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Background: In dogs, the most frequently reported mycosis associated with Aspergillus spp. are respiratory infections. Systemic aspergillosis is uncommon, with reported cases been associated with several Aspergillus species. Aspergillus terreus species complex are ubiquitous organisms, unfrequently associated with local or systemic disease in animals and humans, and treatment of osteomyelitis caused by this species is usually unfavorable., Case Presentation: This case report describes the case of a 5-year-old dog, referred to the Veterinary Hospital of the Faculty of Veterinary Medicine of the University of Lisbon, Portugal, with a history of lameness of the right thoracic limb. Radiographs and CT scan revealed two different lesions on right humerus and radio, which were biopsied. The samples collected were submitted to cytological and histopathological evaluation and bacterial and mycological culture. Environmental samples, including of the surgery room and of the biopsy needle were also evaluated for the presence of fungi. Regarding biopsy samples, bacterial culture was negative, but mycological analysis originated a pure culture of a fungal species later identified as Aspergillus terreus by Sanger sequencing. Results were compatible with histopathologic examination, which revealed periosteal reaction and invasion of hyphae elements. Also, mycological analysis of both environmental samples evaluated were negative. The virulence profile of the fungal isolate was phenotypically characterized using specific media, allowing to reveal its ability to produce several enzymes involved in its pathogenicity, namely lipase, hemolysin and DNAse, corresponding to a Virulence Index (V. Index.) of 0.43. The patient was submitted to itraconazole therapy for 8 weeks. After 3 weeks, the patient showed significant clinical improvement, and after 6 weeks no radiographic signs were observed., Conclusions: Antifungal therapy with itraconazole can contribute to the remission of canine infections promoted by Aspergillus terreus complex with a relevant V. Index., (© 2023. The Author(s).)

Introduction: The changing epidemiology of Clostridioides difficile reflects a well-established and intricate community transmission network. With rising numbers of reported community-acquired infections, recent studies tried to identify the role played by non-human reservoirs in the pathogen's transmission chain. This study aimed at describing the C. difficile strains circulating in canine and feline populations, and to evaluate their genetic overlap with human strains to assess the possibility of interspecies transmission., Methods: Fecal samples from dogs ( n = 335) and cats ( n = 140) were collected from two populations (group A and group B) in Portugal. C. difficile isolates were characterized for toxigenic profile and PCR-ribotyping. The presence of genetic determinants of antimicrobial resistance was assessed in all phenotypically resistant isolates. To evaluate the genetic overlap between companion animals and human isolates from Portugal, RT106 ( n = 42) and RT014/020 ( n = 41) strains from both sources were subjected to whole genome sequencing and integrated with previously sequenced RT106 ( n = 43) and RT014/020 ( n = 142) genomes from different countries. The genetic overlap was assessed based on core-single nucleotide polymorphism (SNP) using a threshold of 2 SNP., Results: The overall positivity rate for C. difficile was 26% (76/292) in group A and 18.6% (34/183) in group B. Toxigenic strains accounted for 50% (38/76) and 52.9% (18/34) of animal carriage rates, respectively. The most prevalent ribotypes (RT) were the toxigenic RT106 and RT014/020, and the non-toxigenic RT010 and RT009. Antimicrobial resistance was found for clindamycin (27.9%), metronidazole (17.1%) and moxifloxacin (12.4%), associated with the presence of the ermB gene, the pCD-METRO plasmid and point mutations in the gyrA gene, respectively. Both RT106 and RT014/020 genetic analysis revealed several clusters integrating isolates from animal and human sources, supporting the possibility of clonal interspecies transmission or a shared environmental contamination source., Discussion: This study shows that companion animals may constitute a source of infection of toxigenic and antimicrobial resistant human associated C. difficile isolates. Additionally, it contributes with important data on the genetic proximity between C. difficile isolates from both sources, adding new information to guide future work on the role of animal reservoirs in the establishment of community associated transmission networks and alerting for potential public health risk., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Alves, Castro, Pinto, Nunes, Pomba, Oliveira, Silveira, Gomes and Oleastro.)

Background: Periodontal disease (PD) in dogs is prompted by the establishment of a polymicrobial biofilm at the tooth surface and a subsequent host inflammatory response. Several strategies may be used for PD control, including dental hygiene home care procedures, like toothbrushing, special diet and chew toys that reduce dental plaque accumulation, or professional periodontal treatments. Aiming at PD control, a biogel composed by nisin and guar-gum was previously developed. This work aimed to establish an in vitro model mimicking the PD-associated biofilms and to evaluate the nisin-biogel inhibitory activity against this polymicrobial biofilm by determining its Minimum Biofilm Inhibitory (MBIC) and Eradication Concentrations (MBEC). Bacterial species tested included Neisseria zoodegmatis CCUG 52598T, Corynebacterium canis CCUG 58627T, Porphyromonas cangingivalis DSMZ VPB 4874, Peptostreptococcus canis CCUG 57081 and an Enterococcus faecalis isolate belonging to a collection of oral bacteria obtained from dogs with PD. Before establishing the biofilm, coaggregation between species was determined by optical density measurement after 2 and 24 hours. Nisin-biogel MBIC and MBEC values regarding the polymicrobial biofilm were determined using a modified version of the Calgary biofilm pin lid device, after confirming the presence of the five bacterial species by Fluorescent In Situ Hybridization., Results: Only 40% of the bacterial dual suspensions were able to coaggregate at 2 hours, but all species tested exhibited a coaggregation percentage higher than 30% at 24 hours. It was possible to establish a 48 h polymicrobial biofilm model composed by the five bacterial species selected. This model was used to determine nisin-biogel MBIC (26.39 ± 5.89 µg/mL) and MBEC (62.5 ± 27.73 µg/mL) values., Conclusions: Our results showed that the nisin-biogel can inhibit and eradicate PD multispecies biofilms. As this in vitro model mimics an in vivo periodontal polymicrobial biofilm, our results reinforce the potential of the application of nisin-biogel for canine PD control.

Periodontal disease (PD) is one of the most frequent and widespread inflammatory diseases in dogs. The onset of PD pathogenesis is prompted by the dental microbial biofilm combined with the animal immune-inflammatory response. Along with local effects, systemic consequences can occur secondary to dental plaque-associated bacteraemia, affecting distant tissues and organs. The main goal of this retrospective study was to access and evaluate the association between PD and systemic consequences (renal, hepatic and cardiac) in a group of 136 dogs. Clinical records of all animals were assessed for general and systemic information, further analysed by general linear model and Odds-Ratio. Animals were organized in two groups, PD group and control group, formed by animals without PD. In this population, increasing age was proven to be a risk factor (OR = 1.04, p < .01) for PD establishment. Small breeds (<10 Kg) were the most prevalent in the PD group, being more susceptible to this disease. On the other hand, no influence of gender or reproductive status in PD progression was observed. Regarding systemic diseases, a statistically significant association (p = .026) was obtained between PD and cardiac disease. Results show that PD can have a significant adverse impact on animals' health, being related with systemic consequences, which may increase morbidity and mortality rates of these animals. PD prevention, with focus on owner's information about this disease and its management, are essential points for an active PD control program., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Periodontal disease (PD) is caused by the development of a microbial biofilm (dental plaque) in the periodontium, affecting approximately 80% of dogs. Several bacterial species present in the canine oral cavity can be implicated in the development of this disease, including Enterococcus spp. To decrease antibiotic administration, a possible control strategy for dog's enterococcal PD may involve the use of the antimicrobial peptide (AMP) nisin. Nisin's inhibitory activity was evaluated against a collection of previously characterized enterococci obtained from the oral cavity of dogs with PD (n = 20), as well as the potential of a guar-gum gel and a veterinary toothpaste as topical delivery systems for this AMP. The Minimum Inhibitory (MIC) and Bactericidal Concentrations (MBC) and the Minimum Biofilm Eradication (MBEC) and Inhibitory Concentrations (MBIC) were determined for nisin and for the supplemented guar-gum gel. For the supplemented veterinary toothpaste an agar-well diffusion assay was used to evaluate its inhibitory potential., Results: Nisin was effective against all isolates. Independently of being or not incorporated in the guar-gum gel, its inhibitory activity on biofilms was higher, with MBIC (12.46 ± 5.16 and 13.60 ± 4.31 μg/mL, respectively) and MBEC values (21.87 ± 11.33 and 42.34 ± 16.61 μg/mL) being lower than MIC (24.61 ± 4.64 and 14.90 ± 4.10 μg/mL) and MBC (63.09 ± 13.22 and 66.63 ± 19.55 μg/mL) values. The supplemented toothpaste was also effective, showing inhibitory activity against 95% of the isolates., Conclusions: The inhibitory ability of nisin when incorporated in the two delivery systems was maintained or increased, demonstrating the potential of these supplemented vehicles to be applied to PD control in dogs.

Background: Canine Distemper Virus (CDV) and Canine Parvovirus (CPV) lead to infections with high mortality rates in dogs. These viruses affect unvaccinated dogs or dogs with incomplete vaccination protocols. Vaccination plays an important role in reducing death rates, preventing clinical cases and controlling the spread of virus However, the efficacy of vaccination might be affected by different factors including vaccine scheduling and the neutralization of the vaccine targets by maternal antibodies. In face of these factors, the main goals of this study are (i) to investigate the antibody responses of puppies undergoing different primary vaccination protocols against CPV and CDV and (ii) to estimate the time until seroreversion in adult dogs unvaccinated for at least 3 years., Results: Antibody protection against CDV and CPV was evaluated in a total of 20 dogs: 5 puppies that initiated immunization at 6 weeks after birth (group A), 8 animals that started vaccination between 8 and 12 weeks of age (group B), and 7 adult dogs that have not been vaccinated for at least 3 years (group C). Blood samples were collected from each animal, with 3 to 4 weeks apart. Antibody responses were measured using indirect ELISA. In the second immunization point, no significant differences were found between the seroconversion of groups A and B for each viral infection (p = 0.81 and 0.20 for CDV and CPV, respectively). In the third immunization, there was evidence for a shorter time to achieve a protective titer against CPV in group B when compared to group A (p = 0.015). Similar evidence was not found for CDV (p-value = 0.41). In Group C, the average time until seroveversion was estimated at 2.86 years and 7.63 years for CDV and CPV, respectively., Conclusion: Vaccine response to CDV and CPV is specific in each individual. Effective immune protection in primary vaccination depends mainly on the initial titer of maternal antibodies acquired by the neonate. Other factors such as environmental exposure, immunization schedules and immune system activity influence the duration of immunity in adult dogs. The variability found reinforces the need to determine individual humoral immunity levels in order to assess vaccine efficacy.

Background: Most of surgical site infections (SSI) are caused by commensal and pathogenic agents from the patient's microbiota, which may include antibiotic resistant strains. Pre-surgical asepsis of the skin is one of the preventive measures performed to reduce SSI incidence and also antibiotic resistance dissemination. However, in veterinary medicine there is no agreement on which biocide is the most effective. The aim of this study was to evaluate the effectiveness of two pre-surgical skin asepsis protocols in dogs. A total of 46 animals were randomly assigned for an asepsis protocol with an aqueous solution of 7.5% povidone-iodine or with an alcoholic solution of 2% chlorhexidine. For each dog, two skin swab samples were collected at pre-asepsis and post-asepsis, for bacterial quantification by conventional techniques and isolation of methicillin-resistant species., Results: Most samples collected at the post-asepsis did not present bacterial growth, both for the animals subjected to the povidone-iodine (74%) or to the chlorhexidine (70%) protocols. In only 9% of the cases a significant bacterial logarithmic reduction was not observed, indicating possible resistance to these agents. Also, the logarithmic reduction of the bacterial quantification from pre- and post-asepsis time, was not statistically different for povidone-iodine (6.51 ± 1.94 log10) and chlorhexidine (6.46 ± 2.62 log10) protocol. From the 39% pre-asepsis swabs which showed bacterial growth in MRSA modified chromogenic agar medium, only one isolate was identified as Staphylococcus aureus and one as S. epidermidis. False positives were mainly other staphylococci species, as well as Enterobacteriaceae., Conclusions: Pre-surgical skin asepsis protocols with povidone-iodine or chlorhexidine showed similar efficacy in the elimination of methicillin resistant bacteria and preventing surgical site infections in dogs undergoing surgery.

Periodontal disease - PD - is one of the most widespread diseases in dogs, but the role of this odontogenic infection in the dissemination of pathogenic bacteria present in the oral mucosa to other animals or pet owners is understudied. Trying to unveil the putative pathogenicity of enterococci present in the gums of dogs diagnosed with PD, thirty-two animals were investigated during routine visits to a private veterinary clinic. Seventy-one enterococci were recovered and characterized regarding species, genomic variability, virulence traits, antimicrobial resistance and biofilm-forming ability. Isolates were mainly identified as Enterococcus faecalis, with the large majority (95%) being able to produce biofilm. Regarding antibiotic resistance, all dog-enterococci were susceptible to ampicillin, amoxicillin/clavulanate, gentamicin-120, imipenem and vancomycin; while distinct levels of resistance were observed for chloramphenicol (10%), erythromycin (20%), streptomycin-300 (35%) and tetracycline (95%). For virulence traits incidence levels of 35% were observed for β-hemolysis and 25% for cylA, 25% for gelatinase and 35% for gelE; 85% harbor efaAfs and ebpABC; while ace, agg and esp are present respectively in 50, 30 and 10% of the dog-enterococci; efaAfm and acm were detected in all the Enterococcus faecium. Overall, the widespread prevalence of PD in dogs, associated with the close contact between companion animals, other animals and humans, may act as source for the dissemination of opportunistic pathogenic bacteria. Hence, aforementioned data on virulence and resistance features, emphasizes the need for active surveillance measures, such as the diagnose of PD in companion animals during routine visits to the veterinary clinic., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

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Periodontal disease is a relevant oral disease in dogs and nisin–biogel has been previously proposed to be used in its control. Enterococci, as inhabitants of the oral cavity with a high genetic versatility, are a reliable bacterial model for antimicrobial studies. Our goal was to evaluate the in vivo influence of the long-term dental application of the nisin–biogel on the virulence and antimicrobial signatures of canine oral enterococci. Twenty dogs were randomly allocated to one of two groups (treatment group—TG with nisin–biogel dental application, or control group—CG without treatment) and submitted to dental plaque sampling at day 0 and after 90 days (T90). Samples were processed for Enterococcus spp. isolation, quantification, identification, molecular typing and antimicrobial and virulence characterization. From a total of 140 enterococci, molecular typing allowed us to obtain 70 representative isolates, mostly identified as E. faecalis and E. faecium. No significant differences (p > 0.05) were observed in the virulence index of the isolates obtained from samples collected from the TG and CG at T90. At T90, a statistically significant difference (p = 0.0008) was observed in the antimicrobial resistance index between the isolates from the TC and CG. Oral enterococci were revealed to be reservoirs of high resistant and virulent phenotypes. [ABSTRACT FROM AUTHOR]

Periodontal disease (PD) is one of the most prevalent oral inflammatory diseases in dogs. PD onset begins with the formation of a polymicrobial biofilm (dental plaque) on the surface of the teeth, followed by a local host inflammatory response. To manage this disease, several procedures focusing on the prevention and control of dental plaque establishment, as well as on the prevention of local and systemic PD-related consequences, are essential. The removal of dental plaque and the inhibition of its formation can be achieved by a combination of dental hygiene homecare procedures including tooth brushing, the application of different oral products and the use of specific diet and chew toys, and regular professional periodontal procedures. Additionally, in some cases, periodontal surgery may be required to reduce PD progression. Associated with these measures, host modulation therapy, antimicrobial therapy, and other innovative therapeutic options may be useful in PD management. Moreover, PD high prevalence and its relation with potential local and systemic consequences reinforce the need for investment in the development of new preventive measures, treatments, and oral procedures to improve the control of this disease in dogs. Knowledge on the specific guidelines and diversity of the available products and procedures are fundamental to apply the most adequate treatment to each dog with PD. [ABSTRACT FROM AUTHOR]

Background: Periodontal disease (PD) is a highly prevalent inflammatory disease in dogs. This disease is initiated by a polymicrobial biofilm on the teeth surface, whose control includes its prevention and removal. Recently, it was shown that nisin displays antimicrobial activity against canine PD-related bacteria. Moreover, guar gum biogel has shown to be a promising topical delivery system for nisin. Methods: In this study we aimed to evaluate the antimicrobial activity of the nisin-biogel in the presence of canine saliva and after a 24-month storage, at different conditions, using a canine oral enterococci collection. We also studied the nisin-biogel cytotoxicity using a Vero cell line and canine primary intestinal fibroblasts. Results: The presence of saliva hampers nisin-biogel antimicrobial activity, and higher nisin concentrations were required for an effective activity. A significant reduction (p ≤ 0.05) in inhibitory activity was observed for nisin-biogel solutions stored at 37 °C, over a 24-month period, which was not observed with the other conditions. The nisin-biogel showed no cytotoxicity against the cells tested at concentrations up to 200 µg/mL. Conclusions: Our results confirmed the potential of the nisin-biogel for canine PD control, supporting the development of an in vivo clinical trial. [ABSTRACT FROM AUTHOR]