Maternal amoxicillin affects piglets colon microbiota: microbial ecology and metabolomics in a gut model

Abstract

The first weeks of life represent a crucial stage for microbial colonization of the piglets’ gastrointestinal tract. Newborns’ microbiota is unstable and easily subject to changes under stimuli or insults. Nonetheless, the administration of antibiotics to the sow is still considered as common practice in intensive farming for pathological conditions in the postpartum. Therefore, transfer of antibiotic residues through milk may occurs, affecting the piglets’ colon microbiota. In this study, we aimed to extend the knowledge on antibiotic transfer through milk, employing an in vitro dedicated piglet colon model (MICODE—Multi Unit In vitro Colon Model). The authors’ focus was set on the shifts of the piglets’ microbiota composition microbiomics (16S r-DNA MiSeq and qPCR—quantitative polymerase chain reaction) and on the production of microbial metabolites (SPME GC/MS—solid phase micro-extraction gas chromatography/mass spectrometry) in response to milk with different concentrations of amoxicillin. The results showed an effective influence of amoxicillin in piglets’ microbiota and metabolites production; however, without altering the overall biodiversity. The scenario is that of a limitation of pathogens and opportunistic taxa, e.g., Staphylococcaceae and Enterobacteriaceae, but also a limitation of commensal dominant Lactobacillaceae, a reduction in commensal Ruminococcaceae and a depletion in beneficial Bifidobactericeae. Lastly, an incremental growth of resistant species, such as Enterococcaceae or Clostridiaceae, was observed. To the authors’ knowledge, this study is the first evaluating the impact of antibiotic residues towards the piglets’ colon microbiota in an in vitro model, opening the way to include such approach in a pipeline of experiments where a reduced number of animals for testing is employed. Key points • Piglet colon model to study antibiotic transfer through milk. • MICODE resulted a robust and versatile in vitro gut model. • Towards the “3Rs” Principles to replace, reduce and refine the use of animals used for scientific purposes (Directive 2010/63/UE). Graphical abstract

The effect of residual antibiotics in food on intestinal microbiota: a systematic review

“…In Nissen's study, the effect of milk with residual antibiotic amoxicillin (1%) on the microbiota of piglets was investigated with a bioreactor model. Receiving milk containing amoxicillin residues did not lead to significant changes in the piglets' intestinal microbiota (Nissen et al, 2022).…”

In vitro assessment of the effect of lactose‐free milk on colon microbiota of lactose‐intolerant adults

“…Enumeration of bacterial groups was made with DNA by qPCR to evidence changes in the microbiota after fermentation following previous protocols (Modesto et al ., 2011; Tanner et al ., 2014; Nissen et al ., 2021d; Tamargo et al ., 2022; Nissen et al ., 2022a; Nissen et al ., 2022b). The changes in the abundances of 6 bacterial targets ( Eubacteria , Firmicutes , Bacteroidetes , Lactobacillales , Bifidobacteriaceae and Enterobacteriaceae ) (Table S1) were assessed by qPCR on QuantStudio 5 System (Applied Biosystem, Thermo Fisher, Waltham, MA, USA).…”

Impact of cooking methods of red-skinned onion on metabolic transformation of phenolic compounds and gut microbiota changes

“…Colonic fermentations were conducted for 24 hours in independent vessels using an in vitro gut model, MICODE [8][9][10]26,27 , obtained by the assembly of Minibio Reactors (Applikon Biotechnology BV, Delft, NL) and controlled by Lucullus PIMS software (Applikon Biotechnology BV, NL). The preparation of the experiments was made according to published procedures [8][9][10]23,27,28 and recently described in details 9 .…”

“…Bacterial DNA was extracted just after sampling (Purelink Microbiome DNA Purification Kit by Invitrogen, Thermo Fisher Scientific, Carlsbad, CA, USA) and quantified (Biodrop, Biochrom Ltd., Cambridge, UK). Enumeration of bacterial groups was made by qPCR to evidence changes in the microbiota after fermentation [31][32][33][34] and expressed as cells/mL (obtained from values in gene copy numbers/mL multiplied by three for multicopy genes, as V3-V4 16S, and by one for monocopy genes, as RecA), following previous protocols [8][9][10]26,27,35 S1) were assessed by qPCR on a QuantStudio 5 System (Applied Biosystem, Thermo Fisher, USA) and SYBR Green I chemistry. The qPCR Prebiotic Index (qPI) was calculated as previously reported 9,10 .…”

“…Enumeration of the bacterial groups was done by qPCR to evidence the changes in the microbiota after fermentation [31][32][33][34] and expressed as cells per mL (obtained from values in gene copy numbers per mL multiplied by three for multicopy genes, as V3-V4 16S, and by one for monocopy genes, as RecA), following previous protocols. [8][9][10]26,27,35 Specifically, 14 different bacterial taxa (Eubacteria, Firmicutes, Bacteroidetes, Lactobacillales, Bifidobacteriaceae, Enterobacteriaceae, Clostridium group I, Clostridium group IV, Bacteroides-Prevotella-Porphyromonas (BPP) group, Atopobium-Collinsella-Eggerthella (ATOP) group, Bifidobacterium longum, Escherichia coli, Faecalibacterium prausnitzii, and Akkermansia muciniphila) (Table S1 †) were assessed by qPCR on a QuantStudio 5 System (Applied Biosystem, Thermo Fisher, USA) and SYBR Green I chemistry. The qPCR prebiotic index (qPI) was calculated as previously reported.…”