveness of the probiotic sources on digestibility and microbial biomass and reduction of methane and ammonia. However, these results need to be confirmed by further experiments. The experimental treatments were: 1- basal diet without probiotic and biochar (control), 2 - basal diet containing probiotic Bacilli (B. coagulans, B. subtilis, and B. licheniformis, at the ratio of 2×1011, 5×109, and 5×109 CFU/g, respectively), 3- basal diet containing probiotic Lactobacilli (L. plantarum, L. rhamnosus, and Enterococcus faecium, at the ratio of 2×109, 2×1010 and 2×1010 CFU/g, respectively), 4- basal diet containing biochar, 5- basal diet containing Bacilli-biochar, 6- basal diet containing Lactobacilli-biochar, and 7- basal diet containing Bacilli-Lactobacilli-biochar. Diet digestibility was determined by the two-stage Tilley and Terry method. In addition, the 24 and 72-h in vitro gas production techniques were conducted. At the end of each incubation, cellulolytic and proteolytic bacteria, as well as protozoa population, enzymatic activity (carboxymethyl cellulose, microcrystalline cellulose, filter paper degrading, and α-amylase), truly digestible substrate (TDS), partitioning factor (PF), microbial biomass production (MBP), methane release, antioxidant capacity, pH, ammonia-N (NH3-N) and volatile fatty acids (VFA) were determined by the standard methods. All in vitro tests were done in three replicates and two batches (runs) in different weeks. Data were analyzed in a completely randomized design using the PROC GLM of SAS. Separate including probiotic Bacilli and biochar in the diet increased the cellulolytic bacteria population and activity of fibrolytic enzymes compared to the control (P<0.05), but these variables were not affected by Lactobacilli. The highest values of these variables were observed with the inclusion of biochar in probiotic Bacilli-containing diets (i.e., Bacilli-biochar and Bacilli-Lactobacilli-biochar treatments). These results were due to probiotics' ability to provide superior growth conditions for useful ruminal microbes, as well as the positive influence of the biochar structure, as a desirable habitat, on establishing, attaching, and developing rumen microorganisms. The protozoa population was not affected by biochar, but it was decreased (P<0.05) in the probiotics-containing diets (Bacilli and Lactobacilli treatments without or with biochar) in comparison to the control. The number of proteolytic bacteria and protease activity were not affected by the experimental treatments. Alpha-amylase activity in 24-h incubation in Basilli and Lactobacilli treatments was higher than the control (P<0.05) but was not affected by biochar. The activity of this enzyme in 72-incubation was lower in the control than in probiotic or biochar treatments (P<0.05). The alpha-amylase activity was the highest in Lactobacilli-biochar and Bacilli-Lactobacilli-biochar groups. Separate inclusion of probiotics and biochar in the diet increased the diet digestibility, degraded substrate, and microbial biomass production (P<0.05). The maximum values of these parameters were detected in the probiotics-biochar diets. The reason for these increases can be related to the improvement of the cellulolytic bacteria population, alpha-amylase, and fibrolytic enzyme activity in the probiotic or biochar groups. The 24-h total antioxidant activity was not affected by the treatments. In 72-hour incubation, the probiotics had no effect on this variable, but a tendency to increase in antioxidant capacity was observed in diets containing biochar (without or with probiotics). The improving effect of biochar on the antioxidant power could be owing to its activity in trapping pollutants, poisons, and adverse factors in the incubation medium. Methane release, NH3-N, and acetate to propionate ratio were decreased, but total VFA was increased by separate use of the probiotics or biochar in the diet, compared to the control (P<0.05). More importantly, the inclusion of biochar in the diets containing the probiotic (i.e., Bacilli-biochar, Lactobacilli-biochar, and Bacilli-Lactobacilli-biochar) resulted in the greatest total VFA and the lowest NH3-N and methane release (P<0.05). One reason for the increased methane and ammonia was the lower protozoa population in the additives groups. The methane decline may also be related to the decreasing effect of the additives on methanogens, and their increasing effect on methanotrophs. Moreover, the improved bacterial biomass production could be considered as another reason for the decreased ammonia; i.e., more ammonia was assimilated into the bacterial protein. The increased VFA production was due to the higher digestibility and degraded substrate in the probiotic and/or biochar groups. Regarding the valuable characteristics of probiotics and biochar and their probable synergistic effects, their simultaneous application resulted in the highest improvement of fermentation variables. Separate use of probiotics Bacilli and Lactobacilli or biochar in the diet improved in vitro ruminal microbial and enzymatic activity and reduced energy (as methane) and nitrogen (as ammonia) losses. More importantly, the addition of biochar to the probiotics-containing diets was a suitable strategy for enhancing the effectiveness of the probiotic sources on digestibility and microbial biomass and reduction of methane and ammonia. However, these results need to be confirmed by further experiments.

Changes in the microbial population of the digestive tract affect the reproductive and neoral systems. Both systems are involved in the process of estrus and ovulation. In this study, the main goal  wasis to investigate the changes in the rumen microbial population in non-breeding seasons during the estrous period for animals that have  signs of estrus and animals that do not show estrus signs.

In order to investigate changes in rumen fluid microbial population in estrus cycle, two groups of 10 animals in estrus and non estrus status were selected.  Once every four days rumen fluid samples were collected for microbial culture and other investigations from both groups. In order to cultivate and separate the colonies, general culture medium for anaerobic bacteria colonies such as Plate count agar, MRS agar (de Man-Rogosa - Sharpe) and starch agar was used. Subtraction of colonies was also done by subtractive cultures or different diagnostic tests to identify colonies. Data analysis was done using SAS software version 1/9. Means were compared with the least square procedure and adjusted for Tukey's test.

The  comparison between the two groups at 5 different times showed that only on the day of estrus, the population of lactic acid producing bacteria was higher than anestrous group (P<0.05). The comparison between two groups of estrus and anestrus  at 5 different times showed that the estrus group had a higher lipolytic bacterial population than the anestrus group in  days of  5, 6 and 17 of estrus cycle (P<0.05), but  at the day of 13th bacterial population in anestrus group  was higher (P<0.05). The comparison between estrus and anestrus groups showed that the amount of the total bacterial population in the estrus group at all the sampling times in the estrus group was higher than anestrus group (P<0.05).

In total, the cultures obtained from the rumen fluid in the estrus group showed that between amylolytic, lipolytic and total colonies of anaerobic bacterial populations in certain days of estrus cycle was different between animals with sign and no sign of estrus.

Nanotechnology brings new solution for improving ruminal fermentation. Meta-analysis study of the effect of nanoparticles on ruminal fermentation can help to better understand how to manipulate ruminal fermentation. After searching and selecting appropriate articles, data related to ruminal fermentation parameters were extracted. Meta-analyses were carried out using the Comprehensive Meta-Analysis package, version 3. The effect sizes of across studies were calculated with fixed and random effect models. Possible publication bias was evaluated with funnel plot and statistical tests. The results of meta-analysis showed that the administration of nanoparticles in the diet, has a positive effect on gas production, total VFAs concentration and apparent dry matter digestibility. Addition of nanoparticles in the diet decrease N-NH3 concentration. The values of I2 for GP and TVFAs indicated moderate and values of I2 for N-NH3 concentration and apparent dry matter digestibility effect size indicated high heterogeneity, respectively. Administration of nanoparticles can improve ruminal fermentation process due to its positive effects on microbial growth, fiber degradation, DM digestibility, TVFAs concentration, and reduce the ratio of acetate to propionate and methane production.

The aim of this study was to investigate the ruminal disappearance (releases or solubility) of mineral elements (calcium, phosphorus, magnesium, sodium and potassium) from feedstuffs (wheat straw, sugar beet pulp, alfalfa, silage and wheat bran) and to compare the disappearance of five types of mineral supplements (mono calcium phosphate, di calcium phosphate, calcium carbonate, oyster shell and limestone). The analysis of the feed section was performed by a completely randomized design five feed and the mineral supplements section was performed. Examination of ruminal release of minerals from feeds showed that except calcium, a large part of minerals in food was released at zero time (P<0.05). Disappearance rate increased with increasing incubation time. Release order of elements in the incubation times was in the form of potassium, phosphorus, calcium, sodium and magnesium and the highest disappearance rate was for potassium and the lowest release rate was for magnesium (P<0.05). In the study of the highest release rate was obtained during 72 hours of incubation for potassium in alfalfa hay (P<0.05). In the other part of the experiment, the highest disappearance was obtained for monocalcium phosphate and the lowest for calcium carbonate in total incubation times (P<0.05). Also, monocalcium phosphate mineral supplements had the highest and calcium carbonate treatment had the lowest disappearance parameters soluble part of dry matter, and significant difference between experimental treatments for effective dry matter degradability at flow rates of 2, 5 and 8 (P<0.05). In general, the amount of potassium disappearance was the highest in all nutrients and in all incubation times, and also monocalcium phosphate had the highest rate of degradability parameters.

 The anaerobic microbial fermentative digestion of feedstuffs in the rumen is not efficient. The gases are considered as waste products of rumen fermentation and also pollutants of the environment. Recent studies indicated that some metal nanoparticles (NPs) were toxic to rumen microbial population and inhibit methane production in anaerobic conditions. Plant extracts can be used to produce cost effective and eco-friendly green nanoparticles. Phlomis cancellata Bunge with Persian names Gushbarre sefid has high medicinal value and antibacterial properties and distributed dramatically in Khorasan, Mazandaran and Golestan. Hence, there is potential of using bio-synthesized nanoparticles in ruminant nutrition. However, there is not enough information regarding the effect of green nanoparticles on runminal condition. Therefore, the present investigation was carried out to study the effect of green silver nanoparticles synthesized via Phlomis cancellata Bunge extract on rumen fermentation in vitro.

Synthesis of sliver nanoparticles was prepared by adding of 1 ml of the aqueous extract to 4.76 mM sliver nitrate solution (pH= 6.84) allowed to react at 77 °C for 24.79 min in the dark to minimize the photo activation of silver nitrate. The color change of solution from yellow to brown after 3 min of incubation is indicative of the bioreduction of Ag+ ions in the solution to Ag°. Effects of increasing the concentration of green silver nanoparticles synthesized via Phlomis cancellata Bunge extract (0, 125 and 250 µg/ml) on rumen fermentation were evaluated using in vitro gas production technique. Gas production volumes were recorded at 3, 6, 9, 12, 24, 48, 72 and 96 h of incubation and then gas production kinetic was estimated. The obtained data from gas production at 24 h after incubation were used for estimation of digestible dry and organic matter, metabolisable energy, short chain fatty acids, partitioning factor, microbial biomass production and microbial biomass production efficiency.

 The increasing level of green silver nanoparticles synthesized via Phlomis cancellata Bunge extract did not significantly affect in vitro potential of gas production and gas production rate. For the cumulative gas production, no significant difference was found among the treatments. The lowest and highest in vitro potential of gas production, gas production rate and cumulative gas production was recorded for treatments 3 and 1, respectively. Silver nanoparticles exhibit unique bacteriostatic and bactericidal properties. At the atomic level, silver has the ability to absorb oxygen and acts as an oxidation catalyst. Atomic oxygen absorbed on the silver surface reacts with the thiol groups surrounding the surface of bacteria and viruses and removes hydrogen atoms. The bacterium loses respiration ability by disruption of the so-called respiratory channel, which results in bacterial death. The apparent in vitro dry matter digestibility and organic matter digestibility and true in vitro organic matter digestibility, were not significantly affected by addition of green silver nanoparticles synthesized via Phlomis cancellata Bunge extract. Addition of green silver nanoparticles synthesized via Phlomis cancellata Bunge extract failed to affect metabolisable energy, short chain fatty acids, partitioning factor, microbial biomass production and microbial biomass production efficiency. However, the lack of significant effect of the synthesized silver nanoparticles on the digestibility may be due to factors such as concentration, surface capacity, size, and other properties of silver nanoparticles. For treatment 1, the cumulative gas production over 24 and 48 h was negatively correlated apparent in vitro dry matter digestibility and organic matter digestibility. There was significant negative correlation between apparent in vitro organic matter digestibility and partitioning factor with cumulative gas production during hours of incubation for treatments 2. No significant correlation was found between apparent in vitro dry matter digestibility and organic matter digestibility and partitioning factor with the volume of cumulative gas produced during different incubation hours in for treatments 3.

 Supplementing of green silver nanoparticles synthesized via Phlomis cancellata Bunge extract could modify the characteristics of gas production and fermentation parameters basal diet and reduce the side effects of the anaerobic microbial fermentation. However, additional microbial studies with different level of green silver nanoparticles are necessary to determine the mode of action. Additionally, further in vivo work is needed to assess the effect of green silver nanoparticles inclusion on animal performance when cattle are fed ingredients commonly used in beef feedlot or dairy diets.

On the contrary, the grain-fed steers received a grain-based regime that served as an efficient source of high-digestible energy. Rumen may function differently in the grass- and grain-fed regimes. Therefore, grain-fed cattle suffer stronger metabolic stress than pasture-fed steers; and they tend to easily have metabolic and infectious diseases. In this study to gain insights into transcriptional regulation in spleen and rumen tissues under two different regimes include grass and grain, the first high expression genes in two different conditions were identified by RNA-seq data analyses. Promoter analysis was performed using a bioinformatics database of Genomatix. Moreover, in this study to the visualization of regulatory networks containing transcription factors and that regulate the genes in the rumen and spleen, were used Cytoscape software. Then, promoter analysis leads to the identification of 31 novel transcription factor activating in the rumen and 10 novel transcription factors candidates in the spleen in cow fed with grass and grains. Results revealed that 10 genes with the highest expression were identified in both rumen and spleen. According to the analysis of the results in the David Database, the processes: reduction of oxidation, regulation of cell proliferation, ion transport, epididymal development ,and ectoderm development were evaluated as significant. The results in this study provide valuable insights into molecular mechanisms in spleen and rumen tissues under two different regimes include grass and grain.

The aim of this study was to determine chemical composition of Gundelia Tournefortii (GT) hay and the effect of replacing GT hay with alfalfa hay and wheat straw on nutrients digestibility, microbial protein synthesis, ruminal and blood parameters. In this experiment, four Kermani mature ram (BW= 55±2.5 kg) were used in a 4×4 Latin Square design with four periods of 21 days. The experimental diets were: 1) control diet (without GT hay), 2) diet containing 10% GT hay, 3) diet containing 20% GT and 4) diet containing 30% GT hay. The organic matter, ether extract and neutral detergent fiber of GT hay were significantly (p < 0.05) higher, while crude protein and acid detergent fiber were lower than alfalfa hay (p < 0.05). The intake and excretion nitrogen, and percentage of retained nitrogen, ruminal ammonia- nitrogen and pH were not affected by experimental diets. The lowest number of total protozoa and its species were obtained with diet containing 30% GT hay (p < 0.05). The concentration of blood glucose in sheep fed diet containing 20% GT was maximum (p < 0.05). In conclusion, the chemical composition of GT hay compared to alfalfa hay and wheat straw showed that this can be used as a feed with the high amount of metabolizable energy and fiber in sheep diet without negative effects on animal performance.

The high levels of concentrate are used for increase gaining in fattening Sheep. This can be caused Acidosis, Liver abscess and Bloat which produce some problems. Using of rumen modifiers can help to prevent of this disorders. The aim of this study was to evaluate the effects of dietary supplementation of malat (Rumalato), maltodextrin and mixture malate-maltodextrin on Sheep performance parameters, Rumen fermentable products and blood metabolites of lambs fed high-concentrate growing diets.

For this purpose, 48 Shall lambs (46.1± 4.1 kg body weight) were randomly assigned to four dietary treatments in a completely randomized design with 12 lambs per treatment for 30 days. The treatment groups were as follows: 1) control: basal diet without any additive, 2) basal diet plus 30 gr maltodextin, 3) basal diet plus 15 gr malate (Rumalato) and; 4) basal diet mixture of 15 gr malate plus 30 gr maltodextrin. Dry matter intake, Body weight gain, Rumen fluid was collected 3-5 hours after morning feeding. Rumen parameters were measured for pH and volatile fatty acid at the end of the experiment. Blood sampling was collected before morning feeding. Blood parameters were defined at the end of experiment for Glucose, Albumin, Cholesterol and Blood urea nitrogen.

Dry matter intake was higher (p < 0.05) in lambs fed maltodextrin (1.86 kg/d) and mixture malate-maltodextrin (1.87 kg/d) than in lambs fed control (1.79 kg/d) or malate diet (1.80 kg/d) (P=0.03). Body weight gain and feed conversion ratio had not significant difference at the end of experiment (P>0.05). But average body weight gain showed tended to increase in lambs fed malate-maltodextrin. The blood glucose was significant between treatments and it was 79 mg/dl in lambs fed maltodextrin and 78 mg/dl in malate-maltodextrin compared to control (63.8 mg/dl) and malate (65 mg/dl) (P=0.04). Other blood parameters had not different. Among of rumen parameters, pH tended to increase in the lambs fed maltodextrin and malate-maltodexrtin compared to other treatments but without significant difference (7.1 and 6.9 to 6.8)(P>0.05). Other rumen fluid parameters such as volatile fatty acids had not significant difference between treatments.

Totally, results of this study showed that adding of maltodextrin and mixture malate-maltodextrin can be improved dry matter intake in fattening sheep and this is effective for body weight gain and feed conversion ratio. However rumen modifiers are effective to rumen pH and prevention of low pH and caused balance in production of rumen fermentable products.

The use of agricultural by-products and waste because of its high value of nutrients, is possible in the preparation of animal feed. About 950 tons of Mentha pulegium pulp is produced annually in Iran. If this pulp dries well, it can be used in animal feed due to its nutritional value. Due to the fact that moisture is a limiting factor for animal feed storage, ensiling can eliminate this limitation. This study was to evaluate of chemical composition of Mentha pulegium pulp silage with wasted date and its feeding effects on - dry matter intake, microbial protein yield and blood parameters in sheep.

400 kg of Mentha pulegium pulp were mixed thoroughly with 30 kg of wasted date at levels of 0 (control), 5, 10 and 15 % (DM basis) and ensiled for 45 days. After determining the chemical composition and sensory assessment of the silages, level of 20 % (DM basis) was used in experimental diets. For determine the effects of Mentha pulegium pulp silage on sheep, four mature ram (with 40 ± 2 live weight) were used in a change-over design with four periods of 21 days. In the 5 days at the end of each period at 0, 3, 6 and 9 h after morning feeding, rumen fluid was sampled from sheep by esophagous tube and were filtered through three layers of cheesecloth. Blood samples were collected at the end of each period and 4 h after morning feeding in 10 mL. To determine the amount of allantoin and microbial protein synthesis, the daily urine produced during 24 hours was collected.

According to the sensory assessment, silages containing of wasted date get score of very good of 20. Adding wasted date to Mentha pulegium pulp increased the DM and ether extract of silages. DM intake and digestibility of DM, crude protein and NDF of experimental diets were not affected by silages of Mentha pulegium pulp with different levels of wasted date. Nitrogen intake, nitrogen excretion and the percentage of nitrogen retention were not affected by experimental diets. Total purine derivatives and microbial protein synthesis did not change. The level of blood cholesterol and albumin were significantly changed by feeding diets containing Mentha pulegiumsilage with wasted date (p < 0.05).

In conclusion, could be used Mentha pulegium pulp silage up to 20 % without additives or with wasted date in sheep diet.

Streptococcus bovis has been considered to be one of the starch utilizers and lactate producers in the rumen. By considering the role of S. bovis as main lactic acid producer, a large amount of biological information about this strain has been published. But there has not been a systematic analysis of metabolic capabilities for S. bovis so far. In the present study, the first genome-scale metabolic model of S. bovis (iStr472) was reconstructed based on the genome annotation of S. bovis B315. The model was analyzed in terms of sensitivity, topology and capabilities for utilization of other substrates. Results revealed that iStr472 comprises 694 reactions, 626 metabolites and 472 genes. The majority of reactions were located on the nucleotides metabolic pathway. The metabolic genes of model estimated as 27.6 % of all coding genes. Comparison of two models (iStr472 and ModelSEED) indicated that iStr472 has a higher accuracy and validity than the ModelSEED. The 16 highly connected metabolites were found in the model. The results of reaction deletion presented that the model has 126 vital reactions essential for the organism's growth. According to the model prediction, production of biomass was inversely influenced by lactate production. The iStr472 is also capable of utilizing fructose as carbon source. Taken together, it would be possible to use the predictions of iStr472 as a tool for better understanding the metabolism and metabolic engineering of S. bovis for reduced lactate production in the rumen.

The aim of this research was to determine effects of different toxin adsorbents on the amount of Diazinon residues and effectiveness of these additives on ruminal parameters and some blood metabolites in Mohabadi lactating goats fed with high white grape pomace-based diets (27.71%). In this study, 20 lactating goats with 50±5 kg body weight were used in a completely randomized design with 4 treatments and 5 replications. Experimental treatments included diets containing: 1- White grape pomace without toxin adsorbent or control group 2- Grape pomace processed with Mycofix-Plus toxin adsorbent 3- Grape pomace processed with Bio-Tox toxin adsorbent 4- Grape pomace processed with Bio-Acid toxin adsorbent. Adding adsorbent compounds of toxins to diets with high levels of grape pomace reduced the amount of Diazinon in the diet (P˂0.05), so the highest amount of Diazinon in control treatment (4.08 mg/kg) and the lowest amount of toxin was observed in treatment group treated with Bio-Tox supplement (0.67 mg/kg). As a result of adding different toxin adsorbents there was an increase in the amount of dry matter intake (DMI), rumen N-NH3, total VFA, propionic acid, glucose and urea concentration (P˂0.05). Adsorbents did not have a significant effect on the Rumen protozoa population (p>0.05). Adding different adsorbents reduced Non-esterified fatty acids (p<0.05). There was not any significant difference among the investigated groups in terms of the concentration of β-hydroxybutyrate (BHBA), total protein and albumin as a result of adsorbent use (p>0.05). As a conclusion, the amount of the adsorbent and deactivating compounds of toxins in diets with high levels of grape pomace reduced the amount of Diazinon and improved feed intake, rumenal and blood parameters in Mahabadi's lactating goats.

According to the Food and Agriculture Organization (FAO) Iran is the largest pistachio (Pistachio vera) producer in the world. Nevertheless, pistachio by-products (PBs) contain a high level of phenolic compounds and tannins, which can affect their utilization by animals. Tannins were primarily considered as anti-nutritional biochemical, however, in recent years, they have been recognized as useful phytochemicals for modulating rumen that they have different effects such as reducing protein degradation in the rumen. The mechanisms by which tannins reduce ruminal degradation of different dietary components are not entirely clear. The present study was conducted to evaluate the effects of PBs tannins on hydrolytic enzyme activities in ruminants.
An experiment was designed in three parts. The first was carried out to examine the effects of PBs tannins on attachment of rumen microorganisms to the substrate. PBs was processed with heat, moisture and poly ethylene glycol to create treatments with different tannin content and similar chemical composition. The treatments were incubated in nylon bags in the rumen for 12 hours and enzyme activities were determined in microbes attached to solid matrix. The second part was carried out to evaluate the effects of PBs tannins on the extracellular enzyme activities (rumen liquor). The PBs aqueous extract containing tannins were added to the culture medium and after 24 hours of incubation at 39 °C, enzyme activities were determined. The aim of the third part was to investigate the effects of PBs tannins on enzyme activities in cell-free extract of rumen liquor (inhibit or enhance the enzyme activities). At this part, after preparation ruminally cell-free extract, the PBs aqueous extract containings tannins were added to the culture medium, and after 24 hours of incubation at 39 °C, enzyme activities were determined.
The highest negative correlations there was between the activity of protease, carboxymethylcellulase (CMCase) and xylanase, with condensed tannins and then total tannins. While there was a positive correlation between amylase activity and various portions of phenolic compounds. In the first part of the experiment, the activities of protease, CMCase and xylanase were increased with decrease the quantity of tannins in treatments, for both micro-organisms that are tightly bound to the washed solid matrix and the fraction containing loosely plus tightly bound micro-organisms, but amylase activity were decreased for two compartments. Moreover the activities of protease, xylanase and amylase were lower in micro-organisms that are tightly bound to the washed solid matrix than the fraction containing loosely plus tightly bound micro-organisms. In the second part of the experiment, the activities of protease, amylase, CMCase and xylanase significantly decreased with increasing the levels of extract, which indicates PBs tannins have negative effects on extracellular enzyme activities (rumen liquor). In the third part by increasing levels of extract the activities of protease, amylase, CMCase and xylanase significantly decreased, which indicates PBs tannins have inhibition effects on hydrolytic enzymes.
Results of this study showed that PBs tannins can restrict proteolytic and fibrolytic enzyme activities in ruminants, which probably do it by reducing the access of microorganisms to substrate, effects on micro-organisms and release enzymes, and effects on the structure of extracellular enzymes and inhibit them.

This study was conducted to evaluate the effects of alfalfa hay (AH) particle size and soybean hull (SH) on physical effective fibre (peNDF), particle size distribution in diets, chewing activity as well as milk composition and production in early lactating dairy cows. Eight lactating Holstein cows averaging 590±33 kg of BW and 47±13 days in milk were assigned in a 4×4 Latin Square design to a 2×2 factorial arrangement of the treatments: alfalfa hay particle size (fine vs. coarse) combined with soy hull (zero or as a substitute for 50% of AH). The peNDF and physically effective factor (pef) values of diets increased by increase in AH particle size and inclusion of SH in diets (P<0.01). Intake of peNDF was increased by increase in the Particle Size (PS) of AH (P=0.08) and SH inclusion (P<0.01) in diets. Milk production was increased by feeding diets containing SH (P=0.04). Milk fat content was increased by increasing PS of AH (P=0.03) and by decrease in SH substitution (P<0.01). Cows fed coarse AH tended to spend more time chewing in comparison with cows fed fine AH (P=0.08). Coarse particles of AH increased total chewing activity based on min/kg of DMI (P<0.05) and tended to increase the eating activity (P=0.09). When chewing activity was expressed as based on NDF intake, ruminating time and total chewing time tended to be increased by coarse AH being fed (P<0.1). Interestingly when chewing activity was reported as based on peNDF intake, the SH inclusion in diet significantly decreased the eating time (P=0.02), total chewing time (P=0.03) and tended to decrease the ruminating time (P=0.08). SH inclusion in the diets increased pH of the feces (P<0.01). With regard to ruminating time and milk fat percentage, the results of the study indicated that fiber in SH can not be physically as effective as that in AH. In diets containing non-forage fiber sources (as SH in this study) using Penn Stat Particles Separator cannot be a reliable tool for an evaluation of some physical properties of diets.