Effects of Oregano Aromatic Water (Oreganum onites L.) Supplementation on Diets of Lactating Dairy cows on Milk Production Performance, Metabolic Health and Antioxidative Defense Mechanism

doi:10.21203/rs.3.rs-1533833/v1

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Effects of Oregano Aromatic Water (Oreganum onites L.) Supplementation on Diets of Lactating Dairy cows on Milk Production Performance, Metabolic Health and Antioxidative Defense Mechanism

https://doi.org/10.21203/rs.3.rs-1533833/v1

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This study aimed to validate the effect of oregano aromatic water (OAW) on blood parameters, oxidative stress, and production and composition of milk when used as a feed additive. Thirty lactating Holstein cows (average 570 kg), average 74 days in milk and 2.44 lactation number, were randomly divided into three groups for a period of 60-days. These groups included: (G1) control ration, (G2) control ration + OAW (10 mg/kg of DM), (G3) control ration + OAW (20 mg/kg of DM). Milk production (MP) were unaffected by OAW doses (P>0.05). The supplementation of OAW did not affect the milk composition, and somatic cell count (SCC) (P>0.05). OAW supplementation did not affect blood biochemical values except ketone and glucose. The glucose value increased as the ketone decreased with OAW supplementation (P<0.05). The hematological blood values were not found to be significant. Supplementing OAW resulted in greater (P<0.05) total antioxidant status in G2 and G3 than G1. The current study's findings indicated that OAW supplementation had no benefits for milk yield and milk composition. It also tended to improve Somatic Cell Counts, udder health, and the antioxidative defense mechanism.

Blood metabolites

Holstein cows

Milk composition

Oregano aromatic water

Oxidative stress

Production performance

For many years, antibiotics have been used as growth factors, especially due to their positive effects on the microflora in the digestive system (Ohya and Sato 1983). However, their use was banned in 2006 due to fact that they leave residues as a result of intensive use and cause the formation of bacteria resistant to antibiotics (Anadon 2006). The pressure from consumer organizations on livestock to reduce the use of antibiotics with customers’ awareness has had a great impact on this topic. After the ban of antibiotics, which were known to have very important roles as growth and health promoters in animal husbandry, it has been revealed that plant extracts and their secondary metabolites can be used as natural alternatives of feed additives for high-yielding cows. Moreover, it has attracted the attention of many researchers in dairy cows for their ability to influence various aspects of ruminant animals and reduce environmental pollution (Abdel Hakim et al. 1989; Huang et al. 2018). Studies have observed that plant extracts and their secondary metabolites support the immune system and antioxidative defense mechanisms as well as their antimicrobial activities (Oh et al. 2017; Braun et al. 2019). It has been suggested that the antimicrobial activity of oregano is due to an increase in the permeability of the bacterial cell wall of thymol and carvacrol (Lambert et al. 2001).

The first lactation period after calving is a critical period strongly related to the health and milk production of a cow. This period, when milk production is high, increases the apparent frequency of diseases such as mastitis, metritis, and ketosis which creates metabolic stress in cows due to increased nutrient requirement, high energy requirement, milk synthesis, and secretion. This metabolic stress leads to increase the animal welfare problems and the loss of yield. In this period, diseases and metabolic stress occur (Sharma et al. 2011; Sordillo and Mavangira 2014). The combined effects of altered nutrient metabolism, nonfunctional inflammatory responses, and oxidative stress characterize metabolic stress. These factors lead to increased metabolic stress and deterioration of the health of cows in the first lactation period (Sordillo and Mavangira 2014). In the first lactation, an imbalance between increased reactive oxygen species and antioxidant defense, which is necessary for the reduction of the accumulation of reactive oxygen species, can cause oxidative stress and disease in cows (Sharma et al. 2011). In this period, the need for safe additives is tried to supply with the use of plant-based additives. However, the different effects of essential oils on animals are indicative of the natural variability in the bioactivity and bioavailability of different essential oils (Giannenas et al. 2011). Silva Filho et al. (2017) stated that essential oils tend to reduce the number of somatic cells in milk and may reduce the risk of mastitis and be effective in preserving the properties of milk. It has been reported by Giller et al. (2020) that thyme, eucalyptus and anise essential oil mixtures have no negative effects on the milk yield and composition of cows.

No studies have been found on the effects of oregano aromatic water, which is a by-product that is released when extracting essential oil, on the performance, metabolic health, and antioxidative defense mechanism of lactating cows. Therefore, in this study, it was aimed to test the hypothesis that whether or not oregano aromatic water improved the performance and health of dairy cows as a by-product without a negative impact on these parameters.

Animals and Rations

Thirty lactating Holstein dairy cows with an average 74 days on milk, and 2.44 lactation numbers, and an average liveweight of 570 kg, were used in this study. The power analysis method was used to determine the number of cows. The highest milk yield average was 31.09 kg/d, the lowest milk yield average was 29.75 kg/d, and the standard deviation was 12.85, and it was found that there should be 8 cows in each group for 95% power. However, 10 cows were included in each group to for better statistical analysis of the results. The experiment lasted for 60 days. The cows were divided into three groups (ten cows each) and were distributed randomly in groups and to allow for valid comparison. Each group had the similar access to feed, clean water and sand sleeping beds. Treatments included: G1) Control group, Basal ration G2) Basal ration + OAW (10 mg/kg of DM), G3) Basal ration + OAW (20 mg/kg of DM). OAW was thoroughly mixed with the vitamin-premix and after the basal ration was distributed to the feeder, it was added on basal ration and presented to the cows. The chemical composition of ration used in the study is presented in Table 1. The farm’s management of feeding and rations for animals has not been intervened in. The cows were fed twice a day at 07:00 and 18:30 directly after milking. The residuals were weighed daily. The crude protein of concentrate and roughage were determined using the Kjeldahl method (AOAC 2000, method: 955.04) and ether extract was obtained using the Soxhlet method (AOAC 2000, method: 948.22). Neutral detergent fiber (NDF) was analyzed by using the ANKOM220 Fibre Analyzer (Ankom Technology, Macedon, NY, USA). The metabolic energy (ME) was calculated based on the data given by the Turkish Standards Institution (1991).

Measurements and Sample Collection.

Milk yields of the cows were monitored by automatic milk counters (Te-Ta Tech. Agric. Com., Izmir, Turkey) for two months (06:00 and 18:00 h). Milk samples were collected twice a week and stored at 4^oC until the analysis. Fat % (Method: IDF 141C:2000), protein % (Method: IDF 141C:2000), lactose % (Method: IDF 141C:2000) compositions and the freezing point (Method: ISO 13366-2:2006/IDF 142:2006) and somatic cell count (ISO 13366-2/AC:2008) of milk were analyzed by using a Bentley B150 milk analyzer (Bentley Combi 150, Bentley Instruments, Inc. Minnesota, Chaska, USA). Analytical portions were determined by the temperature and pressure controlled START D-Microwave digestion system (Milestone Instruments, Sorisole, Italy). The microwave digestion system can reach a maximum power of 1450 W, which can provide a controlled temperature and can operate up to 200 ^oC and 45 bars. Pure water was supplied from Sartorius Arium Ultra-pure water purifier (Sartorius Weighing Tech. GmbH, Goettingen, Germany).

Blood samples were collected from the jugular veins of cows at the end of the study. Biochemical blood parameters were analyzed by Mindray BS120 (Mindray Corporation, Nanshan, China). Blood samples were placed into non-additive tubes for biochemical parameters (Ketone, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP), Gamma-glutamyltransferase(GGT), Lactate dehydrogenase (LDH), Glucose (GLU), Creatinine (CREA), Urea, Total Bilirubin (TBIL), Direct Bilirubin (DBIL), Total Protein (TP)) was transferred to laboratory on ice. Blood samples were centrifuged at 3000 rpm per minute for 10 min. Hematologic parameters were analyzed by ABACUS 38 (Diatron Corporation, Budapest, Hungary) and blood samples were placed into EDTA tubes to measure hematologic parameters.

Oregano Aromatic Water Production

Oregano leaves (500 g) dried in room conditions were placed in a bottle (5L) connected to the concentration of a Clevenger hydrodistillation device with 2.5 liters of water (European Pharmacopoeia 1975). After hydrodistillation, aromatic water was collected by separating it from oregano oil. The essential oil in aromatic water was extracted with n-hexane to determine the odor components using GC/MS (Gas chromatography/Mass spectrometry) device (QP Shimadzu 2010 Plus, Shimadzu Corp. Kyoto, Japan). The identification of the components was carried out using the data given Wiley, NIST and Tutor libraries for retention times of standard substances according to the composition of mass spectra.

Statistical Analysis

The results were analyzed with one-way ANOVA using Minitab Statistical Software Package (Version 17; Minitab Ltd., Coventry, UK) and a Tukey test was used to separate the treatment means.

The mathematical model for one-way ANOVA was given as;

$${Y}_{ij}=\mu + {T}_{i}+ {\epsilon }_{ij}$$

Where, ${Y}_{ij}$ is the j-th observation value in the i-th experiment, $\mu$ is the overall mean, ${T}_{i}$ effect of the i-th experiment, ${\epsilon }_{ij}$ random error terms.

Individual dry matter intakes (DMI) and feed efficiency (FE) could not be determined because the cows were housed in groups. DMI and FE was recorded weekly per experimental group. Therefore, DMI and FE were not calculated statistically. The DMI was found 20.78 for G1, 21.60 for G2 and 25.46 for G3. The FE was found 1.09 for G1, 1.12 for G2 and 1.09 for G3.

Supplementation of OAW showed a tendency to increase in MP. MP increased in parallel with the supplemented OAW doses. OAW did not significantly affect the dry matter (DM), fat, protein, and lactose content in the milk (Table 2). The SCC of milk decreased non-significantly with the supplementation of OAW. SCC decreased as the dose of OAW increased (Table 2).

In the biochemical evaluation of blood parameters, no effect of OAW was observed except for blood ketone concentration and serum glucose level (Table 3). OAW doses significantly reduced ketone concentration while increasing glucose level (P<0.05).

When the blood parameters were examined hematologically, the numerical differences between groups were not found to be significant (Table4).

It was observed that OAW supplementation significantly decreased (P<0.05) the oxidant capacity and increased (P<0.05) the antioxidant capacity (Table 5). The oxidative stress index decreased (P<0.05) with OAW supplementation (Table 5).

Feed additives (such as monensin and essential oils) tend to increase the dry matter and crude protein digestibility of animals and the concentration of volatile fatty acids in the rumen, increasing feed efficiency and nitrogen utilization (Silva et al. 2018). One of the main regulators of DMI, especially in high-yielding dairy cows, is physical occupancy in the rumen. Variations in DMI are particularly related to the fiber content of the diet and its digestibility. The increase in fiber’s digestibility causes a reduction of undigested material in the rumen content, which positively affects DMI by increasing intestinal filling (Benchaar 2021). The OAW we used in our study provided a slight increase in DMI. This increase in DMI indicates that OAW supplementation to the diet provides a slight increase in the palatability of the diets. Some researchers have stated that the essential oils and/or their mixtures supplemented to the diet increase the palatability of diets (Giller et al. 2020), while others have stated the opposite (Hristov et al. 2013; Benchaar 2021). Many researchers have stated that DMI was not affected by herbal feed additives (Meiring 2014; Hashemzadeh-Cigari et al. 2015; Drong 2016; Silva et al. 2018; Prayitno et al. 2019; Stivanin et al. 2019; Braun et al. 2019). However, Hashemzadeh-Cagari et al. (2014) and Kuester (2016) indicated that DMI was affected by herbal feed additives. Milk production is related to DMI and nutrient supply. In parallel with the increase in DMI in dairy cows, an increase occurs in milk production (Hassanat et al. 2017; Giller et al. 2020). However, Benchaar (2021) reported that the supplementation of thyme oil did not change either DMI or milk production in cows. Thyme oil (Benchaar 2021) and mixtures of several different essential oils (Giller et al. 2020) were supplemented to the diets of dairy cows, but the milk fat ratio was not affected as in our study. It has been reported that the reason why milk fat is not affected may be related to the lack of an effect on the molar ratio of acetate, which is the main precursor of fat synthesis by the mammary glands (Benchaar 2021). Like the milk fat ratio, the milk protein, lactose and DM ratios were not affected by the supplementation of AOW. There are researchers reporting that herbal feed additives do not affect milk production and milk compositions (Meiring 2014; Hashemzadeh-Cigari et al. 2015; Drong 2016; Kuester 2016; Nowers 2016; Silva et al. 2018; Prayitno et al. 2019; Stivanin et al. 2019; Braun et al. 2019). However, Hashemzadeh-Cagari et al. (2014) and Braun et al. (2019) reported the opposite of these results. However, perhaps the different results obtained were related to the fact that the researchers studied with different herbal feed additives and dosages.

The SCC values obtained in the study were lower than the legal values for human consumption (< 500.000 cells in mL milk) (De Villiers et al. 2000). SCC is associated with udder health and milk quality. High SSC (above 500.000 cells/mL) is an indicator of low milk quality and unhealthy udder (Hashemzadeh-Cigari et al. 2014). Similarly to our result, Silva Filho et al. (2017) reported that the supplementation of thyme EO tends to reduce SCC. However, Giannenas et al. (2011) stated that the use of essential oil significantly reduces SCC in sheep milk. Many researchers who experimented with herbal products and their essential oils also reported reducing the SCC value (Hashemzadeh-Cigari et al. 2014; Meiring 2014; Moller 2015; Kuester 2016). On the contrary, Giller et al. (2020) reported that the use of thyme oil significantly increased SCC in cows, but this increase did not reach the value indicating mastitis. The large number of compounds present in EO can explain these variable results. Sometimes, the bioactivity of an EO can be attributed to one or two of its major compounds. In some cases, however, the sum of several compounds may be more effective than any single major compound (Bakkali et al. 2008; Giller et al. 2020).

Analysis of hematological and biochemical blood values is the decision making parameters used to obtain information about the animal’s metabolic and health status. Besides, biochemical parameters are important in the evaluation of damage in organs and tissues (Klinkon and Jezek 2012). Because of the negative energy balance in the early lactation dairy cows, almost all are faced with the risk of ketosis. During this period, ketone values increases and glucose values decreases. When the blood ketone value between 1.2–1.4 mmol/L, there is subclinical ketosis (Oetzel 2013). The blood ketone and glucose values of cows at the beginning of the trial were above these reference values. At the end of the experiment, it is thought that the antiketogenic effects of the plant and its extracts may be the reason for the decrease in blood ketone concentration in OAW supplemented groups and increased glucose levels in parallel. Studies have shown that propionate has an antiketogenic effect (Bush and Milligan 1971) and oregano aromatic water increases the concentration of propionate (Ozkaya 2020; Ozkaya et al. 2020). In contrast, Tassoul and Shaver (2009) also reported that the essential oils did not affect the GLU and ketone values of the cows in the early lactation. The different results may be due to the use of different herbal feed additives and dosages. Hashemzadeh-Cagari et al. (2015) observed that the herbal mixtures did not affect the biochemical blood values of cows. No significant differences was found among the means of hematological blood values. This result supported Drong (2016), who stated that essential oil supplementation had no effect on hematological blood values.

Sharma et al. (2011) reported that in the early lactation, cows were under oxidative stress and low antioxidative defense system and this situation increased the sensitivity to diseases such as mastitis and metritis. The results obtained from this study showed that OAW supplementation may support the antioxidative defense systems of cows in early lactation and prevent the effects of oxidative stress. However, However, Hashemzadeh-Cagari et al. (2015) and Mazur et al. (2019) found that supplementing cows with an herbal mixture had no effect on their antioxidative defense system.

Since there has been no published study on the effects of oregano aromatic water supplementation on milk yield, dry matter intake, blood parameters, and somatic cell count, it is difficult to make a reliable comparison with other researchers who used different additives at different doses.

In conclusion, this study showed that the amounts of OAW used did not have a significant negative effect on milk production, milk components, metabolic health and oxidative stress. However, OAW’s tendency to reduce SCC showed that it could have positive effects on udder health. Similarly, the decrease in blood ketone value and increase in glucose value led us to believe that it may be beneficial in preventing ketosis caused by a negative energy balance in the early stages of lactation.

At the same time, OAW showed that it can be used to prevent disease by significantly supporting the antioxidative defense mechanism against oxidative stress, which is thought to be the cause of many diseases. However, the amounts of OAW used in the study were likely too low to show positive effects on milk production and milk quality. The wide variety of EO in plants containing different EO, even within the same species due to geographic location, creates a significant challenge and an uncertainty element that is difficult to control in scientific research with natural EO extracts. In order to better understand the effects of single EO compounds and their combinations on the metabolism and production-related parameters of dairy cows, studies should be conducted to determine which compounds and which defined mixtures of these compounds are promising and in what amounts they should be used.

Acknowledgements The support of the Osmanli farm by allowing it to work is highly appreciated.

Authors’ contributions Serkan Ozkaya: Designed and conducted the experiment, wrote the first draft of the article. Koray Taskin: Helped conduct the experiment by collecting blood from cows. Sabri Erbas: Obtained aromatic water in oregano leaves. Oktay Ozkan: Performed blood analyzes and evaluated the blood results. Wojciech Neja: Conducted data and statistical analyzes and revised the article.

Statement of animal rights All procedures were reviewed and approved by the Akdeniz University, Animal Research and the Local Ethics Committee, Antalya, Turkey (Protocol number: 2015.01.004).

Conflict of interest statement The authors declared that there is no conflict of interest.

Data availability Datasets obtained and/or analyzed during the current study are available from corresponding after upon reasonable request.

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Table 1 Diet ingredients and chemical composition of total mixed ration for dairy cows

	TMR
Chemical profile
Dry Matter g/kg	604.2
Crude ash g/kg	50.60
Crude protein g/kg	152,10
Ether extract g/kg	42.30
Neutral Detergent Fiber g/kg	206.30
Acid Detergent Fiber g/kg	132.00
Acid Detergent Lignin g/kg	27.30
Starch g/kg	146.60
Metabolic Energy kcal kg^-1	1567.93
Feed items (% of dry matter)
Corn silage	47.24
Alfalfa	11.81
Wheat straw	2.36
Commercial concentrated milk feed*	11.81
Grain of maize	9.45
Cottonseed meal	3.54
Sun flower seed meal	3.54
Brewer’s grain	9.45
Vitamin premix	0.18
Salt	0.24
Sodium carbonate	0.14
Calcium carbonate	0.24

TMR: Total mixed ration, *Ingredients: Wheat bran, Corn, Sunflower meal, Barley, Soybean meal, Boncalite, Molasses, Calcium carbonate, Sodium chloride, Vitamin premix

Table 2 Milk production and composition of lactating cows

Items	G1	G2	G3	P
	Mean±SE	Mean±SE	Mean±SE
DIM	69.83±0.60	76.83±3.22	75.83±1.96	0.14
LN	2.67±0.33	2.33±0.33	2.33±0.33	0.73
MP	22.75±0.37	24.12±0.75	26.25±1.11	0.06
Fat, %	3.80±0.05	3.82±0.02	3.85±0.01	0.52
Protein, %	4.37±0.04	4.37±0.04	4.42±0.01	0.52
Lactose, %	4.96±0.06	4.97±0.05	5.04±0.01	0.58
DM, %	14.50±0.12	14.51±0.12	14.58±0.05	0.83
SCC, cell/mL	401000±10017	387333±9821	385667±8373	0.49

DIM: Days in milk, LN: Lactation number, DMI: Dry matter intake, kg/d; MP: Milk production, kg/d; FE: Feed efficiency, kg milk/kg DMI; SCC: Somatic cell count, G1: Control ration, G2: Control ration+10cc/head/day OAW, G2: Control ration+20cc/head/day OAW.

Table 3 Blood serum parameters of lactating cows

Items		G1 Mean±SE	G2 Mean±SE	G3 Mean±SE	P
Ketone	Initial	1.43±0.03	1.47±0.13	1.50±0.00	0.77
	Final	0.83±0.83^a	0.67±0.03^ab	0.57±0.07^b	0.02
ALT	Initial	52.00±5.77	59.67±8.17	70.67±3.84	0.18
	Final	45.67±4.41	35.00±4.04	39.67±1.67	0.19
ALP	Initial	97.70±6.72	63.33±7.54	81.67±7.13	0,32
	Final	119.00±15.30	83.33±4.91	47.30±31.30	0.12
GGT	Initial	32.67±3.67	21.00±4.73	18.33±1.76	0.06
	Final	22.67±2.03	27.67±4.37	14.33±3.33	0.08
GLU	Initial	42.67±1.20	42.33±0.88	43.33±3.67	0.88
	Final	44.33±1.45^b	53.67±4.06^a	57.67±2.60^a	0.01
CREA	Initial	1.84±0.01	1.72±0.14	1.78±0.02	0.60
	Final	1.49±0.28	1.79±0.08	0.92±0.32	0.12
UREA	Initial	18.10±1.24	18.60±0.31	16.87±3.46	0.84
	Final	15.87±2.96	21.20±1.47	9.57±4.87	0,13
TBIL	Initial	0.39±0.30	0.01±0.01	0.15±0.15	0.43
	Final	0.15±0.10	0.04±0.04	0.01±0.01	0.28
TP	Initial	7.91±0.15	6.98±0.24	7.24±0.33	0.10
	Final	6.69±0.25	6.77±0.32	6.61±0.27	0.92

Initial: Beginning of experiment, Final: End of experiment after 60 days, ALT: Alanine transaminase, ALP: Alkaline phosphatase, GGT: Gamma-glutamyltransferase, GLU: Glucose, CREA: Creatinine, TBil: Total bilirubin, TP: Total protein, ^a,b Means in a line with different lower case letters differ significantly

Table 4 Hematologic parameters of lactating cows

Items		G1 Mean±SE	G2 Mean±SE	G3 Mean±SE	P
WBC, 10³/ul	Initial	9.37±0.80	8.08±1.55	8.51±0.18	0.67
	Final	11.82±2.43	8.93±0.66	9.32±0.57	0.37
Lymphocytes, 10³/ul	Initial	4.30±0.70	3.16±0.92	3.86±0.22	0.52
	Final	4.37±0.99	4.01±0.61	3.83±0.23	0.85
Monocytes,10³/ul	Initial	1.06±0.12	1.03±0.16	0.82±0.11	0.40
	Final	2.11±1.00	0.97±0.17	0.93±0.17	0.32
Granulocytes, 10³/ul	Initial	4.03±0.65	3.90±0.68	3.84±0.07	0.97
	Final	5.34±1.31	3.95±0.30	4.57±0.38	0.50
RBC, 10⁶/ul	Initial	6.24±0.31	6.20±0.45	5.99±0.19	0.86
	Final	6.76±0.49	6.49±0.52	6.82±0.21	0.85
Hemoglobin, g/dl	Initial	9.65±0.20	9.80±0.44	8.88±0.25	0.14
	Final	10.55±0.49	10.35±0.38	10.50±0.22	0.93
Hematocrit, %	Initial	27.85±0.71	28.00±1.28	26.18±0.43	0.32
	Final	28.55±1.23	28.03±0.85	28.01±0.78	0.91
MCV, fl	Initial	45.00±1.73	45.50±1.50	43.70±0.85	0.68
	Final	42.75±1.93	43.50±2.25	41.25±1.80	0.73
MCH, pg	Initial	15.53±0.62	15.98±0.81	14.88±0.33	0.48
	Final	15.70±0.60	16.10±0.80	15.45±0.51	0.78
MCHC, g/dl	Initial	34.55±0.21	35.00±0.67	33.95±0.52	0.37
	Final	36.88±0.28	36.93±0.23	37.58±0.34	0.21
PCT, %	Initial	0.15±0.04	0.19±0.04	0.21±0.03	0.62
	Final	0.20±0.03	0.17±0.02	0.19±0.02	0.69
Platelets, 10³/ul	Initial	255.30±54.40	235.00±60.60	313.00±42.00	0.58
	Final	308.30±55.00	247.50±35.50	296.80±34.30	0.58

Initial: Beginning of experiment, Final: End of experiment after 60 days, WBC: White blood cell, RBC: Red blood cell, MCV: Mean corpuscular volume, MCH: Mean corpuscular hemoglobin, MCHC: Mean corpuscular hemoglobin concentration, PCT: Procalcitonin,

Table 5 Effect of OAW on oxidative stress

Items	G1 Mean±SE	G2 Mean±SE	G3 Mean±SE	P
TOC	2.84±0.54^a	0.71±0.25^b	1.19±0.25^b	0.02
TAC	2.26±0.29^b	5.46±1.96^a	5.98±1.75^a	0.02
OSI	0.14±0.04^a	0.01±0.01^b	0.02±0.01^b	0.02

OAW, TOC: Total oxidative capacity, mmol H₂O₂ Eq/L; TAC: Total antioxidant capacity, mmol Trolox Eq/L; OSI: Oxidative stress index, TOC/TAC*10; ^a,b Means in a line with different lower case letters differ significantly

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Effects of Oregano Aromatic Water (Oreganum onites L.) Supplementation on Diets of Lactating Dairy cows on Milk Production Performance, Metabolic Health and Antioxidative Defense Mechanism

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Abstract

Introduction

Material And Methods

Animals and Rations

Oregano Aromatic Water Production

Statistical Analysis

Results

Discussion

Declarations

References

Tables

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