Mycological diversity in whole-plant corn silages inoculated with Lentilactobacillus buchneri or Lentilactobacillus farraginis

Morais, L.B.; Carvalho, B.F.; Batista, L.R.; Schwan, R.F.; Ávila, C.L.S.

doi:10.1590/1678-4162-13023

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Animal Science • Arq. Bras. Med. Vet. Zootec. 76 (1) • Jan-Feb 2024 • https://doi.org/10.1590/1678-4162-13023 copy

Mycological diversity in whole-plant corn silages inoculated with Lentilactobacillus buchneri or Lentilactobacillus farraginis

(Diversidade micológica em silagens de milho planta inteira inoculadas com Lentilactobacillus buchneri ou Lentilactobacillus farraginis)

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

The objective was to investigate the effect of wild lactic acid bacteria (LAB) strains on the fungal and chemical composition of whole-plant corn silage. Three treatments were evaluated: CON-without inoculation, LB-inoculated with Lentilactobacillus buchneri (CCMA 1366), and LF-inoculated with L. farraginis (CCMA 1362). The silages were stored for 29, 103, and 193 d. The highest acetic acid (19.7 g/kg DM) and 1,2-propanediol (6.3 g/kg DM) concentrations were found in the LF, the highest aerobic stability was observed in this silage after 29 d (158.3 h). The yeast population was below the detection level at 29 d in the LF and 103 and 193 d in all silages. Seven genera of filamentous fungi (FF) were identified before ensiling; none of these were found after ensiling. At 29 d, Monascus pilosus was identified in the LB after aerobic exposure but did not produce citrinin. At 103 and 193 d, there was Aspergillus fumigatus predominance. A. flavus was identified at 103 d in the CON and LB; none produced B₁, B₂, G_1, or G₂ toxins. LAB strains modified the silages’ chemical and FF composition, and LF increased aerobic stability and reduced FF’s yeast population and diversity.

Keywords:
Aspergillus; β-Tubulin; filamentous fungi molecular identification; MALDI-TOF-MS; silage quality

Variable	Silage (S)	Storage periods (SP) (days)			Mean	SEM	P-value
Variable	Silage (S)	29	103	193			S	SP	S × SP
Dry matter (g/kg as fed)	CON	276.1	291.4	264.5	277.3	3.37	0.66	0.13	0.30
	LB	269.8	286.6	286.5	281.0
	LF	282.7	281.5	256.2	273.5
	Mean	276.2	286.5	269.1
pH	CON	3.54a*	3.50a	3.43b	3.49	0.004	<0.01	<0.01	<0.01
	LB	3.60a	3.52b	3.46c	3.53
	LF	3.53b	3.63a	3.53b	3.56
	Mean	3.56	3.55	3.47
	P-value for contrasts
	CON vs. LB	0.01	0.60	0.17
	CON vs. LF	0.88	<0.01	<0.01
Water activity	CON	0.9853	0.9893	0.9857	0.9868	0.0004	0.19	0.04	0.77
	LB	0.9843	0.9873	0.9860	0.9859
	LF	0.9873	0.9887	0.9877	0.9879
	Mean	0.9857b	0.9884a	0.9864ab
Lactic acid (g/kg DM)	CON	41.1	59.7	66.3	52.4	1.17	0.05	<0.01	0.31
	LB	48.2	52.5	56.0	52.3
	LF	41.5	47.3	48.2	45.7
	Mean	43.6b	53.2a	53.5a
	P-value for contrasts
	CON vs. LB				0.67
	CON vs. LF				0.01
Acetic acid (g/kg DM)	CON	5.1	9.1	9.7	7.9	0.37	<0.01	0.01	0.22
	LB	11.2	9.7	13.2	11.4
	LF	18.1	20.2	20.8	19.7
	Mean	11.4b	13.0ab	14.6a
	P-value for contrasts
	CON vs. LB				0.01
	CON vs. LF				<0.01
Lactic/ Acetic ratio	CON	8.15a	6.57ab	5.82b	6.84	0.157	<0.01	0.14	0.05
	LB	4.34a	5.45a	4.36a	4.72
	LF	2.29a	2.34a	2.33a	2.32
	Mean	4.92	4.79	4.17
	P-value for contrasts
	CON vs. LB	0.01	0.11	0.02
	CON vs. LF	<0.01	<0.01	<0.01
Butiric acid (g/kg DM)	CON	0.0	0.4	2.9	1.0	0.10	0.87	<0.01	0.99
	LB	0.0	0.5	3.1	1.2
	LF	0.1	0.5	2.9	1.1
	Mean	0.0b	0.5b	2.9a
1,2- propanediol (g/kg DM)	CON	0.0	0.5	0.1	0.3	0.24	<0.01	0.54	0.25
	LB	1.7	2.4	4.2	2.7
	LF	6.6	6.2	6.0	6.3
	Mean	2.8	3.0	3.4
	P-value for contrasts
	CON vs. LB				<0.01
	CON vs. LF				<0.01
Ethanol (g/kg DM)	CON	15.3	15.4	15.4	15.4	0.33	0.26	0.23	0.17
	LB	19.2	16.0	14.9	16.7
	LF	15.9	15.5	16.2	15.8
	Mean	16.8	15.6	15.5

Variable	Silage (S)	Storage periods (SP) (days)		Mean	SEM	P-value
Variable	Silage (S)	29	193	Mean	SEM	S	SP	S × SP
pH (after 24h*)	CON	3.66	3.50	3.57	0.001	0.08	<0.01	0.06
	LB	3.75	3.52	3.64
	LF	3.68	3.59	3.64
	Mean	3.69a**	3.54b
Water activity (after 24h*)	CON	0.9867	0.9800	0.9833	0.0002	0.02	0.02	0.18
	LB	0.9900	0.9833	0.9867¹
	LF	0.9900	0.9900	0.9900²
	Mean	0.9889a	0.9844b
Aerobic stability (h)	CON	37.7b	191.0a	114.3	36.27	<0.01	<0.01	<0.01
	LB	35.3b	194.0a	114.7
	LF	158.3b	>216a	187.2
	Mean	77.1	200.3
	P-value for contrasts
	CON vs. LB	0.96	0.95
	CON vs. LF	<0.01	0.10
Maximum temperature (MT) (°C)	CON	39.1	40.9	40.0	2.19	0.01	0.59	0.35
	LB	40.8	40.1	40.4³
	LF	34.2	30.7	32.5⁴
	Mean	38.0	37.3
Time to reach MT (h)	CON	214.7a	>216.0a	215.3	0.54	<0.01	<0.01	<0.01
	LB	215.3a	214.7a	215.0
	LF	215.3a	207.3b	211.3
	Mean	215.1	212.7
	P-value for contrasts
	CON vs. LB	0.82	0.48
	CON vs. LF	0.82	<0.01

	Silages	Fungi diversity	Population in different media‡ (Log CFU/g)
	Silages	Fungi diversity	DG18	PDA	DRBC
0	Fresh corn	Aureobasidium sp.	4.00	-	4.30
		Beauveria sp.	-	-	4.30
		Epicoccum sp.	4.00	-	-
		Fusarium sp.	4.85	-	4.30
		Pestalotiopsis sp.	-	6.30	-
		Phoma sp.	-	-	4.30
		Simplicillium sp.	4.00	-	-
29	LB	M.* pilosus	3.51	3.51	3.50
29	LB	Not identified	-	2.78	2.60
29 AE†	LB	M. pilosus	3.60	3.78	3.90
29 AE†	LB	Not identified	-	3.30	-
103	CON	A.** fumigatus	4.08	4.19	4.21
		A. flavus	2.78	2.60	-
		Rhizopus oryzae	-	2.30	-
	LB	A. fumigatus	4.45	4.29	4.27
		A. flavus	2.30	-	-
		A. niger	-	2.30	-
		A. micronesiensis	-	-	2.30
		R. oryzae	-	2.30	2.30
	LF	A. fumigatus	4.13	4.21	4.25
193	CON	A. cristatus	2.00	-	-
	CON	A. fumigatus	-	2.00	-
	LB	A. fumigatus	2.00	-	-
	LB	A. giganteus	2.47	-	2.00
193 AE	LB	A. giganteus	-	2.30	2.30

Fungi diversity	Access code*	Collection code**	Culture media
Fungi diversity	Access code*	Collection code**	DG18	PDA	DRBC
Before ensiling
Aureobasidium sp.	KX023301.1	CCDCA 11554, CCDCA 11559	1	0	1
Beauveria sp.	MH231203.1	CCDCA 11558	0	0	1
Epicoccum sp.	MK809036.1	CCDCA 11556	1	0	0
Fusarium sp.	MK841428.1	CCDCA 11555, CCDCA 11557	3	0	1
Pestalotiopsis sp.	MK862236.1		0	1	0
Phoma sp.	KR261531.1		0	0	1
Simplicillium sp.	MH990628.1		1	0	0
After ensiling
Aspergillus cristatus	MK696342.1		1	0	0
Aspergillus flavus	MH781287.1	CCDCA 11571, CCDCA 11582, CCDCA 11583	4	2	0
Aspergillus fumigatus	MH536091.1	CCDCA 11566, CCDCA 11567, CCDCA 11570, CCDCA 11572, CCDCA 11574, CCDCA 11578, CCDCA 11584 a 11586	42	42	43
Aspergillus giganteus	FR775339.1	CCDCA 11579, CCDCA 11580, CCDCA 11587 a 11590	4	5	3
Aspergillus micronesiensis	KP987047.1	CCDCA 11573, CCDCA 11575, CCDCA 11576	0	0	4
Aspergillus niger	MH208814.1	CCDCA 11568	0	1	0
Monascus pilosus	AB607170.1	CCDCA 11560 a 11565	8	12	10
Rhizopus oryzae	KX035094.1	CCDCA 11569	0	2	1
Not identified			0	4	2
Total isolates			65	69	67
Number of different species			9	8	10

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[1] *Corresponding author: carlaavila@ufla.br