Effect of Raw and Boiled Oak Quercus brantii in Performance, Biochemical and Blood Indices, and Proximate Composition of Cyprinus Carpio L.

This study was conducted in fish diseases of Sulaimani to evaluate the effect of boiling oak as compared with raw oak on performance of common carp at 0.05 significant level for all studied parameters. This study was carried out on C. carpio L. for five treatments/ three replicates. The experimental trial consists of five treatments with three duplicates, each containing seven fish: Group 1: Diet without any supplement as control group, Group 2: adding 10g raw oak/ kg diet, Group 3: adding 20g raw oak/kg diet, Group 4: adding 10g boiled oak/kg diet, Group 5: 20 grams of boiling oak / kg diet. The second group with 10g raw oak/ kg diet and T5 with 20g boiled oak/ kg diet was high significantly in the final weight, weight gain, significant and relative growth rate. T3 with 20g raw oak/ kg diet was higher significantly in FCR, T4 with 10g boiled oak/kg diet was higher significantly in FER. RBC *10 12 /L was higher in T4 with 10g boiled oak/ kg diet and T5 with 20g boiled oak/ kg diet were higher significantly, in HGB g/L the T2 with 10g raw oak/ kg diet and T3 with 20g raw oak/kg diet were higher significantly. T3 with 20g raw oak/kg diet was significant in each of the indices MCH pg and MCHC g/L. T4 with 10g boiled oak/kg diet was higher significantly in each of HCT %. The control was significantly higher in PLT *10 9 /L. T5 with 20g boiled oak/ kg diet was significantly high in Sugar and HDL. Total Protein was significantly high in control and T2 with a 10g raw oak/ kg diet. Cholesterol was significantly high in T2, T4, and T5 as compared to control group. GOT and CKI was significant in T3. A Hepatosomatic index was higher significantly in each of T2, T3 and T5. Gill somatic Index was significant in all treatments as compared to T3. Gonadosomatic Index was significant in all treatments as compared to T2 and T4, in the Intestine Weight Index and Intestine Length Index, all treatments were significantly different from the control. In the Intestine Length (Fish Length) Index, T2 was substantially greater. The current study's findings imply that oak acorn powder is a viable feed additive, indicating a starting point for additional research into its potential use as an antioxidant and immune booster in various fish species. This article


Introduction
Aquaculture has evolved rapidly during the last few decades  at an average yearly rate of 8.8% to meet the world's high needs, resulting in intensive and super-intense cultures (1). As a result, a wide range of disorders appeared, and stresses such as overcrowding, transportation, handling, grading, and poor water quality all played a role in the disease epidemic in these cultures (2). Various techniques, such as antibiotics, chemotherapeutics, and vaccinations, have been used to combat these issues. Antibiotics and chemotherapeutics are known to cause resistance in infections and to be damaging to humans and the environment. Vaccines are not a viable solution since they are costly and designed for certain infections (3). The Valonia oak (Quercus aegilops L) is the common native species in the Iraqi Kurdistan region, which is a medium-sized deciduous tree (4). Herbal extracts, which are a type of immunostimulant, have been used to boost the immune system and have been widely utilized to improve defective immunological functioning (5,6,7). This species' extracts contain antibacterial and wound-healing properties, while its tannins have contractive and disinfecting properties (8,9,10), Oak is also utilized as animal feed in various places of Iran due to its low cost and availability. External and interior layers cover oak fruit, with the internal layer known as jaft in the area. Some microorganisms are inhibited by ethanolic and methanolic extracts of Persian oak fruits at varied doses. (10). Quercus brantii, The Brant's oak is a native of Western Asia, including Kurdistan, Iran, Iraq, Syria, and Turkey. The most significant tree species in the Zagros in Iran is Quercus brantii, which covers more than half of the Zagros Forest area. Traditional Iranian medicine makes use of their seed. Fuelwood, charcoal, and timber hardwood are some of the other essential products made from oaks. Though the exact nutrient profile depends on the species of acorn, all are packed with essential nutrients.
Acorns are especially high in potassium, iron, vitamins A and E, and several other important minerals. Plus, these nuts are low in calories. Most of their calories come in the form of healthy unsaturated fats. This study was conducted to assess the effect of oak fruit powder affected common carp growth, hematological, and immunological measures.

Experimental fish
On 105 common carp C. carpio L. collected from local fish ponds in Peramagrun/ Sulaimani/ Iraq, the experiment lasted 60 days. The weight of the fish varied (116.61 gm ± 5.8). 30 days of laboratory pre-acclimation and commercial pellet feeding preceded the real feeding experiments.

Experimental system
In this study, fifteen plastic tanks (70 L) were utilized for five treatments, each with three duplicates. Each tank received proper continuous aeration thanks to fifteen small aquarium air pumps, Luckiness 828 (power: 5 watts, air flow: 3.5L/min), and Chinese air compressors, Hailea ACO-318 (power: 45 watts, air flow: 70L/min), Hailea ACO-328 (power: 55 watts, air flow: 82L/min), Resun ACO-010 (power: 200 watts, air flow: 0.135 m Each replicate was stocked with seven fish. The duplicates were set at random to minimize variations between treatments. To eliminate any remaining feeds and excrement from the system, a daily cleaning by siphoning approach is used. The experimental trial consists of five treatments with three duplicates, each containing seven fish: group 1: Diet without any supplement as a control group, group 2: adding 10g raw oak/ kg diet, group 3: adding 20g raw oak/kg diet, group 4: adding 10g boiled oak/kg diet, group 5: 20 gm boiled oak/kg diet.

Diet formulation
Standard components found in Sulaimani city markets are supplemented with raw and cooked oak powder in the experimental meals. To make dough, combine the ingredients. Then, Kenwood Multi-processors use an electrical mincer to pelletize the material. Three days of room temperature drying were utilized, followed by crushing to get fine particles. Using Albumin and Globulin were higher significant in each of control, T2, and T4 when compared with each other, as biomass twice a day at 9:00 a.m. and 2:00 p.m. Bimonthly, the fish in each tank were weighed together. The feeding amounts were then revised based on the new weights. Growth and feed utilization parameters, biological parameters (indices), Proximate composition, Blood picture and Biochemical measurements were studied.

Growth and feed utilization parameters
The fish were weighed every two weeks for all replicates in order to calculate growth performance metrics. Every two weeks, the feed intake of each treatment was monitored and modified according on the biomass gained.
The following formulae were used to determine weight increase and daily weight gain: "Weight gain (gm. /fish) = W2-W1 Where W1: Fish weight (gm.) at the beginning of the experimental period and W2: Fish weight (gm) at the end of the experimental period. Relative growth rate was calculated according to the method described by Brown, (11) as follows: Relative growth rate (RGR %) = Weight gain/Initial weight x 100 = W2-W1/ W1 x 100 Specific growth rate was calculated according to the method described by Uten, (12) as follows: Specific growth rate (SGR) % = (Ln final body weight-Ln initial body weight)/ experimental period) x 100 = ((Ln W2-Ln W1)/ T) x 100 Feed conversion ratio was calculated as follows: Feed conversion ratio (FCR) = Total feed fed (gm.)/ Total wet weight gain (g). Feed efficiency ratio was calculated as previously described by Uten (12)

Blood and biochemicals parameters
At the end of the experiment, three fish were chosen at random from each replication, weighed, and their length measured, before piercing the caudal peduncle to obtain blood samples, which were collected in heparinized plastic vials and preserved under refrigeration. All blood tests were performed by a hematological analyzing instrument of the type ACCENT 200 specialists to veterinarywhich was made in Poland. The enzyme levels were measured according to the instructions that came with the enzymatic kits. Biochemical parameters include, Aspartate aminotransferase activity (AST), Total proteins, Alanine aminotransferase activity (ALT), Globulin (g/dL), Albumin (g/dL).

Statistical analysis
The experiment will be done using SPSS Version 20's one-way analysis of variance (ANOVA) with fully randomized design (CRD) and general linear models (GLM) approach. At a P value of 0.05, Duncan's test will be performed to compare the means of the treatments. Table 1 show the effect of boiling oak on growth performance in which T2 with 10g raw oak/kg diet and T5 with 20g boiled oak/kg diet were high significantly in final weight, weight gain, significant and relative growth rate.

Results and Discussion
T3 with 20g raw oak/kg diet was higher significantly in Feed conversion ratio, T4 with 10g boiled oak/kg diet was higher significantly (p<0.05) in FER, T2 with 10g raw oak/kg diet and T5 with 20g boiled oak/kg diet were high significantly in Protein ER as seen in table (2). Table (3) showed the effect of boiling and raw oak on RBC *10 12 /L in which T4 with 10g boiled oak/kg diet and T5 with 20g boiled oak/ kg diet were higher significantly, in HGB g/L the T2 with 10g raw oak/ kg diet and T3 with 20g raw oak/kg diet were higher significantly. T3 with 20g raw oak/kg diet was significant in each of the indices MCH pg and MCHC g/L. T4 with 10g boiled oak/kg diet was higher significantly in each of HCT % and MCV fL. The control group had a much higher significantly in PLT *10 9 /L. Table 4 showed that T5 with 20g boiled oak/ kg diet was significantly high in Sugar and HDL. Total Protein was significantly high in control and T2 with 10g raw oak/ kg diet. Cholesterol was significantly high in T2, T4 and T5. Triglyceride was significantly high in T4 with 10g boiled oak/ kg diet. LDL was higher in T2 with 10g raw oak/ kg diet as shown in table (4).
GPT was higher significantly in T3 and T4. GOT and CKI were significant in T3. Albumin and Globulin were higher significant in each of control, T2 and T4 as observed in table (5).
Hepatosomatic index was higher significantly in each of T2, T3 and T5. Gillsomatic Index was significant in all treatments as compared to T3. Gonadosomatic Index was significant in all treatments as compared to T2 and T4. As seen in the table (6), there were no significant changes in the Spleen and Kidney Indexes. The effect of boiling on oak quality in terms of chemical composition is seen in Table (8); there are no significant variations in crude protein, crude lipid, or ash. T3, T4, and T5 had much more moisture than the other treatments. No significant differences seen in meat indices in table (9). Abdel-Tawwab et al., (13) found that Fish in low stocking density (LSD) conditions and fed with the dietary oak acorn powder presented the highest final weight (75.1 g) compared to those in the LSD conditions and fed with the control diet (P < 0.05) with the feed utilization parameters. Supplementation with the dietary oak acorn powder has modified hematological parameters related to stocking density stress effects. The dietary oak acorn powder caused a significant increase in RBCs count, Hb, and Hct values in fish of LSD conditions. Boiling the oak substantially increased the performance of common carp in this investigation. Furthermore, owing to the presence of oak in the diet, feed consumption rose as a consequence of increased need for nutrients during the fish's development or as a result of sensory stimulation and better appetite. Oak contains phenolic compounds, vitamins, minerals, essential oils, and aromatic substances, as well as lipids, amino acids, proteins, carbohydrates and different sterols (10,14), all of which have digestive and stimulatory qualities (15,16). Cheynier (17) found a wide range of phenolic chemicals in Quercus acorns, ranging from simple molecules (e.g., phenolic acids) to polyphenols (e.g., stilbenes, flavonoids, and derived polymers), with antioxidant, antibacterial, anti-inflammatory, and anticarcinogenic properties (18,19). These chemicals improved the overall performance and immunity of fish, consequently enhancing the fish's health and productivity (20). Rashidian et al. (21) observed that feeding rainbow trout, O. mykiss, fingerlings (10.46 g) on diets enriched with alcoholic extract of acorn (Q. brantii) significantly boosted growth performance, feed intake, and digestive enzyme activity as levels of acorn extract rose. In contrast to the findings of the current investigation, Bohlouli et al. (22) fed Persian oak (Q. brantii var. persica) acorn extract to rainbow trout fingerlings (6.25 g) during 8 weeks at 0.5, 1.0, and 2.0 g/ kg diet. They discovered that survival and growth parameters did not differ significantly across treatments (P 0.05). The oak (Q. castaneifolia) leaf extract demonstrated no growth-promoting effects in common carp, according to Paray et al. (23) implying that this extract may not activate digestive enzymes and/or gut absorption. They also related the fish's slowed growth to decreased feed intake. These discrepancies might be due to variances in oak species, fish type, size, and rearing circumstances, among other things. Abdel-Tawwab et al., (13) conclude that dietary oak powder supplementation not only promoted common carp development but also mitigated the stress caused by high stocking density, resulting in improved health. This might assist enhance aquaculture productivity while also improving fish resilience to unfavorable culture conditions.

Conclusion
The current study's findings imply that oak acorn powder is a viable feed additive, indicating a starting point for additional research into its potential use as an antioxidant and immune booster in various fish species.      At P < 0.05, means in the same column with different letters are significantly different