In Vivo Validation of a Nanostructured Electrospun Polycaprolactone Membrane Loaded with Gentamicin and Nano-Hydroxyapatite for the Treatment of Periodontitis.

The aim of this research was to validate the use of a gentamicin (GEN) and nano-hydroxiapatite (nHAP)-loaded polycaprolactone nanostructured membrane (NM) as an innovative, highly efficient, low-cost treatment for periodontitis. We conducted an in vivo study on Wistar rats, in which we induced periodontitis by placing silk ligatures around the first right and left upper molars. The subjects were divided into three groups; the first group received no periodontal treatment, the second group received open flap debridement, and the third group received open flap debridement, together with the positioning of the GEN and nHAP-loaded nanostructured membrane as a treatment. The extent of periodontal regeneration was assessed by the periodontal pocket depth, bleeding on probing, tooth mobility, dental plaque, microbiological analysis, concentration of MMP-8 in saliva, plasma levels of CRP, and histological analysis. The results showed that using open flap debridement with the NM is more efficient, and it significantly reduces the probing depth, extent of bleeding on probing, dental mobility, bacterial plaque, and pathogenic flora. The concentrations of MMP-8 and CRP decrease. The histological analysis demonstrated that NM leads to bone regeneration. Our study indicates that gentamicin and nano-hydroxyapatite embedded in the fiber of the biodegradable membranes might be a promising therapeutic option for periodontitis treatment.

The Role of Wild Boars in the Circulation of Tick-Borne Pathogens: The First Evidence of Rickettsia monacensis Presence.

Most wild mammals can serve as hosts both for tick-borne pathogens (TBPs) and for the ticks themselves. Among these, wild boars, due to their large body size, habitat and life span, show high exposure to ticks and TBPs. These species are now one of the widest-ranging mammals in the world, as well as the most widespread suid. Despite the fact that certain local populations have been decimated by African swine fever (ASF), wild boars are still considered overabundant in most parts of the world, including Europe. Altogether, their long-life expectancy, large home ranges including migration, feeding and social behaviors, wide distribution, overabundance and increased chances of interactions with livestock or humans make them suitable sentinel species for general health threats, such as antimicrobial-resistant microorganisms, pollution and ASF geographical distribution, as well as for the distribution and abundance of hard ticks and also for certain TBPs, such as Anaplasma phagocytophilum. The aim of this study was to evaluate the presence of rickettsial agents in wild boars from two counties in Romania. Among 203 blood samples of wild boars (Sus scrofa ssp. attila) collected during 3 (2019-2022) hunting seasons (September-February), 15 were found positive for tick-borne pathogen DNA. Six wild boars were positive for A. phagocytophilum DNA presence and nine for Rickettsia spp. The identified rickettsial species were R. monacensis (six) and R. helvetica (three). No animal was positive either for Borrelia spp., Ehrlichia spp. or Babesia spp. To the best of our knowledge, this is the first report of R. monacensis in European wild boars, thus adding the third species from the SFG Rickettsia, in the epidemiology of which this wild species may have a role as a reservoir host.

Milk Pathogens in Correlation with Inflammatory, Oxidative and Nitrosative Stress Markers in Goat Subclinical Mastitis.

Goat mastitis is still frequently diagnosed in dairy farms, with serious consequences on milk quality and composition. The aim of this study was to establish correlations between milk microorganisms and biochemical parameters in goats with no signs of clinical mastitis. Thus, 76 milk samples were collected from a dairy goat farm, Carpathian breed, followed by microbiological, molecular (16S rRNA sequencing) and somatic cells analysis, determination of lactate dehydrogenase (LDH), ß-glucuronidase, catalase (CAT), glutathione peroxidase (GPx) activity, total antioxidant capacity (TAC), nitric oxide (NO) and lipid peroxides (LPO) using spectrophotometry and the ELISA method for 8-hydroxy-deoxyguanosine (8-OHdG) as the oxidative DNA damage indicator. Samples positive for bacterial growth showed a significant (p < 0.05) increase in the number of somatic cells, LDH and ß-glucuronidase activity, as well as higher levels of CAT, GPx, NO, LPO and 8-OHdG compared with pathogen-free milk whereas TAC was lower in milk from an infected udder. These findings suggest that subclinical mastitis is associated with increased enzymatic activity and induction of oxidative stress. Nevertheless, changes in biochemical parameters tended to vary depending on the pathogen, the most notable mean values being observed overall in milk positive for Staphylococcus aureus.

In Vitro Antimicrobial Effect of Novel Electrospun Polylactic Acid/Hydroxyapatite Nanofibres Loaded with Doxycycline.

The present study aimed to assess the in vitro antimicrobial effects of a novel biomaterial containing polylactic acid (PLA), nano-hydroxyapatite (nano-HAP) and Doxycycline (Doxy) obtained by electrospinning and designed for the non-surgical periodontal treatment. The antimicrobial activity of two samples (test sample, PLA-HAP-Doxy7: 5% PLA, nano-HAP, 7% Doxy and control sample, PLA-HAP: 5% PLA, nano-HAP) against two periodontal pathogens­Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis­was assessed using the Kirby−Bauer Disk Diffusion Susceptibility Test and compared with the effect of four antibiotics used as adjuvants in periodontal therapy: Amoxicillin, Ampicillin, Doxy and Metronidazole. The test sample (embedded with Doxy) showed higher inhibitory effects than commonly used antibiotics used in the treatment of periodontitis, while the control sample showed no inhibitory effects. Moreover, significant differences were observed between the inhibition zones of the two samples (p < 0.05). The Doxy-loaded PLA nanofibres had an antimicrobial effect against the periodontal pathogens. Based on these results, the novel biomaterial could be a promising candidate as adjuvant for the non-surgical local treatment in periodontitis.

Antimicrobial Properties of Bacterial Cellulose Films Enriched with Bioactive Herbal Extracts Obtained by Microwave-Assisted Extraction.

The use of bacterial cellulose (BC) as scaffold for active biofilms is one of the most interesting applications, especially for the biomedical and food industries. However, there are currently few studies evaluating the potential of incorporating herbal extracts into various biomaterials, including BC. Thus, the aim of this study is to report a screening of the total phenolic content and antioxidant and antimicrobial activity of ethanolic extracts of oregano, rosemary, parsley, and lovage. At the same time, the bioactive potential of BC enriched with the four ethanolic extracts is described. Microwave-assisted extraction was used to extract bioactive compounds from the four selected herbs. The physical, mechanical, structural, and chemical properties of BC were also assessed. Next, BC was enriched with the extracts, and their effect against Escherichia coli, Staphylococcus aureus, and Candida albicans was evaluated. The results showed that the bioactivity of the herbs varied significantly, with rosemary extract being the most bioactive. The BC films possessed good mechanical properties, and a three-dimensional network fibrillar structure appropriate for ethanolic-extract incorporation. The BC samples enriched with rosemary extracts had the highest antibacterial activity against S. aureus, while E. coli. and C. albicans seemed to be resistant to all extracts, regardless of herbs.

Prevalence of Antimicrobial Resistant Bacteria from Conjunctival Flora in an Eye Infection Prone Breed (Saint Bernard).

The conjunctival bacterial resident and opportunistic flora of dogs may represent a major source of dissemination of pathogens throughout the environment or to other animals and humans. Nevertheless, contamination with bacteria from external sources is common. In this context, the study of the antimicrobial resistance (AMR) pattern may represent an indicator of multidrug resistant (MDR) strains exchange. The present study was focused on a single predisposed breed-Saint Bernard. The evaluated animals were healthy, but about half had a history of ocular disease/treatment. The swabs collected from conjunctival sacs were evaluated by conventional microbiological cultivation and antimicrobial susceptibility testing (AST). The most prevalent Gram-positive was Staphylococcus spp.; regardless of the history, while Gram-negative was Pseudomonas spp.; exclusively from dogs with a history of ocular disease/treatment. Other identified genera were represented by Bacillus, Streptococcus, Trueperella, Aeromonas and Neisseria. The obtained results suggest a possible association between the presence of mixed flora and a history of ocular disease/treatment. A high AMR was generally observed (90%) in all isolates, especially for kanamycin, doxycycline, chloramphenicol and penicillin. MDR was recorded in Staphylococcus spp. and Pseudomonas spp. This result together with a well-known zoonotic potential may suggest an exchange of these strains within animal human populations and the environment.

In Vitro Antibacterial Susceptibility of Different Pathogens to Thirty Nano-Polyoxometalates.

Due to their unique properties, nano-polyoxometalates (POMs) can be alternative chemotherapeutic agents instrumental in designing new antibiotics. In this research, we synthesized and characterized "smart" nanocompounds and validated their antibacterial effects in order to formulate and implement potential new drugs. We characterized thirty POMs in terms of antibacterial activity-structure relationship. The antibacterial effects of these compounds are directly dependent upon their structure and the type of bacterial strain tested. We identified three POMs that presented sound antibacterial activity against S. aureus, B. cereus, E. coli, S. enteritidis and P. aeruginosa strains. A newly synthesized compound K6[(VO)SiMo2W9O39]·11H2O (POM 7) presented antibacterial activity only against S. aureus (ATCC 6538P). Twelve POMs exerted antibacterial effects against both Gram-positive and Gram-negative strains. Only one POM (a cluster derivatized with organometallic fragments) exhibited a stronger effect compared to amoxicillin. New studies in terms of selectivity and specificity are required to clarify these extremely important aspects needed to be considered in drug design.

Efficacy of Natural Formulations in Bovine Mastitis Pathology: Alternative Solution to Antibiotic Treatment.

INTRODUCTION: Bovine mastitis is an inflammatory disease of the udder that causes important economic losses in the animal breeding and dairy product industries. Nowadays, the conventional livestock antibiotic treatments are slowly being replaced by alternative treatments. In this context, the main aim of this study was to evaluate the efficacy of natural products in alternative treatment of bovine mastitis. MATERIAL AND METHODS: Two natural formulations with previously suggested in vitro antimicrobial effect were tested in vivo on mastitic cows. Animals with a positive diagnosis for mastitis (n = 20) were divided into three treatment groups: two groups (n = 8) were administered formulations of propolis, alcoholic extracts of Brewers Gold and Perle hops, plum lichen, common mallow, marigold, absinthe wormwood, black poplar buds, lemon balm, and essential oils of oregano, lavender, and rosemary designated R4 and R7 (differing only in the latter being more concentrated) and one group (n = 4) a conventional antibiotic mixture. In vivo efficacy of treatments was evaluated by somatic cell and standard plate counts, the treatment being considered efficacious when both parameters were under the maximum limit. RESULTS: R7 was effective in the most cases, being therapeutically bactericidal in six out of eight cows, while R4 gave good results in three out of eight cows, and conventional antibiotics cured one out of four. CONCLUSION: These results suggest the possible therapeutic potential of these natural products in bovine mastitis.

Medicinal Plants Based Products Tested on Pathogens Isolated from Mastitis Milk.

Bovine mastitis a major disease that is commonly associated with bacterial infection. The common treatment is with antibiotics administered intramammary into infected quarters of the udder. The excessive use of antibiotics leads to multidrug resistance and associated risks for human health. In this context, the search for alternative drugs based on plants has become a priority in livestock medicine. These products have a low manufacturing cost and no reports of antimicrobial resistance to these have been documented. In this context, the main objective of this study was to determine the antimicrobial effect of extracts and products of several indigenous, or acclimatized plants on pathogens isolated from bovine mastitis. A total of eleven plant alcoholic extracts and eight plant-derived products were tested against 32 microorganisms isolated from milk. The obtained results have shown an inhibition of bacterial growth for all tested plants, with better results for Everniaprunastri, Artemisiaabsinthium, and Lavandulaangustifolia. Moreover, E.prunastri, Populus nigra, and L. angustifolia presented small averages of minimum inhibitory and bactericidal concentrations. Among the plant-derived products, three out of eight have shown a strong anti-microbial effect comparable with the effect of florfenicol and enrofloxacin, and better than individual plant extracts possibly due to synergism. These results suggest an important anti-microbial effect of these products on pathogens isolated from bovine mastitis with a possible applicability in this disease.