Characterization of an Enterococcus faecium strain in a murine mastitis model.

AIM: The aim of the study was to characterize phenotypically and genotypically Enterococcus faecium strains collected from bovine mastitis milk and to evaluate one of them for its virulence in a murine mastitis model. METHODS AND RESULTS: A total of five E. faecium isolates were collected from cows with subclinical mastitis. EF-7A showed resistance to antibiotics tested, it presented alpha haemolysin and did not present gelatinase activity. It yielded cyA, efafm and gelE1 genes and it could be characterized as a moderate biofilm producer. It was able to internalize in MAC-T cells and 1×108 colony forming unit ml-1 was able to establish an intramammary infection in mice. The strain could be recovered from liver, kidney and blood samples. RAPD profiles showed different bands with respect to the inoculated strain. Histopathology analyses showed different grades of polymorphonuclear neutrophils infiltration in mammary glands. CONCLUSION: This is the first report that studied E. faecium strain in a lactating mouse model of mastitis and showed that the experimental inoculation was able to stimulate an inflammatory response resulting in mastitis. Results contribute to a better understanding of intramammary infections caused by E. faecium. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation shows that mice represent a valuable model for the study of the mastitis pathogenesis caused by E. faecium considering the high costs of using cows for mastitis research.

In Vitro and In Vivo Cytogenotoxic Effects of Hot Aqueous Extract of Achyrocline satureioides (Lam.) DC.

In this work we extend the toxicological studies of hot aqueous extract of A. satureioides (As-HAE) evaluating cytotoxic and apoptotic effects on human peripheral blood mononuclear cells (PBMCs). We also determine genotoxic action of this extract in vivo. In addition, the extract was chemically characterized. Finally, we established a comparison with previous data of cold aqueous extract. The As-HAE induced cytotoxicity on PBMCs determined by trypan blue dye exclusion (CC50 = 653 µg/mL) and MTT (CC50 = 588 µg/mL) assays being more toxic than cold extract. However, As-HAE as well as cold extract did not induce apoptosis measured by Hoechst 33258 staining, TUNEL assay, and DNA fragmentation analysis. The in vivo micronucleus test showed that As-HAE exerted cytogenotoxic effects on bone marrow of mice, contrary to what was observed with cold extract. The chemical study of As-HAE allowed identifying the flavonoids found in cold extract: luteolin, quercetin, and 3-O-methylquercetin, but at higher concentrations. We suggest that toxic effects induced by As-HAE could be due to high concentrations of these flavonoids. Given that As-HAE is the most used in folkloric medicine, its administration should be controlled in order to prevent potential cell damage.

Polyphenols as possible bioprotectors against cytotoxicity and DNA damage induced by ochratoxin A.

The present study aimed to investigate the protective effects of luteolin (L), chlorogenic acid (ChlA) and caffeic acid (CafA) against cyto-genotoxic effects caused by OTA. Vero cells and rat lymphocytes were used and viability was measured by neutral red uptake, MTT and trypan blue dye exclusion method. L (50 and 100µg/mL), ChlA (100 and 200µg/mL) and CafA (10-50µg/mL) reduced the damage induced by OTA (10µg/mL) on both cells type shown a good protective effect. The comet and micronucleus tests in Balb/c mice were performed. ChlA (10mg/kg bw) reduced OTA (0.85mg/kg bw)-induced DNA damage on blood and bone marrow cells, CafA (10mg/kg bw) showed protective effect only in blood cells and luteolin (2.5mg/kg bw) failed to protect DNA integrity on cells. In conclusion, polyphenols tested reduced the toxicity caused by OTA on different target cells with good protective effect, being ChlA the compound that showed the best effects.