Effect of thymol and Enterocin M administration on biochemical, antioxidant and immunological parameters, small intestinal morphology and microbiota in rabbits.

Figures & data

Table 1. Ingredients and chemical composition (g/kg DM) of experimental diets.

Ingredients	g/kg DM	Composition	g/kg DM
Dehydrated lucerne meal	324.35	Dry matter (g/kg)	900.98
Dry malting sprouts	135.15	Organic compounds	749.42
Oats	117.13	Nitrogen free extract	400.26
Wheat bran	81.09	Neutral detergent fibre	317.95
Barley	72.08	Acid detergent fibre	187.49
Extracted sunflower meal	49.55	Crude fiber	160.23
Extracted rapeseed meal	49.55	Crude protein	159.12
Dried distillers grain with solubles	45.05	Cellulose	147.00
Premix^a	15.32	Hemicellulose	130.45
Limestone	9.01	Starch	119.88
Sodium chloride	2.70	Ash	62.35
Total	900.98	Fat	29.80
		Metabolic energy, MJ/kg DM	9.91

^a The vitamin-mineral premix provided per kg of complete diet: Retinyl acetate 5.16 mg, Cholecalciferol 0.03 mg, Tocopherol 0.03 mg, Thiamine 0.8 mg, Riboflavin 3.0 mg, Pyridoxine 2.0 mg, Cyanocobalamin 0.02 mg, Niacin 38 mg, Folic acid 0.6 mg, Calcium 1.8 mg, Iron 70 mg, Zinc 66 mg, Copper 15 mg, Selenium 0.25 mg.

DM: dry matter.

Table 2. Effects of Enterocin M and thymol on biochemical parameters in serum of rabbits (n = 8).

Parameter	Experimental group				SD	p - value
	Control	Ent M	Thymol	Ent M with Thymol		Ent M	Thymol	Interaction
AST (µkat/L)	0.33	0.41	0.29	0.29	0.12	.4249	.1074	.3859
ALT (µkat/L)	0.61^ab	0.88^a	0.78^a	0.44^b	0.27	.6286	.1116	.0009
GMT (µkat/L)	0.07^a	0.12^a	0.09^a	0.08^a	0.04	.1503	.6642	.0264
ALP (µkat/L)	1.87^a	2.71^a	2.66^a	2.27^a	0.60	.2457	.3713	.0037
Triglycerides (mmol/L)	1.51^ab	2.98^a	2.17^ab	1.32^b	1.15	.4065	.1822	.0038
Cholesterol (mmol/L)	1.06^ab	2.04^a	1.60^ab	0.87^b	0.77	.6068	.1890	.0011
Total protein (g/L)	53.84^ac	65.65^ab	68.24^b	50.38^c	10.43	.2635	.8708	<.0001
Urea (mmol/L)	4.64^a	6.28^a	6.02^a	5.73^a	1.51	.5190	.04040	.0014
Creatinine (µmol/L)	75.33	76.33	95.14	83.50	23.15	.5340	.1230	.4608

^{a ,b}Means within a row with different superscript letters differed significantly (p < .05). Data are presented as mean ± standard deviation (SD).

Ent M: enterocin M; AST: aspartate aminotransferase; ALT: alanine aminotransferase; GMT, gamma glutamyl transferase; ALP: alkaline phosphatase.

Table 3. Effects of Enterocin M and thymol on antioxidant parameters and activity of lactate dehydrogenase in blood and tissues of rabbits (n = 8).

Parameter	Experimental group				SD	p-value
	Control	Ent M	Thymol	Ent M with Thymol		Ent M	Thymol	Interaction
Blood
GPx (µkat/g Hb)	2.20	2.42	1.54	3.13	1.53	.0966	.9610	.2038
MDA (nmol/mL)	0.33	0.35	0.31	0.34	0.03	.0548	.1730	.6087
Liver
MDA (nmol/g protein)	87.21^a	65.85^b	62.87^b	78.57^ab	15.70	.5475	.2219	.0005
LDH (µkat/g protein)	16.76	17.50	17.89	19.24	2.77	.3020	.1600	.7611
Kidney
MDA (nmol/g protein)	164.10^ab	230.00^a	148.20^b	215.20^a	51.36	.9696	.2925	<.0001
LDH (µkat/g protein)	16.02	16.04	16.87	19.01	2.83	.2641	.0536	.2729
Muscle
MDA (nmol/g protein)	30.00	31.89	35.29	32.53	6.43	.8570	.2258	.3396
LDH (µkat/g protein)	23.89	23.73	24.44	22.24	6.34	.6512	.8568	.6958

^{a ,b}Means within a row with different superscript letters differed significantly (p < .05). Data are presented as mean ± standard deviation (SD).

Ent M: enterocin M; GPx: glutathione peroxidase; MDA: malondialdehyde; LDH: lactate dehydrogenase.

Table 4. Effects of Enterocin M and thymol on jejunal organ morphometric indices, and immunoglobulin A; and phagocytic activity in blood of rabbits (n = 8).

Parameter	Experimental group				SD	p-value
	Control	Ent M	Thymol	Ent M with Thymol		Ent M	Thymol	Interaction
Vh (µm)	691.40	706.20	695.80	704.20	14.26	.0823	.8503	.6160
Cd (µm)	181.00	175.00	176.80	178.40	8.27	.5782	.9191	.3415
Vh/Cd	3.82	4.04	3.94	3.95	0.14	.0567	.9074	.0927
IgA (µg/g)	11.46	52.02	29.84	61.85	25.74	.0002	.0880	.5912
PA (%)	62.63	63.51	66.00	66.13	3.08	.6169	.0051	.7109

Data are presented as mean ± standard deviation (SD).

Ent M: enterocin M; Vh: villi height; Cd: crypt depth; Vh/Cd: villus height to crypt depth ratio; IgA: immunoglobulin A; PA: phagocytic activity.

Table 5. Effects of Enterocin M and thymol on organic acids concentration in caecum of rabbits (n = 8).

Parameter	Experimental group				SD	p-value
	Control	Ent M	Thymol	Ent M with Thymol		Ent M	Thymol	Interaction
Acetic acid (mmol/L)	6.18^a	7.57^a	8.83^a	6.97^a	1.65	.7115	.1305	.0226
Lactic acid (g/100g)	16.64	15.78	19.92	22.16	4.46	.6994	.0142	.3903
Butyric acid (mmol/L)	1.56	1.82	2.19	1.56	0.60	.4941	.4918	.1063
Propionic acid (mmol/L)	0.43	0.63	0.57	0.46	0.18	.6083	.8497	.0576

Data are presented as mean ± standard deviation (SD).

Ent M: enterocin M.

Table 6. Effect of Enterocin M and thymol on bacterial counts in appendix, caecum and faeces of rabbits (log 10 CFU/g, n = 8).

Parameter	Experimental group				SD	p-value
	Control	Ent M	Thymol	Ent M with Thymol		Ent M	Thymol	Interaction
Enterococcus sp.
appendix	1.68	2.40	1.99	3.00	1.46	.2153	.5033	.8353
caecum	1.27	0.90	0.90	1.53	0.60	.6303	.6303	.084
faeces	2.61	1.82	2.86	3.20	1.15	.6459	.1120	.2635
LAB
appendix	2.60	2.55	2.62	3.09	1.03	.6840	.5741	.6014
caecum	3.02	3.66	2.71	3.20	0.72	.0844	.2252	.8060
faeces	3.66	3.54	3.41	3.62	0.35	.7302	.5161	.2205
CoNS
appendix	2.45	2.05	2.48	1.99	0.72	.2001	.9671	.9061
caecum	3.24	3.51	3.04	3.60	0.74	.2391	.8784	.6680
faeces	3.25	3.14	3.27	3.03	0.35	.1818	.7212	.6067
CoPS
appendix	2.70	3.21	4.29	2.88	1.30	.4283	.2653	.0998
caecum	2.75	2.89	2.14	2.31	0.61	.5433	.0329	.9445
faeces	3.23	3.13	3.10	3.78	0.63	.2992	.3402	.3402
Coliform bacteria
appendix	1.18	1.83	3.15	2.12	1.43	.7629	.0887	.1954
caecum	2.19	3.51	3.17	3.69	1.37	.1455	.3497	.5156
faeces	5.73	2.62	4.96	3.57	1.57	.0002	.8450	.1798

Data are presented as mean ± standard deviation (SD).

Ent M: enterocin M; LAB: lactic acid bacteria; CoNS: coagulase-negative staphylococci; CoPS: coagulase-positive staphylococci; sp.: species.

Data availability statement

Data available on request from the authors.