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Pathologic and Microbiologic Aspects of Pet Psittacine Infected by Escherichia Coli and Salmonella Typhimurium1

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Pathologic and Microbiologic Aspects of Pet Psittacine Infected by Escherichia Coli and Salmonella Typhimurium1 Pesq. Vet. Bras. 37(4):379-384, abril 2017 DOI: 10.1590/S0100-736X2017000400012 Pathologic and microbiologic aspects of pet psittacine infected by Escherichia coli and Salmonella Typhimurium1 Raul A.S. Siqueira2*, William C. Maciel2, Ruben H. Vasconcelos2, Windleyanne G.A. Bezerra2, Elisângela S. Lopes2, Débora N. Machado2, Marcel F. de Lucena2 and Ricardo B. de Lucena3 ABSTRACT.- Siqueira R.A.S., Maciel W.C., Vasconcelos R.H., Bezerra W.G.A., Lopes E.S., Ma- chado D.N., Lucena M.F. & Lucena R.B. 2017. Pathologic and microbiologic aspects of pet psittacine infected by Escherichia coli and Salmonella Typhimurium. Pesquisa Veteriná- ria Brasileira 37(4):379-384. Laboratório de Estudos Ornitológicos, Universidade Estadual do Ceará, Itaperi, Fortaleza, CE 60740-000, Brazil. Email: [email protected] The role of Escherichia coli in healthy microbiota of psittacine is controversial, and the presence of Salmonella sp. indicates possible disease. Therefore, this study aimed to iden- tify the presence of E. coli and Salmonella spp. in a psittacine pet that died in Fortaleza, Brazil, correlating pathogenicity aspects of the isolates through the evaluation of lesions and antimicrobial susceptibility. Psittacine pets sent to the Laboratory of Ornithological Studies, State University of Ceará, that died in 2014 and 2015 were necropsied. Fragments of liver, kidneys, intestine, lung, heart, spleen and brain were collected for microbiologi- cal and histopathological analyses. Scores were attributed to lesions and isolated strains submitted to antimicrobial susceptibility test. From the seventy necropsied birds, ninete- en were positive for E. coli and one for Salmonella Typhimurium. Congestive lesions and groupslymphoplasmocitic of antibiotics inflammatory was observed. infiltrate In conclusion, were observed according varying to the from results, light E. to coli moderate strains and werethe Salmonella the main findings. In the analyzed strains, multidrug resistance against different pets, and multidrug resistance may hinder treatments with antibiotics used in avian pet medicine. Typhimurium isolate produced significant lesions in the psittacine INDEX TERMS: Escherichia coli, Salmonella Typhimurium, colibacillosis, salmonellosis, Psittacidae, anatomopathological features, antimicrobial susceptibility test. RESUMO.- [Aspectos patológicos e microbiológicos de dos ao Laboratório de Estudos Ornitológicos da Universi- Psittaciformes de companhia infectados por Escherichia dade Estadual do Ceará durante o período de 2014 a 2015. coli e Salmonella Typhimurium.] A participação de Esche- No momento da necropsia foram coletados fragmentos de richia coli na microbiota saudável de Psicittaciformes e a de Salmonella spp. já indica possível doença. O objetivo deste posterior processamento microbiológico e histopatológico. estudo foi pesquisar a presença de E. coli e Salmonella spp. Asfígado, lesões rins, foram intestino, graduadas pulmão, e as coração, cepas isoladasbaço e encéfalo submetidas para em psittaciformes de companhia na cidade de Fortaleza/ a antibiograma. Das setenta aves necropsiadas, dezenove Ceará, traçando os aspectos de patogenicidade destas cepas foram positivas para E. coli e apenas uma para Salmonella através das lesões e da sensibilidade antimicrobiana. Foram - necropsiados os psittaciformes de companhia encaminha- rio linfoplasmocitário variaram de leve a moderado, e foram asTyphimurium. principais lesões As lesões encontradas. de congestão Nas e cepasinfiltrado analisadas inflamató foi 1 Received on November 6, 2015. constatada multiresistência a diferentes grupos de antibió- Accepted for publication on July 13, 2016. ticos testados. De acordo com os achados, pode-se concluir 2 Laboratório de Estudos Ornitopatlógicos, Universidade Estadual do que os isolados de E. coli e Salmonella Typhimurium produ- Ceará (UECE), Av. Silas Munguba 1700, Fortaleza, CE 60740-000, Brazil. *Corresponding author: [email protected] 3 Laboratório de Patologia Animal, Universidade Federal da Paraíba, Areia, PB 58397-000, Brazil. antibióticosziram lesões usados significativas na clínica em de psittaciformes aves de companhia. em Fortaleza, Brasil, e a multirresistência pode dificultar o tratamento com 379 380 Raul A.S. Siqueira et al. TERMOS DE INDEXAÇÃO: Escherichia coli, Salmonella Typhimu- Table 1. Absolute Frequency and Relative Frequency from rium, colibacilose, Salmonelose, Psittacidae, características ana- species of seventy five psittacine birds necropsied tomopatológicas. Species Frequency of birds AF RF INTRODUCTION AA 13 17,33 Escherichia coli is an anaerobic Gram negative bacillary AC 1 1,33 bacterium, which is a potential pathogen of birds, named AR 2 2,67 Avian Pathogenic E. coli (APEC) when are responsible for AZ 1 1,33 avian colibacillosis. This disease may develop in different ER 1 1,33 ET 11 14,67 forms, such as: colisepticemia, coligranuloma, cellulitis, ae- MT 25 33,33 rossacullitis, sinusitis, pericarditis, peritonitis, salpingitis, NH 19 25,33 hepatitis, panoftalmitis and osteomyelitis; however res- PH 2 2,67 piratory disease followed by septicemia and death is the AF = Absolute frequency, RF = Relative frequency, AA = Amazona aestiva, most frequent process (Janben et al. 2001). The genus Sal- AC = Agapornis roseicollis, AR = Ara chloptera, AZ = Amazona amazôni- monella has two species, S. bongori and S. enterica, which ca, ER = Eclectus rotates, ET = Eupsittula cactorum, MT = Melopsittacus presents six subspecies and 2759 serotypes which cause undulates, NH = Nymphicus hollandicus, PH = Psephotus haematonotus. three distinct diseases: pullorum disease, caused by Salmo- agar. After incubation, a single colony with morphological char- serotype Pullorum; Fowl typhoid, caused by nella enterica acteristics compatible with E. coli was selected and submitted to Salmonella enterica serotype Gallinarum; and paratyphoid the following biochemical tests: triple-sugar-iron (TSI) agar, sul- infection, caused by any of the remaining serotypes (Seo - et al. 2000). Both Salmonella sp. and E. coli are not regu- es-Proskauer, methyl red, urease broth and malonate. In addition, lar members of the intestinal microbiota of psittacine and, afite-indole-motility single colony from (SIM) each agar, plate lysine-iron-agar with morphological (LIA), aspects citrate, Vogsim- therefore isolating these bacteria from asymptomatic or ilar to Salmonella sp. was selected and submitted to TSI, urease immunosupressed individuals indicates a possibility of di- and LIA, followed by rapid slide agglutination test, whenever the sease (Marietto-Gonçalves & Almeida 2010, Marietto-Gon- biochemical results indicated the presence of this pathogen. If the çalves et al. 2010). serological test was positive, the isolate was submitted to the En- Antimicrobial resistance is the mechanism by which terobacteria Laboratory of the Oswaldo Cruz Institute Foundation bacteria may overcome the activity of antibiotics, which ac- All the isolates were maintained in nutrient agar in refriger- cording to WHO (World Health Organization) mainly occurs (Fiocruz)ator until tothe be antimicrobialconfirmed and susceptibility serotyped. test, which was per- due to the indiscriminate use of these drugs important for formed with the isolates being diluted until a turbidity equivalent human and veterinary medicine (Tortora et al. 2012). The- to 1.0 in McFarland scale was achieved. Then, the inoculum was re is scarce information about antimicrobial resistance and streaked in plate with Mueller-Hinton agar, to which discs con- diseases in pet birds, however there are reports involving taining the following antibiotics were added: amoxicillin (10µg), free-living birds as potential disseminators of E. coli and - Salmonella sp. resistant to cefalosporins, ampicillin, strep- - tomycin, sulfoxazole and tetracycline isolated from passe- mycinampicillin (10µg), (10µg), tetracycline azithromycin (30µg) (15µg), and sulfazotrim ciprofloxacin (25µg). (5µg), After dox rines and columbids (Andrés et al. 2013, Poirel et al. 2012, incubation,ycycline (30µg), inhibition enrofloxacin zone diameters (5µg), fosfomycin were measured (200µg), and genta com- Lopes et al. 2014). This study aimed to perform a survey pared to standards previously established (CLSI 2012). Multidrug resistance was considered whenever an isolate was resistant to at of and spp. from pet psittacine that died E. coli Salmonella least two antibiotics of different groups and in all incubation steps during one year and to describe the mains lesions in the afore mentioned, the conditions in bacteriological incubator were affected organs attributing scores, but also to identify the temperature of 37°C and duration 24h. Histopathological procedure was performed at the Animal Pathology Laboratory of the Federal University of Paraíba, where antimicrobialMATERIALS susceptibility ANDprofiles METHODS of the isolates. - slides of 5μm were prepared and stained with hematoxylin-eosin to to or that died in the Laboratory of Ornithological Studies from be evaluated for the presence of hemosiderosis (HEMO), inflamma AugustSeventy 2014 five psittacine to July 2015 pets were from submitted Fortaleza, to Brazil necropsy delivered to identify dead Lesions were observed and to each a mild (+), moderate (++) or tory infiltrate (INFINT), congestion (CNGT) and necrosis (NECRO). the presence of E. coli and Salmonella sp. This study was approved marked (+++) score was assigned according to the intensity. by the Ethics Committee of the Use of Animals of the State
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