فصلنامه تحقیقات تولیدات دامی
سال دوازدهم شماره 3 (پاییز 1402)

The use of medicinal plants and their derivatives can stimulate feed consumption, increase daily weight, feed conversion ratio, increase shelf life, improve the health and function of the digestive system. It seems that the use of an optimal mixture of several medicinal plants in the diet has positive effects on performance and carcass quality of broiler chickens in comparison to each one. Nowadays, with the popularization of ready-made feed in raising broiler chickens, many breeders tend to add food additives in drinking water. Therefore, it is a question that adding these compounds in feed or drinking water makes a difference. Three commercial plant additives that are used today are Coxan, O.X. Plant, and Entex. Coxan contains oregano (with the active ingredient of menthol) and garlic (with the active ingredients of allin and allicin), O.X. Plant contains savory (with the active ingredients of carvacrol and thymol), thyme (with the active ingredients of thymol and carvacrol) and red pepper oleoresin (with the active ingredient of capsaicin), Entex contains cinnamon (with the active ingredient of cinnamaldehyde), and garlic and eucalyptus (with the active ingredient of cineol). The aim of this research was to compare the effects of using these commercial plant additives (in water and feed) on growth performance, intestinal microflora and morphology and immune response of broilers.
A total of 336 Ross 308 broilers were examined in a completely randomized design with seven treatments, four replications, and 12 chickens per replication. Experimental treatments included: control (without phytogenic in feed or water), Coxan in feed (300 mg/kg) and in water (200 mL/1000 L), O.X. Plant in feed (200 mg/kg) and in water (135 mL/1000 L) and Entex in feed (500 mg/kg) and water (350 mL/1000 L). Daily weight gain, daily feed intake, daily water intake and feed conversion ratio were measured. At 42 days of age, two birds were selected from each experimental unit and after slaughter, the length of the intestinal components was measured separately (duodenum, jejunum, and ileum). To determine the microbial population, samples were taken from the ileum of chickens. EMB culture medium was used to determine Escherichia coli population and MRS culture medium was used for Lactobacillus bacteria. To check the humoral immune response, 0.1 mL of 25% sheep red blood cell solution in PBS was injected into the breast muscle of chickens on the 12th and 29th days of rearing. Blood was taken from the chickens on the 28th, 35th and 42nd days of breeding and the levels of Anti-SRBC, and immunoglobulins G and M were calculated. Antibody titer against Newcastle disease was measured by HI method.
The effect of additives in water and feed on average daily feed consumption, daily weight gain, feed conversion ratio and daily water consumption was not significant in the whole period. Since the amount of water consumed by the chickens did not change, it can be concluded that the additives added to the water in the examined amounts do not have a spicy or unpleasant taste that would cause the birds to refuse to drink water. The relative length of duodenum, jejunum, cecum and colon was not affected by experimental treatments. However, the relative length of the ileum was lower in the chickens that received Entex in the feed compared to the control group. No significant difference was observed in the relative length of different parts of the intestine in the treatments that received herbal additives either in feed or water. The relative length of duodenum, jejunum, cecum, and colon, villus length, villus area, crypt depth, thickness of lamina propria, thickness of muscular layer, and thickness of advantis layer were not affected by experimental treatments. The chickens that consumed 500 mg/kg of Entex herbal additive of feed had a lower villus width in the ileum region compared to the control group chickens. The population of Escherichia coli bacteria in the ileum of chickens that had received the addition of O.X. Plant and Entex in the feed decreased compared to the control group. The population of Escherichia coli bacteria in the Entex treatment in the feed was also reduced compared to the Coxan treatment in the feed. The population of Lactobacillus bacteria in the ileum of chickens that received Coxan in water, O.X. Plant in feed and water, and Entex in feed increased compared to the control group. There was no significant difference in the population of Escherichia coli and Lactobacillus bacteria in the treatments that received herbal additives either in feed or in water. The use of plant essential oils in poultry feed, while improving the microbial population by increasing the number of lactobacilli, by improving the morphological characteristics of the intestine, probably improves the ability of digestion and absorption in the digestive system and improves the growth efficiency of broiler chickens. Antibody levels against SRBC were higher at 28, 35, and 42 days in all chickens consuming herbal additives in water. The performance of animals is significantly influenced by the state of health and safety of the animal. A weak or stressed immune system causes weight loss when dealing with infectious diseases, so the use of immune system stimulating substances can increase performance by improving the immune status. In raising poultry, it is important to strengthen the immune system to prevent the occurrence of diseases.
The results of this research showed that the commercial plant additives of Coxan, O.X. Plant, and Entex had no significant effect on growth performance, relative length and intestinal morphology. The population of opportunistic bacteria Escherichia coli was significantly reduced compared to the control group with the addition of O.X. Plant and Entex in the diet. In general, it can be concluded that adding tested herbal additives not only did not have a negative effect on the drinking water consumption of broiler chickens, but also using them in water improved the immune responses of broilers more than adding them in feed. A total of 336 Ross 308 broilers were examined in a completely randomized design with 7 treatments, 4 replications, and 12 chickens per replication. Experimental treatments included: control (without phytogenic in feed or water), Coxan in feed (300 mg/kg) and in water (200 mL/1000 L), O.X. Plant in feed (200 mg/kg) and in water (135 mL/1000 L) and Entex in feed (500 mg/kg) and water (350 mL/1000 L). Daily weight gain, daily feed intake, daily water intake and feed conversion ratio were measured. At 42 days of age, 2 birds were selected from each experimental unit and after slaughter, the length of the intestinal components was measured separately (duodenum, jejunum, ileum). In order to determine the microbial population, samples were taken from the ileum of chickens. EMB culture medium was used to determine Escherichia coli population and MRS culture medium was used for Lactobacillus bacteria. To check the humoral immune response, 0.1 ml of 25% sheep red blood cell solution in PBS was injected into the breast muscle of chickens on the 12th and 29th days of rearing. Blood was taken from the chickens on the 28th, 35th and 42nd days of breeding and the levels of Anti-SRBC, immunoglobulin G and M were calculated. Antibody titer against Newcastle disease was measured by HI method. The effect of additives in water and feed on average daily feed consumption, daily weight gain, feed conversion ratio and daily water consumption was not significant in the whole period. Since the amount of water consumed by the chickens did not change, it can be concluded that the additives added to the water in the examined amounts do not have a spicy or unpleasant taste that would cause the birds to refuse to drink water. The relative length of duodenum, jejunum, cecum and colon was not affected by experimental treatments. However, the relative length of the ileum was lower in the chickens that received Entex in the feed compared to the control group. No significant difference was observed in the relative length of different parts of the intestine in the treatments that received herbal additives either in feed or water. The relative length of duodenum, jejunum, cecum, and colon, villus length, villus area, crypt depth, thickness of lamina propria, thickness of muscular layer, thickness of advantis layer were not affected by the experimental treatments. The chickens that consumed 500 mg/kg of Entex herbal additive of feed had a lower villus width in the ileum region compared to the control group chickens. The population of Escherichia coli bacteria in the ileum of chickens that had received the addition of O.X. Plant and Entex in the feed decreased compared to the control group. The population of Escherichia coli bacteria in the Entex treatment in the feed was also reduced compared to the Coxan treatment in the feed. The population of Lactobacillus bacteria in the ileum of chickens that received Coxan in water, O.X. Plant in feed and water and Entex in feed increased compared to the control group. There was no significant difference in the population of Escherichia coli and Lactobacillus bacteria in the treatments that received herbal additives either in feed or in water. The use of plant essential oils in poultry feed, while improving the microbial population by increasing the number of lactobacilli, by improving the morphological characteristics of the intestine, probably improves the ability of digestion and absorption in the digestive system and improves the growth efficiency of broiler chickens. Antibody levels against SRBC were higher at 28, 35 and 42 days in all chickens consuming herbal additives in water. The performance of animals is significantly influenced by the state of health and safety of the animal. A weak or stressed immune system causes weight loss when dealing with infectious diseases, so the use of immune system stimulating substances can increase performance by improving the immune status. In raising poultry, it is important to strengthen the immune system to prevent the occurrence of diseases. The results of this research showed that the commercial plant additives Coxan, O.X. Plant and Entex had no significant effect on growth performance, relative length and intestinal morphology. The population of opportunistic bacteria Escherichia coli was significantly reduced compared to the control group with the addition of O.X. Plant and Entex in the diet. In general, it can be concluded that adding tested herbal additives not only does not have a negative effect on the drinking water consumption of broiler chickens, but using them in water improves the immune responses of broilers more than adding them in feed.

Ascites is one of the most common metabolic syndromes in today's fast-growing broilers and is associated with rapidly growing tissues and characterized by accumulation of lymph fluid in the peritoneal spaces. Effective factors in the occurrence of ascites syndrome include free radicals in the body such as superoxide, hydroxide, and hydrogen peroxide. By reducing the capacity of the body's antioxidant systems, free radicals make the bird susceptible to various diseases. Free radicals produced in the body by damaging the cell membrane lead to cell death and ultimately tissue damage. Therefore, by increasing cell protection from such injuries, one of the common causes of ascites and heart failure abnormalities can be prevented. In addition, free radicals from oxygen derivatives reduce the half-life of nitric oxide (vasodilating agent), causing a decrease in the ability of vasodilation and providing the basis for the occurrence of ascites. Therefore, it is suggested that the use of antioxidants can prevent ascites and improve performance in broilers. Medicinal plants are among the rich sources of natural antioxidants that do not have the harmful effects of antibiotics and synthetic antioxidants. For this reason, the use of medicinal plant extracts is an important step in animal nutrition to increase the immunity of poultry. The mentioned extracts have strong antioxidant, antibacterial, and digestive properties. Probably, the positive effects of medicinal plants are due to the active compounds found in plant extracts such as menthol, thymol, and carvacrol. This study aimed to investigate the effects of ethanolic extract of sage on the performance, antioxidant status, and blood parameters of broiler chickens under induced ascites.
A total of 450 one-day-old chickens (Ross 308) were reared in the form of a completely randomized design with six treatments and five replications (15 chickens in each replication). Experimental treatments include 1. Positive control group (without inducing ascites and fed with basic diet), 2. Negative control group (inducing ascites and fed with basic diet), 3 and 4. Vitamin C group (inducing ascites with levels of 1000 and 2000 ppm vitamin C), 5 and 6- Sage group (ascites induction with 1000 and 2000 ppm of sage extract). To induce ascites, water containing 1200 mg of sodium (three grams per liter of sodium salt) was provided to the chickens from the 15th day of the experiment. Growth performance parameters including feed intake, weight gain, and feed conversion ratio were calculated for the total period. On the last day of the experiment (42 d), two birds were randomly selected from each cage and after sampling from the wing vein, killed and the ascites index was calculated as the ratio of the weight of the right ventricle to the total ventricles. The mortalities were collected as soon as they were observed and after weighing to correct the feed conversion ratio, were necropsied to investigate the cause of death. Blood and biochemical parameters such as the number of red and white blood cells, hemoglobin, hematocrit, heterophil and lymphocyte, serum triglyceride, and cholesterol, as well as serum antioxidant parameters including the level of malondialdehyde and the activity of antioxidant enzymes including glutathione peroxidase, superoxide dismutase, and catalase were measured. In addition, liver enzymes present in the serum including alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and alkaline phosphatase (ALP) were also measured.
The results showed that induction of ascites increased the ratio of the right ventricle to total ventricles, mortality due to ascites, feed consumption, feed conversion ratio, and body weight loss (P<0.05). Administering sage extract improved the mentioned traits and vitamin C also reduced the ratio of right ventricle to total ventricles, losses due to ascites (P<0.05). Sage extract and vitamin C moderated the increasing effects of ascites induction on heterophiles, triglyceride, and cholesterol and increased red blood cells, hematocrit, and hemoglobin compared to the negative control treatment (P<0.05). In addition, the induction of ascites decreased the activity of glutathione peroxidase, superoxide dismutase, and catalase enzymes and increased serum malondialdehyde, experimental treatments, especially sage extract, improved the antioxidant status compared to the negative control treatment (P<0.05). Also, the induction of ascites increased the serum levels of aspartate aminotransferase, alanine aminotransferase, gammaglutaryl transferase, and alkaline phosphatase enzymes. The addition of experimental supplements, especially sage, decreased the serum level of the mentioned enzymes (P<0.05).
The results of this research showed that the use of sage extract compared to vitamin C has better results on performance, increasing antioxidant power, and reducing stress of broilers with ascites.

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.

Rumen fermentation manipulation aims to maximize the feed efficiency and increase the usefulness of rearing ruminants; in simpler words, the goal of rumen manipulation is to increase processes in rumen fermentation that are beneficial for the host animal and to reduce, change, or eliminate inefficient or harmful processes in rumen fermentation. In this regard, trying to use natural products such as medicinal plants has been widely accepted. The desire and demand of consumers to use healthy meat and livestock products has increased the research to search for natural compounds and additives of plant origin that are beneficial for animal health. The positive effects of medicinal plants on the process of microbial fermentation and reduction of greenhouse gases through their active compounds (such as anthole in fennel or thymoquinone in black seed) have been shown. The present study was designed in two experiments to evaluate the effect of different levels of Foeniculum vulgare and Nigella sativa powder on rumen fermentation parameters and protozoa population of Sanjabi sheep by in vitro and in vivo methods.
The present study was designed and carried out in two experiments, in vitro and in vivo. In the first experiment (in vitro), 18 Sanjabi lambs (four to six months of age) with an average weight of 30.8 ± 6.7 kg were randomly divided into three groups with six replicates. Treatments included: control (basal diet without additives), fennel (basal diet plus 20 grams of fennel per kilogram of concentrate), and black seed group (basal diet plus 20 grams of black seed per kilogram of concentrate). In the second experiment (in vitro), 0, 5, 25, and 50 mg of fennel and black seed powder were added to the rumen liquor collected from six Sanjabi lambs (the control group of the first experiment). In both experiments, rumen liquor was taken through the esophageal tube. Gas production, pH, ammonia nitrogen concentration, total volatile fatty acids, in vitro digestibility of organic matter, and protozoa count were measured. In the in vivo experiment, the amount of excrement of sheep was measured by installing nets under the boxes of the animals. Data analysis was done using SAS statistical software. The normality of the counting data (population of protozoa) was first checked by a non-parametric Kolomogorov-Smirnov test and then analysis was done. The experimental design used in this research was completely randomized and for the first experiment, it was repeated measurements in time. Duncan's multiple range test was used to compare the mean of the treatments.
The results of the in vivo experiment showed that the use of fennel and black seed increased the amount of digested organic matter in the rumen significantly compared to the control (P<0.05). In the in vivo experiment, all studied protozoa populations decreased in all treatments (P<0.05). In the in vitro experiment, the addition of fennel caused a significant decrease in gas production (P˂0.0001), organic matter digestibility, and microbial mass at the levels of 25 and 50 mg compared to the control (P<0.05). Ammonia nitrogen concentration at all levels of black seed and the level of 50 mg of fennel had a significant decrease compared to the control (P<0.05). Total protozoa population and Entodinium spp. subfamily was affected by both fennel and black seed treatments in vitro and showed a significant decrease (P˂0.0001). The results of this study showed that black seed and fennel have anti-protozoa properties and can reduce different protozoa populations both in vitro and in vivo. Also, low levels of fennel can improve the fermentation efficiency by increasing the degraded organic matter and reducing gas production. In addition, the results obtained from this research showed that fennel and black seed medicinal plants had the potential to manipulate and change the process of rumen fermentation, and their anti-protozoal effects were evident and significant in both experiments. However, the effects of these plants on the fermentation process in vitro and in vivo did not follow the same process in most of the fermentation parameters, and in vitro results cannot be fully and reliably generalized to the conditions inside the rumen of live sheep.
The levels of fennel and black seed used in this study can reduce different protozoa populations and improve the process of rumen fermentation, although more researches are needed with different levels of these herbs to get more results.

The first step in designing livestock breeding programs is to decide on appropriate breeding objectives. The most significant factor in the inefficiency of livestock breeding programs is the undesired breeding goals, which causes the selection pressure to be applied to the wrong traits. The production system, market conditions, and flock competence cause the breeding goals to be different from each other. This research was conducted to investigate the production system and cost-benefit analysis of Kurdi sheep flocks in the mountainous regions of Kurdistan province under the conditions of village breeding, as well as estimate the economic values and relative importance of traits and determine the breeding goals of this breed.
In an entire production period, seven flocks of Kurdi sheep of the mountain type were covered and investigated to collect detailed information about the breeding system. The parameters required for analysis in the used model include flock structure, production variables, management variables, feed consumption variables, management costs, marketing costs, fixed costs, and prices gathered directly from the covered flocks, market, and scientific sources. The collected data were used to estimate the economic values, economic weights, and relative importance of the traits and determine the breeding goals. In this study, the economic value of traits was calculated using a deterministic static model, and the total annual profit of the flock was calculated by deducting the costs from the system's revenues under survey. The economic values of each trait were defined as a change in the profit obtained per one-unit increase in the desired trait while keeping other traits constant at the population average. The economic weights of traits were also calculated by multiplying the economic value of each trait by the genetic standard deviation of that trait. After determining the relative importance of the traits, the traits present in the breeding goals were determined in order.
The study of the dynamics of Kurdi sheep flocks, and their reproduction and survival characteristics in the conditions of rural farming showed that the survival rate of ewes, pregnancy rate, frequency of lambing, and litter size were 97%, 84%, 20%, and 1.08, respectively, and the rates of lamb survival until weaning and after that were 85% and 96%, respectively. In this way, the number of weaned and salable lambs per head of breeding ewe were 0.90 and 0.54, respectively. The cost-benefit analysis of sheep flocks showed that the costs of feeding, labor, veterinary services, fixed, transportation of animal feed, and marketing were 55.7%, 21.5%, 15.6%, 4.7%, 1.8%, and 0.7% of the total costs, respectively. Meat was the first with 99.5%, and wool with 0.5% was the second source of flocks’ income. The profit from each head of breeding ewe per year was 11313953 Rials, and the ratio of income to cost and the ratio of cost to income were 1.38 and 0.73, respectively. The absolute economic values were positive for all investigated traits except for the live weight traits of the replacements, and the culled mature ewes. Calculation of economic weight and relative importance of traits showed that reproductive traits, survival traits, and productive traits were important, respectively. The traits of pregnancy rate and litter size were the most important, and the live weight of replacements and wool production were the least important in increasing profitability or reducing costs.
The efficiency of each head of breeding ewe compared to costs was calculated as 38%, which has increased by about 3% compared to the last four years. These figures showed that the profit from each head of breeding ewe is noticeable, and the policy of freeing the price of livestock inputs in recent years has not had a significant effect on the profitability of Kurdi sheep breeding in rural conditions. Of course, the number of salable lambs per head of breeding ewe as the net reproduction efficiency in this study was low, and with its improvement, the economic efficiency will also be better.  By examining each trait, it was determined that the breeding goals of this breed, in order of importance, included pregnancy rate, litter size, lamb survival until weaning, lambing frequency, lamb survival from weaning to sale, lamb live weight at sale, ewe live weight at maturity (with a negative value), ewe survival, replacements’ weight (with a negative value) and wool production. This ranking should be considered in breeding programs. In an entire production period, seven flocks of Kurdi sheep of the mountain type were covered and investigated to collect detailed information about the breeding system. The parameters required for analysis in the used model include flock structure, production variables, management variables, feed consumption variables, management costs, marketing costs, fixed costs, and prices gathered directly from the covered flocks, market, and scientific sources. The collected data were used to estimate the economic values, economic weights, and relative importance of the traits and determine the breeding goals. In this study, the economic value of traits was calculated using a deterministic static model, and the total annual profit of the flock was calculated by deducting the costs from the system's revenues under survey. The economic values of each trait were defined as a change in the profit obtained per one-unit increase in the desired trait while keeping other traits constant at the population average. The economic weights of traits were also calculated by multiplying the economic value of each trait by the genetic standard deviation of that trait. After determining the relative importance of the traits, the traits present in the breeding goals were determined in order. The study of the dynamics of Kurdi sheep flocks, and their reproduction and survival characteristics in the conditions of rural farming showed that the survival rate of ewes, pregnancy rate, frequency of lambing, and litter size were (97%), (84%), (20%), and (1.08), respectively, and the rate of Lamb survival until weaning and after that were (85%) and (96%), respectively. In this way, the number of weaned and salable lambs per head of breeding ewe were (0.90) and (0.54), respectively. The cost-benefit analysis of sheep flocks showed that the costs of feeding, labour, veterinary services, fixed, transportation of animal feed, and marketing were (55.7%), (21.5%), (15.6%), (4.7%), (1.8%), and (0.7%) of the total costs, respectively. The meat was the first, with (99.5%), and produced wool, with (0.5%), was the second source of flocks’ income. The profit from each head of breeding ewe per year was (11313953) Rials, and the ratio of income to cost and the ratio of cost to income were (1.38) and (0.73), respectively. The absolute economic values were positive for all investigated traits except for the live weight traits of the replacements, and the culled mature ewes. Calculation of economic weight and relative importance of traits showed that reproductive traits, survival traits, and productive traits were important, respectively. The traits of pregnancy rate and litter size were the most important, and the live weight of replacements and wool production were the least important in increasing profitability or reducing costs. The efficiency of each head of breeding ewe compared to costs was calculated as (38%), which has increased by about (3%) compared to the last four years. These figures show that the profit from each head of breeding ewe is noticeable, and the policy of freeing the price of livestock inputs in recent years has not had a significant effect on the profitability of Kurdi sheep breeding in rural conditions. Of course, the number of salable lambs per head of breeding ewe as the net reproduction efficiency in this study was low, and with its improvement, the economic efficiency will also be better. By examining each trait, it was determined that the breeding goals of this breed, in order of importance, include pregnancy rate, litter size, lamb survival until weaning, lambing frequency, lamb survival from weaning to sale, lamb live weight at sale, ewe maturity live weight (with a negative value), ewe survival, replacements weight (with a negative value) and wool production. This ranking should be considered in breeding acts and programs.

Artificial and natural selection not only increases the frequency of new-useful mutations but also remains some signals throughout the genome. Since these regions often control economically important traits, identifying and tracking these regions is the most important subject in animal genetics. Also, natural and artificial selection related to adaptation and economic traits, such as litter size, results in changes at the genomic level which leads to the appearance of selection signatures. Several tests including the linkage disequilibrium-based approach, site frequency spectrum, and population differentiation-based approach have been developed to explore the footprints of selection in the genome. Domestication and selection have significantly changed the behavioral and phenotypic traits in modern domestic animals. The selection of animals by humans left detectable signatures on the genome of modern sheep. The identification of these signals can help us to improve the genetic characteristics of economically important traits in sheep. Over the last decade, interest in the detection of genes or genomic regions that are targeted by selection has been growing. Identifying signatures of selection can provide valuable insights about the genes or genomic regions that are or have been under selection pressure, which in turn leads to a better understanding of genotype-phenotype relationships. One of the best ways to understand physiological processes is to analyze gene regulation networks. Identification of genes involved in economic traits as molecular markers in breeding is of special importance. Gene regulation networks enable the researcher to study all of the genes together. This study aimed to identify selection signature regions and candidate genes related to adaptation and the number of lambs born.
To identify the signatures of selection in Iranian native sheep and Egyptian breeds, genomic information of 96 native sheep (including 96 Zandi) and 107 Egyptian sheep (including 59 Barki and 48 Rahmani) were used. The genomic information of foreign breeds was extracted from the Dryad database (https://dryad.com/articles/dataset). To determine the genotype of the samples, Illumina Bead Chip 50K was used. Quality control was conducted using the Plink software. The markers or individuals were excluded from the further study based on the following criteria: unknown chromosomal or physical location, call rate <0.95, missing genotype frequency >0.05, minor allele frequency (MAF) < 0.05, and a P-value for Hardy–Weinberg equilibrium test less than 10-6. After the quality control of the data, the hapFLK statistical method, with hapFLK v1.4 software, was used to identify selection signatures. The genomic version of the Oar_v4.0 database in NCBI was used for detecting the genomic position of single nucleotide polymorphisms (SNPs) in the sheep genome. Candidate genes were identified by SNPs located at 0.1% upper range of hapFLK using BioMart software in ensemble 109. Then, using the PANTHER database, the general biological function of the genes was checked. At this stage, it is assumed that genes that belong to a functional class can be considered as a group of genes that have some specific and common characteristics, and the QTLs in the selected region were extracted using the Animal Genome database, and the genes were compared with other research. GeneCards (http://www.genecards.org) and UniProtKB (http://www.uniprot.org) databases were also used to interpret the function of the obtained genes.
Based on the results of hapFLK, by comparing the Zandi population with Egyptian breeds (Barki and Rahmani), seven genomic regions on chromosomes 1, 2 (three regions), 10, 25, and 26 were identified. Candidate genes of DNAJB4, FNDC3B, GULP1, ACVR1, and FGF9 were in these regions. Further investigation using bioinformatics tools showed these genomic regions overlapped with the immune system, adaptation, litter size, and lipid and muscle metabolism.
The results of this study may provide an important source to facilitate the identification of genomic regions and then, the genes affecting economically important traits in the sheep industry. However, it will be necessary to carry out more association and functional studies to demonstrate the implications of these genes. Therefore, in subsequent studies with more samples and more breeds of domestic and wild sheep in Iran, a better understanding of candidate genes for important economic traits in domestic and wild species would be achieved.

Considering the production of new silkworm hybrids by prominent countries in the sericulture industry such as China and the import of these hybrids to the country in recent years, it is necessary to update these evaluations so that correct decisions can be made to choose the most suitable type of hybrid in terms of functional traits. Therefore, such comparisons are done annually and the results are sent to the Iran Sericultural Corporation- Silk Worm Research Center as an executive body for final decision. The present study aimed to investigate the performance of eight Chinese commercial silkworm hybrids, including Suju×Minghu (S×M), Minghu×Suju (M×S), Baiyue×Qiufeng (B×Q), Qiufeng×Baiyue (Q×B), BaiyueB×QiufengA (BB×QA), QiufengA×BaiyueB (QA×BB), 874×873, and 874×873. Hybrids 872×871 and 872×871 also were among the studied hybrids, but due to the decrease in the performance of cocoon-related traits compared to other hybrids that had a big difference in terms of the obtained values, they were removed and excluded from the final analysis. The desired hybrids for 14 traits including best cocoons weight produced per 26,000 larvae (BCW), total cocoons weight produced per 26,000 larvae (TCW), number of cocoons per liter (NCPL), weight of cocoons per liter (WCPL), percentage of pupa viability (PPV), mean weight of a cocoon (MWC), mean weight of a cocoon shell (MWCS), mean of cocoon shell percentage (MCSP), mean weight of a male cocoon (MWMC), mean weight of a male cocoon shell (MWMCS), mean of male cocoon shell percentage (MMCSP), mean weight of a female cocoon (MWFC), mean weight of a female cocoon shell (MWFCS), and mean of female cocoon shell percentage (MFCSP) were reared in the spring of 2022 after hatching and according to standard methods and under the same conditions in Iran Silk Research Center and their performance was recorded. Each silkworm hybrid was reared in three replications of 200 larvae (total number of observations including 600 records per hybrid for all traits) in a completely randomized design. The analysis of the obtained data was done by SAS software by using the procedure of generalized linear model (GLM), and the average performance of the traits was compared to each other using Tukey's range test at the probability level of P<0.05. The results of the analysis of variance for the studied traits showed that the effect of the hybrid type for most of the studied traits including best cocoons weight produced per 26,000 larvae (BCW), total cocoons weight produced per 26,000 larvae (TCW), number of cocoons per liter (NCPL), the weight of cocoons per liter (WCPL), mean weight of a cocoon (MWC), mean weight of a cocoon shell (MWCS), mean of cocoon shell percentage (MCSP), mean weight of a male cocoon (MWMC), mean weight of a male cocoon shell (MWMCS), mean of male cocoon shell percentage (MMCSP), mean weight of a female cocoon (MWFC), mean weight of a female cocoon shell (MWFCS) and mean of female cocoon shell percentage (MFCSP) was significant (P<0.05). In the meantime, only traits of the percentage of pupa viability (PPV) as the most important index related to survival and the mean of male cocoon shell percentage (MMCSP) were not affected by the hybrid type (P<0.05). The average performance of the hybrids showed a significant difference (P<0.05) for all the traits except for the percentage of pupa viability (PPV) and the mean of cocoon shell percentage (MCSP). The BB×QA hybrid showed a higher performance than other hybrids for six traits including the best cocoons weight produced per 26,000 larvae (BCW), the total cocoons weight produced per 26,000 larvae (TCW), the weight of cocoons per liter (WCPL), the mean weight of a cocoon (MWC), the mean weight of a male cocoon (MWMC), and the mean weight of a female cocoon (MWFC) (P<0.01). Also, the M×S hybrid showed a higher performance than other hybrids for the five traits of the mean weight of a cocoon shell (MWCS), mean of cocoon shell percentage (MCSP), mean weight of a male cocoon shell (MWMCS), mean weight of a female cocoon shell (MWFCS), and the mean of female cocoon shell percentage (MFCSP) (P<0.05). In addition, the M×S hybrid showed the lowest value compared to other hybrids for the number of cocoons per liter (NCPL), which the lower values of this trait are preferred (P<0.01).  Based on the results of the present research, the priority of importing and rearing silkworm eggs in 2023 is suggested for BB×QA and M×S hybrids. Although the performance of these two hybrids with other hybrids, except for the 874×873 and 874×873 hybrids, is not significant in most of the examined traits, they can still be considered as next-import priorities. The desired hybrids for 14 traits including: best cocoons weight produced per 26,000 larvae (BCW), total cocoons weight produced per 26,000 larvae (TCW), number of cocoons per liter (NCPL), weight of cocoons per liter (WCPL), percentage of pupa viability (PPV), mean weight of a cocoon (MWC), mean weight of a cocoon shell (MWCS), mean of cocoon shell percentage (MCSP), mean weight of a male cocoon (MWMC), mean weight of a male cocoon shell (MWMCS), mean of male cocoon shell percentage (MMCSP), mean weight of a female cocoon (MWFC), mean weight of a female cocoon shell (MWFCS) and mean of female cocoon shell percentage (MFCSP) were reared in the spring of 2022 after hatching and according to standard methods and under the same conditions in Iran Silk Research Center and their performance was recorded. Each silkworm hybrid was reared in 3 replications of 200 larvae (total number of observations including 600 records per hybrid for all traits) in a completely randomized design. The analysis of the obtained data was done by SAS software (version 9.4) by using the procedure of generalized linear models (GLM) and the average performance of the traits was compared to each other using Tukey's range test at the probability level of P<0.05. The results of the analysis of variance for the studied traits show that the effect of the hybrid type for most of the studied traits including best cocoons weight produced per 26,000 larvae (BCW), total cocoons weight produced per 26,000 larvae (TCW), number of cocoons per liter (NCPL), the weight of cocoons per liter (WCPL), mean weight of a cocoon (MWC), mean weight of a cocoon shell (MWCS), mean of cocoon shell percentage (MCSP), mean weight of a male cocoon (MWMC), mean weight of a male cocoon shell (MWMCS), mean of male cocoon shell percentage (MMCSP), mean weight of a female cocoon (MWFC), mean weight of a female cocoon shell (MWFCS) and mean of female cocoon shell percentage (MFCSP) is significant (P<0.05). In the meantime, only traits of the percentage of pupa viability (PPV) as the most important index related to survival and the mean of male cocoon shell percentage (MMCSP) were not affected by the hybrid type (P<0.05). The average performance of the hybrids showed a significant difference (P<0.05) for all the traits except for percentage of pupa viability (PPV) and the mean of cocoon shell percentage (MCSP). The hybrid BB*QA showed a higher performance than other hybrids for six traits including the best cocoons weight produced per 26,000 larvae (BCW), the total cocoons weight produced per 26,000 larvae (TCW), the weight of cocoons per liter (WCPL), the mean weight of a cocoon (MWC), the mean weight of a male cocoon (MWMC) and the mean weight of a female cocoon (MWFC) (P<0.01). Also, the M*S hybrid showed a higher performance than other hybrids for the five traits of mean weight of a cocoon shell (MWCS), mean of cocoon shell percentage (MCSP), mean weight of a male cocoon shell (MWMCS), mean weight of a female cocoon shell (MWFCS), and the mean of female cocoon shell percentage (MFCSP) (P< 0.05). Also, the M*S hybrid showed the lowest value compared to other hybrids for the number of cocoons per liter (NCPL); the lower values of which are preferred (P<0.01). Based on the results of the present research, the priority of importing and rearing silkworm eggs in 2023 is suggested for BB*QA and M*S hybrids. Although the performance of these two hybrids with other hybrids, except for the 874×873 and 874×873 hybrids, is not significant in most of the examined traits, and they can still be considered as next import priorities.