Impact of dietary-nucleotides and Saccharomyces cerevisiae-derivatives on growth-performance, antioxidant-capacity, immune-response, small-intestine histomorphometry, caecal-Clostridia, and litter-hygiene of broiler-chickens treated with florfenicol

Figures & data

Table 1. Feed ingredients and nutrient contents of broiler diets from 1 to 33 days of the experiment.

Ingredients %	Control basal diet (G1)
	Starter	Grower	Finisher
Yellow corn	57.43	61.23	65.93
Corn gluten meal (60% CP)	3.00	2.00	2.00
Soybean meal (46% CP)	36.0	32.20	27.00
Soy-oil	0.00	1.20	2.0
Di-calcium phosphate	0.90	0.80	0.50
Limestone	1.40	1.30	1.25
Common salt	0.35	0.35	0.35
Sodium bicarbonate	0.15	0.15	0.15
DL-Methionine	0.20	0.20	0.20
L-Lysine HCl	0.20	0.20	0.25
UNIKE® Plus *	0.05	0.05	0.05
Phytase	0.01	0.01	0.01
Xylanase	0.01	0.01	0.01
Vitamin-Mineral premixes **	0.30	0.30	0.30
Total (%)	100	100	100
Chemical composition
ME (Kcal/kg)	3010.06	3105.84	3201.51
Crude protein (%)	23.11	21.04	19.05
Crude fat (%)	2.57	3.83	4.73
Crude fibre (%)	2.36	2.30	2.22
Calcium (%)	1.02	0.95	0.86
Available phosphorus (%)	0.50	0.47	0.41

*Toxin binder (Adisseo, France).

**Vitamin-mineral premix per Kg of diet: 1200000 IU vit. A, 350,000 IU vit. D3, 4000 mg vit. E, 250 mg vit. B1, 800 mg vit. B2, 600 mg vit. B6, 3.2 mg vit. B12, 450 mg vit. K3, 4.5 g nicotinic acid, 1.5 g Ca pantothenate, 120 mg folic acid, 5 mg biotin, 55 mg choline chloride, 3 g Fe, 2 g Cu, 10 g Mn, 8 g Zn, 120 mg I, 40 mg Co.

Table 2. Composition of NutriFix® (Agromed GMBH Company, Austria).

Composition	Quantity
Brewer’s yeast (Saccharomyces cerevisiae)	960 g
Mannan oligosaccharide	100 g
Beta glucan (1,3&1,6)	125 g
Diatomaceous earth	Up to 1 kg

Table 3. Influence of dietary Nutrifix® on performance parameters of broiler chickens.

	Body weight (g)					Weight gain (g)					Feed intake (g)					FCR (g/g)
Groups	D 7	D 14	D 21	D 28	D 33	D 7	D 14	D 21	D 28	D 33	D 7	D 14	D 21	D 28	D 33	D 7	D 14	D 21	D 28	D 33
G1	198	547	1105	1576	1905^b	152	349	558	471	329	147	471	682	861	731	0.97	1.35	1.22	1.84	2.30
G2	200	565	1144	1700	2059^a	154	365	580	556	359	148	470	724	893	721	0.96	1.29	1.25	1.63	2.13
G3	210	560	1163	1681	2059^a	164	350	603	518	378	166	479	730	864	715	1.02	1.37	1.21	1.69	1.89
G4	205	572	1162	1664	2058^a	159	367	591	502	394	150	459	726	895	744	0.94	1.25	1.23	1.79	1.90
SEM^1	3.42	6.72	10.07	22.36	21.88	3.42	6.15	8.84	17.54	18.09	4.37	3.60	13.14	10.13	14.63	0.02	0.02	0.02	0.06	0.12
P-value	0.667	0.665	0.123	0.216	0.001	0.667	0.691	0.369	0.435	0.679	0.395	0.297	0.606	0.543	0.928	0.347	0.278	0.971	0.635	0.644

^abMean values with different superscripts in the same column indicate significant differences (Tukey’s test; p ≤ 0.05).

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

FCR, Feed Conversion Ratio (g of feed/g of weight gain).

¹SEM:Pooled standard error of means.

Table 4. Influence of dietary Nutrifix® on cumulative growth performance parameters of broiler chickens (days 1–33).

Groups	Body weight (g)	Daily gain (g)	Feed intake (g)	FCR (g/g)	EPEF	Mortality (%)
G1	1905^b	56^b	2892	1.56^a	332	10.00
G2	2059^a	61^a	2956	1.47^b	389	10.00
G3	2059^a	61^a	2954	1.47^b	392	9.33
G4	2058^a	61^a	2974	1.48^b	386	10.00
SEM^1	21.88	0.66	22.13	0.01	9.32	1.38
P-value	0.001	0.001	0.643	0.003	0.129	0.998

^a,bMean values with different superscripts in the same column indicate significant differences (Tukey’s test; p ≤ 0.05).

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

FCR, Feed Conversion Ratio (g of feed/g of weight gain).

EPEF: European Production Efficiency Factor= (liveability × live weight (kg)/(age in days × FCR) × 100.

¹SEM:Pooled standard error of means.

Table 5. Influence of dietary Nutrifix® on Carcass traits of broiler chickens.

Groups	Dressing (%)	Breast (%)	Thigh (%)	Drum (%)	Abdominal fat (%)
G1	71.36	33.50	25.52	12.85	1.18
G2	73.86	32.25	25.47	12.89	1.42
G3	72.89	33.81	25.00	12.81	0.93
G4	73.07	33.28	24.68	13.07	1.17
SEM^1	0.41	0.35	0.21	0.12	0.15
P-value	0.180	0.446	0.454	0.891	0.710

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

¹SEM:Pooled standard error of means.

Table 6. Effect of diet supplementation with NutriFix® on the histomorphometry of the small intestine of broiler chickens medicated with florfenicol.

	Duodenum			Jejunum			Ileum
Groups	Villi length	Crypt depth	V/C	Villi length	Crypt depth	V/C	Villi length	Crypt depth	V/C
G1	1702.78^b	114.91	14.88^b	1053.19^b	88.23^b	12.23^a	580.94^b	76.51^b	7.61^b
G2	1566.86^c	102.91	15.25^b	905.83^c	132.15^a	6.85^b	785.78^a	110.14^a	7.13^b
G3	1888.57^a	105.37	17.92^a	1428.85^a	123.11^a	11.65^a	840.13^a	106.42^a	7.91^ab
G4	1989.42^a	105.49	19.10^a	1035.29^b	109.71^b	9.44^ab	854.59^a	100.89^a	8.48^a
SEM^1	44.84	6.01	0.87	43.63	6.18	0.82	26.54	3.90	0.29
P-value	<0.0001	0.241	<0.0001	<0.0001	<0.0001	<0.0001	<0.0001	<0.0001	0.002

^a,b,cMean values with different superscripts in the same column indicate significant differences (Tukey’s test; p ≤ 0.05).

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

¹SEM:Pooled standard error of means.

V/C: Villus crypt ratio.

Table 7. Effect of diet supplementation with NutriFix® on blood biochemical indices of broiler chickens medicated with florfenicol.

	Protein profile				Lipid profile		Glucose (mg/dl)
Groups	Total protein (g\dl)	Albumin (g\dl)	Globulin (g/dl)	A/G ratio	Cholesterol (mg\dl)	TAG (mg\dl)
G1	3.99^b	1.82^b	2.10^ab	0.89	154.62^a	55.75^a	228.25^b
G2	4.55^a	2.36^a	2.24^ab	1.08	137.75^b	41.13^b	234.25^b
G3	3.46^c	1.85^b	1.57^b	1.21	160.50^a	49.87^a	256.50^a
G4	4.43^a	2.19^a	2.51^a	1.00	159.00^a	52.75^a	242.13^ab
SEM^1	0.87	0.05	0.67	0.05	2.66	1.43	2.80
P-value	0.0001	0.0001	0.030	0.125	0.004	0.0001	0.001

^a,b,cMean values with different superscripts in the same column indicate significant differences (Tukey’s test; p ≤ 0.05).

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

¹SEM:Pooled standard error of means.

Table 8. Effect of diet supplementation with NutriFix® on total and differential leucocytic counts of broiler chickens medicated with florfenicol.

Groups	TLC	Heterophiles (%)	Lymphocytes (%)	Monocytes (%)	Eosinophils (%)	Basophils (%)	H/L ratio
G1	22.16^a	17.00^b	71.16^a	8.83^ab	2.50	0.67	0.24^b
G2	23.16^a	18.17^b	69.33^ab	8.85^b	3.00	0.66	0.26^b
G3	18.17^b	24.00^a	61.00 ^c	11.16^a	3.50	0.68	0.39^a
G4	20.17^a	19.00^b	67.83^b	9.83^ab	2.50	1.00	0.28^b
SEM^1	0.47	0.71	0.88	0.36	0.18	0.14	0.02
P-value	0.0001	0.0001	0.0001	0.024	0.120	0.801	0.0001

^a,b,cMean values with different superscripts in the same column indicate significant differences (Tukey’s test; p ≤ 0.05).

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

¹SEM:Pooled standard error of means.

Figure 1. Effect of diet supplementation with Nutrifix on serum total antioxidant capacity (TAC) of broiler chickens (day 33). (G1), control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days. ^a,b,c,d mean values with different letters on the same columns indicate significant differences (Tukey’s test; p ≤0.05).

Table 9. Effect of diet supplementation with NutriFix® on the phagocytic assay of broiler chickens medicated with florfenicol.

	Phagocytosis %		Phagocytic Index (PI)
Groups	Day 27	Day 33	Day 27	Day 33
G1	27.67^c	31.33^c	5.10^c	5.43^c
G2	58.33^a	66.00^a	7.57^a	9.30^a
G3	29.33^c	31.67^c	4.07^d	4.27^d
G4	42.00^b	49.00^b	5.93^b	6.73^b
SEM^1	3.82	4.37	0.39	0.57
P-value	<0.0001	<0.0001	<0.0001	<0.0001

^a,b,c,dMean values with different superscripts in the same column indicate significant differences (Tukey’s test; p ≤ 0.05).

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

¹SEM:Pooled standard error of means.

Table 10. Effect of diet supplementation with NutriFix® on NDV vaccinal antibody titres of broiler chickens medicated with florfenicol.

	Mean HI titre (log2)
Groups	Day 27	Day 33
G1	3.67	4.50
G2	3.33	6.33
G3	3.00	5.00
G4	4.00	6.40
SEM^1	0.15	0.35
P-value	0.077	0.126

(G1), Control group (basal diet); (G2), basal diet +250 g NutriFix®/Ton of feed; (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

¹SEM:Pooled standard error of means.

Figure 2. Effect of diet supplementation with Nutrifix® on the caecal clostridial count of broiler chickens (day 33). Control (G1), control group (basal diet); NutriFix (G2), basal diet +250 g NutriFix®/Ton of feed; Florfenicol (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; NutriFix+ florfenicol (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days.

Figure 3. Effect of diet supplementation with nutrifix® on litter moisture % of broiler chickens (day 33). Control (G1), control group (basal diet); NutriFix (G2), basal diet +250 g NutriFix®/Ton of feed; Florfenicol (G3), basal diet + florfenicol (25 mg/Kg body weight) in drinking water for 5 days; NutriFix+ florfenicol (G4), basal diet +250 g NutriFix®/Ton of feed + florfenicol (25 mg/Kg body weight) in drinking water for 5 days. Asterisk (*) indicates significance at p ≤0.05.

Data availability statement

The generated data of the current study are offered on a reasonable request.