Advanced platelet-rich fibrin (A-PRF) induces more proliferation and migration of fibroblasts compared with standard PRF, but it being freshly prepared prior to it being applied is necessary. Therefore, this study aimed to determine the effect of lyophilized A-PRF on growth factor release and cell biological activity. Blood samples were collected from six dogs and processed for fresh and lyophilized A-PRF. The growth factors released included transforming growth factor beta-1 (TGF-β1), vascular endothelial growth factor-A (VEGFA), and platelet-derived growth factor-BB (PDGF-BB), and the fibroblast proliferation as well as wound closure enhancement of both products were compared. The results showed that TGF-β1, PDGF-BB, and VEGFA were continually released from lyophilized A-PRF for over 72 h. Lyophilized A-PRF released significantly more accumulated VEGEA and a tendency to release more TGF-β1 at 72 h as well as VEGFA at 24 h and 72 h than fresh A-PRF. Moreover, lyophilized A-PRF increased fibroblast proliferation and induced a significantly faster wound closure than the control, while no significant difference between fresh and lyophilized A-PRF was found. In conclusion, the lyophilization of canine A-PRF can preserve the release of growth factors and has similar biological activities to a fresh preparation. This encourages the substitution of lyophilized A-PRF instead of fresh A-PRF in regenerative treatments in which the stability of the product is concerned.

Background: Vibrio species are common foodborne pathogens that cause gastrointestinal tract inflammation. Multidrug resistance (MDR) in Vibrio spp. is a global health concern, especially in aquaculture systems and food chain systems. This study aimed to detect Vibrio contamination in food collected from 14 markets in Nakhon Si Thammarat, Thailand, and determine their antibiotic susceptibility. Methods: One hundred and thirty-six food samples were investigated for Vibrio contamination. All isolates were tested for antibiogram and biofilm-forming ability. Moreover, the ceftazidime or cefotaxime resistance isolates were additionally investigated for extended-spectrum β-lactamase (ESBL) producers. The isolates were additionally examined for the presence of antibiotic resistance genes. The ESBL-suspected isolates with moderate-to-high biofilm-forming ability were further analyzed for their whole genome. Results: The prevalence of Vibrio contamination in food samples was 42.65%, with V. parahaemolyticus demonstrating the highest prevalence. Most isolates were resistant to β-lactam antibiotics, followed by aminoglycosides. The overall MDR of isolated Vibrio was 18.29%, with an average multiple antibiotic resistance (MAR) index of 16.41%. Most isolates were found to have β-lactam resistance-related genes (bla_TEM) for 41.46%, followed by aminoglycoside resistance genes (aac(6′)-Ib) for 18.29%. Most Vibrio showed moderate to strong biofilm-forming ability, particularly in MDR isolates (92.86%). Two ESBL-suspected isolates, one V. parahaemolyticus isolate and one V. navarrensis, were sequenced. Interestingly, V. parahaemolyticus was an ESBL producer that harbored the bla_CTX-M-55 gene located in the mobile genetic element region. While V. navarrensis was not ESBL producer, this isolate carried the bla_AmpC gene in the region of horizontal gene transfer event. Remarkably, the Inoviridae sp. DNA integration event was present in two Vibrio genomes. Conclusions: These findings impact the understanding of antibiotic-resistant Vibrio spp. in food samples, which could be applied for implementing control measures in aquaculture farming and food safety plans.

Background and Aim: Bacillus cereus and Staphylococcus aureus cause foodborne intoxication in humans and animals. Pathogens can produce biofilms controlled by the quorum sensing system. The study aimed to investigate the antibacterial, antibiofilm, and anti-quorum sensing activities of Coffea canephora P. ex Fr. (Robusta coffee) extracts against B. cereus and S. aureus. Materials and Methods: Ethanol extracts of fruit peels and seeds of Robusta coffee were tested for antibacterial activity against B. cereus and S. aureus using a broth microdilution assay. Reduction of the biofilm formation and elimination of the viability of mature biofilm-grown cells of B. cereus and S. aureus were determined. Inhibition of quorum sensing activity in Chromobacterium violaceum by the extracts was investigated using the disk diffusion method and flask incubation assay. Results: Fresh fruit peel extract showed the strongest antibacterial activity against B. cereus and S. aureus with minimum inhibitory concentration (MIC) values of 2 and 4 mg/mL, respectively. However, the extracts did not inhibit Escherichia coli, avian pathogenic E. coli, and Pseudomonas aeruginosa at 8 mg/mL. Significant inhibition of biofilm formation at 1/2 × MIC of the fresh peel extract was detected in B. cereus (56.37%) and S. aureus (39.69 %), respectively. At 8 × MIC of the fresh peel extract, a significant elimination of the mature biofilm viability was detected in B. cereus (92.48%) and S. aureus (74.49%), respectively. The results showed that fresh and dried peel fruit extracts at 1/2 × MIC significantly reduced violacein production with the highest percentage inhibition ranging from 44.53 to 47.48% at 24 h (p ≤ 0.05). Conclusion: The results of the present study suggest the potential therapeutic benefits of Robusta coffee extracts in inhibiting the growth, biofilm, and quorum sensing of both B. cereus and S. aureus. The results put forward an alternative strategy to control the foodborne intoxications caused by both pathogens.

The increasing prevalence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli ( E. coli ) highlights the growing issue of multidrug resistance within the food processing chain. This study aimed to determine the prevalence, antimicrobial resistance profiles, molecular genetic structures, and genetic diversity of ESBL-producing E. coli isolated from pork, cutting boards, and knives. A total of 90 samples were collected from 10 local markets in Southern Thailand. ESBL-producing E. coli were identified in 9 samples (10%), with knife samples showing the highest contamination rate (5/90, 5.55%). The ESBL-producing E. coli isolates exhibited high-level resistance to ampicillin (100%), ceftriaxone (88.89%), and cefpodoxime (88.89%), followed by tetracycline (77.78%). Multidrug resistance was detected in 88.89% (8/9) of ESBL-producing E. coli isolates and 28.26% (13/46) of all E. coli isolates. All presumptive ESBL-producing isolates carried ESBL resistance genes (9/9, 100%), with bla _CTX-M-1 being the most prevalent (6/9, 66.67%). Additionally, the tetA gene was frequently detected in ESBL-producing E. coli (6/9, 66.67%) and in all E. coli isolates (16/46, 34.78%). Overall, E. coli O157:H7 was identified in 5 isolates (10.87%). The genetic relatedness analysis revealed that five ESBL-producing E. coli isolates were closely related to E. coli ATCC 23502. Our findings confirm a high prevalence of ESBL-producing E. coli carrying resistance genes in knife samples, underscoring the importance of proper sanitary handling practices to minimize microbial contamination in pork retail shops.

Platelet-rich fibrin (PRF), which has been shown to promote wound and bone regeneration, has demonstrated antimicrobial properties against periodontal pathogens. However, in veterinary medicine, no study has determined the antimicrobial effects of canine platelet-rich fibrin (cPRF). Therefore, this study aimed to determine the antimicrobial effect of cPRF against E. coli and S. pseudintermedius found in dogs’ wounds and against the standard strain S. aureus. Additionally, the mechanism of the existing antibacterial activity of cPRF, which involves the formation of reactive oxygen species (ROS), was tested. Blood samples from six dogs were processed for cPRF. The antimicrobial properties of three groups (growth control, cPRF, and drug control) were evaluated at 0.5, 4, 8, and 24 h using a time–kill assay. The killing mechanisms involving ROS were evaluated using horseradish peroxidase (HRP) to suppress ROS production in PRF (PRF-SR). Subsequently, tests for antimicrobial properties and ROS generation were compared to those of the growth control and cPRF groups. The results showed that cPRF had significant antimicrobial properties against E. coli but no antimicrobial properties against S. pseudintermedius. After the ROS suppression, PRF-SR did not show an antimicrobial property against E. coli. Moreover, cPRF-treated bacteria exhibited significantly greater intracellular ROS than PRF-SR. In conclusion, canine PRF showed an antimicrobial effect against E. coli, and its antibacterial mechanism was related to releasing ROS.