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.

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.

A Pandora's box in feline medicine: presenting signs and surgical outcomes in 58 previously hoarded cats with chronic otitis media-interna.

OBJECTIVES: The aim of the present study was to report clinical findings, surgical complications and outcomes for previously hoarded cats treated surgically for otitis media-interna (OMI) and to investigate the risk factors for complications and poor outcomes. METHODS: A retrospective study was conducted of 58 cats from an institutional hoarding environment that underwent ventral bulla osteotomy (VBO). RESULTS: Inappetence was uncommon at presentation (9/58, 16%) compared with pruritus/alopecia (50%), nasopharyngeal signs (45%), otitis externa (OE) (79%) and otitis interna (OI) (ataxia ± head tilt/head excursions) in 40%. Purulent aural discharge occurred in 36% and polyps in 26%. The tympanic bulla wall was moderately or severely thickened radiographically in 38/108 (35%) ears. Cultures were positive for Streptococcus equi subspecies zooepidemicus in 26/48 (54%) cats. Of the 58 cats, 40 (69%) had complications after the first VBO and 19/30 (63%) after the second. Of 101 complications, 56 (55%), from 27/88 (31%) surgeries, were considered serious, including life-threatening perioperative complications in seven, OI in eight, prolonged anorexia in six and worsening of pruritus/alopecia in nine cases. Three cats developed xerostomia (dry mouth) after the second VBO. Pruritus/alopecia, nasopharyngeal signs, OE and purulent aural discharge resolved in a statistically significant proportion of cats but persisted in some. Full resolution of OI was uncommon. OI preoperatively, and surgery performed by a generalist (vs specialist) surgeon, were risk factors for OE at recheck (OI: odds ratio [OR] 4.35; 95% confidence interval [CI] 1.21-15.70; P = 0.02; surgery: OR 3.64; 95% CI 1.03-12.87; P = 0.045). No other prognostic indicators were identified. No variables tested were significantly associated with risk of serious complications or euthanasia. CONCLUSIONS AND RELEVANCE: Surgical management of chronic OMI was successful in most cases but was not benign and not always beneficial. The analysis was unable to identify clinically helpful outcome predictors. Optimal management of chronic feline OMI remains a challenge, particularly for animal shelters. Less invasive approaches and chronic medical management require further investigation.

Refractory otitis media and otitis externa in a koala (Phascolarctos cinereus) treated with total ear canal ablation and lateral bulla osteotomy.

BACKGROUND: To the authors' knowledge, this is the first report fully describing the surgical and medical management of otitis media and otitis externa in the koala (Phascolarctos cinereus) treated by total ear canal ablation and lateral bulla osteotomy. CASE REPORT: An adult male koala captured as part of a monitoring project in South East Queensland was diagnosed with chlamydial cystitis. Purulent discharge from the right ear was also observed; diagnostic imaging was consistent with otitis media and otitis externa. Yokenella regensburgei was repeatedly cultured from the site. Clinical signs resolved with topical antibiotic therapy; however, recrudesced following cessation of antibiotics. A total ear canal ablation and lateral bulla osteotomy was performed, followed by an extended period of systemic antibiotic therapy. Mild facial nerve paresis was observed for 4 weeks postoperatively and resolved spontaneously. The koala remained clinically healthy for the 6 months it was monitored following release. CONCLUSION: Total ear canal ablation combined with lateral bulla osteotomy appears to be an appropriate surgical intervention for otitis media combined with otitis externa refractory to medical management in the koala. Transient postoperative facial nerve paresis is a possible complication, as documented in other species. To the authors' knowledge this is the first case of Yokenella regensburgei infection outside of humans and American alligators (Alligator mississippiensis); the clinical significance of this pathogen in the koala remains unknown.

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.

Managing recurrent otitis externa in dogs: what have we learned and what can we do better?

Recurrent otitis externa is a common problem in dogs. Topical treatment for each flare is successful in the short term, but repeated cycles of inflammation and infection lead to chronic inflammatory changes, pain and aversion, and antimicrobial resistance. These make the flares more frequent and harder to control. Eventually, the changes become irreversible and require a total ear canal ablation/lateral bulla osteotomy or ablative laser surgery. Most ear canal surgery is avoidable if recurrent otitis is properly managed at an earlier stage. This requires a different mindset and approach to these cases, taking advantage of recent research and clinical findings. Most importantly, clinicians must appreciate that all recurrent ear infections in dogs are secondary. To achieve a good long-term outcome, it is essential that all the underlying factors in each case are diagnosed and managed using the primary, secondary, predisposing, and perpetuating framework. This means that the primary condition must be diagnosed and managed, the secondary infection treated, predisposing risks identified and corrected, and the perpetuating factors reversed. Treatment is in 2 phases: induction to get the ears in remission and then long-term maintenance therapy to prevent relapses. Treatment should be appropriate to each dog but will typically involve ear cleaning, topical antimicrobial therapy, and topical or systemic glucocorticoids. Novel treatments for infection and inflammation will offer additional options in the future. Understanding the triggers for recurrent otitis in dogs will help clinicians plan effective management regimens that will make a huge difference to the quality of life of their patients and their owners.

Perioperative complications and outcomes following partial ear canal ablation and lateral bulla osteotomy in pet rabbits: 20 cases (2009-2021).

OBJECTIVE: To investigate the perioperative complications and outcomes of rabbits undergoing partial ear canal ablation and lateral bulla osteotomy for treatment of chronic otitis. MATERIALS AND METHODS: Surgical logbooks were searched to identify rabbits that underwent partial ear canal ablation and lateral bulla osteotomy between December 2009 and September 2021. Data collected included signalment, history, clinical signs, historical and current medical management, physical examination findings, preoperative diagnostic imaging and aural bacterial culture results, surgical procedure information, duration of hospitalisation, complications and final outcome. RESULTS: Twenty rabbits with chronic, medically unresponsive otitis underwent 27 partial ear canal ablation and lateral bulla osteotomies. Ten rabbits had pre-existing neurologic deficits and 17 had an ear base abscess at the time of surgery. A total of 13 (65%) rabbits developed at least one complication, either in the perioperative period or between discharge and recheck. Three of these rabbits (15%) underwent revision surgery for persistent or recurrent ear base abscess. Complete resolution of clinical signs ultimately occurred in 12 (60%) rabbits. Three (15%) rabbits showed improved but not completely resolved clinical signs (three facial asymmetry). Five (25%) rabbits died of causes potentially related to chronic otitis and/or partial ear canal ablation and lateral bulla osteotomy; two of these rabbits had undergone revision surgery. CLINICAL SIGNIFICANCE: Although this study reports a high complication rate (65%), most complications resolved, with 15 (75%) rabbits having a good to excellent final outcome. Since the majority of rabbits in this study had chronic and severe ear disease, further studies are required to determine if surgical intervention earlier in the disease process could result in a lower complication rate.

Otitis media and interna with or without polyps in cats: association between meningeal enhancement on postcontrast MRI, cerebrospinal fluid abnormalities, and clinician treatment choice and outcome.

OBJECTIVES: The aim of this study was to evaluate the association between meningeal enhancement (MgE) and cerebrospinal fluid (CSF) analysis results, their individual association with bacteriology results from affected ear samples and whether these test results influenced clinicians' therapeutic choice in cats with otitis media and interna (OMI). METHODS: This was a multicentre retrospective study carried out over an 8-year period. Cats diagnosed with OMI, with or without a nasopharyngeal polyp, leading to peripheral vestibular signs were included. Only cats for which MRI with postcontrast T1-weighted sequences and CSF analyses available were included. Cats with intra-axial MRI lesions or empyema were excluded. RESULTS: Fifty-eight cats met the inclusion criteria. MgE was reported in 26/58 cases, of which nine had an abnormal CSF result (increased total nucleated cell count [TNCC] or total protein); 32/58 cases had no MgE, of which 10 showed abnormal CSF results. There was no association between bacteriology results (external ear canal or bulla) and MgE or abnormal CSF results. CSF abnormalities were statistically significantly more common in acute cases (n = 16/37) than in chronic cases (n = 3/21; Fischer's test P = 0.04). Prednisolone was prescribed in 10/16 cases with increased TNCC. Among the 42 cases with normal TNCC, 15 received prednisolone and 13 received non-steroidal anti-inflammatory drugs. Various antimicrobial drugs were prescribed in 53/58 cats. Duration of antimicrobial treatment was similar, regardless of positive bacterial culture (5.58 vs 4.22 weeks), abnormal CSF (5.83 vs 4.76 weeks) or MgE (5.33 vs 4.90 weeks). CONCLUSIONS AND RELEVANCE: No association was found between the CSF and MgE results. Furthermore, no association was found between MgE, CSF or bacteriology findings. In addition, abnormal CSF results might lead the clinician to treat with corticosteroids, but they did not have any impact on duration of antimicrobial treatment. CSF abnormalities were seen significantly less frequently in chronic cases. The outcome tended to be poorer when MgE was detected on MRI.

In vitro antimicrobial activity of cold atmospheric microwave plasma against bacteria causing canine skin and ear infections.

BACKGROUND: Cold atmospheric plasma (CAP) is a new generation medical therapeutic option for bacterial infections. CAP causes physical cell wall rupture and DNA damage, therefore making it highly useful in the treatment of various conditions such as skin infections. HYPOTHESIS/OBJECTIVES: The antimicrobial activity of cold atmospheric microwave plasma (CAMP) against major strains in canine skin infections was tested and the difference in antimicrobial activity between the antibiotic-resistant and antibiotic-susceptible strains of Staphylococcus pseudintermedius was evaluated. METHODS AND MATERIALS: American Type Culture Collection (ATCC) strains (Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli) and clinical isolates identified as methicillin-resistant S. pseudintermedius (n = 27) and methicillin-susceptible S. pseudintermedius (n = 13) were exposed to CAMP for 10 s, 30 s and 60 s. Afterwards, the bacterial survival rate was confirmed. RESULTS: Gram-negative bacteria (P. aeruginosa and E. coli) were more susceptible than Gram-positive bacteria (S. aureus and S. pseudintermedius) for the same duration of CAMP exposure. Only the Gram-negative bacteria were completely killed after 60 s exposure. In S. pseudintermedius isolates, CAMP exposure had similar antibacterial effects regardless of antibiotic resistance. CONCLUSIONS AND CLINICAL IMPORTANCE: CAMP has sufficient antimicrobial activity against major bacterial strains that cause pyoderma and otitis externa in dogs, and may be an alternative therapeutic option for S. pseudintermedius skin infections, for which antibiotics often are ineffective because of antimicrobial resistance in clinical veterinary medicine.

Chronic Otitis in Cats: Clinical management of primary, predisposing and perpetuating factors.

PRACTICAL RELEVANCE: Chronic otitis can be one of the most frustrating diseases to manage for a small animal practitioner. While it occurs less commonly in the cat than the dog, it is no less challenging. The purpose of this review is to discuss the common and uncommon causes of chronic otitis in the cat within the clinical framework used for diagnosis and treatment. The focus is on diseases that affect the ear canal, rather than those restricted to the pinnae. CLINICAL CHALLENGES: Otitis is multifactorial, which complicates management. A common clinical mistake is to focus solely on treating the infection present. Only by addressing all factors will a clinician successfully control chronic otitis. For the purposes of this review, the authors have adopted the established model of separating primary, predisposing and perpetuating causes of otitis. Primary factors are those that directly cause otitis (inflammation); predisposing factors are those that put the patient at risk for development of otitis; and perpetuating factors are those that result in ongoing clinical signs of otitis or that prevent clinical resolution. AUDIENCE: This review is aimed at veterinarians who treat cats and particularly those with an interest in feline dermatology and otology. EQUIPMENT: While many practitioners rely on a hand-held otoscope, a video-otoscope can be very helpful for the diagnosis and treatment of chronic otitis. EVIDENCE BASE: This review presents up-to-date information regarding the diagnosis and treatment of chronic otitis in cats, with emphasis on the most recent peer-reviewed literature.

Carbon dioxide laser surgery for chronic proliferative and obstructive otitis externa in 26 dogs.

BACKGROUND: Some dogs with chronic otitis externa (OE) develop proliferation of the tissues surrounding the opening of the external ear canal, resulting in obstruction. Traditionally total ear canal ablation with bulla osteotomy (TECABO) has been recommended. OBJECTIVES: To evaluate the efficacy of a novel treatment using carbon dioxide (CO2 ) laser surgery and to describe the histopathological features of chronic proliferative and obstructive OE. ANIMALS: Twenty-six dogs were included, 16 with bilateral and 10 with unilateral disease (42 ears were treated). Dogs with nonpatent horizontal ear canal or macroscopic calcification of the ear canal were excluded. For histopathological evaluation, tissue samples were collected from 11 dogs (17 ears). METHODS AND MATERIALS: Hyperplastic tissue around the canal opening and within the vertical ear canal was dissected and ablated using a CO2 laser. Biopsy samples were evaluated for sebaceous and ceruminous gland hyperplasia, epidermal hyperplasia, inflammation and fibrosis. RESULTS: Following CO2 laser surgery there was a good or excellent outcome with substantial resolution of proliferative changes in 39 of 42 ears from 24 of 26 dogs. One surgery was sufficient in 21 dogs and three dogs had two surgeries. Two dogs had recurrence of proliferative tissue after one surgery and underwent TECABO. Two dogs had no recurrence of proliferative tissue after surgery, yet had persistent luminal infection and underwent TECABO. The remainder of the dogs were effectively medically managed long-term following surgery. Histologically, eight ears had a predominantly sebaceous gland response, three had a predominantly ceruminous response and six had a mixed glandular pattern. Epidermal hyperplasia, inflammation and fibrosis varied from mild to severe. CONCLUSIONS AND CLINICAL RELEVANCE: Carbon dioxide laser surgery is an effective treatment of proliferative OE causing obstruction of the ear canal opening and vertical canal, and should be considered as an alternative to TECABO whenever possible.

Experience level as a predictor of entry into the hypotympanum during feline total ear canal ablation and lateral bulla osteotomy.

OBJECTIVES: The study hypotheses were as follows: (1) owing to the unique anatomy of the feline middle ear, the hypotympanum would be entered in less than 100% of cats during total ear canal ablation and lateral bulla osteotomies (TECA-LBOs); and (2) incomplete penetration of the septum and subsequent failure to enter the hypotympanum is more likely to occur in surgeries performed by a novice surgeon when compared with an experienced surgeon and may be under-recognized. METHODS: Head CT was performed in 12 feline cadavers to confirm absence of gross ear disease. A novice surgeon and an experienced surgeon were randomly assigned to perform TECA-LBO on the left or right ear. Surgeons were blinded to each other's surgical technique. CT of cadavers was performed after the procedure. Successful penetration of the septum, entry into the hypotympanic cavity and amount of bone removed in bulla osteotomy, quantified via CT, were compared between the novice surgeon and experienced surgeon. RESULTS: The novice surgeon entered the hypotympanum in 3/12 (25%) procedures, compared with 9/12 (75%) procedures performed by the experienced surgeon. The experienced surgeon performed a larger osteotomy than the novice surgeon (3301 mm vs 1376 mm, P <0.0023). Regardless of surgeon experience, more bone was removed in surgeries in which the hypotympanum was entered. CONCLUSIONS AND RELEVANCE: Our results underscore the need for familiarity with feline middle ear anatomy when performing TECA-LBOs. Postoperative CT is recommended for novice surgeons to confirm entry into the hypotympanum.

Azithromycin concentration in severely inflamed canine external ear canals - a case series.

OBJECTIVE: To determine azithromycin concentration in severely inflamed canine external ear canals. MATERIAL AND METHODS: Five dogs of various breeds and ages with severe and chronic otitis externa underwent ear canal reconstruction surgery. A single oral dose of azithromycin at 10 mg/kg was administered 12 to 24 hours prior to surgery. Tissue samples were collected from the excised external ear canals and azithromycin concentration was determined using a liquid chromatography-tandem mass spectrometry method. RESULTS: Azithromycin concentrations ranging from 11.4 to 107.0 µg/g (mean 59.2 ± 44.6 µg/g, median 50.9 µg/g) were detected in the chronically infected external ear canal tissue 12 to 24 hours after administration. CLINICAL SIGNIFICANCE: Little information exists on antibiotic concentrations in pathological tissues of dogs. Macrolides are known to concentrate in skin tissue. In light of the present results, investigation of clinical efficacy of azithromycin in chronic canine otitis externa is warranted.

Otoscopic evaluation of epithelial remnants in the tympanic cavity after total ear canal ablation and lateral bulla osteotomy.

OBJECTIVE: To determine the ability to detect the presence of epithelial remnants after total ear canal ablation (TECA) and lateral bulla osteotomy (LBO) with endoscopy and to identify the most common locations of epithelial remnants after tympanic curettage. STUDY DESIGN: Experimental study. ANIMALS: Five fresh canine cadavers with no gross evidence of middle ear disease. METHODS: Ten TECA-LBO were performed by four surgeons. After tympanic curettage, a 1.9-mm rigid 30° endoscope was inserted into the rostral, caudal, dorsal, ventral, and medial sections of the tympanic cavity. Three observers evaluated otoscopic images for epithelial remnants in each compartment. The median distribution of epithelial remnants was calculated for each section of the tympanic cavity with a three-dimensional tympanic cavity model. RESULTS: Epithelial remnants were identified in at least one of the five areas of the tympanic cavity after each TECA-LBO. The rostral section contained the most epithelial remnants (35.6%), while the medial section contained the least amount (1.8%). CONCLUSION: Use of a 1.9-mm rigid endoscope was an effective method to evaluate all sections of the tympanic cavity after curettage in TECA-LBO. Epithelial remnants were consistently found after TECA-LBO, especially in the rostral compartment. CLINICAL SIGNIFICANCE: Intraoperative endoscopy should be considered to improve removal of epithelium after initial TECA-LBO or revision surgeries.

Patulous Eustachian tube and palatine defect in a Dachshund with chronic unilateral otitis externa and otitis media.

BACKGROUND: Patulous Eustachian tube (pET) is a rare dysfunction of the Eustachian tube described in humans. It is characterized by failure of the ET to close, resulting in unrestricted passage of air, sound and material between the nasopharynx and the middle ear. OBJECTIVE: To report a case of pET associated with otitis in a dog. ANIMAL: A 6-year old-female spayed Dachshund dog. METHODS AND MATERIALS: Otoscopic examination, cytological evaluation, culture and susceptibility, computerized tomography (CT), video-otoscopic flushing and surgery. RESULTS: Left ear otoscopic examination revealed erythema, purulent frothy discharge, ceruminous gland hyperplasia, stenosis and a partial tear of the tympanum. Cytological evaluation from the left external canal showed neutrophils, macrophages, rods and cocci. Aerobic culture showed predominantly multidrug-resistant Pseudomonas aeruginosa. The CT findings of the left ear included chronic changes in the external canal, marked lysis of the tympanic bulla and marked dilation of the ET. During video-otoscope flushing, saline drained through the mouth. Bilateral incomplete hypoplasia of the soft palate was noted. Total ear canal ablation and bulla osteotomy with ET dissection were curative. Histopathological findings were compatible with chronic otitis externa (OE) and media. CONCLUSION AND CLINICAL IMPORTANCE: To the best of the authors' knowledge, this is the first case of pET described in animals. The ET dysfunction and palatine defect were likely the cause of the otitis in this dog. Clinicians should investigate pET in animals with signs of OE characterized by frothy liquid and food fragments in the ear canal in addition to sneezing after drinking water.

In vitro antibacterial activity of the manuka essential oil from Leptospermum scoparium combined with Tris-EDTA against Gram-negative bacterial isolates from dogs with otitis externa.

BACKGROUND: The increasing prevalence of antimicrobial resistance among bacteria in dogs with otitis externa has led to a need for novel therapeutic agents. HYPOTHESIS/OBJECTIVE: To examine the antibacterial effects of manuka oil combined with ethylenediaminetetraacetic acid-tromethamine (Tris-EDTA) against Gram-negative bacteria isolates from dogs with otitis externa. METHODS AND MATERIALS: A total of 53 clinical isolates including Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae ssp. pneumoniae and Proteus mirabilis. Antimicrobial susceptibility was determined using disk diffusion; the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of manuka essential oil, with or without Tris-EDTA, were investigated. RESULTS: A total of 44 isolates were resistant to at least one antibiotic and 19 strains were multidrug-resistant, with resistance to at least one agent in three or more antimicrobial classes. The MICs and MBCs of manuka oil alone were ≥1% (v/v) and ≥2% (v/v), respectively. There was no antimicrobial effect of Tris-EDTA (1.125:0.3 mg/mL) without manuka oil. However, the combination of manuka oil with Tris-EDTA significantly decreased the MICs (ranging from 0.06% to 0.5%, v/v; P < 0.001) and MBCs (ranging from 0.06% to 1%, v/v; P < 0.001). There also was no significant difference between multidrug-resistant and nonresistant bacterial isolates in terms of the antimicrobial activity of manuka oil with Tris-EDTA. CONCLUSIONS AND CLINICAL IMPORTANCE: The study findings suggest that manuka oil, especially when combined with Tris-EDTA, may be a promising alternative therapeutic option for Gram-negative otic pathogens. Clinical studies are needed to assess potential for in vivo ototoxic effects and efficacy.

Partial vertical ear canal resection in two cats.

CASE DESCRIPTION: An 18-year-old domestic medium-hair cat (cat 1) and a 16-year-old domestic shorthair cat (cat 2) were evaluated because of obstructive skin lesions involving the perimeter of the left external auditory canal. CLINICAL FINDINGS: Otitis externa was present in affected ears secondary to obstructive soft tissue growths involving the outer margin of the external auditory canal and outer third of the vertical ear canal. Histologic examination of a preoperative biopsy sample revealed multiple ulcerated ceruminous gland adenomas in the affected ear of cat 1. Histologic examination of the submitted tissue from cat 2 confirmed ceruminous cystomatosis with surface colonization of yeast compatible with Malassezia spp. TREATMENT AND OUTCOME: Both cats underwent partial resection of the upper third of the affected vertical ear canal and associated diseased skin. The incised margin of the pinna was sutured to the margin of the remaining portion of the vertical ear canal with absorbable sutures. Both cats were disease free over a 12-month (cat 1) or 10-month (cat 2) follow-up period. Cat 1 later developed a small ceruminous gland adenocarcinoma in the adjacent rostrolateral margin of the vertical ear canal 1 year after surgery; the mass was resected, and the patient was free of recurrence 4 months later. CLINICAL RELEVANCE: Partial resection was an effective alternative to complete vertical ear canal resection for lesions involving the upper third of the vertical ear canal in these cats; the partial resection procedure was deemed simpler to perform and less traumatic to the cat. Functional and cosmetic results were excellent, with preservation of the overall anatomy of the external auditory canal.

Antibacterial effect of N-acetylcysteine in combination with antimicrobials on common canine otitis externa bacterial isolates.

BACKGROUND: Approved treatments for canine otitis externa are limited in variety and may contain ototoxic ingredients. With bacterial resistance an ongoing concern, it would be ideal if non-ototoxic agents combined with antibiotics resulted in a synergistic effect, requiring lower antibiotic concentrations to treat infections. Evidence of synergism and antagonism between N-acetylcysteine (NAC) and various antibiotic classes has been reported; the present research group was interested in examining these interactions. HYPOTHESIS/OBJECTIVES: To determine if NAC, an otoprotective and antimicrobial compound, has synergistic activity when combined with enrofloxacin or gentamicin in vitro against bacterial isolates causing canine otitis externa. ANIMALS: Twenty-two isolates from canine clinical cases of otitis externa were identified and tested, including seven Staphylococcus pseudintermedius, 12 Pseudomonas aeruginosa and three Corynebacterium spp. isolates. METHODS AND MATERIALS: Each isolate was grown on blood agar for 24 h and transferred to Mueller-Hinton broth (MHB), with a final concentration of 5 × 105 cfu/mL. Each well was inoculated with 50 µL of bacterial suspension. N-acetylcysteine was diluted in MHB to a starting concentration of 160 mg/mL. Enrofloxacin and gentamicin were diluted to 64 µg/mL. Individual and checkerboard serial microdilution assays were performed in triplicate with negative controls for all isolates tested. RESULTS: Interactions observed for NAC and enrofloxacin were synergistic (4.5%), indifferent (77.3%) or antagonistic (18.2%). Interactions observed for NAC and gentamicin were synergistic (4.5%), indifferent (45.5%) or antagonistic (50%). CONCLUSIONS AND CLINICAL RELEVANCE: Most interactions between NAC and enrofloxacin or gentamicin were indifferent or antagonistic at the concentrations tested in vitro.

Genotyping of Malassezia pachydermatis disclosed genetic variation in isolates from dogs in Colombia / Genotipagem de Malassezia pachydermatis revelou variação genética em isolados de cães na Colômbia

Malassezia pachydermatis is a lipophilic and lipid-dependent yeast mostly isolated from animals' skin; hence, it is regarded as a zoophilic species causing otitis externa in dogs. Aspects associated with its epidemiology and pathogenicity is a matter of interest. This study aimed to conduct a molecular characterization of 43 isolates of M. pachydermatis obtained from dogs with otitis externa. For this purpose, the 5.8S internal transcribed spacer 2 (ITS2) and D1/D2 26S rRNA regions were amplified, sequenced and analyzed using restriction fragment length polymorphism (RFLP) with AluI, CfoI, and BstF5I endonucleases. Phylogenetic analyses revealed that these isolates grouped with the sequence types I, IV and V, previously proposed for M. pachydermatis. Interestingly, we found a new polymorphic RFLP pattern using BstF5I, these isolates were associated with the sequence types IV and V, nevertheless an association between polymorphic RFLP patterns, and fosfolipase activity or canine population data was not observed. These findings underline the genetic diversity of M. pachydermatis and provide new insights about the epidemiology of this species in the analyzed population.(AU)

Malassezia pachydermatis é uma levedura lipofílica e dependente de lipídios, principalmente da pele de animais. Sendo, por essa razão, considerada uma espécie zoofílica e causadora de otite externa em cães. Neste sentido, aspectos associados à sua epidemiologia e patogenicidade constituem um tema de interesse científico. O objetivo deste estudo foi realizar a caracterização molecular de 43 isolados de M. pachydermatis obtidos a partir de cães com otite externa. Para esta propósito, foram amplificadas, sequenciadas e analisadas com enzimas de restrição as regiões do gene 5.8S, do espaçador interno transcrito 2 (ITS2) e D1/D2 do 26S do rRNA pelo método RFLP, com as endonucleases AluI, CfOI e BstF5I. Análises filogenéticas revelaram que os isolados se agruparam com as sequências tipo I, IV e V de M. pachydermatis como já descrito anteriormente. De maneira interessante, se observou um novo RFLP polimórfico utilizando BstF5I. Os isolados que mostraram esse padrão foram associados com os padrões IV e V. No entanto, não foi observada associação entre padrões polimórficos de RFLP e atividade de fosfolipase ou dados da população canina. Estes resultados demonstram a diversidade genética de M. pachydermatis e fornecem novas perspectivas sobre a epidemiologia destas espécies na população analisada.(AU)

Antimicrobial activity of thyme oil, oregano oil, thymol and carvacrol against sensitive and resistant microbial isolates from dogs with otitis externa.

BACKGROUND: Multidrug-resistant pathogens present a major global challenge in antimicrobial therapy and frequently complicate otitis externa in dogs. HYPOTHESIS/OBJECTIVES: In vitro efficacy of oregano oil, thyme oil and their main phenolic constituents against bacterial and fungal isolates associated with canine otitis externa were investigated. It was hypothesized that the main phenolic components would have greater antimicrobial activity compared to the relative essential oil. METHODS AND MATERIALS: Antimicrobial susceptibility testing was performed using broth microdilution with spot-plating technique to determine minimum inhibitory and bactericidal/fungicidal concentrations (MICs, MBCs and MFCs). A time-kill kinetics assay was performed to confirm the bactericidal and fungicidal activity of the oils and their phenolic constituents. One hundred bacterial and fungal isolates, including meticillin-susceptible Staphylococcus pseudintermedius (n = 10), meticillin-resistant S. pseudintermedius (n = 10), ß-haemolytic Streptococcus spp. (n = 20), Pseudomonas aeruginosa (n = 20; including 10 isolates resistant to one or two antimicrobials), Proteus mirabilis (n = 20) and Malassezia pachydermatis (n = 20) from dogs with otitis externa were used. RESULTS: Oregano oil, thyme oil, carvacrol and thymol exhibited antibacterial activity against all bacterial and fungal isolates tested. MIC90 values ranged from 0.015 to 0.03% (146-292 µg/mL) for the Gram-positive bacteria and P. mirabilis. For P. aeruginosa and M. pachydermatis, MIC90 values ranged from 0.09 to 0.25% (800-2,292 µg/mL). CONCLUSIONS AND CLINICAL SIGNIFICANCE: Oregano oil, thyme oil, carvacrol and thymol showed good in vitro bactericidal and fungicidal activity against 100 isolates from dogs with otitis externa, including some highly drug-resistant isolates. These essential oils and their main phenolic constituents have the potential to be further investigated in vivo for the treatment of canine otitis externa.