ERDRP-0519 inhibits feline coronavirus in vitro.

BACKGROUND: Coronaviruses (CoVs) are major human and animal pathogens and antiviral drugs are pursued as a complementary strategy, chiefly if vaccines are not available. Feline infectious peritonitis (FIP) is a fatal systemic disease of felids caused by FIP virus (FIPV), a virulent pathotype of feline enteric coronavirus (FeCoV). Some antiviral drugs active on FIPV have been identified, but they are not available in veterinary medicine. ERDRP-0519 (ERDRP) is a non-nucleoside inhibitor, targeting viral RNA polymerase, effective against morbilliviruses in vitro and in vivo. RESULTS: The antiviral efficacy of ERDRP against a type II FIPV was evaluated in vitro in Crandell Reese Feline Kidney (CRFK) cells. ERDRP significantly inhibited replication of FIPV in a dose-dependent manner. Viral infectivity was decreased by up to 3.00 logarithms in cell cultures whilst viral load, estimated by quantification of nucleic acids, was reduced by nearly 3.11 logaritms. CONCLUSIONS: These findings confirm that ERDRP is highly effective against a CoV. Experiments will be necessary to assess whether ERDRP is suitable for treatment of FIPV in vivo.

In vitro virucidal activity of mouthwashes on SARS-CoV-2.

OBJECTIVES: The objective of the study was to evaluate the in vitro virucidal activity of commercial mouthwashes against SARS-CoV-2 and variants of concern. MATERIALS AND METHODS: Antiviral activity was assessed at different time intervals, based on common use of these products by titrating residual viral infectivity on Vero E6 cells. RESULTS: All the mouthwashes were effective to reduce the infectious titers of SARS-CoV-2 and its tested variants. Mouthwashes Listerine® Cool Mint milder taste and Listerine® Cavity Protection milder taste reduced the infectious viral titer by up to 3.9 log10 after 30 s, while mouthwash Cetilsan® Sugar Free was able to reduce the viral titer by 2.2-2.9 log10 at all tested time intervals. Mouthwash Curasept® ADS DNA Intensive treatment was less effective to decrease viral infectivity (0.7-2.2 log10 TCID50/ml at all tested time intervals). Interestingly, the Gamma variant appeared more resistant to treatment in vitro with the different mouthwashes. CONCLUSIONS: In this study, we were able to assess the ability of different mouthwashes to in vitro decrease the infectivity of SARS-CoV-2 and its variants, and we observed that Gamma variant of concern was more resistant to treatment with mouthwashes.

Virucidal activity in vitro of mouthwashes against a feline coronavirus type II.

Transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can occur through saliva and aerosol droplets deriving from the upper aerodigestive tract during coughing, sneezing, talking, and even during oral inspection or dental procedures. The aim of this study was to assess in vitro virucidal activity of commercial and experimental mouthwashes against a feline coronavirus (FCoV) strain. Commercial and experimental (commercial-based products with addition of either sodium dodecyl sulfate (SDS) or thymus vulgaris essential oil (TEO) at different concentrations) mouthwashes were placed in contact with FCoV for different time intervals, that is, 30 s (T30), 60 s (T60), and 180 s (T180); subsequently, the virus was titrated on Crandell Reese Feline Kidney cells. An SDS-based commercial mouthwash reduced the viral load by 5 log10 tissue culture infectious dose (TCID)50 /50 µl at T30 while a cetylpyridinium (CPC)-based commercial mouthwash was able to reduce the viral titer of 4.75 log10 at T60. Furthermore, five experimental mouthwashes supplemented with SDS reduced the viral titer by 4.75-5 log10 according to a dose- (up to 4 mM) and time-dependent fashion.

Alveolar osteitis associated with methicillin-resistant Staphylococcus epidermidis.

A critical point in dentistry is the empiric prescription of broad-spectrum antibiotics that could increase the levels of antimicrobial resistance. Alveolar osteitis is one of the most common post-op- erative complications in which antibiotic use is controversial. A 35-year-old female, with pain in the right mandibular region and treated with cefixime, was diagnosed with cracked tooth syndrome and pulpitis. The tooth was extracted and a massive purulent bleeding drainage was observed. Irrigation of the socket and a new therapy with azithromycin were done. Bacteriological analysis, a specific mecA gene PCR for the methicillin resistance, and the antimicrobial susceptibility test were per- formed on the bacterial isolate. A Staphylococcus epidermidis isolate was methicillin-resistant and showed resistance to erythromycin, azithromycin, clarithromycin, and sulfamethoxazole + trimeth- oprim. After 7 days, intraoral examination showed a complete resolution. The aim of this report is to suggest that systemic antibiotics may provide insufficient efficacy during alveolar osteitis, especially when caused by a multidrug-resistant organism.

Dentinal tubule penetration following ultrasonic, sonic, and single-cone technique of a biosealer: An ex vivo study.

Background: The endodontic space is complex, and using a single-cone obturation technique fails to ensure a complete filling. Introduction: This study aimed to investigate the effect of ultrasonic activation, sonic activation, and single-cone technique of a biosealer on its dentinal tubular penetration. Materials and Methods: In the experiment, single-root mandibular premolars were randomly assigned to three groups (n = 20): group A, ultrasonic activation; Group B, sonic activation; and Group C, single-cone technique. Penetration of the fluorescently labeled biosealer was investigated using a confocal laser scanning microscope. Results: The data were statistically analyzed using Kruskal-Wallis and Mann-Whitney tests (P = 0.05). The highest penetration of biosealer was observed in Group A, followed by Group B (P < 0.05). Conclusions: Dentinal tubule penetration of biosealer was significantly improved by ultrasonic and sonic activation techniques.

Mechanism of surfactant interactions with feline coronavirus: A physical chemistry perspective.

HYPOTHESIS: Surfactants are inexpensive chemicals with promising applications in virus inactivation, particularly for enveloped viruses. Yet, the detailed mechanisms by which surfactants deactivate coronaviruses remain underexplored. This study delves into the virucidal mechanisms of various surfactants on Feline Coronavirus (FCoV) and their potential applications against more pathogenic coronaviruses. EXPERIMENTS: By integrating virucidal activity assays with fluorescence spectroscopy, dynamic light scattering and laser Doppler electrophoresis, alongside liposome permeability experiments, we have analyzed the effects of non-ionic and ionic surfactants on viral activity. FINDINGS: The non-ionic surfactant octaethylene glycol monodecyl ether (C10EO8) inactivates the virus by disrupting the lipid envelope, whereas ionic surfactants like Sodium Dodecyl Sulfate and Cetylpyridinium Chloride predominantly affect the spike proteins, with their impact on the viral membrane being hampered by kinetic and thermodynamic constraints. FCoV served as a safe model for studying virucidal activity, offering a faster alternative to traditional virucidal assays. The study demonstrates that physicochemical techniques can expedite the screening of virucidal compounds, contributing to the design of effective disinfectant formulations. Our results not only highlight the critical role of surfactant-virus interactions but also contribute to strategic advancements in public health measures for future pandemic containment and the ongoing challenge of antimicrobial resistance.

Symptomatic and asymptomatic apical periodontitis associated with red complex bacteria: clinical and microbiological evaluation.

In this study, the association of red complex (RC) bacteria that include Treponema denticola, Tannerella forsythia and Porphyromonas gingivalis with acute, exacerbated or chronic apical periodontitis was evaluated. Seventy-one patients with periapical disease were evaluated by clinical examination and microbiological samples obtained from the root canals were analyzed by a polymerase chain reaction assay. Twenty-one (29.6%) samples were positive for RC bacteria, with T. denticola, T. forsythia and P. gingivalis being detected in 14 (19.7%), 10 (14.1%) and 6 (8.5%) samples, respectively. RC bacteria were mainly associated with acute apical periodontitis (29.2%) and phoenix abscess (63.2%), while they were only sporadically detected (7.1%) in patients with chronic apical periodontitis. Generally, RC bacteria were associated with pain and a higher frequency of intracanalar/intrasulcular pus drainage. Involvement of RC bacteria in symptomatic periapical disease should be suspected in the presence of particularly severe clinical pain and pus drainage.

10-year Historical Prospective Cohort Study of Calcium Phosphate-Blasted Acid-Etched Titanium Implants Placed in Different Ridges.

Purpose: To evaluate the survival rate and marginal bone level (MBL) of calcium phosphate-blasted acid-etched titanium implants placed in a cohort of patients with different ridges after a follow-up period of at least 10 years. Materials and Methods: A total of 61 patients with a minimum implant follow-up of 10 years were selected for this historical prospective, best clinical practice cohort study. Between 2009 and 2012, 121 titanium implants were placed using a flap, flapless, or postextractive technique. The implant placement timing was performed according to pre-extractive diagnosis and divided into immediate (immediately after tooth extraction with an absence of infection), early (within 2 to 3 months with an acute periapical lesion), delayed (6 to 12 months from extraction with a large periapical infection), or late (> 12 months from extraction with healed edentulous ridges). All implants were loaded after 3 months with provisional and definitive cemented restorations. Periapical radiographs were taken before implant insertion and at 3, 6, 12, 24, 36, 48, 96, and 120 months (T3, T6, T12, T24, T36, T48, T96, and T120, respectively). The MBL was calculated in single blind by an additional examiner. Linear logistic regression was performed to analyze statistically significant differences in relation to different operative variables at all evaluation times. Multilevel mixed logistic regression was made to evaluate the factors associated to MBL at 10 years (T120). Results: After 10 years, 47 patients and 92 implant restorations were analyzed, showing that 88 implants (95.6%) survived and 4 implants (4.4%) failed. The cumulative drop-out rate was 22.1%. Loosening and/or mobility was observed in a total of 9 abutments (9.7%) during the observational time. No other complications were reported. Implants placed with a flapless technique revealed a similar MBL to those placed with a flap technique. No significant differences were observed between the surgical techniques at T96 and T120. Immediate and early implants revealed a more stable MBL than both delayed and late implants up to T48. At longer evaluation times (T96 and T120), the MBL values were not statistically significant (P > .05). Narrower diameter implants (3.5 mm) revealed a higher bone loss when compared to the 4.1-mm- and 5.0-mm-diameters, especially in the first year from implant insertion (from T3 to T12) and at longer follow-up (T36 and T48). After that, the difference was reduced. Multilevel analysis showed that none of these variables appear to significantly influence MBL at 120 months. Conclusions: MBL was not influenced by surgical technique or implant placement timing after 10 years. Maintaining a strict occlusal and hygiene control created the conditions to preserve bone integrity and achieve a high implant survival rate.

A One Health Perspective on Canine Coronavirus: A Wolf in Sheep's Clothing?

Canine coronavirus (CCoV) is a positive-strand RNA virus generally responsible for mild-to-severe gastroenteritis in dogs. In recent years, new CCoVs with acquired pathogenic characteristics have emerged, turning the spotlight on the evolutionary potential of CCoVs. To date, two genotypes are known, CCoV type I and CCoV type II, sharing up to 96% nucleotide identity in the genome but highly divergent in the spike gene. In 2009, the detection of a novel CCoV type II, which likely originated from a double recombination event with transmissible gastroenteritis virus (TGEV), led to the proposal of a new classification: CCoV type IIa, including classical CCoVs and CCoV type IIb, including TGEV-like CCoV. Recently, a virus strictly correlated to CCoV was isolated from children with pneumonia in Malaysia. The HuPn-2018 strain, classified as a novel canine-feline-like recombinant virus, is supposed to have jumped from dogs into people. A novel CoV of canine origin, HuCCoV_Z19Haiti, closely related to the Malaysian strain was also detected in a man with fever after travel to Haiti, suggesting that infection with Malaysian-like strains may occur. These data and the emergence of highly pathogenic CoVs in humans underscore the significant threat that CoV spillovers pose to humans and how we should mitigate this hazard.

Old and Novel Enteric Parvoviruses of Dogs.

Parvovirus infections have been well known for around 100 years in domestic carnivores. However, the use of molecular assays and metagenomic approaches for virus discovery and characterization has led to the detection of novel parvovirus species and/or variants in dogs. Although some evidence suggests that these emerging canine parvoviruses may act as primary causative agents or as synergistic pathogens in the diseases of domestic carnivores, several aspects regarding epidemiology and virus-host interaction remain to be elucidated.

Traditional and Recent Root Canal Irrigation Methods and Their Effectiveness: A Review.

According to contemporary dental standards, the primary goal of endodontic therapy is the chemo-mechanical cleaning of the complex root canal system. Watering root canals with approved solutions and activating them are essential parts of this operation. This review outlines various irrigant activation methods for root canal therapy. Specifically, a comparison among the methods of manual dynamic activation, sonics (subsonic, sonic, and ultrasonic), internal heating, and lasers, was conducted. The results in this work were gathered using Scopus, Web of Science, Google Scholar, and PubMed databases by searching the following keywords: sodium hypochlorite, cleaning, activation, and irrigation methods. The present work concluded that the use of irrigant activation has a greater benefit than its absence. Regardless, it is impossible to point to a single effective activation method.

RETRACTED: Iandolo et al. Traditional and Recent Root Canal Irrigation Methods and Their Effectiveness: A Review. Clin. Pract. 2023, 13, 1059-1072.

The Clinics and Practice Editorial Office retracts the article titled "Traditional and Recent Root Canal Irrigation Methods and Their Effectiveness: A Review" [...].

Bacterial Periodontitis in Horses: An Epidemiological Study in Southern Italy.

Equine periodontal disease (EPD) is a painful oral inflammatory syndrome characterized by multifactorial pathogenesis. Although it is well known that bacterial proliferation and consequent gingivitis are caused by the decomposition process of food residues, in hypsodont species, the pathogenetic role and the different bacterial species involved in the progression of EPD must be fully clarified. This study aimed to investigate the association of bacteria, including the complex red bacteria (RCB), with EPD, and to evaluate possible EPD risk factors. Bacterial species, including Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis (belonging to the RCB), Fusobacterium nucleatum, Veilonella parvula, and Prevotella intermedia, were investigated in 125 oral swabs from healthy and EPD-affected horses using real-time multiplex PCR. Subsequently, possible risk factors (i.e., age, gender, and breed of the animals and type of feed used) were evaluated using univariate and multivariate analyses. Tannerella spp. and Treponema spp. were detected in a significantly higher proportion of horses affected by EPD than in healthy animals, although pathogens belonging to RCB were detected in low number of horses. At the same time, none of the investigated pathogens was significantly associated with a particular stage of disease severity. Horses aged older than 20 years were at higher risk of EPD. The high rate of coinfection, statistically associated with EPD, supports the hypothesis that EPD is a complex syndrome characterized by the possible simultaneous involvement of several pathogens and an increased risk depending on the animal's age. Constant oral hygiene is the best prevention to prevent and treat the disease, especially in old animals.

Assessing Contact Time and Concentration of Thymus vulgaris Essential Oil on Antibacterial Efficacy In Vitro.

The overuse and misuse of antibiotics can pose the risk of spreading mutant strains that show antimicrobial resistance (AMR), with negative impacts on the management of bacterial infections and economic implications for healthcare systems. The research and development of natural antibacterial agents could be a priority in the next years to improve a number of effective antibacterial molecules and to reduce the AMR phenomenon and its development. The present study identified the most effective concentration and contact time of Thymus vulgaris L. essential oil (TEO) to obtain bactericidal effects in vitro against different Gram-positive and Gram-negative bacterial strains. Six clinically isolated (wild types) bacterial strains, (Citrobacter freundii, Enterococcus feciorum, Proteus mirabilis, Acinetobacter cioffi, Pseudomonas putrefaciens and Klebsiella pneumoniae) and two ATCCs (Staphylococcus aureus and Streptococcus mutans) were tested after 1 min, 3 min and 5 min of contact with TEO. The preliminary results on S. aureus after 24 h of incubation revealed a TEO concentration of 9.28 mg/mL (w/v) that completely inhibited bacteria growth, keeping cell viability. The total suppression of bacterial growth at all tested contact times was observed for all tested bacterial strains, and the results were confirmed after 48 h of incubation. Bacterial growth suppression was confirmed even with the presence of organic components. These preliminary results showed the in vitro antimicrobial efficacy of TEO against different Gram-positive and Gram-negative bacterial strains. Future studies are necessary to confirm the reproducibility of these results even on other strains and to define the exact molecular mechanisms of EOs in order to consider TEO as a valid alternative to classic antibiotic therapies and subsequently to reduce the occurrence of AMR.

Analysis of oral microbiota in non-vital teeth and clinically intact external surface from patients with severe periodontitis using Nanopore sequencing: a case study.

Periodontal diseases include a wide range of pathological conditions, damaging the supporting structures of the teeth. Origin and propagation of periodontal disease is believed to be caused by dysbiosis of the commensal oral microbiota. The aim of this study was to evaluate the presence of bacteria in the pulp cavity of teeth with severe periodontal disease with clinically intact external surface. Periodontal (P) and endodontic (E) tissue samples of root canals from six intact teeth of three patients were sampled for analysis of microbial population using Nanopore technology. Streptococcus was the predominant genus in E samples. Porphyromonas (33.4%, p = 0.047), Tannerella (41.7%, p = 0.042), and Treponema (50.0%, p = 0.0064) were significantly more present in P than in E samples. Some samples (E6 and E1) exhibited a remarkable difference in terms of microbial composition, whilst Streptococcus was a common signature in samples E2 to E5, all which were obtained from the same patient. In conclusion, bacteria were identified on both the root surface and the root canal system, thus demonstrating the possibility of bacteria to spread directly from the periodontal pocket to the root canal system even in the absence of crown's loss of integrity.

Endodontic Microbial Communities in Apical Periodontitis.

INTRODUCTION: Apical periodontitis (AP) represents an inflammatory condition of peri-radicular tissues due to invasion and colonization of bacteria in the root canals. Primary apical periodontitis (PAP) is associated with untreated necrotic root canal and can be efficiently treated with endodontic treatment to remove bacteria. Persistent/secondary apical periodontitis (SAP) is a perpetual periapical lesion due to unsuccessfully treated root canals after an initial apparent healing of the tooth. The aim of the study was evaluating the microbial communities associated with root canals using Nanopore sequencing. METHODS: Seventeen samples from the root canals of 15 patients with AP were Polymerase Chain Reaction-amplified for 16s ribosomal DNA gene and sequenced. Information regarding the presence or absence of AP symptoms, PAP and SAP, and periapical index of patients were recorded. RESULTS: Firmicutes, Bacteroidetes, and Actinobacteria were the most abundant phyla detected and Phocaeicola, Pseudomonas, Rothia, and Prevotella were the most prominent genera. In samples of patients with AP symptoms, the most frequent detected genera were Cutibacterium, Lactobacillus, Pseudomonas, Dialister, Prevotella, and Staphylococcus. In PAP samples, the most represented genera were Cutibacterium, Lactobacillus, Pseudomonas, and Prevotella, whilst in SAP cases were Cutibacterium, Prevotella, Atopobium, Capnocytophaga, Fusobacterium, Pseudomonas, Solobacterium, and Streptococcus. CONCLUSIONS: The results provide additional information on the microbiota of root-canals. These data evidence the complexity of the microbiota and the relationship with many clinical and endodontic conditions. Future studies must evaluate these conditions and identify their role in inducing bone damage and local and systemic disease, aiming to better elucidate the relationship between microbes and endodontic pathologies.

Intracellular persistence of Leishmania tarentolae in primary canine macrophage cells.

Leishmania tarentolae is a non-pathogenic species first isolated from geckoes in the Mediterranean basin. The finding that dogs test positive against both Leishmania infantum and L. tarentolae raises questions regarding the ability of the latter species to persist and adapt to new hosts. This study aimed to evaluate in vitro the capability of L. tarentolae to colonize, survive and persist in canine primary monocyte-derived mononuclear cells. Monocytes were isolated from dog whole blood samples and placed in 24-well plates for differentiation into macrophages and for incubation with L. tarentolae field-isolated strains (RI-325 and SF-178) and laboratory (LEM-124) strain; the parasite burden was assessed at different time points post-infection. The L. infantum laboratory strain (MON-1) was used as control. Infection parameters were evaluated by microscopy, counting the number of amastigotes/200 infected cells, and by duplex real-time PCR from supernatants and detached cells. Similar to L. infantum, L. tarentolae strains developed into round-shaped amastigote-like forms, with higher infection rates detected at 4 h followed by an overall decrease until 48 h. RI-325 presented also a higher infection rate at 72 h. Data showed that L. tarentolae strains infect and persist inside in vitro primary canine mononuclear cells, opening new perspectives for further laboratory studies.

Identification of new astroviruses in synanthropic squamates.

Astroviruses have been identified in a wide variety of animal species and are associated with gastro-intestinal disease in humans. Pathologies due to extra-intestinal localization are known in different hosts. We report the detection of astroviruses in synanthropic squamate reptile species (Podercis siculus and Tarentola mauritanica). Fecal samples were collected from 100 squamates from urban and peri-urban areas of three regions in South Italy and tested for the presence of astroviruses using a broadly reactive (pan-astrovirus) RT-PCR protocol targeting the RNA-dependent RNA polymerase. Astrovirus RNA was detected in 11% of the samples and for six strains a 3 kb-long fragment at the 3' end of the genome was sequenced, obtaining information on the complete capsid-encoding ORF2 sequence. Viral RNA was also detected in the brain of one of the positive animals. The sequences generated from the astrovirus strains shared low nucleotide identities in the ORF2 (< 43.7%) with other known reptilian astrovirus sequences, hinting to the massive genetic diversity of members of this viral family. Based on the partial RdRp gene of the sequenced strains, however, we observed species-specific patterns, regardless of the geographic origin of the animals, and we also identified a possible inter-species transmission event between geckoes and lizards.

Investigating the genetic diversity of CRESS DNA viruses in cats identifies a novel feline circovirus and unveils exposure of cats to canine circovirus.

Circular replication-associated protein (Rep)-encoding single stranded (CRESS) DNA viruses include Circoviruses which have been found in several animal species and in human specimens. Circoviruses are associated with severe disease in pigs and birds and with respiratory and gastrointestinal disorders and systemic disease in dogs. In cats there are only a few anecdotical studies reporting CRESS DNA viruses. In this study, a total of 530 samples (361 sera, 131 stools, and 38 respiratory swabs) from cats, were screened for the presence of CRESS DNA viruses. Overall, 48 (9.0%) of 530 samples tested positive using a pan-Rep PCR. A total of 30 Rep sequences were obtained. Ten sequences of fecal origin were tightly related to each other (82.4-100% nt identity) and more distantly related to mongoose circoviruses (68.3 to 77.2% nt identity). At genome level these circoviruses displayed the highest nt identity (74.3-78.7%) to mongoose circoviruses thus representing a novel circovirus species. Circoviruses from different animal hosts (n = 12) and from humans (n = 8) were also identified. However, six Rep sequences were obtained from serum samples, including canine circoviruses, a human cyclovirus and human and fish-associated CRESS DNA viruses. The presence of these viruses in the sera would imply, to various extent, virus replication in the animal host, able to sustain viremia. Overall, these findings indicate a wide genetic diversity of CRESS DNA viruses in cats and warrant further investigations.

Antibacterial and Biofilm Production Inhibition Activity of Thymus vulgaris L. Essential Oil against Salmonella spp. Isolates from Reptiles.

Salmonellosis is an infectious disease affecting both animals and humans. Antimicrobial resistant (AMR) and biofilm-producing Salmonella spp., frequently detected in reptiles (who can then act as asymptomatic carriers for warm-blooded animals), have developed resistance to biocides; this represents a warning for the emergence of biocide/antimicrobial cross-resistance. The aim of this study was to evaluate the efficacy of Thymus vulgaris L. essential oil (TEO) in inhibiting bacterial growth and biofilm production of Salmonella spp., which had been isolated from wild reptiles housed in a Zoo in Italy. The resistance profile against different classes of antibiotics showed that all the isolates were susceptible to the tested antibiotics, despite the presence of several AMR genes. All the isolates were also tested with aqueous solutions of TEO at different dilutions (5% to 0.039%). Interestingly, TEO proved effective both in inhibiting bacterial growth at low dilutions, with MIC and MBC values ranging between 0.078% and 0.312%, and in inhibiting biofilm production, with values ranging from 0.039% to 0.156%. TEO demonstrated effective bioactivity against the biofilm producer Salmonella spp., proving to be a valid disinfectant for the prevention of salmonellosis from reptiles, a possible source of infection for humans exposed to the reptiles' environment.