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. 2022 Jan 17;11(1):117.
doi: 10.3390/antibiotics11010117.

Effect of Cephalosporin Treatment on the Microbiota and Antibiotic Resistance Genes in Feces of Dairy Cows with Clinical Mastitis

Lei Dong  1   2   3 , Lu Meng  1   2 , Huimin Liu  1   2 , Haoming Wu  1   2 , Martine Schroyen  3 , Nan Zheng  1   2 , Jiaqi Wang  1   2
Affiliations

Effect of Cephalosporin Treatment on the Microbiota and Antibiotic Resistance Genes in Feces of Dairy Cows with Clinical Mastitis

Lei Dong et al. Antibiotics (Basel). .

Abstract

Antibiotics are frequently used to treat dairy cows with mastitis. However, the potential effects of β-lactam antibiotics, such as cephalosporins, on the fecal microbiome is unknown. The objective was to investigate the effects of ceftiofur and cefquinome on the fecal microbiota and antibiotic resistance genes of dairy cows with mastitis. The fecal samples were collected from 8 dairy cows at the following periods: the start day (Day 0), medication (Days 1, 2, and 3), withdrawal (Days 4, 6, 7, and 8), and recovery (Days 9, 11, 13, and 15). 16S rRNA gene sequencing was applied to explore the changes in microbiota, and qPCR was used to investigate the antibiotic resistance genes. The cephalosporin treatment significantly decreased the microbial diversity and richness, indicated by the decreased Shannon and Chao 1 indexes, respectively (p < 0.05). The relative abundance of Bacteroides, Bacteroidaceae, Bacteroidales, and Bacteroidia increased, and the relative abundance of Clostridia, Clostridiales, Ethanoligenens, and Clostridium IV decreased at the withdrawal period. The cephalosporin treatment increased the relative abundance of β-lactam resistance genes (blaTEM and cfxA) at the withdrawal period (p < 0.05). In conclusion, the cephalosporin treatment decreased the microbial diversity and richness at the medication period, and increased the relative abundance of two β-lactam resistance genes at the withdrawal period.

Keywords: antibiotic resistance genes; cephalosporin; dairy cow mastitis; fecal microbiota.

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Conflict of interest statement

The authors have no conflict of interest to declare.

Figures

Figure 1
Figure 1
The rarefaction analysis performed using mothur. The rarefaction curve based on the species diversity showed sufficient coverage for the sequences. The first letter of the number (A–H) represented the different cows. X0, X (X1, X2, X3), Y (Y1, Y3, Y4, Y5), and Z (Z1, Z3, Z5, Z7) indicate the periods Day 0, medication, withdrawal, and recovery, respectively.
Figure 2
Figure 2
(a) Chao 1 richness index of samples from four different periods. (b) Shannon diversity index of samples from four different periods. (c) Simpson diversity index of samples from four different periods. Different lowercase letters within each sampling group represent significantly different means (p < 0.05). X0, X, Y, and Z indicate the periods Day 0, medication, withdrawal, and recovery, respectively.
Figure 2
Figure 2
(a) Chao 1 richness index of samples from four different periods. (b) Shannon diversity index of samples from four different periods. (c) Simpson diversity index of samples from four different periods. Different lowercase letters within each sampling group represent significantly different means (p < 0.05). X0, X, Y, and Z indicate the periods Day 0, medication, withdrawal, and recovery, respectively.
Figure 3
Figure 3
Alterations in bacterial genera abundance in the feces at different periods. (a) Selected genera belonging to a different phylum with significantly different abundances in the feces. (b) Lefse analysis histogram; different colors represent different groups. The horizontal axis is the LDA score obtained after LDA analysis, and the vertical axis is the group of microorganisms with significant effects.
Figure 3
Figure 3
Alterations in bacterial genera abundance in the feces at different periods. (a) Selected genera belonging to a different phylum with significantly different abundances in the feces. (b) Lefse analysis histogram; different colors represent different groups. The horizontal axis is the LDA score obtained after LDA analysis, and the vertical axis is the group of microorganisms with significant effects.
Figure 4
Figure 4
Proportion of the antibiotic resistance genes (a) blaTEM and (b) cfxA compared with the 16S rRNA gene. Different lowercase letters suggest significantly different means (p < 0.05). X0, X, Y, and Z indicate the periods Day 0, medication, withdrawal, and recovery, respectively.
Figure 5
Figure 5
Timeline for fecal sampling. Sampling days are displayed above the black lines. The antibiotic treatments are noted at Days 1, 2, and 3. X0, X, Y, and Z indicate the periods Day 0, medication, withdrawal, and recovery, respectively.
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