Adverse food reactions in dogs due to antibiotic residues in pet food: a preliminary study.

In the last decades, adverse food reactions have increased considerably in dogs and cats. In this study we report on the possible onset of food intolerances symptoms, including otitis, diarrhoea, generalised anxiety, and dermatitis in a cohort of 8 dogs consuming commercial diets. All dogs received an organic chicken-based diet for 15 days. We performed analysis of blood biochemical parameters, kibble composition, and oxytetracycline (OTC) serum concentration before and after 15 days of organic chicken-based diet supplementation. We hypothesised that a chronic intake of contaminated food enhanced by the presence of nanoparticle aggregates might be at the base of the onset of pharmacologic or idiopathic food intolerances. At the end of the evaluation period, an overall significant reduction of otitis, diarrhoea, generalised anxiety, and dermatitis was observed. Biochemical analyses indicate a significant increase in the alkaline phosphatase, from 41 to 52.5 U/L, after 15 days (••p <0.01), while a significant decrease in Gamma-glutamyl transferase and urea, from 9.37 to 6.25 U/L and from 32.13 ± 8.72 to 22.13 ± 7.8 mg/dL, respectively, was observed (•p <0.05). A significant decrease, from 0.22 to 0.02 µg/mL, in mean OTC serum concentration was also observed (••p <0.01). Composition analysis revealed the presence of OTC, calcium, aluminium, silicon, and phosphorous nanoparticle aggregates. Further research on a wider sample size would help to confirm the hypothesis proposed here.

In vitro efficacy of a honey-based gel against canine clinical isolates of Staphylococcus pseudintermedius and Malassezia pachydermatis.

BACKGROUND: Staphylococcus pseudintermedius and Malassezia pachydermatis are important agents in canine pyoderma and otitis. HYPOTHESIS/OBJECTIVES: Determine the in vitro efficacy of a honey-based gel (HBO) against meticillin-susceptible S. pseudintermedius (MSSP), meticillin-resistant S. pseudintermedius (MRSP) and M. pachydermatis, by minimum bactericidal concentration (MBC), minimum fungicidal concentration (MFC) and time-kill assay (TKA). Efficacy of the product's honey component (HO) also was evaluated. METHODS: Sixty S. pseudintermedius and 10 M. pachydermatis canine isolates were selected. All isolates were tested against serial dilutions of an HBO containing 40% HO (40%, 20%, 10%, 5% and 2.5% w/v) and HO alone (undiluted, 40%, 20%, 10%, 5% and 2.5% w/v). Microbroth assay followed by subculture was used to determine MBC and MFC. The same protocol was applied after product exposure to catalase. A well-diffusion assay for S. pseudintermedius was used to generate inhibition zones. A TKA for 10 isolates of S. pseudintermedius and 10 isolates of M. pachydermatis was performed. RESULTS: MBC was 20% w/v (5-20% w/v) for HBO and HO. HBO had lower MBC values when compared to HO (P = 0.003). No statistical difference was observed between MSSP/MRSP isolates (HBO P = 0.757, HO P = 0.743). Only HO was affected by catalase (P = 0.015). MFC for HBO was 10% w/v (5-10% w/v) and 40% w/v for HO (20-≥40% w/v). All isolates were killed after 4 h of exposure. CONCLUSIONS AND CLINICAL IMPORTANCE: Staphylococcus pseudintermedius and M. pachydermatis are susceptible to the HBO and these results can be used for future clinical trials.

Identification of VIM-2 metallo-ß-lactamase-producing Pseudomonas aeruginosa isolated from dogs with pyoderma and otitis in Korea.

BACKGROUND: Pseudomonas aeruginosa is a challenging pathogen cultured from cases of acute and chronic canine otitis and sometimes in cases of deep pyoderma. The spread of antimicrobial resistance, especially carbapenem resistance, is a serious therapeutic challenge worldwide. HYPOTHESIS/OBJECTIVES: To investigate the identification and characterization of resistant P. aeruginosa clinical canine isolates. MATERIALS: Clinical isolates (n = 80) were collected from dogs with pyoderma (n = 18) and otitis (n = 62) in Korea. METHODS: Antimicrobial susceptibility was determined using agar dilution and using Clinical and Laboratory Standards Institute guidelines for recording susceptibility for human Pseudomonas isolates; genetic relatedness of isolates was investigated by multilocus sequence typing (MLST) and SpeI macrorestriction analysis. The class 1 integrons were amplified and sequenced using primer walking. RESULTS: Most isolates were susceptible to colistin (97.5%), polymyxin B (96.3%), ciprofloxacin (81.3%) and meropenem (80.0%); whereas resistance to aztreonam (80%), piperacillin (52.5%), piperacillin/tazobactam (41.3%) and cefepime (37.5%) was high; 12 carbapenem-nonsusceptible isolates (15%) were detected. MLST revealed 45 different sequence types (STs) and macrorestriction analysis detected 55 distinct pulsotypes (PTs), which were divided into 25 clonal groups. Among carbapenem-nonsusceptible isolates, 10 (83.3%) were VIM-2-producing strains. Nine VIM-2-producing isolates were identified as ST1047 and harboured the same 2.8 kb class 1 integron. One remaining isolate was ST1203 with 2.1 kb class 1 integron. CONCLUSIONS AND CLINICAL IMPORTANCE: This study demonstrated the diversity of the phenotype and genotype of clinical P. aeruginosa isolates from dogs with pyoderma and otitis. The identification of VIM-2-producing P. aeruginosa in dogs is alarming and warrants further surveillance.

Effect of an injectable trace mineral supplement containing selenium, copper, zinc, and manganese on immunity, health, and growth of dairy calves.

The objective of this study was to evaluate the effect of 2 subcutaneous injections of a multimineral preparation, each containing 60 mg of zinc, 10mg of manganese, 5mg of selenium, and 15 mg of copper at 3 and 30 d after birth on immunity, health, and growth of dairy calves during the preweaning period. The study was conducted in upstate New York in 2 commercial dairy farms. A total of 790 Holstein heifer calves were randomly allocated at birth into 1 of 2 treatments: trace mineral supplement (TMS) treated or control. Blood samples were collected at 3, 14, and 35 d after birth to evaluate glutathione peroxidase (GPx) activity, superoxide dismutase (SOD) activity, haptoglobin, and neutrophil and monocyte function. Incidence of diseases and average daily gain was evaluated in the first 50 d of life. At 14 d of life, TMS-treated calves had increased neutrophil activity compared with control calves. Moreover, TMS-treated calves had greater GPx activity on d 14 after birth than control calves. The TMS treatment reduced the incidence of diarrhea (TMS=41.7% vs. control=49.7%) and combined incidence of pneumonia or otitis or both (TMS=41.7% vs. control=49.1%). Additionally, GPx was greater for calves diagnosed with otitis at d 35 after birth. However, calves diagnosed with pneumonia had decreased GPx activity at d 35 after birth. Serum SOD and haptoglobin concentrations were not affected by treatment or disease. Moreover, no effects were observed on average daily gain and survivability between TMS-treated and control calves during the preweaning period. Supplementation with trace minerals at 3 and 30 d of life increased neutrophil function and GPx activity and reduced the incidence of health disorders.

Atividade antifúngica do óleo essencial de Origanum vulgare frente a Malassezia pachydermatis / Antifungal activity of Origanum vulgare essential oil against Malassezia pachydermatis

Objetivou-se com este estudo avaliar a atividade antifúngica in vitro do óleo essencial de Origanum vulgare frente a isolados clínicos de Malassezia pachydermatis. As folhas secas de O. vulgare foram adquiridas de distribuidor comercial com certificado de qualidade e origem e encaminhadas para extração do óleo essencial e cromatografia. Para realização do teste in vitro, foi utilizada a técnica de microdiluição em caldo (CLSI M27A3) com modificações para fitofármacos e M. pachydermatis. O óleo essencial de orégano foi testado nas concentrações de 28 a 0,87mg/mL diluído em caldo Sabouraud com 1% de tween 80. Todos os isolados foram testados em duplicata. Na análise cromatográfica do óleo essencial, foram identificados 12 compostos, sendo timol, a-terpineno e 4-terpineol os compostos majoritários. A CIM e a CFM dos 42 isolados de M. pachydermatis variaram de <0,87 a 7mg/mL, com valores de CIM50 e CIM90 de 1,18 e 3,28mg/mL, respectivamente. Com este estudo foi possível concluir que M. pachydermatis é sensível ao óleo essencial de orégano mesmo em concentrações baixas. Dessa maneira, o óleo essencial de orégano apresenta-se como promissor na bioprospecção de novos fármacos para o tratamento das otites e dermatites na clínica de pequenos animais...

The aim of this study was to evaluate the in vitro antifungal activity of essential oil of Origanum vulgare against clinical isolates of Malassezia pachydermatis. The dried leaves of O. vulgare were purchased from a commercial distributor with certified quality and origin and referred for essential oil extraction and chromatography. The technique for in vitro testing was microdilution (CLSI M27A3) with modifications to phytochemicals and M. pachydermatis. The essential oil of O. vulgare was tested at concentrations from 28 to 0.87mg/mL in Sabouraud broth diluted with 1% of tween 80. All isolates were tested in duplicate. In the chromatographic analysis of the essential oil 12 compounds were identified, and thymol, α-terpinene, 4-terpineol were the major compounds. The MIC and the MFC of the 42 isolates of M. pachydermatis ranged from <0.87 to 7mg/mL with MIC50 and MIC90 values of 1.18 and 3.28 mg/mL, respectively. With this study it was concluded that M. pachydermatis is sensible to O. vulgare essential oil even at low concentrations. Thus, the essential oil of O. vulgare is presented as bioprospecting in the promising new drugs for the treatment of otitis and dermatitis in small animal clinic...

In vitro and in vivo activity of a killer peptide against Malassezia pachydermatis causing otitis in dogs.

In order to overcome the limitations inherent in current pharmacological treatments for Malassezia pachydermatis, the cause of otitis externa in dogs, the efficacy of a killer decapeptide (KP) was evaluated in vitro and in vivo. Sixteen dogs with naturally occurring M. pachydermatis otitis externa were enrolled, and the in vitro fungicidal activity of KP was evaluated using yeasts recovered from these animals. The therapeutic activity was evaluated in four groups of four animals each. The dogs were topically treated with KP (150 µl, 2 mg/ml) three times per week (group A) or every day (group B), treated with a scramble peptide every day (group C), or left untreated (group D). Assessment of clinical signs (pruritus, erythema, and lichenification and/or hyperpigmentation), expressed as mean of the total clinical index score (mTCIS), the population size of M. pachydermatis at the cytological examination (mean number of yeast cells at 40× magnification [mYC]), and culture testing (mean number of log10 CFU/swab [mCFU]), were conducted daily from the first day of treatment (T0) until two consecutive negative cultures (mCFU ≤ 2). KP showed an in vitro fungicidal effect against M. pachydermatis isolates, with an MFC90 value of 1 µg/ml. The mTCIS, mYC and mCFU were negative only in animals in group B after T8. Daily administration of KP for 8 days was safe and effective in controlling both clinical signs and the population size of M. pachydermatis causing otitis externa, thus offering an alternative to the currently available therapeutic or prophylactic protocols for recurrent cases of Malassezia otitis in dogs.

Atividade antimicrobiana do oleorresina de copaíba (Copaifera reticulata) frente a Staphylococcus coagulase positiva isolados de casos de otite em cães / Antimicrobial activity of copaiba oil (Copaifera reticulata) against coagulase positive Staphylococcus of canine otitis

The aim of this study was to investigate the antimicrobial activity of the oleoresin Copaifera reticulata Ducke against Staphylococcus coagulase positive (SCP) isolated from otitis externa in dogs. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the oleoresin were determined by broth microdilution method. In addition, we verified the antimicrobial susceptibility profile of the isolates of SCP by agar diffusion method. Eight classes of antimicrobial were used to calculate the multidrug resistance. The chemical composition of the oleoresin was performed by gas chromatography coupled to the mass spectrometry (GC/MS), and β-caryophyllene, β-bisabolene, and (E)-α-bergamotene were the main compounds found. The copaiba oleoresin showed a MIC90 of 0.164mg/mL and a CBM90 of 1.3mg/mL. The multidrug resistance was found in 27% of the strains tested. The results suggest that copaiba oleoresin has bacteriostatic and bactericidal activity even in multidrug-resistant coagulase-positive strains.

O objetivo do presente trabalho foi investigar o potencial antimicrobiano do oleorresina de Copaifera reticulata Ducke em isolados de Staphylococcus coagulase positiva (SCP) provenientes de casos de otite externa em cães. O método de microdiluição em caldo foi utilizado para determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM) de oleorresina de copaíba. Em adição, foi determinado o perfil de suscetibilidade aos antimicrobianos dos isolados de SCP pelo método de difusão em ágar. Oito classes de antimicrobianos foram usadas para o cálculo de multirresistência antimicrobiana. A determinação da composição química do oleorresina de copaíba foi realizada por cromatografia em fase gasosa acoplada à espectrometria de massas (GC/MS), sendo que β-cariofileno, β-bisaboleno e (E)-α-bergamoteno foram os compostos majoritários. O oleorresina de copaíba demonstrou CIM90 de 0,164mg/mL e CBM90 de 1,31mg/mL. A multirresistência foi verificada em 27% das cepas testadas. Os resultados sugerem que o oleorresina de copaíba exerceu atividade bacteriostática e bactericida mesmo em cepas multirresistentes de Staphylococcus coagulase-positiva.

Isolation of Staphylococcus schleiferi from healthy dogs and dogs with otitis, pyoderma, or both.

OBJECTIVE: To determine the frequency of isolation and susceptibility patterns of Staphylococcus schleiferi from healthy dogs and dogs with otitis, pyoderma, or both that had or had not received antimicrobial treatment. DESIGN: Prospective study. ANIMALS: 50 dogs. PROCEDURE: Dogs were allocated to 1 of 4 groups: healthy dogs (n=13), dogs without otitis but with pyoderma (10), dogs with otitis but without pyoderma (11), and dogs with otitis and pyoderma (16). Bacteriologic culture of ear swab specimens was performed in all dogs. Bacteriologic culture of skin swab specimens was also performed in dogs with concurrent pyoderma. Isolates were identified as S schleiferi subsp schleiferi or S schleiferi subsp coagulans on the basis of growth and biochemical characteristics. RESULTS: S schleiferi was not isolated from any dogs with pyoderma only. Staphylococcus schleiferi subsp schleiferi was isolated from the ears of 2 healthy dogs, and the skin and ears of 2 dogs and the skin of 1 dog with otitis and pyoderma. Staphylococcus schleiferi subsp coagulans was isolated from the ears of 3 dogs with otitis only, and the ears of 6 dogs and the skin of 2 dogs with otitis and pyoderma. One of the S schleiferi subsp schleiferi isolates from ears, 2 of the S schleiferi subsp coagulans isolates from ears, and 1 of the S schleiferi subsp coagulans isolates from the skin were resistant to methicillin. One methicillin-resistant isolate from the ears and 1 from the skin were also resistant to fluoroquinolones. CONCLUSIONS AND CLINICAL RELEVANCE: S schleiferi subsp schleiferi was detected in healthy dogs and dogs with otitis and pyoderma. Methicillin-resistant and -susceptible S schleiferi subsp schleiferi and S schleiferi subsp coagulans were detected as the predominant organisms in dogs with otitis.

Effect of emu oil on auricular inflammation induced with croton oil in mice.

OBJECTIVE: To determine the acute anti-inflammatory effects of topically applied emu oil. ANIMALS: 96 male CD-1 mice assigned randomly to 4 groups, each comprising 24 mice. PROCEDURE: To induce auricular inflammation, 50 microl of a solution comprising 10 microl of croton oil dissolved in 1 ml of acetone was applied to the inner surface of the left auricle (pinna). One hour later, 3 or 5 microl of emu oil (low- and high-dose groups, respectively) or 5 microl of porcine oil (oil-control) was applied to the left pinna. Control mice remained untreated. Six mice per group were euthanatized 3, 6, 12, and 24 hours after induction of inflammation. Specimens of auricular tissue (ear plugs) were obtained, using a 6-mm biopsy punch. Magnitude of swelling was calculated as the weight difference between left (inflamed) and right (noninflamed) ear plugs; degree of edema was determined as the difference between wet and dry weights of the left ear plug. RESULTS: Magnitude of swelling was significantly reduced at 6 and 12 hours in mice treated with emu or porcine oil, compared with controls. The greatest reduction in swelling was detected in the high-dose emu group at 6 hours. Compared with controls, degree of edema was significantly reduced at 6 hours only in the high-dose group, whereas by 12 hours, all groups treated with oils had significantly less edema than controls. At 24 hours, magnitude of swelling and degree of edema did not differ among groups. CONCLUSION: Topically applied emu oil significantly reduced severity of acute auricular inflammation induced by croton oil in mice.

Potentiating effect of EDTA-Tris on the activity of antibiotics against resistant bacteria associated with otitis, dermatitis and cystitis.

Possible synergistic effects of the combination of EDTA-tromethamine (EDTA-Tris) and three antimicrobial agents (cephaloridine, kanendomycin and enrofloxacin) against resistant Gram-positive and Gram-negative bacteria are reported. Bacteria were isolated from eight cases of chronic otitis externa, five cases of chronic dermatitis and four cases of recurrent cystitis in dogs which had previously been treated with one of the three antibiotics without success. Animals exposed to EDTA-tromethamine plus the antibiotic recovered completely within 10 days, and were controlled clinically and bacteriologically for 180 days. Local irrigation with EDTA-tromethamine solution was well tolerated and no side effects were recorded.