Beneficial Effects of Adding a Mixture of Medicinal Herbs to Quail Diet on some Physiological and Biochemical Blood Parameters

This study was conducted in the poultry field of the Department of Animal production / College of Agriculture / University of Al-Qadisiyah to know the effect of using a mixture of herbs (licorice, thyme, turmeric and red pepper) on some physiological and biochemical characteristics of quail. 120 quail were used in this study. Which were randomly distributed in four treatments (2 replicates for each treatment, 15 birds for each replicate). The mixture of herbs was added to the diet in equal proportions and with different concentrations.(T1) A mixture of medicinal herbs was used at 2% of the ration (T2) A mixture of medicinal herbs was used at 4% of the ration (T3) A mixture of medicinal herbs was used at 6% of the ration (CONTROL), The mixture of medicinal herbs was used with a percentage of 0% of the feed. Feed and water were provided freely to the birds throughout the experiment period.The effect of a mixture of herbs (licorice, thyme, turmeric and red pepper) on some physiological characteristics such as the number of red cells (RBC) and white blood cells (WBC), types of white blood cells, the packed cell volume (PCV), hemoglobin concentration (Hb) and biochemical traits such as glucose, total protein and cholesterol concentration was studied. The results indicated that there were no significant differences between the experimental treatments (medical herbal mixture) compared with the control treatment in both eosinophil and basophil white blood cells, as well as the percentage of total protein. A significant increase (p<0.05) in (red blood cell count(RBC), packed cell volume (PCV), hemoglobin percentage(Hb), white blood cell count(WBC), and white lymphocyte count, in the experimental treatments (medical herbal mixture) compared with the control treatment. There was a significant decrease in both glucose and cholesterol concentration in the experimental treatments (medical herbal mixture) compared with the control treatment. From this study we conclude that the use of the medical herbal mixture led to the improvement of some of the studied traits.


Introduction
The world has recently tended to rely on medicinal herbs as an alternative to chemical antibiotics, because they contain antioxidants and are free from side effects that harm health through the occurrence of oxidative stress [1][2][3].Researchers were able to extract the substance effective in all medicinal plants, whether this substance is present in the leaves, fruits, stems or roots [4]., where these medicinal plants have a positive role in improving the productive, physiological and immune qualities and increasing the body's resistance to diseases through their possession effective compounds that inhibit harmful oxidative processes caused by free radicals and turn them into stable compounds that are unable to interact with body cells [1].These medicinal plants have been used in animal diets or added to water for the purpose of improving the productive and health status of animals [5].The licorice plant (Licorice Roots) is considered one of the medicinal antioxidant plants that improves the productive qualities and metabolic processes of the cells of the body [6] and is characterized by containing many vitamins, minerals and effective chemical compounds, especially the compound ( Glycyrrhinzin is a sweet-tasting compound [7].When this compound enters the body, it is decomposed into (Glycyrrhizinic acid), which is similar in structure to cortisone, which is highly effective against inflammation and oxidation factors due to the similar structural structure of both compounds [8].It is also characterized by containing flavonoids and phenolic compounds [9], which are effective against diseases and oxidative factors by eliminating free radicals [10].The thyme plant (Thymus vulgaris L.) is a herbal and medicinal plant whose effectiveness lies in the fact that it contains active substances such as thymon and carvacol [11] and is also characterized by being an antioxidant [12] as well as having a role in improving increasing the weight, improving the quality of carcasses and reducing the percentage of deaths in poultry [13].The red pepper plant is also one of the commonly used medicinal plants because of its many properties and benefits.It is rich in flavonoids, which are considered antioxidants.It also contains vitamins A and C, as well as vitamin B. It is also rich in mineral elements such as iron, copper and potassium.And many active substances such as Capsaicine) [14].Capsaicine is one of the effective compounds against bacterial, viral and fungal diseases [15].In addition, the turmeric plant (Curcuma longa), which belongs to the ginger family, is a tropical plant that contains curcumin, which is used in many medicines in the treatment of liver disease , anorexia [16] and antivirals [17].In recent years, turmeric powder has been used as additives in poultry diets to improve production performance and improve the work of the immune system.Because of the active substances found in the composition of turmeric, it has the advantage of having an anti-oxidant, anti-fungal and coccidian effect [18].Therefore, this study aimed to know the effect of adding a mixture of medicinal herbs to the quail diet on some physiological and biochemical characteristics of the blood.

Material and Methods
This study was conducted in the field of the College of Agriculture / University of Al-Qadisiyah and laboratories in Mosul Ethical approval No. um.VET.2021.5., using 120 quail birds, which were divided into four treatments, and each treatment was divided into two replicates, with (15) birds for each replicate.The transactions were divided as follows: T1:.A mixture of medicinal herbs was used by 2% of the diet.T2: A mixture of medicinal herbs was used by 4% of the diet.T3: A mixture of medicinal herbs was used by 6% of the diet.. CONTROL: The control treatment is free of a mixture of medicinal herbs, 0% of the diet.Quail birds were fed on the final diet shown in Table No. (1), which was prepared according to [19] and contains different concentrations of the above-mentioned medicinal herbs mixture.The birds were left for a week for the purpose of adaptation and then the experiment began, which lasted for six weeks and feed and water were provided freely throughout the experiment period.The additives (medicinal herbs), which are licorice plant, thyme plant, turmeric plant and red pepper were obtained from local herbs.Then it was mixed manually with a small amount of forage, then the quantity was increased with good mixing until reaching the required consistency between the feed materials included in the diet of each treatment.After the mixing was completed, it was filled with labeled bags, each one according to its treatment and the required ratios of 2%, 4% and 6%, and closed tightly for the purpose of preserving the additives.On the last day of the experiment, 5 birds were selected from each treatment and blood was drawn from the brachial vein in tubes containing EDTA.Physiological characteristics such as Red blood cells (RBC), White blood cells (WBC) and its types, determination of hemoglobin (Hb) and hematocrit (PCV) according to the method of Wintrobe hematocrit, Neubauer hematocytometer and cyanomethaemoglobin [20] were measured.As for the measurement of vital characteristics such as (cholesterol and blood sugar) using a special kit from (Randox) company, as for the total protein concentration, it was measured using the Biuret method [21].Complete random design (CRD) was used to study the effect of different treatments on physiological and biochemical characteristics, and the significant differences between the mean were compared by Duncan's test [22] and the ready-made statistical program (SPSS 17) was used.

Results and Discussion
The results shown in Table (2) indicate that there are significant differences for the studied traits (the number of red blood cells(RBC), the packed cell volume(PCV), the hemoglobin percentage(Hb), the number of white blood cells(WBC), and the number of white lymphocytes, between the experimental treatments, which include (a mixture of medicinal herbs with different concentrations) 2%, 4% and 6% compared with the control treatment, where the third treatment( T3) showed a significant superiority over all treatments (T2,T1) compared to the control diet (CONTROL) and these results agreed with the addition of red pepper powder with what was reached [23].Which showed that the addition of red pepper powder caused a significant increase in the physiological characteristics of the blood under the influence of heat stress, and the significant increase of the experimental treatments compared with the control treatment may be attributed to the role of the mixture of medicinal herbs in increasing the metabolism of nutrients and optimizing them, as well as the role of each of licorice , thyme, turmeric and red pepper as anti-oxidant agents because they contain volatile phenolic compounds such as flavonoids and their ability to reduce the concentration of malondialdehyde (MAD) in cells by eliminating free radicals that cause the emergence of oxidative stress in cells (ROS), which leads to an increase in the number of red blood cells and packed cell volume and reduce the process of cell death apoptosis [1,24] While the third treatment and the second treatment (T2,T3) showed the least significant differences in the percentage of white monocytes and neutrophils compared with the first treatment and the control (CONTROL,T1).The reason for this may be due to the role of the licorice plant because it contains many compounds and substances effective chemicals, vitamins and mineral elements, and among those active compounds, including the so-called Glycyrrhizin, which is one of the saponins compounds [7], which decomposed inside the body to Glycyrrhizinic acid, which is similar to cortisone in its effectiveness against inflammation and against factors different stresses due to the similarity of their structural structure [8],or the reason may be due to the role of flavonoid compounds and their isoflavones similar to them (Flavonoids and Isoflavonoids) found in the mixture of medicinal herbs (licorice, thyme, turmeric and red pepper) and phenolic compounds [9].Which has an important role against some diseases and prolongs the effectiveness of vitamin C and acts as an antioxidant agent against oxidative stress factors [25] by eliminating free radicals and inhibiting the oxidation of lipids in the membranes of different cells of the body [10,26].
The results shown in Table (3) showed that there were significant (p<0.05)differences between the three treatments (T3,T2,T1).Herbs mixture, 2% herbal mixture, 4% herbal mixture 6%, respectively compared with control treatment free of the herbal mixture 0% in the percentage of glucose concentration, where the both treatment (CONTROL) and the first (T1) (herbal mixture 2%) showed the highest significant difference, which amounted to (297.00) and (304.50)mg / 100 ml plasma, respectively.While the third treatment (herbs mixture 6%) (T3) recorded the least significant differences, which amounted to (230.50) mg / 100 ml plasma.
The reason for the decrease may be attributed to the active ingredients in these extracts, including alkaloids, saponins and tannins, which in turn lead to stimulating the peripheral use of sugar by adipose and muscle tissue, directly or indirectly, by increasing insulin sensitivity with a simultaneous decrease in the process of building glucose [27,28].Or, this may be attributed to the fact that these active substances stimulate beta cells in the pancreas, as well as enhancing or stimulating the secretion of a greater amount of insulin in response to the rise in glucose in the blood and increasing the representation of glucose through the glycolysis pathway, as well as increasing the entry of glucose into cells through increased transporters of glucose on the plasma membrane [29,30].In addition, the reason for the decrease in the level of glucose may be due to flavonoids, which can play a role effective in curbing the process of glucose formation in the liver as well as inhibiting the process of formation of glucose from non-carbohydrate sources (Gluconeogenesis) by inhibiting the enzyme glucose-6-phosphate in the liver [31].Also, the third treatment (T3) (the mixture of medicinal herbs 6%) showed the least significant difference in cholesterol concentration, which amounted to (180.60) compared to the other treatment, the control (CONTROL), which amounted to (309.05).The reason for lowering the cholesterol level is attributed to the fact that these extracts contain flavonoids.Which acts as an antioxidant, which is characterized by its ability to reduce cholesterol level and promote its metabolism [32,33].Or, the reason may be due to the increase in the effectiveness of the 7-α-hydroxylase enzyme responsible for converting cholesterol into bile acids.In addition, some research has indicated the possibility of the active ingredients to inhibit the enzyme Hydroxymethyl Glutary Reductase responsible for the synthesis of cholesterol [34].While there were no significant differences in the concentration of total protein between the three treatments (T1,T2,T3).Herbal mixture 2% , herbal mixture 4%, herbal mixture 6%, respectively compared with the control treatment (CONTROL) free of mixture herbs 0% , and this result agreed with the findings of both researchers [35,36], they did not show any significant differences in the concentration of total protein when adding turmeric powder to the diet of broilers and Japanese quail on this result also agreed with [37] when he using licorice plant added to the diet of female quail, while this study did not agree with what was reached by [38] who noticed a significant increase in total protein in blood serum when adding licorice powder in drinking water.Perhaps the reason for maintaining the level of total protein in the blood serum is the low level of malondialdehyde (MDA) and high glutathione in the blood serum, which give an indication of the inhibition of protein catabolism by the process of gluconeogenesis, because this mixture of medicinal herbs, including licorice plant, contains: many flavonoid and phenolic antioxidant compounds that reduce the secretion of corticosterone and thus maintain the protein level [39].

Conclusions
In the our current study, it is accomplished that the addition of the mixture of medicine herbs to the quail diet at a rate ranging between 4% to 6% led to an improvement in some physiological and biochemical characteristics.

Table 1 .
Ingredient of finisher diet-fed birds of the study (according to NRC 1994).

Table 2 .
Mean± standard error of mean (SEM) of some physiological traits of the birds fed on rations of different levels of (mixture of herbs).
a,b,c: values bearing different superscripts within same row are significantly different (p < 0.05).

Table 3 .
Mean± standard error of mean (SEM) of some biochemical traits of the birds fed on rations of different levels of (mixture of herbs).,c,d: values bearing different superscripts within same row are significantly different (p < 0.05).