Novel Biocement/Honey Composites for Bone Regenerative Medicine.

New biocements based on a powdered mixture of calcium phosphate/monetite (TTCPM) modified with the addition of honey were prepared by mixing the powder and honey liquid components at a non-cytotoxic concentration of honey (up to 10% (w/v)). The setting process of the cements was not affected by the addition of honey, and the setting time of ~4 min corresponded to the fast setting calcium phosphate cements (CPCs). The cement powder mixture was completely transformed into calcium-deficient nanohydroxyapatite after 24 h of hardening in a simulated body fluid, and the columnar growth of long, needle-like nanohydroxyapatite particles around the original calcium phosphate particles was observed in the honey cements. The compressive strength of the honey cements was reduced with the content of honey in the cement. Comparable antibacterial activities were found for the cements with honey solutions on Escherichia coli, but very low antibacterial activities were found for Staphylococcus aureus for all the cements. The enhanced antioxidant inhibitory activity of the composite extracts was verified. In vitro cytotoxicity testing verified the non-cytotoxic nature of the honey cement extracts, and the addition of honey promoted alkaline phosphatase activity, calcium deposit production, and the upregulation of osteogenic genes (osteopontin, osteocalcin, and osteonectin) by mesenchymal stem cells, demonstrating the positive synergistic effect of honey and CPCs on the bioactivity of cements that could be promising therapeutic candidates for the repair of bone defects.

Detection of Microbiota during the Fermentation Process of Wine in Relation to the Biogenic Amine Content.

Viticulture is one of the traditional industries in Slovakia, where there are six wine-growing regions: Malokarpatska, Southern Slovakia, Central Slovakia, Nitra, Eastern Slovakia, and Tokaj. This study focuses on the detection of microbiota in soil samples, grape leaves and berries, and samples taken from fermenting must and young wine (the variety Tramín cervený) in relation to the detected concentrations of biogenic amines during the fermentation process. In the examined samples, the number of yeasts and molds (from 3.8 to 6.8 log cfu/g or mL) and TVC (from 3.7 to 6.5 log cfu/g or mL) were determined via culture examination. At the same time, the number of LAB (from ˂3.0 to 4.4 log cfu/g or mL) was determined, which was the highest on day 4 of the must fermentation process and was related to the detected of the highest concentration of biogenic amines (histamine and tyramine) on day 6 in the investigated must samples using the UHPLC system. Mycobiota species were identified by MALDI-TOF MS, PCR, ITS-PCR-RFLP, and PCR sequencing of the amplified products. The study confirmed the presence of the yeasts Saccharomyces cerevisiae, Metschnikowia pulcherrima, Hanseniospora uvarum, Pichia kudriavzevii, Pichia kluyveri, Pichia fermentas, Torulaspora delbrueckii, and Candida tenuis. At the same time, the presence of molds (Cladosporium herbarum, Cladosporium cladosporioides, Penicillium granulatum, Penicillium mononematosum, Botritis cinerea, and Penicillium glabrum) was also confirmed in soil samples, leaves, grape berries, and fresh grape must. The study confirmed the reduction in the species diversity of the microbiota during the must fermentation process, which resulted in decreases in the concentrations of the monitored biogenic amines in the early stages of the must fermentation process and young wine of the variety Tramín cervený.

Detection of Resistant and Enterotoxigenic Strains of Staphylococcus warneri Isolated from Food of Animal Origin.

The topic of this work is the detection of antimicrobial resistance to Staphylococcus warneri strains and the genes encoding staphylococcal enterotoxins. It is considered a potential pathogen that can cause various-mostly inflammatory-diseases in immunosuppressed patients. The experimental part of the paper deals with the isolation of individual isolates from meat samples of Oryctolagus cuniculus, Oncorhynchus mykiss, Scomber scombrus, chicken thigh, beef thigh muscle, pork thigh muscle, and bryndza cheese. In total, 45 isolates were obtained and subjected to phenotypic (plasma coagulase activity, nuclease, pigment, hemolysis, lecithinase, and lipase production) and genotypic analyses to confirm the presence of the S. warneri species. The presence of genes encoding staphylococcal enterotoxins A (three isolates) and D (six isolates) was determined by PCR. Using the Miditech system, the minimum inhibitory concentration for various antibiotics or antibiotics combinations was determined, namely for ampicillin; ampicillin + sulbactam; oxacillin; cefoxitin; piperacillin + tazobactam; erythromycin; clindamycin; linezolid; rifampicin; gentamicin; teicoplanin; vancomycin; trimethoprim; chloramphenicol; tigecycline; moxifloxacin; ciprofloxacin; tetracycline; trimethoprim + sulfonamide; and nitrofurantoin. Resistance to ciprofloxacin and tetracycline was most common (73%). At the same time, out of a total of 45 isolates, 22% of the isolates were confirmed as multi-resistant. Isolates that showed phenotypic resistance to ß-lactam antibiotics were subjected to mecA gene detection by PCR.

Effects of vacuum packaging conditions on the quality, biochemical changes and the durability of ostrich meat.

In this study, the effects of vacuum packaging conditions on the concentrations of lactic acid, malondialdehyde, pH, microbial and sensory analysis were determined during chilled storage of ostrich meat. Meat was packed as follows: vacuum packed from 1st day (VP-1), vacuum packed from 3rd day (VP-3) and non-vacuum packed (NVP). Analysis were performed at 1st, 7th, 14th, 21st day after slaughter. Meat consisted of 74.69% water, 2.29% fats, 20.95% proteins. Package conditions had significant effect on the pH (NVP: 6.54 on the 14th day, VP-1: 6.05 and VP-3: 6.07 on the 21th day p<0.001), amount of malondialdehyde (NVP: 8.62mg/kg on the 14th day; VP-1: 1.95 and VP-3: 2.55 on the 21th day; p<0.001) and total microbial count (NVP: 7.4 log CFU/g on the 14th day; VP-1: 6.7 and VP-3: 6.8 on the 21th day p<0.01). Based on these results we can assess that vacuum packed from 1st day is necessary for the 21 days storage of ostrich meat.

Species identification and antimicrobial resistance of coagulase-negative staphylococci isolated from the meat of sea fish.

UNLABELLED: Seventy-eight isolates of staphylococci obtained from the meat of Theragra chalcogramma, Scomber scombrus, and Clupea harengus were identified and tested in this study. 16S rDNA sequence specific for the genus Staphylococcus was detected in all isolates with the help of PCR method. All of 78 isolates were coagulase-negative, and DNAse activity was only confirmed in 4 of them. The following species of staphylococci were identified using MALDI-TOF mass spectrometry: S. warneri (52%), S. epidermidis (33%), S. haemolyticus (6.4%), S. pasteuri (3.8%), S. sciuri (1.2%), S. capitis (1.2%), and S. hominis (1.2%). Antimicrobial resistance to 7 antibiotics was determined in each isolate with the help of agar dilution method. In general, resistance against ampicillin was observed in majority of isolates (87%). On the contrary, the best sensitivity of CoNS was determined to gentamicin (96%). Only 1 S. warneri strain showed resistance to cefoxitin. Furthermore, 83% of staphylococcal isolates were simultaneously resistant to 2 or more antibiotics. PRACTICAL APPLICATION: This study confirmed the need of monitoring antimicrobial resistance in coagulase-negative staphylococci not only in the meat of slaughter animals but also in retail marine fish. The results showed that MALDI-TOF mass spectrometry is useful, accurate, and rapid method for species identification of food pathogens including Staphylococcus spp.