Antibacterial effects of chitosan-based hydrogels containing Trachyspermum ammi essential oil on pathogens isolated from dogs with otitis externa.

BACKGROUND: Growing antibiotic resistance has made treating otitis externa (OE) increasingly challenging. On the other hand, local antimicrobial treatments, especially those that combine essential oils (EOs) with nanoparticles, tend to be preferred over systemic ones. It was investigated whether Ajwain (Trachyspermum ammi) EO, combined with chitosan nanoparticles modified by cholesterol, could inhibit the growth of bacterial pathogens isolated from OE cases in dogs. In total, 57 dogs with clinical signs of OE were examined and bacteriologically tested. Hydrogels of Chitosan were synthesized by self-assembly and investigated. EO was extracted (Clevenger machine), and its ingredients were checked (GC-MS analysis) and encapsulated in chitosan-cholesterol nanoparticles. Disc-diffusion and broth Micro-dilution (MIC and MBC) examined its antimicrobial and therapeutic properties. RESULTS: Staphylococcus pseudintermedius (49.3%) was the most common bacteria isolated from OE cases, followed by Pseudomonas aeruginosa (14.7%), Escherichia coli (13.3%), Streptococcus canis (9.3%), Corynebacterium auriscanis (6.7%), Klebsiella pneumoniae (2.7%), Proteus mirabilis (2.7%), and Bacillus cereus (1.3%). The investigation into the antimicrobial properties of Ajwain EO encapsulated in chitosan nanoparticles revealed that it exhibited a more pronounced antimicrobial effect against the pathogens responsible for OE. CONCLUSIONS: Using chitosan nanoparticles encapsulated with EO presents an effective treatment approach for dogs with OE that conventional antimicrobial treatments have not cured. This approach not only enhances antibacterial effects but also reduces the required dosage of antimicrobials, potentially preventing the emergence of antimicrobial resistance.

Efficacy and safety of a hydrocortisone aceponate-containing ear spray solution in dogs with erythemato-ceruminous otitis externa: A randomised, multicentric, single-blinded, controlled trial.

BACKGROUND: Erythemato-ceruminous otitis externa (ECOE) is frequently seen in dogs affected with an allergic skin disease, with recurrent secondary bacteria and yeast overgrowths (detected on cytological examination). OBJECTIVES: The objective of the study was to compare the efficacy and safety of an ear spray containing only hydrocortisone aceponate glucocorticoid diester (HCA) to a control product (CTRL), an approved otic formulation containing prednisolone-miconazole-polymyxin combination, in dogs with ECOE. ANIMALS: In total, 97 and 104 dogs with ECOE were respectively randomly assigned to the tested ear treatment product group (HCA) or the commercially available ear treatment control product group (CTRL). MATERIALS AND METHODS: Dogs were treated for 7-14 days, as needed. At Day (D)0, D7, D14, D28 and D42, Otitis Index Score-3, hearing test, pruritus and pain visual analogue scales, and cytological scores were graded. The overall response to treatment also was assessed. RESULTS: All clinical parameters decreased rapidly and in a similar way without any significant difference at any time between treatment groups. A good-to-excellent response to treatment was seen in >90% of dogs of both groups as early as D14. The treatment was considered safe in all dogs. CONCLUSIONS AND CLINICAL RELEVANCE: A 7- to 14-day ear topical application of HCA alone to dogs with ECOE accompanied with bacterial and/or fungal (yeast) overgrowth was safe and led to no statistical difference in improvement of clinical scores relative to the CTRL combination. Based on these results, it may be necessary to reconsider the routine use of antimicrobial drugs such as antibiotics and antifungals as a first-line treatment for ECOE that is likely to have been caused by an allergic reaction.

Evaluation of plasma canine C-reactive protein concentrations in dogs with otitis media, healthy dogs and dogs with chronic otitis externa.

BACKGROUND: Otitis is characterised by inflammation of one or more of the structures of the ear. At present, to confirm or exclude otitis media (OM), it is most often necessary to perform a computed tomography (CT) scan or magnetic resonance imaging. Inflammation is an immune defence response found in many conditions that can be detected and tracked by measuring biological markers of inflammation as the Canine C-reactive protein (CRP). OBJECTIVES: The objective of this study was to determine whether CRP measurement is useful as an adjunctive diagnostic tool in dogs with otitis and whether elevated concentrations correlated with disease severity/presence of OM. ANIMALS: Twenty-four client-owned dogs were recruited over 1 year. MATERIALS AND METHODS: The dogs were divided into three groups: chronic or recurrent otitis externa (CO), otitis media (OM) and H (healthy). The dogs with otitis underwent a CT scan of the head, measurement of the plasma CRP concentration and evaluation of a 0-3 Otitis Index Score 3 (OTIS3 score). RESULTS: No dog (0%) in group H had an increased CRP value, compared to 20% in the CO group (one of five dogs) and 23% in the OM group (3 of 13 dogs). Plasma CRP concentrations show a statistically significant positive relationship with the OTIS3 score (p = 0.04). CONCLUSION AND CLINICAL RELEVANCE: Plasma CRP concentration is not reliable as a discriminatory tool in cases of otitis, although there is a trend for elevation in cases with more severe disease. However, a larger study may provide a statistically more reliable correlation between the severity of OM and CRP concentrations.

Bilateral pyogranulomatous otitis externa with putative cartilage destruction in a dog: A severe case of auricular chondritis?

Auricular chondritis of unknown cause was suspected in a 10-year-old male Bolognese dog with a five-month history of painful bilateral nodular and ulcerative pyogranulomatous dermatitis of the pinnae with putative auricular cartilage destruction. Pain and lesions resolved with immunosuppressive doses of prednisolone, yet the condition resulted in deformity of both pinnae and external canals.

Une chondrite auriculaire d'étiologie inconnue est suspectée chez un bichon bolonais mâle de 10 ans qui présente depuis 5 mois une dermatite pyogranulomateuse nodulaire et ulcéreuse bilatérale douloureuse du pavillon de l'oreille avec une destruction présumée du cartilage auriculaire. La douleur et les lésions disparaissent avec des doses immunosuppressives de prednisolone, mais l'affection entraîne une déformation des deux pavillons et des conduits auriculaires externes.

Suspeitou­se de condrite auricular de causa desconhecida em um cão macho Bolonhês de 10 anos de idade com um histórico de cinco meses de dermatite piogranulomatosa ulcerativa e nodular bilateral no pavilhão auricular com suposta destruição de cartilagem auricular. A dor e as lesões resolveram com doses imunossupressoras de prednisolona apesar de a etiologia ter resultado na deformidade de ambas as orelhas e condutos auditivos.

Se sospechó la existencia de una condritis auricular de causa desconocida en un perro boloñés de 10 años con historia de 5 meses de duración de una dermatitis nodular ulcerativa piogramulomatosa y bilateral en las orejas con posible destrucción del cartílago auricular. El dolor y las lesiones se resolvieron con dosis inmunosupresoras de prednisolona pero la enfermedad produjo deformación de ambas orejas y de los canales auriculares externos.

A pilot study of antimicrobial effects and ototoxicity of a Norway spruce (Picea abies) resin-based canine otic rinse product.

BACKGROUND: Norway spruce (Picea abies) resin-based products are used in human medicine. A resin-based otic rinse also could be useful in supportive care of canine otitis externa (COE), yet information on its antimicrobial effect against canine pathogens or ototoxicity is lacking. OBJECTIVES: To investigate the antimicrobial properties and ototoxicity of a commercial resin-based otic product. MATERIALS AND METHODS: Antimicrobial effect was evaluated using a standardised challenge test on Staphylococcus pseudintermedius, Corynebacterium auriscanis, Pseudomonas aeruginosa, Escherichia coli, Malassezia pachydermatis, and Streptococcus halichoeri strains to measure reduction in growth after 24 h exposure to the product. Effect on cell morphology was investigated by exposing S. pseudintermedius, C. auriscanis, P. aeruginosa and M. pachydermatis to the product in 20% and 100% (v/v) concentrations for 6, 24 and 48 h, and evaluating cells by transmission (TEM) and scanning (SEM) electron microscopy. An in vitro microbial kill-rate assay also was performed. Auditory brain stem response test, clinical evaluation and postmortem histological evaluation of ear canals were undertaken on experimental guinea pigs treated with the test product or saline controls. RESULTS: The product showed >log 5 growth reduction for all strains in the challenge test. TEM and SEM images showed clear changes in the cells' inner structures and deterioration of cells, and 100% (v/v) test product exposure induced microbial killing in 1-2 h. Ototoxicity was not detected in guinea pigs. CONCLUSIONS AND CLINICAL RELEVANCE: The product may be an option in supportive care of COE because of antimicrobial effects and lack of ototoxic properties in a guinea pig model.

Physicochemical properties of otic products for Canine Otitis Externa: comparative analysis of marketed products.

BACKGROUND: Otitis externa is a commonly diagnosed dermatological disorder in canines. The pathogens primarily involved in canine otitis externa (COE) include Staphylococcus pseudintermedius, Pseudomonas aeruginosa, Proteus mirabilis, and Malassezia pachydermatis. As COE tends to be superficial, medications delivered topically are often effective and practical in managing the condition. As such, there is a wide variety of approved topical products currently available in the market. The efficacy of topical dosage forms can be dependent on various factors such as the pharmacology of active constituents and the physicochemical properties of the formulation, including pH, viscosity, spreadability, and bio-adhesion. Currently, there is a lack of published literature available on the optimal properties of topical COE products. In this study, we compared the physicochemical properties of nine commercially available otic veterinarian products in Australia used clinically to manage COE. RESULTS: Based on our comparative analysis, the pH (6.26 ± 0.04) of an aqueous-based product was similar to a healthy dog's external auditory canal. Products containing polymers exhibited higher viscosity and bio-adhesion. Spreadability was inversely related to viscosity and Osurnia ® a product with high viscosity demonstrated the lowest spreadability. Aqueous-based otic products showed better syringebility whereas oil-based systems required higher force to expel the products. Variability in droplet size was noted. Derm Otic, Baytril Otic, and Aurizon Ear Drops had the lower standard deviation which indicates they would give a more consistent dose. CONCLUSIONS: Findings from this work provide considerations for industry researchers or formulation scientists working in the area of otic dosage formulations.

Evaluation of hypochlorous acid as an ear flush in dogs with chronic otitis externa.

BACKGROUND: Chronic otitis externa (OE) in dogs frequently requires anaesthetised ear flushing. OBJECTIVES: To evaluate hypochlorous acid as an ear flushing and antimicrobial agent in dogs with chronic OE. ANIMALS: Twenty dogs with chronic OE caused by the same organisms bilaterally. MATERIALS AND METHODS: One ear was flushed under anaesthesia with hypochlorous acid, the other with saline solution. Subsequently, the ear flushed with hypochlorous acid was cleaned with the same solution twice daily for 2 weeks, the other ear with a commercial ear cleaner. An ear medication containing miconazole, polymyxin B and prednisolone was used once daily in both ears. Clinical scores were determined before the flush. Ear cytological results were obtained, a hearing test was conducted before and after the ear flush, and a culture was taken directly after flushing. Ears were evaluated after 2 weeks of therapy. RESULTS: Yeast was present in the ears of 11, cocci in one and a mixed infection in eight dogs. Five ears were negative on culture after flushing with hypochlorous acid, one after the saline flush. Clinical and cytological scores decreased significantly with both solutions after 2 weeks of treatment. There was no difference between treatments in any of the scores at any time point between treatments and in the results of the hearing test before and after the flushing procedure. Adverse effects were not seen. CONCLUSIONS AND CLINICAL RELEVANCE: Hypochlorous acid is a suitable cleaning solution for canine OE.

Comparison of the quantity and antimicrobial activity of host defence peptides in ear canals between healthy and atopic dogs: A preliminary study.

BACKGROUND: Host defence peptides (HDPs) are involved in cutaneous immune defence. The secretion of HDPs in the ears of healthy normal and noninfected atopic dogs has not been measured. HYPOTHESES/OBJECTIVES: The aim of this study was to quantify the concentrations of ß-defensins (cBD3)-like and cathelicidin (cCath) HDPs in ears of healthy and atopic dogs without infectious otitis, additionally to evaluate the antimicrobial effect of the HDPs obtained. ANIMALS: Ten healthy and 20 atopic dogs with mild inflammatory, noninfectious otitis were included. MATERIALS AND METHODS: Absence of infection was determined by cytological evaluation, and the severity of clinical signs, if present, was assessed by a previously validated score (Otitis Externa Scoring System for Clinical Study, OTIS-3). The left ear canal of each dog was rinsed with 2 mL of sodium phosphate buffer. The solution obtained was analysed by enzyme-linked immunosorbent assay to quantify HDPs. Additionally, aural secretions were incubated with two concentrations (5 × 105 and 5 × 104 colony-forming units/mL) of Staphylococcus pseudintermedius and the bacterial density measured after 24 h of incubation. Data were statistically analysed. Significance was set as p ≤ 0.05. RESULTS: There was a significantly lower concentration of HDPs from atopic ears when compared with those from normal healthy dogs (cBD3-like: p = 0.0007; cCath: p = 0.049). There was minimal to variable antimicrobial activity in the aural secretions of both groups. CONCLUSIONS AND CLINICAL RELEVANCE: This study showed, for the first time, that the ear canals of atopic dogs with mild noninfectious otitis contain a lower concentration of cBD3-like and cCath HDPs than those of healthy dogs with normal ears. A consistent antimicrobial activity was not present in the aural secretions from either group.

Comparison between clinical evaluations and laboratory findings and the impact of biofilm on antimicrobial susceptibility in vitro in canine otitis externa.

BACKGROUND: In canine otitis externa (OE), biofilm-producing bacteria are frequently present but biofilm may be underdiagnosed clinically. HYPOTHESIS/OBJECTIVES: The study aimed to investigate an association between clinical and cytological findings with bacteriological data from dogs with OE, to establish, through Environmental Scanning Electron Microscope (ESEM) examination, whether the presence of biofilm in vivo can be predicted and to evaluate the impact of biofilm on antimicrobial susceptibility tests. MATERIALS AND METHODS: Fifty-six dogs showing clinical signs of OE were enrolled. One cotton swab each was collected for ESEM, bacterial culture and susceptibility testing and for cytology. Staphylococcus pseudintermedius (n = 42, 48.8%) and Pseudomonas aeruginosa (n = 26, 30.2%) were tested for their ability to form biofilm. Minimum Inhibitory Concentrations (MIC), Minimal Biofilm Inhibitory Concentrations (MBIC) and Minimal Biofilm Eradication Concentrations (MBEC) towards enrofloxacin, gentamicin, polymyxin B and rifampicin were determined. RESULTS: Pseudomonas aeruginosa was positively associated with the biofilm clinical evaluation (p < 0.01) and neutrophils (p < 0.05), nuclear streaks (p < 0.01) and rods bacteria (p < 0.01) on cytology. S. pseudintermedius was associated with a low presence of neutrophils. There was a statistical correlation between clinical and cytological biofilm presence (p ≤ 0.01), but none with the biofilm production assay nor ESEM biofilm detection. No differences were found comparing the results of MIC and MBIC. MBEC results showed higher values than MIC and MBIC for all antimicrobials tested (p ≤ 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: Biofilm presence in OE was often underdiagnosed. Even if there is no specific clinical or cytological pattern related to biofilm, its presence should always be suspected.

The stability and in vitro antibacterial efficacy of enrofloxacin and gentamicin solutions against Staphylococcus pseudintermedius over 28 days.

BACKGROUND: Many clinicians prepare compounded otic solutions to treat otitis externa (OE). Research evaluating the stability and antimicrobial efficacy of these solutions is limited. HYPOTHESIS/OBJECTIVES: This study determined the chemical stability and in vitro bactericidal efficacy of compounded solutions of enrofloxacin and gentamicin during storage for 28 days. MATERIALS AND METHODS: Solutions of enrofloxacin (10 mg/mL, 1%) and gentamicin (3 mg/mL, 0.3%) were prepared with normal saline and 1 mg/mL dexamethasone. Solutions were stored at room temperature (25°C) for 0, 14 and 28 days. The chemical stability of the antibiotics and dexamethasone were determined using liquid chromatography tandem mass spectrometry in triplicate. Efficacy assessment was made with 10 isolates of Staphylococcus pseudintermedius obtained from dogs with OE. Serial 10-fold dilutions of the bacteria with the compounded solutions were prepared and the colony count results were converted into colony-forming units (cfus). The mean cfu/mL and cfu/mL reduction rates were compared between Day (D)0, D14 and D28. All of the antimicrobial testing solutions were performed in triplicate. RESULTS: Chromatography showed that both antibiotics and dexamethasone were stable for 28 days. No significant differences were observed in the antibiotic bactericidal efficacy of stored solutions at D0, D14 or D28. CONCLUSIONS AND CLINICAL RELEVANCE: Solutions of 1% enrofloxacin and 0.3% gentamicin in normal saline with 0.1% dexamethasone maintained chemical stability and bactericidal efficacy over 28 days. These solutions can be considered as alternatives to commercial preparations for treatment of canine OE when indicated.

Lateral ear canal resection and bulla osteotomy with marsupialization to treat otitis media in rabbits: forty-eight procedures.

OBJECTIVE: Describe lateral ear canal resection and bulla osteotomy with marsupialization (LECARBOM) in rabbits with otitis media (OM), and report outcomes, complications, bacteria cultured from middle ears, and their antimicrobial susceptibility testing (AST) results. STUDY DESIGN: Retrospective clinical case series; single referral hospital. ANIMALS: Forty-two rabbits with naturally occurring OM. METHODS: Medical records (2011-2021) of rabbits with CT-confirmed OM undergoing surgery were reviewed for outcomes, complications, bacteria cultured from middle ears, and AST. RESULTS: Surgery was performed on 48 ears, and outcomes determined 21 days postoperatively. All rabbits survived the procedure. Otitis externa resolved in all lop-eared rabbits. Of 29 ears with OM-associated head tilt, eight (28%) resolved, nine (31%) improved, seven (24%) remained unchanged and five (17%) worsened. Outcome was not associated with head tilt duration nor OM severity. Postoperative complications occurred in 12 (25%) cases, seven of which had wound margin dehiscence that healed by 21 days. The most frequent bacteria isolated were Pasteurella multocida (16%), Bordetella bronchiseptica (14%) and Staphylococcus aureus (14%). Bacteria were sensitive to azithromycin, marbofloxacin or enrofloxacin and resistant to penicillin. CONCLUSION: LECARBOM is a well-tolerated surgical procedure to treat OM. It also improved or stabilized 83% of cases with head tilt and resolved all cases with otitis externa. CLINICAL SIGNIFICANCE: LECARBOM with administration of an AST-sensitive antibiotic, or azithromycin when no AST is performed, should be effective in rabbits with OM. Postsurgically, it resolves or alleviates most secondary conditions such as otitis externa, head tilt and facial nerve paralysis.

High rates of multidrug resistance in bacteria associated with small animal otitis: A study of cumulative microbiological culture and antimicrobial susceptibility.

The etiology of otitis in dogs and cats is multifactorial and complex, involving bacterial and fungal pathogens. As empiric antimicrobial prescription is a common practice when treating such cases, antimicrobial resistance may represent a complicating factor. The aim of this study was to describe microbiological features and susceptibility profiles of pathogens associated with 142 cases of external otitis, comprising 138 dogs and 4 cats.. The specimens were processed to identify bacterial and fungal etiologies following standard microbiological methods. Antimicrobial susceptibility was determined in vitro against 15 antibiotics and 3 antifungals. Further, Staphylococcus spp. isolates were screened for the detection of ß-lactamase enzymes using cefinase paper discs. Pseudomonas spp. and isolates from Enterobacteriaceae family were screened for colistin (Polymyxin E) resistance and for the mcr-1-mediated colistin resistance gene by PCR. The presence of mixed cultures of Enterobacteriaceae, Pseudomonas spp. and Staphylococcus spp., and co-infections with Malassezia spp., emphasizes the polymicrobial etiology of external otitis in small animals. Emerging rates of multidrug resistance observed in almost 50% of the isolates may alert for a near future of challenging veterinary cases unresponsive to first-line antimicrobials. In addition, these results highlight a potential public health concern of multidrug resistant bacteria, given the proximity of pets and their owners. This study addressed central aspects of external otitis, providing microbiologists and clinicians updated information on the etiology and treatment of challenging cases of multidrug resistant bacteria. It also provides priceless surveillance value in monitoring resistant bacteria in small animals.

Determination of amikacin stability at 1% and 3% concentrations in four topical solutions over a 56-day period.

BACKGROUND: Anecdotally, amikacin has been added to compounded topical preparations for the management of canine bacterial otitis externa. However, the stability of amikacin within these solutions is unknown. HYPOTHESIS/OBJECTIVES: The purpose of this study was to determine the stability of amikacin at 10 and 30 mg/mL concentrations in four topical solutions over a 56 day period. We hypothesised that amikacin would maintain chemical stability within the various solutions. METHODS AND MATERIALS: Amikacin was formulated to 10 and 30 mg/mL (1% and 3%) concentrations within four topical solutions: tris-EDTA (TrizEDTA Aqueous Flush) (TE); 0.15% chlorhexidine gluconate and tris-EDTA (TrizCHLOR Flush) (TC); 0.9% NaCl (NA); and 0.9% NaCl + 2 mg/mL dexamethasone (ND). Samples were made in duplicate and stored at room temperature (25°C) for 0, 7,14, 21, 28 and 56 days. Amikacin content was quantified, in triplicate, by ultrahigh-performance liquid chromatography tandem mass spectrometry. RESULTS: The recovered amikacin concentrations for the 10 mg/mL solutions ranged from 10 to 13.5 mg/mL (mean 11.5 mg/mL) with the exception of NA sample 2 at Day (D)0 (9.4 mg/mL) and D7 (9.2 mg/mL). The recovered amikacin concentrations for the 30 mg/mL solutions ranged from 30 to 40.2 mg/mL (mean 35.7 mg/mL). No significant difference was seen between the amikacin concentrations at D0 compared to D56 for all solutions except 10 mg/mL TE (P < 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: Amikacin maintained stability within TE, TC, NA and ND over 56 days except when formulated at 10 mg/mL within TE.

Effect of topical florfenicol, terbinafine and mometasone furoate ear drops on adrenocortical function in healthy small breed dogs with bilateral otitis externa.

BACKGROUND: Testing for hyperadrenocorticism is commonly pursued in adult dogs with dermatological disease, and adrenocortical suppression has been well-documented following the use of topical corticosteroids in otic preparations. An otic suspension that contains florfenicol, terbinafine and mometasone furoate, and lasts for 30 days after a single application, frequently is used to treat canine otitis externa (OE). This medication was shown to cause adrenocortical suppression on Day (D)2 postadministration and two weeks after two applications two weeks apart. HYPOTHESIS/OBJECTIVES: The objective of this study was to determine if topical florfenicol/terbinafine/mometasone furoate causes adrenocortical suppression in healthy, small-breed dogs with bilateral OE at D28 postapplication. ANIMAL: Seven client-owned dogs weighing <10 kg diagnosed with non-Pseudomonas bilateral OE. MATERIALS AND METHODS: Cortisol was measured pre- and post-adrenocorticotropic (ACTH) stimulation on D0. Topical florfenicol/terbinafine/mometasone furoate was applied in both ears. Dogs were reassessed on D28, and cortisol was measured pre- and post-ACTH stimulation. RESULTS: The median pre- and post-ACTH cortisol concentrations on D28 were 2.5 µg/dL (range 2.0-5.0 µg/dL) and 14.3 µg/dL (range 11.5-23.1 µg/dL), respectively. There was no significant difference (P = 0.85) between post-ACTH cortisol concentrations from D0 to D28. CONCLUSIONS AND CLINICAL RELEVANCE: Results demonstrated no evidence of adrenocortical suppression, suggesting that there is no need to delay adrenocortical function testing in dogs treated with topical florfenicol/terbinafine/mometasone furoate when applied as per the manufacturer's recommendations.

Effect of an ear cleaner instillation containing lipacids in a model of re-acidification of the external auditory canal in dogs.

BACKGROUND: In humans, the acidic pH of the ear canal plays a protective role against infection and a change towards alkalinity of the external auditory canal (EAC) is a local factor in the progression of acute to chronic otitis externa (OE). The use of acidic preparations alone for treatment of OE without concurrent antibiotic use is well-documented in humans. In dogs, only one study has investigated the EAC pH in healthy dogs and in dogs with OE, and investigations to understand the role of EAC pH in the pathogenesis of canine OE are lacking. HYPOTHESIS/OBJECTIVES: To obtain physiological EAC pH values in beagle dogs. To develop a model of re-acidification of the EAC in dogs and to investigate how an acidic solution may accelerate the return to a physiological pH. ANIMALS: Ten healthy beagle dogs in a laboratory setting. MATERIALS AND METHODS: A model of re-acidification of the EAC was developed by instillation of a pH 10.1 phosphate-buffered saline (PBS) solution and the subsequent acidic effect of an ear cleaner containing lipacids was evaluated in this model. RESULTS: Mean physiological EAC pH was 6.12 (± 0.36). EAC re-acidification took up to 9 h in this model. Mean pH values dropped immediately to 6.38 (± 0.27) on ears treated with an acidic ear cleaner. No abrupt drop was observed of the mean pH values for the control ears. CONCLUSION AND CLINICAL IMPORTANCE: This study confirms that physiological EAC pH in dogs is acidic. This model of re-acidification of the EAC pH allows investigations on acidic properties of topical ear products in healthy ears.

Contexte - Chez l'homme, le pH acide du conduit auditif joue un rôle protecteur contre l'infection et l'évolution vers l'alcalinité du conduit auditif externe (CAE) et est un facteur local de progression de l'otite externe (OE) aiguë à chronique. L'utilisation de préparations acides seules pour le traitement de l'OE sans utilisation concomitante d'antibiotiques est bien documentée chez l'homme. Chez les chiens, une seule étude a étudié le pH du CAE chez les chiens en bonne santé et chez les chiens atteints d'OE, et les recherches pour comprendre le rôle du pH du CAE dans la pathogenèse de l'OE canine font défaut. Hypothèses/objectifs - Obtenir des valeurs physiologiques de pH du CAE chez des chiens beagle. Développer un modèle de réacidification du CAE chez le chien et étudier comment une solution acide peut accélérer le retour à un pH physiologique. Animaux - Dix chiens beagle de laboratoire en bonne santé. Matériels et méthodes - Un modèle de réacidification du CAE a été développé par instillation d'une solution saline tamponnée (PBS) à pH 10,1 et l'effet acide ultérieur d'un nettoyant pour oreilles contenant des lipacides a été évalué dans ce modèle. Résultats - Le pH physiologique moyen du CAE était de 6,12 (± 0,36). La réacidification du CAE a pris jusqu'à 9 h dans ce modèle. Les valeurs moyennes du pH chutent immédiatement à 6,38 (± 0,27) sur les oreilles traitées avec un nettoyant auriculaire acide. Aucune chute brutale n'a été observée des valeurs moyennes de pH pour les oreilles témoins. Conclusion et importance clinique - Cette étude confirme que le pH physiologique du CAE chez le chien est acide. Ce modèle de réacidification du pH du CAE permet des investigations sur les propriétés acides des produits topiques auriculaires dans des oreilles saines.

Introducción; en humanos, el pH ácido del canal auditivo juega un papel protector contra la infección y un cambio hacia la alcalinidad del canal auditivo externo (EAC) es un factor local en la progresión de la otitis externa (OE) aguda a crónica. El uso de preparaciones ácidas solas para el tratamiento de la OE sin el uso concomitante de antibióticos está bien documentado en humanos. En perros, solo un estudio ha investigado el pH de EAC en perros sanos y en perros con OE, y faltan investigaciones para comprender el papel del pH de EAC en la patogenia de la OE canina. Hipótesis/objetivos - Obtener valores de pH fisiológico de EAC en perros beagle. Desarrollar un modelo de reacidificación del EAC en perros e investigar cómo una solución ácida puede acelerar el retorno a un pH fisiológico. Animales- diez perros beagle sanos en un laboratorio. Materiales y métodos- se desarrolló un modelo de reacidificación del EAC mediante la instilación de una solución salina tamponada con fosfato (PBS) de pH 10,1 y se evaluó en este modelo el efecto ácido subsiguiente de un limpiador de oídos que contenía lípidos. Resultados - El pH fisiológico medio del EAC fue de 6,12 (± 0,36). La reacidificación de EAC tomó hasta 9 h en este modelo. Los valores medios de pH cayeron inmediatamente a 6,38 (± 0,27) en los oídos tratados con un limpiador de oídos ácido. No se observó una caída abrupta de los valores medios de pH para los oíds de control. Conclusión e importancia clínica- este estudio confirma que el pH fisiológico de EAC en perros es ácido. Este modelo de reacidificación del pH de EAC permite realizar investigaciones sobre las propiedades ácidas de los productos tópicos para el oído en oídos sanos.

Contexto - Em humanos, o pH ácido do conduto auditivo exerce uma grande função protetora contra infecções e a alcalinização do conduto auditivo externo (CAE) é um fator local de progressão de otite externa (OE) aguda para crônica. A utilização unicamente de formulações ácidas para o tratamento de OE sem a utilização concomitante de antibióticos é bem documentada em humanos. Em cães, apenas um estudo investigou o pH do CAE em cães saudáveis com OE, e são escassas as pesquisas investigando o papel do pH do CAE na patogênese da OE. Hipótese/objetivos - Se obter os valores fisiológicos do pH dos CAEs de cães Beagle. Desenvolver um modelo de re-acidificação do CAE em cães e investigar como uma solução ácida pode acelerar o retorno ao pH fisiológico original. Animais - Dez cães Beagle saudáveis de laboratório. Materiais e Métodos - Um modelo de re-acidificação do CAE foi desenvolvido por instilação de solução salina tamponada com fosfato (PBS) com um pH 10,1 e o efeito acidificante subsequente de um limpador de ouvido contendo ácidos lipídicos foi avaliado neste modelo. Resultados - O pH fisiológico médio do CAE foi de 6,12 (± 0,36). A re-acidificação do CAE levou até 9h neste modelo. Os valores médios de pH caíram imediatamente para 6,38 (± 0,27) nas orelhas tratadas com um limpador otológico ácido. Não foi observada queda abrupta dos valores médios de pH para as orelhas controle. Conclusão e importância clínica - Este estudo confirmou que o pH fisiológico do CAE de cães é ácido. Este modelo de re-acidificação do pH do CAE permite investigações sobre as propriedades acidificantes de produtos otológicos tópicos para orelhas saudáveis.

Brachycephaly in French bulldogs and pugs is associated with narrow ear canals.

BACKGROUND: Brachycephalic dog breeds have multiple skull malformations which may lead to anatomical changes in the external auditory canal. It is our frequent observation that in the otoscopic examination of the external ear in these breeds we are unable to visualise the tympanic membrane as a consequence of extreme narrowing of the proximal ear canal. Additionally brachycephalic dogs reportedly are predisposed to otitis externa (OE) and otitis media. OBJECTIVES: To characterizse the transition of the cartilaginous ear canal to the bony meatus acusticus externus using computed tomography (CT) and to investigate a possible association with OE in brachycephalic dogs. MATERIALS AND METHODS: Seventy-five client-owned dogs [pugs (n = 20), French bulldogs (n = 55)] were included and assessed for OE using an owner questionnaire and otoscopic and cytological examinations. In dorsal plane CT scans, the diameter of the porus acusticus externus was measured using novel methodology. The results were compared with a normocephalic control group without preexisting otological disorders. RESULTS: Brachycephalic dogs had a significantly smaller porus acusticus externus diameter (2.6 mm) than normocephalic dogs (5.0 mm). Of the brachycephalic dogs, 32% had OE yet this was not statistically significantly related to the diameter of the porus acusticus externus. Middle ear effusion (44%) and narrowing of the external ear canal (82.6%) were significantly more frequent in brachycephalic dogs. Only five of 150 eardrums could be visualised otoscopically. CONCLUSIONS AND CLINICAL RELEVANCE: Malformation of the porus acusticus externus causes severe stenosis of the proximal ear canal in brachycephalic dogs. A connection between stenosis of the external auditory canal and OE could not be confirmed.

Contexte - Les races de chiens brachycéphales présentent de multiples malformations crâniennes qui peuvent entraîner des modifications anatomiques du conduit auditif externe. Nous observons fréquemment que lors de l'examen otoscopique de l'oreille externe chez ces races, nous sommes incapables de visualiser la membrane tympanique en raison d'un rétrécissement extrême du conduit auditif proximal. De plus, les chiens brachycéphales seraient prédisposés à l'otite externe (OE) et à l'otite moyenne. Objectifs - Caractériser la transition du conduit auditif cartilagineux au méat osseux externe par tomodensitométrie (TDM) et étudier une éventuelle association avec l'OE chez les chiens brachycéphales. Matériels et méthodes - Soixante-quinze chiens de propriétaires [carlins (n = 20), bouledogues français (n = 55)] ont été inclus et évalués pour l'OE à l'aide d'un questionnaire du propriétaire et d'examens otoscopiques et cytologiques. Dans les tomodensitogrammes du plan dorsal, le diamètre du porus acusticus externus a été mesuré à l'aide d'une nouvelle méthodologie. Les résultats ont été comparés à un groupe témoin normocéphale sans trouble otologique préexistant. Résultats - Les chiens brachycéphales avaient un diamètre de porus acusticus externus significativement plus petit (2,6 mm) que les chiens normocéphales (5,0 mm). Parmi les chiens brachycéphales, 32 % avaient une OE, mais cela n'était pas statistiquement lié de manière significative au diamètre du porus acusticus externus. L'épanchement de l'oreille moyenne (44 %) et le rétrécissement du conduit auditif externe (82,6 %) étaient significativement plus fréquents chez les chiens brachycéphales. Seuls cinq des 150 tympans ont pu être visualisés par otoscopie. Conclusions et pertinence clinique - La malformation du porus acusticus externus provoque une sténose sévère du conduit auditif proximal chez le chien brachycéphale. Un lien entre la sténose du conduit auditif externe et l'OE n'a pas pu être confirmé.

Contexto - Raças de cães braquicefálicos apresentam múltiplas malformações cranianas que podem levar a alterações anatômicas no conduto auditivo externo. Frequentemente, observamos que na avaliação otoscópica da orelha externa destas raças não conseguimos visualizar a membrana timpânica como uma consequência do estreitamento extremo do conduto auditivo proximal. Além disso, cães braquicefálicos são sabidamente predispostos à otite externa (OE) e otite média. Objetivos - Caracterizar a transição do conduto auditivo cartilaginoso para o meato acústico ósseo externo utilizando tomografia computadorizada (TC) e investigar uma possível associação com OE em cães braquicefálicos. Materiais e métodos - Setenta cães de clientes [pugs (n=20), buldogues franceses (n = 55)] foram inclusos e avaliados para OE utilizando um questionário para os proprietários, citologia e otoscopia. Nas TCs em plano dorsal, o diâmetro do poro acústico externo foi mensurado utilizando uma metodologia nova. Os resultados foram comparados com um grupo de cães normocefálicos controle sem alterações otológicas pré-existentes. Resultados - Os cães braquicefálicos apresentaram o diâmetro do poro acústico externo (2,6mm) significativamente menor que os cães normocefálicos (5,0mm). Dos cães braquicefálicos, 32% apresentavam OE, mas não houve relação significativa com o diâmetro do poro acústico externo. Presença de efusão na orelha média (44%) e estreitamento do conduto auditivo externo (82,6%) foram significativamente mais frequentes em cães braquicefálicos. Apenas cinco de 150 tímpanos puderam ser visualizados por otoscopia. Conclusões e relevância clínica - Malformações do poro acústico externo causa estenose grave do conduto auditivo proximal em cães braquicefálicos. Não foi possível comprovar a conexão entre a estenose do conduto auditivo externo e OE.

Introducción- las razas de perros braquicéfalos tienen múltiples malformaciones craneales que pueden provocar cambios anatómicos en el conducto auditivo externo. Con frecuencia observamos que en el examen otoscópico del oído externo en estas razas no podemos visualizar la membrana timpánica como consecuencia del estrechamiento extremo del conducto auditivo proximal. Además, según la literatura al respecto, los perros braquicéfalos están predispuestos a padecer otitis externa (OE) y otitis media. Objetivos - Caracterizar la transición del canal auditivo cartilaginoso al meato acusticus externus óseo mediante tomografía computerizada (CT) e investigar una posible asociación con OE en perros braquicefálicos. Materiales y métodos- setenta y cinco perros de propietarios particulares [carlinos (n = 20), bulldogs franceses (n = 55)] fueron incluidos y evaluados por posible OE mediante un cuestionario para propietarios y exámenes otoscópicos y citológicos. En tomografías computerizadas del plano dorsal, el diámetro del porus acusticus externus se midió utilizando una metodología novedosa. Los resultados se compararon con un grupo control normocefálico sin trastornos otológicos preexistentes. Resultados- los perros braquicefálicos tenían un diámetro del porus acusticus externus significativamente más pequeño (2,6 mm) que los perros normocefálicos (5,0 mm). De los perros braquicefálicos, el 32 % tenía OE, pero esto no se relacionó estadísticamente de manera significativa con el diámetro del porus acusticus externus. La efusión del oído medio (44 %) y el estrechamiento del conducto auditivo externo (82,6 %) fueron significativamente más frecuentes en perros braquicefálicos. Solo cinco de 150 tímpanos pudieron visualizarse por otoscopia. Conclusiones y relevancia clínica - La malformación del porus acusticus externus causa estenosis severa del canal auditivo proximal en perros braquicefálicos. No se pudo confirmar una conexión entre la estenosis del conducto auditivo externo y la OE.

Changes in the ear canal microbiota of dogs with otitis externa.

AIMS: Otitis externa (OE), one of the most common ear diseases in dogs, is caused by bacterial pathogens such as Staphylococcus sp. To understand the network of microbial communities in the canine ear canal affected with OE, we performed a cross-sectional study using next-generation sequencing. METHODS AND RESULTS: Ear swab samples were collected from 23 OE-affected and 10 healthy control dogs, and the 16S rRNA gene sequenced using Illumina MiSeq. The otic microbiota in the OE-affected dogs showed significantly decreased alpha diversity compared to controls. The community composition also differed in the affected group, with significantly higher relative abundance of the phylum Firmicutes and the genus Staphylococcus (P = 0·01 and 0·04 respectively). Contrary to our expectations, the severity of the disease did not impact the otic microbiota in OE-affected dogs. CONCLUSIONS: The ear canal microbiota of OE-affected dogs is distinct from that of healthy dogs, irrespective of disease status. SIGNIFICANCE AND IMPACT OF THE STUDY: This study, one of the few detailed analyses of the otic microbiota, can provide practical information for the appropriate treatment of canine OE.

Preliminary study of the stability of dexamethasone when added to commercial veterinary ear cleaners over a 90 day period.

BACKGROUND: Topical corticosteroids are commonly used in the management of allergic otitis externa to diminish inflammation. A common strategy is to make compounded solutions of dexamethasone in ear cleaner. HYPOTHESIS/OBJECTIVES: The objective of this study was to determine the stability of dexamethasone when added to four commercial ear cleaners (ec): designated ecA, ecB, ecC and ecD. METHODS AND MATERIALS: Two concentrations (0.1 and 0.25 mg/mL) of dexamethasone were formulated for each cleaner solution from a 2 mg/mL solution and stored in the original manufacturers' bottles at two temperatures: room (22 ËšC) and refrigerated (4 ËšC). Samples were evaluated in triplicate, using liquid chromatography-tandem mass spectrometry at 10 time points over 90 days. The mean and standard deviation were calculated for each time point. RESULTS: A solution was considered stable if the dexamethasone value remained >90% of the target concentration. All dexamethasone solution values were stable to 90 days, except two solutions for ecA; the 0.25 mg/mL dexamethasone concentration was only stable to 14 (4 ËšC) and 21 days (22 ËšC). CONCLUSIONS AND CLINICAL IMPORTANCE: These results provide preliminary evidence in support of pharmaceutical stability data for dexamethasone when included in the above compounded solutions at the noted concentrations and temperatures.

Effect of an anti-inflammatory pomegranate otic treatment on the clinical evolution and microbiota profile of dogs with otitis externa.

BACKGROUND: Canine otitis externa (OE) is a common disease characterised by inflammation of the epithelial tissue of the external ear canal. Secondary infections are frequent, and Malassezia pachydermatis and Staphylococcus pseudintermedius are routinely isolated and treated with antifungal and antibiotic compounds. HYPOTHESIS/OBJECTIVES: To analyse the otitis ear microbiome before and after a treatment with prednisolone plus pomegranate or antimicrobial drugs ANIMALS: 15 dogs with nonpurulent OE. METHODS AND MATERIALS: A 30 day, double-blinded, multicentre, randomized and controlled parallel-group (1:1) trial was conducted in 15 dogs with nonpurulent OE, following two different topical treatments (prednisolone plus pomegranate versus prednisolone plus antibiotic and antifungal drugs). On days (D)0, D15 and D30, serum and skin otic samples were collected, and clinical examination and microbiome analysis (bacteria and fungi) were performed. Results were compared with validated otitis clinical scores to assess the effectiveness of both treatments. RESULTS: Nine bacterial and four fungal families were detected during the three time-points tested. An increase in fungal diversity (Shannon index) and composition was the most significant change observed after both treatments. At treatment D15 and D30, the reduction in clinical signs was statistically significant in both treatment groups (P ≤ 0.05). Prednisolone plus pomegranate cleanser treatment was able to control the clinical signs of otitis as well as the bacterial and fungal overgrowth. CONCLUSIONS AND CLINICAL IMPORTANCE: Mild otitis cases associated with microbial overgrowth may be managed with topical antiseptic and anti-inflammatory agents without the need for antibiotic and/or antifungal compounds.

In vitro and in vivo evaluation of a new phytotherapic blend to treat acute externa otitis in dogs.

Canine otitis externa is frequently encountered in veterinary practice, caused by primary factors with bacteria and yeast overgrowth acting as secondary and perpetuating factors. The pharmacological support includes anti-inflammatory, antimicrobials, and antimycotic drugs, but therapeutic failure and antimicrobial resistance are leading to alternative strategies based on phytotherapic products. This study aimed to evaluate an essential oil blend (Otogen® ) to treat otitis externa in dogs. The experimental design was divided in: (a) an in vitro approach, based on the European Normative UNI EN 1275:2006, to assess the efficacy of the product against the most frequently isolated microorganisms during otitis externa. (b) an in vivo part, 12 owned dogs presenting with acute otitis externa were enrolled. A significant growth reduction (>99.9%) of Malassezia pachydermatis and Candida albicans after 15 min of contact and Pseudomonas aeruginosa after 1 h of incubation was recorded. For Staphylococcus pseudintermedius, 50% of growth reduction were appreciated after 15 min. Results obtained in vivo after 7 days of blend administration, noted a significant improvement of all the considered parameters (most important were head shaking, erythema, and scraping). The results obtained may support the usefulness of the tested phytotherapic blend to manage acute otitis externa in dogs.