Detection and Characterisation of Lactobacillus spp. in the Bovine Uterus and Their Influence on Bovine Endometrial Epithelial Cells In Vitro

Abstract

Bacterial infections and inflammation of the uterus are common in dairy cattle after parturition. In particular, pathogenic bacteria that cause endometritis have been the focus of research in cattle reproduction in the last ten years. The aim of the present study was to identify commensal lactobacilli in the bovine uterus and to examine their influence on the synthesis of pro-inflammatory factors in bovine endometrial epithelial cells in vitro. Lactobacillus species were isolated from healthy bovine uteri and further characterised. Bovine endometrial epithelial cells in the second passage (n = 5 animals) were co-cultured with the autochthonous isolates L. buchneri, L. ruminis and L. amylovorus as well as with a commercially available L. vaginalis in different multiplicities of infection (MOI = 1, 5 and 10, respectively). Endometrial epithelial cells cultured without bacteria served as controls. At distinct points in time (2, 4 and 6 h) total RNA was extracted from co-cultured epithelial cells and subjected to reverse transcription quantitative PCR of pro-inflammatory factors. Furthermore, the release of such factors by co-cultured epithelial cells was measured by ELISA or EIA after 24 and 48 h. L. ruminis and L. amylovorus induced increased interleukin (IL) IL1A, IL6, IL8 and prostaglandin-endoperoxide synthase 2 mRNA levels and the release of IL8 and prostaglandin F2α in endometrial epithelial cells compared with control cells. In contrast, L. buchneri did not significantly influence the expression and release of these factors. Toll-like receptors 2 and 6 transcripts were found unchanged in co-cultured and untreated epithelial cells in vitro. However, endometrial epithelial cells of each animal showed individual differences in the response to bacterial load. These results suggest that Lactobacillus species are present in the bovine uterus, revealing immunomodulatory properties.

Antiviral effects of Lactobacillus ruminis SPM0211 and Bifidobacterium longum SPM1205 and SPM1206 on rotavirus-infected Caco-2 cells and a neonatal mouse model

“…Recently, some researchers isolated L. ruminis strains from various species and reported the character of them (Forde et al, 2011;Gärtner et al, 2015;O'Donnell et al, 2015). However, there was no report about the antiviral activity of L. ruminis.…”

Detection and characterization of Lactobacillus spp. in the porcine seminal plasma and their influence on boar semen quality

“…An aliquot (2 mL) of each ejaculate was frozen with 400 μl glycerol, transported to the laboratory in liquid nitrogen, and stored at -80°C until microbial analyses. Samples were cultured aerobically or anaerobically on Rogosa SL agar and LBS agar (both Sigma-Aldrich, Steinheim, Germany) at 37°C for up to 72 h to detect lactobacilli as described previously [ 27 ]. All isolates were stored at -80°C in MRS broth containing 15% (v/v) glycerol.…”

“…Only colonies with a negative catalase reaction were used for culturing in MRS broth. For a first screening, a PCR was carried out using a Lactobacillus -specific primer pair [ 37 ] (for: ; rev: ; synthesized by Eurofins MWG, Ebersberg, Germany) based on the 16S ribosomal ribonucleic acid (16S rRNA) gene as previously described [ 27 ]. The forward and reverse sequences of the PCR products were obtained by a commercial sequence service (GATC Biotech, Konstanz, Germany) followed by comparing the resulting sequences with the NCBI database to identify the specific bacterial species.…”

“…buchneri (now registered as L . buchneri DSM 32407) up to a multiplicity of infection (MOI) of 10 did not affect the viability of epithelial cells [ 27 ]. In addition, the mRNA expression or release of pro-inflammatory factors was not influenced for up to 6 h and 48 h, respectively.…”

Nachweis von Weissella spp. in Viertelgemelksproben von zwei Milchkühen mit klinischer Mastitis

“…und Weissella (W.) spp. ließen sich natürlich vorkommend aus dem bovinen Euter isolieren (4,13,33). Dass andererseits einzelne Lakto-, Aero-und Enterokokkenspezies überdies Infektionen der Milchdrüse bedingen können, wurde bereits beschrieben (28,32).…”