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Bacteria Isolated from the Lower Respiratory Tract of Sheep and Their Relationship to Clinical Signs of Sheep Respiratory Disease1 Mariane F

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Bacteria Isolated from the Lower Respiratory Tract of Sheep and Their Relationship to Clinical Signs of Sheep Respiratory Disease1 Mariane F Pesq. Vet. Bras. 39(10):796-801, October 2019 DOI: 10.1590/1678-5150-PVB-6315 Original Article Livestock Diseases ISSN 0100-736X (Print) ISSN 1678-5150 (Online) PVB-6315 LD Bacteria isolated from the lower respiratory tract of sheep and their relationship to clinical signs of sheep respiratory disease1 Mariane F. Franco2, Natália C. Gaeta2* , Mario A.R. Alemán2, Priscilla A. Mellville3, Jorge Timenetsky4, Mário F.A. Balaro5 and Lilian Gregory2 ABSTRACT.- Franco M.F., Gaeta N.C., Alemán M.A.R., Mellville P.A., Timenetsky J., Balaro M.F.A. & Gregory L. 2019. Bacteria detected from the lower respiratory tract of sheep and their relationship to clinical signs of sheep respiratory disease. Pesquisa Veterinária Brasileira 39(10):796-801. Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. E-mail: [email protected] was studied the bacterial etiology of respiratory disease and the clinical signs of 99 female andRespiratory male sheep diseasesbreed in arethe amongstates of the São most Paulo important (SP) and diseases Rio de Janeiro in sheep (RJ), flocks. Brazil. Herein After bacteria and Mycoplasma spp. were searched, as well as their association with the clinical physicalstatus and examination clinical signs of animals, of sheep tracheobronchial with respiratory flushing disease. samples The main were observed obtained. signs The were:usual tachypnea (75%), increase of rectal temperature (09.4%), mucopurulent/purulent nasal discharge (21.9%), cough (25%), dyspnea (31.2%), changes of lung sounds at auscultation (87.5%) and chest percussion (28.1%) in pneumonic sheep. Non-fermenting gram-negative bacteria and Bacillus sp. were the most isolated bacteria. Microorganisms of the Mollicutes detection of M. mycoides subsp. capri classof São were Paulo, molecularly Brazil. (PCR) detected in 33.3% of the animals. In addition, the specific was described for the first time in sheep from the state INDEX TERMS: Bacteria, lower respiratory tract, sheep, clinics, sheep respiratory disease, Mycoplasma spp., pneumonia, respiratory disease, bacterioses. RESUMO.- [Bactérias detectadas no trato respiratório pesquisa teve como objetivo determinar a etiologia bacteriana inferior de ovinos e a relação com as manifestações clínicas da doença respiratória e sua relação com sinais clínicos em da doença respiratória ovina.] A doença respiratória é uma ovinos criados nos estados de São Paulo e Rio de Janeiro, Brasil. das doenças mais importantes em rebanhos ovinos. Esta Noventa e nove ovelhas machos e fêmeas dos Estados de São Paulo (SP) e Rio de Janeiro (RJ) foram estudadas. Após o exame 1 Received on May 6, 2019. Accepted for publication on June 4, 2019. A presença de bactérias aeróbias e Mycoplasma spp. foram 2 Departamento de Clínica Médica, Faculdade de Medicina Veterinária físico,estudados, amostras assim de como lavagem a associação traqueobrônquica entre os microrganismos foram obtidas. e Zootecnia (FMVZ), Universidade de São Paulo (USP), Av. Prof. Orlando e estado clínico e sinais clínicos de doença respiratória em Marques de Paiva 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil. *Corresponding author: [email protected] ovinos. As principais manifestações clínicas observadas 3 Departamento de Medicina Preventiva e Saúde Animal, Faculdade de Medicina foram: taquipneia (75%), alta temperatura retal (09,4%), Veterinária e Zootecnia (FMVZ), Universidade de São Paulo (USP), Av. Prof. secreção nasal mucopurulenta/purulenta (21,9%), tosse Orlando Marques de Paiva, 87, Cidade Universitária, São Paulo, SP 05508-270. (25%), dispneia (31,2%), sons pulmonares alterados na 4 Laboratório de Micoplasmas, Instituto de Ciências Biomédicas, Universidade ausculta (87,5%) e na percussão torácica (28,1%) em ovelhas de São Paulo (USP), Av. Prof. Lineu Prestes 2415, Cidade Universitária, São Paulo, SP 05508-000. Bacillus sp. foram as bactérias mais isoladas. Microrganismos 5 Unidade de Pesquisa Experimental em Caprinos e Ovinos, Setor de Ovinos e Caprinos, Fazenda Escola de Cachoeiras de Macacu (FECM), Universidade pneumônicas.da classe Mollicutes Bactérias foram gram-negativas detectados não molecularmente fermentadoras e Federal Fluminense (UFF), Rod. João Goularte 1300-1306, Setenta, Cachoeiras (PCR) em 33,3% dos ovinos. Além disso, descreve-se pela de Macacu, RJ 28680-000, Brazil. primeira vez no estado de São Paulo, Brasil, a detecção do 796 Bacteria isolated in the lower respiratory tract of sheep and their relationship to clinical signs of sheep respiratory disease 797 M. mycoides subsp. capri na espécie ovina utilizando a reação disease and their relationship to clinical signs of sheep de polimerase em cadeia. respiratory disease. TERMOS DE INDEXAÇÃO: Bactérias, trato respiratório inferior, ovinos, ovelhas, clínica, doença respiratória ovina, Mycoplasma spp., MATERIALS AND METHODS pneumonia, bacterioses. All procedures were carried out in agreement with the guidelines of the Committee of Ethics on Animal Use (Protocol no. 3498060716). INTRODUCTION In this cross-sectional research, 99 male and female sheep from Sheep respiratory disease is one of the most important diseases sampling12 flocks forfrom convenience. the States of São Paulo (SP=6) and Rio de Janeiro due to its economic losses. In New Zealand, pneumonia in (RJ=6) were enrolled in this study, according to a non-probabilistic inlambs sheep cost flocks NZ$28.1 (Marcondes million in et 2008 al. 2011, (Goodwin-Ray Radostits et al. 2008).2017) Animals were classified as healthy and unhealthy (showing From 1996 to 2010, 10% of diagnosed diseases in lambs in the clinical signs of respiratory disease) after physical exam (Benesi et al. “Universidade Federal do Mato Grosso do Sul” were pneumonic 2013, Gaeta et al. 2018). Sheep were considered unhealthy if related (Almeida et al. 2013). The incidence of pneumonia show at least two of the following parameters: respiratory rate in sheep and other respiratory diseases in Argentina was more than 30 respiratory movements per minute, coughing, 57.9% (Suárez & Busetti 2009). In Tanzania, 31.4% of lungs rectal temperature higher than 40°C and abnormal sound on had pneumonic lesions in slaughter (Mellau et al. 2010). thoracic auscultation. Recently, McRae et al. (2016) detected pneumonic lesions Tracheobronchial wash samples were obtained from both in 28% of lambs at slaughter in New Zealand. Important risk factors related to respiratory disease of trichotomy and antisepsis using 70% alcohol, chlorhexidine and ruminants are the environmental high temperature and humidity, iodopovidonehealth and pneumonic were performed sheep (Anton in the third& Mayayo part of2007). the trachea. Briefly, increased animal density, stress, dust, poor ventilation, and An Intracath (BD, USA) was introduced by tracheocentesis and parasites (Goodwin-Ray et al. 2008, Taylor et al. 2010, Scott 20 to 50mL of sterile saline 0.9% was instilled, recovering up to 2011). Some viruses are considered primary pathogens, changing the epithelial cells, and interfering in the immune 5mL. An aliquot (500µL) was added to Stuart transport media and response (Bosch et al. 2013), leading to secondary bacterial stored at 4°C until the isolation of bacteria. Another aliquot was infections. According to Sharp & Nettleton (2007) acute added to transport media for Mycoplasma spp. containing glycerol respiratory disease is not caused by virus infections alone. and stored in liquid nitrogen. Mannheimia (M.) haemolytica and Pasteurella (P.) multocida are important respiratory pathogens in sheep broth and the cultures in agar plates were performed by plating 10µL of theseTracheal samples suspensions in MacConkey were inoculatedand Blood agarin Brain (5%). Heart The Infusioncultures Pasteurellaceae, order Pasteurellales (Griffin 2010). These were incubated for 24 to 72 hours at 37°C in aerobic atmosphere. flocksbacteria (Radostits are part ofet theal. 2017). respiratory Both tractbacteria microbiota belong to in family sheep The obtained colonies were noted for their morphology, Gram stained (Viana et al. 2007) and as stated before, immunosuppression and screened for biochemical tests (Staphylococcus sp.: catalase and viral infections predispose to respiratory disease caused test, sensitivity to bacitracin and furazolidone and oxidase test; by both species (Bosch et al. 2013, Viana et al. 2007). Almost Streptococcus sp.: catalase test, sensitivity to bacitracin, growth at 10% of pneumonic sheep were positive for P. multocida in 65% NaCl, bile-esculin test and growth at 45°C; Bacillus sp.: catalase lungs in Nigeria (Odugbo et al. 2006). Ten sheep died during test; Klebsiella sp. and non-fermenting bacteria: triple sugar iron an outbreak of M. haemolytica associated pneumonia in the State of Minas Gerais in 2009 (Araujo et al. 2009). growth, production of indole, urease and fenilalanine desaminase, Mycoplasma spp. belongs to class Mollicutes, order oxidation-fermentation test, motility test, lysine descarboxylation Mycoplasmatales and family Mycoplasmataceae (Razin test, use of citrate and malonate, lactose and glucose fermentation, nitrate reduction, Voges-Proskauer test and Methyl-red test) (Murray et al. 1999). &detected Hayflick in healthy2010). and They diseased are the ruminants smallest (Ayling organisms et al. 2004,with Mycoplasma spp. isolation was performed using solid and liquid selfKumar-replication et al. 2011, (Razin Oliveira & etHayflick
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