Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells

<i>Lactococcus lactis</i> (<i>L. lactis</i>) is a pathogenic Gram-positive, catalase-negative coccobacillus (GPCN) associated with bovine mastitis. In this study, nine strains of <i>L. lactis</i> were successfully isolated and characterized from 457 milk samples f...

Full description

Saved in:
Bibliographic Details
Main Authors: Tiancheng Wang, Fan Wu, Tao Du, Xiaodan Jiang, Shuhong Liu, Yiru Cheng, Jianmin Hu
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Microorganisms
Subjects:
Online Access:https://www.mdpi.com/2076-2607/13/7/1674
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849418099275595776
author Tiancheng Wang
Fan Wu
Tao Du
Xiaodan Jiang
Shuhong Liu
Yiru Cheng
Jianmin Hu
author_facet Tiancheng Wang
Fan Wu
Tao Du
Xiaodan Jiang
Shuhong Liu
Yiru Cheng
Jianmin Hu
author_sort Tiancheng Wang
collection DOAJ
description <i>Lactococcus lactis</i> (<i>L. lactis</i>) is a pathogenic Gram-positive, catalase-negative coccobacillus (GPCN) associated with bovine mastitis. In this study, nine strains of <i>L. lactis</i> were successfully isolated and characterized from 457 milk samples from cows with clinical mastitis in China. All isolates exhibited a high degree of susceptibility to marbofloxacin and vancomycin. A series of molecular and cell biological techniques were used to explore the biological characteristics and pathogenicity of these isolates. The virulence gene profiles of the isolates were analyzed using whole genome resequencing combined with polymerase chain reaction (PCR) to elucidate the differences in virulence gene expression between isolates. To provide a more visual demonstration of the pathogenic effect of <i>L. lactis</i> on bovine mammary epithelial cells, an in vitro infection model was established using MAC-T cells. The results showed that <i>L. lactis</i> rapidly adhered to the surface of bovine mammary epithelial cells and significantly induced the release of lactate dehydrogenase, suggesting that the cell membranes might be damaged. Ultrastructural observations showed that <i>L. lactis</i> not only adhered to MAC-T cells, but also invaded the cells through a perforation mechanism, leading to a cascade of organelle damage, including mitochondrial swelling and ribosome detachment from the endoplasmic reticulum. The objective of this study was to provide strong evidence for the cytotoxic effects of <i>L. lactis</i> on bovine mammary epithelial cells. Based on this research, a prevention and treatment strategy for <i>L. lactis</i> as well as major pathogenic mastitis bacteria should be established, and there is a need for continuous monitoring.
format Article
id doaj-art-acc48a094abf47b4be93f64fbdc6300b
institution Kabale University
issn 2076-2607
language English
publishDate 2025-07-01
publisher MDPI AG
record_format Article
series Microorganisms
spelling doaj-art-acc48a094abf47b4be93f64fbdc6300b2025-08-20T03:32:32ZengMDPI AGMicroorganisms2076-26072025-07-01137167410.3390/microorganisms13071674Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T CellsTiancheng Wang0Fan Wu1Tao Du2Xiaodan Jiang3Shuhong Liu4Yiru Cheng5Jianmin Hu6College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, ChinaCollege of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China<i>Lactococcus lactis</i> (<i>L. lactis</i>) is a pathogenic Gram-positive, catalase-negative coccobacillus (GPCN) associated with bovine mastitis. In this study, nine strains of <i>L. lactis</i> were successfully isolated and characterized from 457 milk samples from cows with clinical mastitis in China. All isolates exhibited a high degree of susceptibility to marbofloxacin and vancomycin. A series of molecular and cell biological techniques were used to explore the biological characteristics and pathogenicity of these isolates. The virulence gene profiles of the isolates were analyzed using whole genome resequencing combined with polymerase chain reaction (PCR) to elucidate the differences in virulence gene expression between isolates. To provide a more visual demonstration of the pathogenic effect of <i>L. lactis</i> on bovine mammary epithelial cells, an in vitro infection model was established using MAC-T cells. The results showed that <i>L. lactis</i> rapidly adhered to the surface of bovine mammary epithelial cells and significantly induced the release of lactate dehydrogenase, suggesting that the cell membranes might be damaged. Ultrastructural observations showed that <i>L. lactis</i> not only adhered to MAC-T cells, but also invaded the cells through a perforation mechanism, leading to a cascade of organelle damage, including mitochondrial swelling and ribosome detachment from the endoplasmic reticulum. The objective of this study was to provide strong evidence for the cytotoxic effects of <i>L. lactis</i> on bovine mammary epithelial cells. Based on this research, a prevention and treatment strategy for <i>L. lactis</i> as well as major pathogenic mastitis bacteria should be established, and there is a need for continuous monitoring.https://www.mdpi.com/2076-2607/13/7/1674bovine clinical mastitis<i>Lactococcus lactis</i>antimicrobial resistancepathogenicityvirulence gene
spellingShingle Tiancheng Wang
Fan Wu
Tao Du
Xiaodan Jiang
Shuhong Liu
Yiru Cheng
Jianmin Hu
Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells
Microorganisms
bovine clinical mastitis
<i>Lactococcus lactis</i>
antimicrobial resistance
pathogenicity
virulence gene
title Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells
title_full Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells
title_fullStr Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells
title_full_unstemmed Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells
title_short Virulence, Antibiotic Resistance and Cytotoxic Effects of <i>Lactococcus lactis</i> Isolated from Chinese Cows with Clinical Mastitis on MAC-T Cells
title_sort virulence antibiotic resistance and cytotoxic effects of i lactococcus lactis i isolated from chinese cows with clinical mastitis on mac t cells
topic bovine clinical mastitis
<i>Lactococcus lactis</i>
antimicrobial resistance
pathogenicity
virulence gene
url https://www.mdpi.com/2076-2607/13/7/1674
work_keys_str_mv AT tianchengwang virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells
AT fanwu virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells
AT taodu virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells
AT xiaodanjiang virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells
AT shuhongliu virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells
AT yirucheng virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells
AT jianminhu virulenceantibioticresistanceandcytotoxiceffectsofilactococcuslactisiisolatedfromchinesecowswithclinicalmastitisonmactcells