Revista Cienﬁca, FCV-LUZ / Vol. XXXV Recibido:25/11/2024 Aceptado:01/02/2025 Publicado: 15/04/2025 hps://doi.org/10.52973/rcfcv-e35578 UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico 1 of 7 of Laurel (Laurus nobilis L.) Esse Bioefficacy Lipopolysaccharide-Induced Inﬂammaon in Japanese Quails Bioeficacia del aceite esencial de laurel (Laurus nobilis L.) sobre la inflamación inducida por lipopolisacáridos en codornices japonesas ¹Hatay Mustafa Kemal University, Faculty of Veterinary, Department of Biochemistry, 31040, Antakya, Turkey. ²Hatay Mustafa Kemal University, Faculty of Veterinary, Zootechnical department, 31040, Antakya, Turkey. ³Necmen Erbakan University, Faculty of Veterinary, Department of Pathology, 42310, Konya, TURKEY. ⁴Fırat University, School of Medicine, Department of Biostascs and Medical Informacs,31040, Elazığ, TURKEY. ⁵Hatay Mustafa Kemal University, Faculty of Veterinary, Department of Animal Nutrion, 31040, Antakya, Turkey. *Correspondence Author: mmisgor@gmail.com ABSTRACT The potenal use of natural plants and essenal oils as feed addives in poultry farming is a popular research area. In the presented study, the eﬀects of laurel essenal oil on growth and feed conversion performances, slaughter-carcass characteriscs oxidave stress index and histopathological parameters in Japanese quails in LPS-induced inﬂammaon model were invesgated. The study groups were established as Control (C), laurel essenal oil (LEO) group, lipopolysaccharide (LPS) group, laurel essenal oil together with lipopolysaccharide (LEO+LPS) group. The live weights on the 19th, 26th and 33rd days were higher in the LEO+LPS and in the LEO group on the 36th day. Feed ulizaon was the worst in the LPS and the best in the LEO group. Non-eviscerated carcass yield was lower in the LPS group compared to the control, and the gizzard weight was higher in the LEO, LEO+LPS and LPS groups. The OSI value was observed as highest in the LPS group and the lowest in the LEO group. In addion, OSI value in the LEO group reduced importantly comparing with LEO+LPS group. Liver OSI values did not show any signiﬁcant diﬀerence in all groups. Histopathologically, no signiﬁcant diﬀerence was observed, in terms of fay liver, congeson, degeneraon necrosis and cell inﬁltraon. The addion of laurel increased degeneraon, necrosis and desquamaon and cell inﬁltraon in the lamina epithelialis in the intesne. In the intesnes, cell inﬁltraon was signiﬁcantly increased in the LPS group compared to the control group. In addion, eosinophilic accumulaons were detected in the brain in the LPS group. As a result, it was determined that laurel essenal oil improved live weight and feed conversion rate, made a signiﬁcant contribuon to balancing the oxidant-anoxidant capacity rao, and showed signiﬁcant bioacvity, especially in terms of turning the negave eﬀect of LPS into posive. Key words: Laurel; inﬂammaon; growth performance; poultry feed; quails RESUMEN El uso potencial de plantas naturales y aceites esenciales como adivos alimentarios en la avicultura es un área de invesgación popular. En el estudio presentado, se invesgaron los efectos del aceite esencial de laurel en el crecimiento y el rendimiento de conversión alimencia, las caracteríscas de la carcasa de sacriﬁcio, el índice de estrés oxidavo y los parámetros histopatológicos en codornices japonesas en el modelo de inﬂamación inducida por LPS. Los grupos de estudio se establecieron como Control (C), grupo de aceite esencial de laurel (LEO), grupo de lipopolisacárido (LPS), grupo de aceite esencial de laurel junto con lipopolisacárido (LEO + LPS). Los pesos vivos en los días 19, 26 y 33 fueron mayores en el grupo LEO + LPS y en el grupo LEO en el día 36. La ulización del alimento fue la peor en el grupo LPS y la mejor en el grupo LEO. El rendimiento de la carcasa no eviscerada fue menor en el grupo LPS en comparación con el control, y el peso de la molleja fue mayor en los grupos LEO, LEO + LPS y LPS. El valor de OSI se observó como más alto en el grupo LPS y el más bajo en el grupo LEO. Además, el valor de OSI en el grupo LEO se redujo de manera importante en comparación con el grupo LEO + LPS. Los valores de OSI del hígado no mostraron ninguna diferencia signiﬁcava en todos los grupos. Histopatológicamente, no se observó ninguna diferencia signiﬁcava en términos de hígado graso, congesón, degeneración, necrosis e inﬁltración celular. La adición de laurel aumentó la degeneración, la necrosis y la descamación y la inﬁltración celular en la lámina epitelial en el intesno. En los intesnos, la inﬁltración celular aumentó signiﬁcavamente en el grupo LPS en comparación con el grupo de control. Además, se detectaron acumulaciones eosinolicas en el cerebro en el grupo LPS. Como resultado, se determinó que el aceite esencial de laurel mejoró el peso vivo y la tasa de conversión alimencia, hizo una contribución signiﬁcava para equilibrar la relación de capacidad oxidante-anoxidante y mostró una bioacvidad signiﬁcava, especialmente en términos de converr el efecto negavo de LPS en posivo. Palabras clave: Laurel; inﬂamación; rendimiento del crecimiento; alimento para aves de corral; codornices ntials Oil on Mehmet Mustafa Işgör 1* , Sema Alaşahan 2 , Altuğ KüÇükgül 1 , Özgür Kanat 3 , Betül Dağoğlu Hark 4 , Süleyman Ercüment Önel 5

Laurel essential oil inflammation Japanese quails / Işgör et al. UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico INTRODUCTION The producon principle of livestock enterprises is generally based on gaining live weight quickly with less feed in a short me. Therefore, feed addives are used to improve the feed conversion rate and the quality of animal products, to ensure the sustainability of animal health and to reduce product costs. Recent studies are carried out to detect the eﬀects of herbal seeds, leaves and oils added to the feed at diﬀerent rates as feed addives on growth and feed evaluaon performances in poultry and on some blood and ssue characteriscs [1 , 2]. The development of resistance as a result of the widespread use of anbiocs in poultry farming has led researchers to ﬁnd alternave soluons. Considering the negave eﬀects of anbiocs, the use of organic acid and essenal oil mixtures obtained from natural sources in poultry, alone or in combinaon, has gained popularity [3 , 4]. For example, it was determined that the feed conversion rao of broiler chickens in the groups containing probiocs, organic acids and essenal oil mixtures including thyme oil, laurel oil, sage oil, myrtle oil, fennel oil and citrus oil was signiﬁcantly beer than the control and organic acid groups [5]. It is reported that the laurel plant, which is in the maquis ﬂora (Aristotelia chilensis), has a long lifespan, is approximately two meters tall, and has dense branches. The fragrant leaves of this plant, which grows widely in countries with a Mediterranean climate, including Türkiye, are rich in bioacve components and are known to have signiﬁcant an-inﬂammatory potenal [6]. In addion, it has been reported that the laurel plant, which is a plant of the Lauraceae family, has anmicrobial and anoxidant eﬀects. The dominant components of the volale oil obtained from the laurel plant are expressed as 1.8-cineole, α-terpinene, and sabinene [7]. It is stated that the addion of sage (Salvia triloba L.) and laurel (Laurus nobilis L.) oil to quail diets numerically changes live weight, feed intake, feed ulizaon, and carcass characteriscs [8 , 9]. Lipopolysaccharides (LPS) are molecules with endotoxin properes located in the outer membrane of gram-negave bacteria [10]. It is possible to experimentally create a bacterial infecon model in chickens using LPS, a powerful inﬂammatory agent [11]. It has been stated that LPS applicaon caused decrease in body weight gain and feed conversion eﬃciency in chickens and protecon against this negave eﬀect will be provided by oil diets [11]. It has also been stated that LPS smulates the expression of inﬂammatory cytokines in chickens and that diﬀerent oil diets have an ameliorang eﬀect on this situaon. Therefore, regulang the immune system in poultry through diﬀerent diets aims to alleviate the decreasing performance of the animals [12]. In this context, the purpose of this study was to determine the eﬀects of adding essenal oil obtained from laurel leaf as a natural feed addive to the feed and growth performances, slaughter-carcass characteriscs, some blood parameters, liver and intesnal histopathological parameters in quails with Lipopolysaccharide (LPS)-induced inﬂammaon. MATERIAL AND METHODS Animal groups Ethical permission was obtained from Hatay Mustafa Kemal University Animal Experiments Local Ethics Commiee to conduct this study (Approval no: 2022/07-04). In the study, 40 quail chicks (Coturnix coturnix) at 5 d of age were used. The study consisted of four groups and the animals were divided into groups such that the inial live weights (Ohaus NV622, USA) of the quails were weighed such that there were no signiﬁcant diﬀerences in average inial live weights among the groups. Animals were divided into 4 groups as follows; Control (C, commercial chick starter feed only), LEO group (commercial chick starter feed + 200 mg/kg laurel essenal oil), LEO+LPS group (commercial chick starter feed + 200 mg/kg laurel essenal oil in feed + 0.25 mL Lipopolysaccharide in water), LPS group (commercial chick starter feed + 0.2 mL lipopolysaccharide in water). The study connued unl the animals reached 36 d of age. Feed and water were given to the animals ad libitum. Faening performance A total of 40 quails in 4 groups were individually weighed each week to determine their live weights. Aﬅer determining the last live weight weighing at 36 d, and then carcass characteriscs were determined by slaughtering animals in each group. Feed intake and the feed conversion rate The amount of feed given to quails in all groups at the beginning of the week and remaining at the end were weighed to determine weekly feed consumpon. In addion, feed conversion rate was calculated using live weight gain and feed consumpon amounts. Carcass characteriscs A total of 40 (4x10) quails of mixed sex (female-male) from each group were slaughtered aﬅer cervical dislocaon. Carcass characteriscs were determined as slaughter live weight, no eviscerated hot carcass weight, eviscerated hot carcass weight, thigh weight, breast+back+neck+wing weight, liver weight, heart weight, gizzard weight and abdominal fat weight. Biochemical analysis Total anoxidant status (TAS) and total oxidant status (TOS) from blood (serum) and liver (homogenate) samples taken from quails were analyzed according to the method of Erel [13] by ready-made commercial kits (Rel Assay, Turkey). The ﬁndings obtained by using TOS (µmol H 2 O 2 Eq/L)/TAS (mmol Trolox Eq/L) ×100 formulaon was given as oxidave stress index (OSI) values. Histopathological examinaon Following the necropsies of the liver, intesne and brain ssues of the quails, samples were taken and ﬁxed in 10% formalin soluon. They were rounely dehydrated in alcohol series and polished in xylene, and paraﬃn blocks were obtained. 5 μm thick secons were taken from the ssues using a microtome (Leica RM2235, Germany). Tissue samples were 2 of 7

Revista Cienﬁca, FCV-LUZ / Vol. XXXV UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico stained with Hematoxylin-Eosin [14]. They were examined with the help of a light microscope (Olympus BX50-F4, Tokyo, Japan) and photographed by using an imaging system (Olympus DP12- BSW, Tokyo, Japan). In histopathological examinaon of HE-stained secons, liver congeson, necrosis, mononuclear cells, heterophil granulocyte inﬁltraon, and lipidosis were evaluated. In the evaluaon of histopathological ﬁndings in the liver, liver secons were photographed at 200x magniﬁcaon to detect inﬂammatory foci. The number of inﬂammatory foci was determined by taking the average of inﬂammatory foci detected in 10 diﬀerent areas of each liver secon [15]. Lesions in the liver were scored as described by Timbermont et al. [16]: no lesion (0), mild lesion (1), moderate lesion (2), and severe lesion (3). The severity of the degree of hepatocellular vacuolizaon, characterized by small or large droplets of lipidosis in the liver, was evaluated by Tro et al. [17] as deﬁned, no fat vacuoles (0), lipidosis with small and large drops containing less than half of the hepatocytes (1), lipidosis containing more than half of the hepatocytes (2) and diﬀuse lipidosis containing small and large drops of fat vacuoles (3). Lesions such as hyperplasia, mononuclear cell inﬁltraon and degeneraon, necrosis and desquamaon in the lamina epithelialis in the small intesnes were scored and evaluated according to their presence and severity, as done by Kanat and Ortatatlı [18]. In each secon, 10 diﬀerent areas were selected at 200x magniﬁcaon and the cells in these areas were counted and their averages were calculated. The lesion was graded as no (0), mild lesion (1), moderate lesion (2) and severe lesion (3). Stascal analysis IBM SPSS Stascs 22 package program was used to perform stascal analyses of the data obtained from the study. In the study, Comparison of groups in terms of faening performance and blood parameters was done by One-way Anova and the diﬀerences were determined by the Duncan test. In addion, the Kruskal-Wallis test was used to compare the groups for histopathological features related to the liver and intesne, and the Mann Whitney U Test was used to determine the diﬀerent groups. RESULTS AND DISCUSSION According to the data obtained in the study, a numerical diﬀerence (P>0.05) was found when the control group was compared with the other groups in terms of weekly live weight throughout the study. The live weights on the 19 th , 26 th and 33 rd d were found to be higher in the LEO+LPS group and on the 36 th day in the LEO group (P>0.05) (TABLE I). Also, weekly feed consumpon was highest between the days 5-12 in the control grup (P<0.001), between the days 12-19 and the d 26-33 in the LEO+LPS group (P<0.001) and between the days 19-26 in the LPS group. Feed ulizaon was the worst in the LPS group (P<0.001) and the best in the LEO group (P<0.001). Accordingly, the addion of laurel essenal oil to quail feed and LPS to its water did not aﬀect the live weight of quails compared to the control group. However, in the laurel added groups, numerical diﬀerences were detected between the groups in terms of live weight and live weight gains (TABLE I). The live weight of quails improved numerically in the LEO and LEO+LPS groups compared to the LPS group. In other words, in the LEO+LPS group, laurel supplementaon to the feed had a posive eﬀect on live weight and live weight gain. Similarly, there are studies reporng that the addion of diﬀerent products of the laurel plant to feed has a posive eﬀect on growth promoon [19 , 20]. It is also stated that laurel (Laurus nobilis L.) leaves are used to increase gastric ﬂuid to stop digesve disorders in poultry [21]. TABLE I. Weekly live weight and producon performance of quails in the groups Days Control LEO LEO+LPS LPS SEM P Weekly live weight (g) Start (5 days) 30.70 30.18 30.77 30.08 0.746 0.982 12 74.99 72.80 74.63 70.69 1.353 0.664 19 125.29 122.87 127.08 122.47 2.017 0.835 26 171.45 170.42 171.99 169.56 2.457 0.986 33 196.47 198.73 203.12 196.48 3.546 0.899 36 201.52 217.17 212.34 208.87 3.767 0.522 Live weight gain (g) 5-12 44.29 42.63 43.86 40.61 1.386 0.786 12-19 50.30 50.06 52.44 51.79 2.049 0.971 19-26 46.16 47.56 44.92 47.09 3.167 0.992 26-33 25.02 28.31 31.13 26.92 3.401 0.934 Feed Consumpon (g) 5-12 98.00a 96.00b 96.00b 94.00c 0.004 0.001 12-19 136.00c 128.00d 152.00a 140.00b 0.005 0.001 19-26 166.00d 178.00c 180.00b 194.00a 0.005 0.001 26-33 196.00c 188.00d 224.00a 218.00b 0.005 0.001 Feed conversion rao, g/g 5-12 2.21c 2.25b 2.18d 2.31a 0.000 0.001 12-19 2.70b 2.56c 2.90a 2.70b 0.000 0.001 19-26 3.60d 3.74c 4.01b 4.12a 0.000 0.001 26-33 7.84b 6.64d 7.20c 8.10a 0.001 0.001 LEO: laurel essenal oil, LPS: lipopolysaccharide and LEO+LPS: laurel essenal oil+lipopoly- saccharide. a,b,c,d : Groups with diﬀerent leers in the same column are diﬀerent from each other (P<0.05) The ﬁndings of this study suggest that although the addion of laurel essenal oil to quail feed did not signiﬁcantly increase the live weight of quails compared to the control group, it is correct to state that there was a tendency for the live weight to be improved with the addion of laurel essenal oil to the feed of quails, especially those to which LPS was added to their water. In the study, it was determined that the feed consumpon and feed conversion rates of quails in the groups where laurel essenal oil was added to the feed were signiﬁcantly diﬀerent from the control group, and the feed conversion rate was improved in the groups laurel essenal oil added to the feed. While feed consumpon in the LEO group decreased compared to the LEO+LPS and LPS groups, the feed conversion rate improved only in the LEO and LEO+LPS groups (TABLE I). Therefore, the 3 of 7

Laurel essential oil inflammation Japanese quails / Işgör et al. UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico results show that the addion of laurel essenal oil to the feed is important to improve growth performance, especially in stress groups in terms of turning the negave eﬀect into a posive one. In similar studies, it has been reported that laurel essenal oil reduces feed consumpon and increases feed ulizaon [22 , 23]. In studies by Çabuk et al. [24] and Namdeo et al. [25], it was reported that broiler chickens fed with laurel leaf, myrtle leaf, thyme, sage leaf, citrus peel and fennel seed essenal oils showed signiﬁcant improvements in feed conversion rao. It was determined that the non-eviscerated carcass yield was lower in the LPS group than in the control group (P<0.05), and the gizzard weight was higher in the LEO, LEO+LPS and LPS groups than in the control group (P<0.01) (TABLE II). TABLE II. Slaughter, carcass and internal organ characteriscs Characteriscs Control LEO LEO+LPS LPS SEM P Slaughter weight (g) 200.45 218.24 208.86 200.04 4.133 0.370 Non-eviscerated carcass weight (g) 150.38 165.32 156.40 148.93 3.093 0.244 Non-eviscerated carcass yield (%) 75.04 ab 75.76a 74.87 ab 74.47 b 0.159 0.049 Breast+back+neck+wing weight (g) 77.66 82.24 77.07 73.70 1.369 0.196 Thigh weight (g) 45.36 46.11 44.36 43.49 0.887 0.743 Heart weight (g) 1.92 1.93 1.80 1.83 0.049 0.715 Liver weight (g) 4.56 6.61 5.72 5.46 0.293 0.122 Gizzard weight (g) 4.34 b 5.14 a 5.52 a 5.47 a 0.129 0.009 Abdominal fat weight (g) 2.37 3.10 2.33 2.09 0.190 0.284 LEO: laurel essenal oil, LPS: lipopolysaccharide and LEO+LPS: laurel essenal oil+lipopolysaccharide. a, b : Groups with diﬀerent leers in the same row are diﬀerent from each other (P<0.05) Addion of laurel essenal oil to the feed did not aﬀect slaughter weight, non-eviscerated carcass weight, non- eviscerated carcass yield, breast+back+neck+wing weight, thigh weight, heart weight, liver weight and abdominal fat weight compared to the control group. However, in the LPS group, non- eviscerated carcass yield was found to be the lowest. In terms of gizzard weight, laurel essenal oil and LPS supplemented groups were signiﬁcantly higher than the control group (TABLE II). Similar to the results of this study, Al-Rubaee [19] reported a posive eﬀect of adding laurel leaf ﬂour to the feed on carcass weight and yield in quails. However, the high gizzard weight determined as a result of the study is not consistent with the statement of Gwaad and Gwaad [26] that there is no diﬀerence in terms of liver and gizzard weight. According to the ﬁndings, it was determined that the blood OSI value increased signiﬁcantly in the LPS group compared to the control group. In the LEO group including laurel essenal oil only this value decreased signiﬁcantly compared to the control group (TABLE III). It was also found that laurel essenal oil added to the feed in the LEO+LPS group caused the OSI value to decrease by 27% compared to the LPS group (p<0.01). However, no stascally signiﬁcant diﬀerence was detected between the control group and the applicaon groups in terms of liver OSI value (P>0.05). TABLE III. Blood and liver oxidave stress index values of quails in the study groups Characteriscs Control LEO LEO+LPS LPS SEM P Blood OSI value 4.45bc 2.54c 7.34ab 10.08a 0.732 0.009 Liver OSI value 11.02 9.02 9.17 9.32 0.311 0.120 LEO: laurel essenal oil, LPS: lipopolysaccharide and LEO+LPS: laurel essenal oil+lipopoly- saccharide, OSI: oxidave stress index Oxidave stress increases as a result of increased producon of free radicals in the body or inadequate eliminaon due to disrupon of the oxidant-anoxidant balance. Oxidave stress levels of poultry can be negavely aﬀected by various factors such as heat, feed changes, and drug use [27]. Therefore, it has been reported that the addion of supplementary nutrients, especially natural plants and essenal oils obtained from them to the feed has signiﬁcant eﬀects in prevenng the negave eﬀects of stress factors [28 , 29]. In the study conducted by D’Alessandro et al. [30], it was reported that although the mixture of olive, laurel and rosemary leaf powders did not aﬀect body weight and egg quality, it had posive eﬀects on oxidave stress and inﬂammaon parameters. In another study, it was reported that the addion of sage and bay leaf oils to quail diets increased serum anoxidant capacity, and that laurel oil had a signiﬁcant anoxidant eﬀect in reducing oxidave stress [31]. In addion, it has been reported that the addion of laurel leaves to the diets of Japanese quails has an eﬀect on triglyceride, palmic and oleic acid levels in quail eggs without adversely aﬀecng performance and egg quality [32]. Oxidave stress index is one of the main parameters used to determine the level of oxidave stress that develops due to the increase in free radicals or the decrease in anoxidant capacity. In this study, it was observed that OSI values were reduced in the group to which laurel oil was added compared to the control group. In addion, it was determined that OSI values were signiﬁcantly reduced in the group to which LPS was given together with laurel compared to the group to which only LPS was applied. In conclusion, it was observed that the addion of laurel essenal oils to the diets of Japanese quails showed a signiﬁcant anoxidant acvity in maintaining or improving the anoxidant-oxidant balance. In some animals in the control, LPS, LEO and LEO+LPS groups, the livers were observed to be macroscopically enlarged to varying degrees, pale yellowish white in color and friable in consistency. Gas accumulaon was observed in the small intesnes of a few quails. No pathological changes were 4 of 7

Revista Cienﬁca, FCV-LUZ / Vol. XXXV UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico observed in the brain. Histopathological examinaon of the liver showed dissociaon, enlargement of sinusoids, congeson, varying degrees of small-drop and large-drop fat vacuoles in hepatocytes, hydropic degeneraon, necrosis, mononuclear cell and heterophil granulocyte inﬁltraons. It was determined that hydropic degeneraon, necrosis and cell inﬁltraons were more intense around the vena centralis and portal area (FIG. 1A-B). Hyperplasia, especially in the epithelium, degeneraon, necrosis and desquamaon in the lamina epithelialis, mononuclear cell inﬁltraons in the propria and a small number of heterophil granulocytes were observed in the intesnes (FIG. 1C). Histopathologically, edema, hyperemia, and a pinkish-red accumulaon in the neuropil ssue were detected in the brains of 2 quails in the LPS group (FIG. 1D). Congeson, degeneraon and necrosis, lipidosis, cell inﬁltraons in the liver, hyperplasia, cell inﬁltraons in the small intesnes, degeneraon, necrosis and desquamaon in the lamina epithelialis and level scores of the quails in the experimental groups are stascally indicated in TABLE IV. FIGURE 1. Histopathological analysis of liver, intesne and brain ssues of quails. A) Con- geson, small and large droplets of lipidosis in hepatocytes, liver. B) 1. Hydropic degenera- on and necrosis (star) in hepatocytes, liver. C) Degeneraon, necrosis and desquamaon in lamina epithelialis, intesne. D) Accumulaon of eosinophilic structure (arrows) brain. HE staining. bar= 100 µm TABLE IV. Descripve stascs and p value for liver and intesnal characteriscs Tissue Lesions Control LEO LEO+LPS LPS P value Liver Congeson 1.00(0-2) 1.00(1-3) 1.00(0-2) 1.00(1-2) 0.465 Degeneraon and necrosis 1.00(0-2) 0.50(0-2) 0.00(0-2) 0.00(0-2) 0.288 Lipidosis 1.00(0-3) 2.00(0-3) 1.00(0-2) 2.00(1-3) 0.135 Cell inﬁltraon 0.00(0-2) 1.00(0-2) 1.00(0-1) 1.00(0-3) 0.376 İntestinal Hyperplasia 2.00(2-2) 2.00(1-3) 2.00(2-3) 2.00(1-3) 0.287 Degeneraon and necrosis in the lamina epithelialis 0.00(0-1) 1.00(0-1) 1.00(0-1) 0.00(0-1) 0.041 Cell inﬁltraon 0.00(0-2) 1.00(0-1) 1.00(0-1) 1.00(0-2) 0.016 LEO: laurel essenal oil, LPS: lipopolysaccharide and LEO+LPS: laurel essenal oil+lipopolysaccharide The diﬀerence in terms of liver congeson, necrosis, lipidosis level and cell inﬁltraons in the control, LEO, LEO+LPS and LPS groups is not stascally signiﬁcant (P>0.05). When intesnal hyperplasia was examined, the diﬀerence between the groups was not stascally signiﬁcant (P>0.05), while the degeneraon, necrosis and desquamaon levels and cell inﬁltraon in the lamina epithelialis between the groups were signiﬁcant (P<0.05). According to the mulple comparisons, the diﬀerence for the LPS and LEO+LPS groups was found to be stascally signiﬁcant (P<0.05). This signiﬁcance is in the direcon of the LPS group being lower than the LEO+LPS group. Other pairwise group comparisons were not found to be signiﬁcant. In the mulple comparison for intesnal cell inﬁltraon, the diﬀerence between the control group (0.00(0-2)) and the LPS group (1.00(0-2)) was stascally signiﬁcant (P<0.05). Other mulple comparisons were not found to be signiﬁcant. In this study, despite the nutrional diﬀerences between the groups, no signiﬁcant diﬀerence was observed histopathologically in terms of the level of lipidosis, congeson, degeneraon, necrosis and cell inﬁltraon in the liver. It was determined that the addion of laurel increased degeneraon, necrosis and desquamaon and cell inﬁltraon in the lamina epithelialis in the intesne. Cell inﬁltraon was signiﬁcantly increased in the groups to which LPS was added compared to the control group (TABLE IV). Studies on the pathological lesions caused by laurel in organs are quite limited. The liver plays a central role during metabolism and is an organ that produces bile, which helps in the breakdown of fats. In bird species, fats, especially triglycerides, accumulate in adipocytes and hepatocytes. Hepac lipogenesis causes steatosis in the liver when it exceeds the capacity of lipoprotein secreon. Researchers have also reported that heat stress causes vacuolar steatosis in the liver [33 , 34]. Some researchers have reported that steatosis, degeneraon and necrosis in hepatocytes, hemorrhage and hematomas in the parenchyma, ﬁbrosis and cell inﬁltraon in portal areas occur [17 , 35]. In the presented study, hydropic degeneraon, mulfocal necrosis, steatosis and cell inﬁltraon in hepatocytes were observed, along with hemorrhage in the liver of only one quail in the LEO+LPS group. Hematomas and ﬁbrosis were not observed. As reported by Malik et al. [36], fats are a high energy source and although depleon of fats in cells occurs during periods when growth is very rapid, fats may begin to accumulate in cells when the growth rate of animals slows down with growth. Similar to what Jacobsen et al. [37] reported, it was not possible to determine whether the fay liver observed in this study was due to the fat storage characteriscs of the livers of bird species or due to the laurel in the diet. 5 of 7

Laurel essential oil inflammation Japanese quails / Işgör et al. UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico It has been reported that quails are resistant to infecons and do not have any obvious symptoms. It has been reported that the intesnal content may change from seromucous to dark brown depending on the infecon [38 , 39]. In the present study, although no other ﬁndings other than gas were observed macroscopically in the small intesnes, histopathological, it was observed that cell inﬁltraon in the LPS group was signiﬁcantly increased compared to the control group. Degeneraon, necrosis and desquamaon in the epithelium were signiﬁcantly observed in the LPS and LEO+LPS groups compared to the control group. It is thought that the histopathological lesions observed were caused by exposure to a fay diet with the addion of laurel as well as infecon. No previous arcle has been found regarding the accumulaons of eosinophilic structure in the brain in the LPS group. CONCLUSION In the study, histopathological changes were observed in the liver, intesne and brain ssue between the groups, but it was determined that they were not stascally signiﬁcant. However, the intensity of cell inﬁltraon in the intesnes, especially in the LPS group, is remarkable when compared to the other groups. Again, the presence of eosinophilic accumulaons in the brain ssue is another remarkable ﬁnding. In the light of all these ﬁndings, it can be said that Laurel has anoxidant and faening performance eﬀects in the experimental inﬂammaon model, but its an-inﬂammatory acvity is limited. However, further and detailed studies are needed to fully elucidate the issue. Conﬂict of interests The authors declare that there is no conﬂict of interest with the publicaon of this manuscript. BIBLIOGRPHICS REFERENCES [1] Kazak F, Çimrin T, Alaşahan S. 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