Department of Microbiology, Universidade Estadual de Londrina, C. Postal 10.011, Londrina, PR, 86057-970, Brazil.
Embrapa Soja, C. Postal 4006, Londrina, PR, 86081-981, Brazil.
Braz J Microbiol. 2022 Mar;53(1):267-280. doi: 10.1007/s42770-021-00649-2. Epub 2022 Jan 1.
The use of inoculants carrying diazotrophic and other plant growth-promoting bacteria plays an essential role in the Brazilian agriculture, with a growing use of microorganism-based bioproducts. However, in the last few years, some farmers have multiplied microorganisms in the farm, known as "on farm" production, including inoculants of Bradyrhizobium spp. for soybean (Glycine max L. Merrill.) and Azospirillum brasilense for corn (Zea mays L.) or co-inoculation in soybean. The objective was to assess the microbiological quality of such inoculants concerning the target microorganisms and contaminants. In the laboratory, 18 samples taken in five states were serial diluted and spread on culture media for obtaining pure and morphologically distinct colonies of bacteria, totaling 85 isolates. Molecular analysis based on partial sequencing of the 16S rRNA gene revealed 25 genera of which 44% harbor species potentially pathogenic to humans; only one of the isolates was identified as Azospirillum brasilense, whereas no isolate was identified as Bradyrhizobium. Among 34 isolates belonging to genera harboring species potentially pathogenic to humans, 12 had no resistance to antibiotics, six presented intrinsic resistance, and 18 presented non-intrinsic resistance to at least one antibiotic. One of the samples analyzed with a shotgun-based metagenomics approach to check for the microbial diversity showed several genera of microorganisms, mainly Acetobacter (~ 32% of sequences) but not the target microorganism. The samples of inoculants produced on farm were highly contaminated with non-target microorganisms, some of them carrying multiple resistances to antibiotics.
在巴西农业中,使用携带固氮菌和其他植物生长促进菌的接种剂起着至关重要的作用,微生物生物制品的使用也在不断增加。然而,在过去几年中,一些农民在农场中繁殖微生物,称为“农场内”生产,包括用于大豆(Glycine max L. Merrill.)的 Bradyrhizobium spp. 和用于玉米(Zea mays L.)的 Azospirillum brasilense 的接种剂,或在大豆中进行共同接种。本研究的目的是评估此类接种剂的微生物质量,包括目标微生物和污染物。在实验室中,从五个州采集了 18 个样本,进行连续稀释并在培养基于获得纯的和形态上不同的细菌菌落,共获得 85 个分离株。基于 16S rRNA 基因部分序列的分子分析显示了 25 个属,其中 44%的属含有对人类潜在致病的物种;仅鉴定出一个分离株为 Azospirillum brasilense,而没有分离株被鉴定为 Bradyrhizobium。在属于对人类潜在致病物种的 34 个分离株中,有 12 个对抗生素没有耐药性,6 个具有固有耐药性,18 个至少对一种抗生素具有非固有耐药性。用基于高通量测序的宏基因组学方法分析的一个样本用于检查微生物多样性,显示了几种属的微生物,主要是醋酸杆菌属(~32%的序列),但没有目标微生物。农场内生产的接种剂样本受到非目标微生物的严重污染,其中一些携带多种抗生素耐药性。