Effects of Bacillus subtilis C-3102 addition on nutrient digestibility, faecal characteristics, blood chemistry and faecal Lactobacilli spp., Enterococci spp., and Escherichia coli in healthy dogs

Figures & data

Table 1. Ingredient and chemical composition of dry extruded food, %.

Ingredient composition
Poultry meal	22
Barley	26
Maize	10
Maize gluten meal	10
Rice	26.6
Potassium chloride	0.5
Salt	0.1
Vitamin/mineral pre-mixture^a	0.4
Sunflower oil	2.5
Beef tallow	1.9
Determined Chemical Composition
Dry matter, %	91.5
Ash, %	5.28
Acid hydrolysed ether extract, %	9.21
Crude fibre, %	6.45
Crude protein, %	18.45
Metabolizable energy (ME), kcal/kg^b	3242.5
Total dietary fibre, %	12.54
Insoluble dietary fibre, %	9.28
Soluble dietary fibre, %	3.26
Calcium, %	1.3
Phosphorus, %	0.85
Sodium, %	0.35
Vit-A vitIU/kg	12 000
Vit-D IU/kg	1100
Vit-E mg/kg	60

^aVitamin-mineral premixture (mg/kg): biotin 250 mg, folate 2 mg, niacin 50 mg, pantothenic acid 20 mg, riboflavin 8 mg, thiamine 4 mg, Vit. B6 5 mg, Vit. B12 25 mg, Vit. A 3.6 mg, cholecalciferol 125 mg, Vit. E 50 mg, Mn proteinate 70 mg, Zn proteinate 100 mg, Fe proteinate 70 mg, Cu proteinate 14 mg, Iodine 1 mg, Co proteinate 350 mg, Se proteinate 140 mg, Mo proteinate 700 mg, Mg oxide 35.000 mg.

^bCalculated using the NRC (2006) equations.

Table 2. Apparent nutrient digestibility coefficients of the groups, % (Mean  ±  SEM).

	CON	BS1	BS2	p Value
DMD	71.52 ± 1.19^b	75.52 ± 1.44^a	75.06 ± 0.57^a	0.031
OMD	78.11 ± 0.92^b	83.33 ± 1.15^a	83.57 ± 0.65^a	0.012
EED	90.78 ± 0.55	91.28 ± 1.22	92.29 ± 0.92	0.589
CFD	61.60 ± 1.02^b	64.94 ± 0.88^a	65.92 ± 0.99^a	0.001
CPD	81.84 ± 0.73^b	85.78 ± 0.97^a	85.30 ± 0.58^a	0.001

DMD: dry matter digestibility; OMD: organic matter digestibility; EED: fat digestibility; CFD: crude fibre digestibility; CPD: crude protein digestibility; CON: control; BS1: B. subtilis supplementation group 1 receiving 1 × 10⁹ CFU/g of B. subtilis; BS2: B. subtilis supplementation group 2 receiving 2 × 10⁹ CFU/g of B. subtilis; SEM: standard error of the mean;^abcvalues in the same row that do not share the same superscripted letter differ significantly (p < 0.05).

Table 3. Faecal parameters of the control and B. subtilis groups of dogs (Mean  ±  SEM).

	CON	BS1	BS2	p Value
Faecal consistency score	3.78 ± 0.13^b	4.15 ± 0.14^a	4.23 ± 0.12^a	0.041
Faecal pH	7.11 ± 0.15^a	6.81 ± 0.08^b	6.78 ± 0.11^b	0.001
Faecal dry matter	45.59 ± 0.57^b	50.86 ± 0.69^a	51.37 ± 0.43^a	0.001
Faecal output (g as-is/day)	101.97 ± 5.72	96.86 ± 7.39	95.97 ± 6.66	0.372
Faecal NH3 (µmol/g DM)	0.71 ± 0.35^a	0.64 ± 0.50^a	0.49 ± 0.15^b	0.011
Acetate (µmol/g DM)	153.15 ± 24.13^b	204.45 ± 25.88^a	195.37 ± 32.01^a	0.001
Propionate (µmol/g DM)	92.93 ± 23.24^b	118.11 ± 26.74^a	104.92 ± 30.87^a	0.001
Butyrate (µmol/g DM)	18.01 ± 2.09	21.31 ± 4.53	20.49 ± 1.99	0.068
Isobutyrate (µmol/g DM)	2.26 ± 0.19^b	3.61 ± 0.72^a	1.91 ± 0.21^b	0.001
Isovalerate (µmol/g DM)	2.42 ± 0.15	3.02 ± 0.20	1.87 ± 0.06	0.074
Valerate (µmol/g DM)	0.76 ± 0.11	0.52 ± 0.04	0.65 ± 0.1	0.186
Total SCFAs (µmol/g DM)	254.11 ± 11.61^b	343.83 ± 13.41^a	320.78 ± 16.31^a	0.001
Total BCFAs (µmol/g DM)	6.37 ± 0.39^ab	7.79 ± 0.76^a	5.26 ± 0.42^b	0.012

CON: basal diet; BS1: CON + 1 × 10⁹ CFU/g of B. subtilis; BS2: CON + 2 × 10⁹ CFU/g of B. subtilis; SCFAs: short-chain fatty acids; BCFAs: branched-chain fatty acids;^a,b,cvalues in the same row that do not share the same superscripted letter differ significantly (p < 0.05).

Table 4. Effects of dietary supplementation with Bacillus subtilis on faecal microbiota in dogs, log₁₀ CFU/g.

Group	Dose (gram)	Time (day)	n	Lactobacillus spp.	E. coli	Enterecoccus spp.
CON	0	zero	6	1.40±0.2	1.28±0.17	2.04±0.1
		30	6	1.93±0.15	1.50±0.12	2.27±0.21
P				0.068	0.181	0.109
BS1	2	zero	6	1.33±0.07	1.27±0.16	2.19±0.22
		30	6	2.36±0.13	1.86±0.1	3.46±0.04
P				0.024	0.042	0.031
BS2	4	zero	6	1.54±0.54	1.31±0.05	2.75±0.05
		30	6	3.14±0.56	1.51±0.15	3.58±0.32
P				0.028	0.061	0.021
CON (day 30)			6	1.93^b	1.50	2.27^b
BS1(day 30)			6	2.36^b	1.86	3.46^a
BS2 (day 30)			6	3.14^a	1.51	3.58^a
P				0.012	0.112	<0.001
Tukey HSD
SEM				0.19	0.07	0.11
Dose				<0.001	0.342	<0.001
Time				<0.001	0.245	<0.001
dose*time				<0.001	0.064	<0.001

CON: basal diet; BS1: CON + 1 × 10⁹ CFU/g of B. subtilis; BS2: CON + 2 × 10⁹ CFU/g of B. subtilis; SEM: standard error of the mean; ^a,bvalues with different superscripted letters are significantly different (p < 0.05).

Table 5. Effects of dietary supplementation with Bacillus subtilis on blood profiles of dogs.

Group	Time (day)	N	Cholesterol, mg/dL	Triglycerides, mg/dL	WBCs, 10^3/µL	Lymphocytes^c, %	Monocytes^c, %	Granulocytes^c, %	RBCs, 10^3/µL
CON	zero	6	223.16±32.1	84.33±3.8	8.37±0.61	25.87±1.20	3.81±0.15	59.34±1.4	7.57±1.67
	30	6	219.5±16.09	87.83±4.85	8.66±0.54	26.16±1.15	3.51±0.11	58.98±1.05	8.01±1.56
		p-value	0.865	0.114	0.563	0.665	0.255	0.842	0.291
BS1	zero	6	228.83±14.1	93.16±2.91	7.8±0.52	27.05±0.1	3.66±0.1	60.65±2.40	8.08±0.78
	30	6	203.84±15.5	88.1±3.1	7.97±0.65	27.1±0.1	3.76±0.12	62.98±1.93	7.66±0.58
		p-value	0.264	0.17	0.856	0.971	0.152	0.401	0.678
BS2	zero	6	232.33±12.8	87.1±4.01	8.81±0.66	21.62±0.7	2.98±0.1	60.51±1.27	6.7±0.4
	30	6	204.1±10.5	82.1±4.23	10.65±0.53	24.23±6	3.33±0.2	59.8±1.02	9.72±0.3
		p-value	0.119	0.269	0.039	0.027	0.67	0.945	0.001
CON	30	6	219.5±16.09	87.83±4.82	8.66±0.54^b	26.16±1.15	3.51±0.1	58.98±1.05^b	8.01±0.63^b
BS1	30	6	203.84±15.5	81.1±7.11	7.97±0.65^b	27.1±0.51	3.76±0.12	62.98±1.13^a	7.66±0.58^b
BS2	30	6	204.1±10.5	80.1±10.5	10.65±0.55^a	24.23±6.01	3.33±0.2	59.8±1.02^ab	9.72±0.3^a
		p-value	0.684	0.178	0.017	0.081	0.095	0.043	0.048
		Reference range	136–392	23–102	6.0–17.0	10.0–30.0	2.0–10.0	50.0–80.0	5.5–8.5

^a,bValues with different superscripted letters are significantly different (p < 0.05); RBCs, red blood cells; WBCs: white blood cells; ^cvalues are presented as a percentage of the total WBCs count; CON, basal diet; BS1, CON + 1 × 10⁹ CFU/g of B. subtilis; BS2, CON + 2 × 10⁹ CFU/g of B. subtilis.

NRC 2006. Nutrient requirements of dogs and cats. National Academies Press.

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Data availability statement

The data presented in this study are available from the corresponding author upon reasonable request.