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REDVET Rev. Electrón. vet. http://www.veterinaria.org/revistas/redvet 2017 Volumen 18 Nº 9 - http://www.veterinaria.org/revistas/redvet/n090917.html

REDVET - Revista electrónica de Veterinaria - ISSN 1695-7504

Antibiotic resistance of pathogenic bacteria isolated from dogs at a veterinary clinic in Callao, Peru - Resistencia a los antibióticos de bacterias patógenas aisladas de canes en una clínica veterinaria del Callao, Perú Daniel Ángel Luján-Roca1,2* │ Ivanhoe Saavedra-Espinoza2 │ Luz Milagros Luján-Roca2 1

Post-graduate program in Infectious Diseases and Tropical Medicine, School of Medicine, Universidade Federal de Minas Gerais, Minas Gerais, Brazil. 2 Clínica Veterinaria La Colonial, Callao, Peru. *Contacto: [email protected]

ABSTRACT The purpose of this study was to determine the antibiotic resistance pattern in bacterial pathogen strains isolated from dogs. Isolates were collected from various body sites of diseased dogs from 2003 to 2012 at a private veterinary clinic in Callao, Peru. Bacterial species were identified by standard microbiological techniques and susceptibility to antibiotics was determined by the disc diffusion method. A total of 81 strains were evaluated; the most prevalent species were Staphylococcus intermedius group (SIG), Pseudomonas aeruginosa and Escherichia coli. The antibiotic susceptibility test indicated resistance of these bacteria to some antibiotics: SIG was resistant to trimethoprim/sulfamethoxazole (31%) and enrofloxacin (23%), P. aeruginosa was resistant to cephalexin (86%) and clindamycin (76%), and E. coli was resistant to clindamycin (78%) and trimethoprim/sulfamethoxazole (75%). In general, P. aeruginosa and E. coli demonstrated the highest levels of resistance, and amikacin showed the best susceptibility. It is advisable to evaluate a larger groups of samples and to determine the molecular patterns of resistance to select the

Antibiotic resistance of pathogenic bacteria isolated from dogs at a veterinary clinic in Callao, Peru http://www.veterinaria.org/revistas/redvet/n090917/091757.pdf


appropriate antibiotic therapy for the treatment of major diseases seen in the field of small animal veterinary medicine. Keywords: Antibiotic resistance, bacterial pathogens, dogs.

RESUMEN El propósito de este estudio fue determinar el patrón de resistencia a antibióticos en cepas de patógenos bacterianos aisladas de perros. Los aislados se recolectaron de varias zonas del cuerpo de perros enfermos de 2003 a 2012 en una clínica veterinaria privada del Callao, Perú. Las especies bacterianas se identificaron mediante técnicas microbiológicas estándar y la susceptibilidad a los antibióticos se determinó mediante el método de difusión en disco. Se evaluaron un total de 81 cepas; Las especies más prevalentes fueron el grupo Staphylococcus intermedius (SIG), Pseudomonas aeruginosa y Escherichia coli. La prueba de susceptibilidad a los antibióticos indicó resistencia de estas bacterias a algunos antibióticos: el SIG fue resistente a trimetoprima/sulfametoxazol (31%) y enrofloxacina (23%), P. aeruginosa fue resistente a cefalexina (86%) y clindamicina (76%) y E. coli fue resistente a clindamicina (78%) y trimetoprima/sulfametoxazol (75%). En general, P. aeruginosa y E. coli demostraron los niveles más altos de resistencia, y la amikacina mostró la mejor susceptibilidad. Es recomendable evaluar grupos más grandes de muestras y determinar los patrones moleculares de resistencia para seleccionar la terapia antibiótica apropiada en el tratamiento de enfermedades graves observadas en el campo de la medicina veterinaria de animales pequeños. Palabras clave: Resistencia antibiótica, patógenos bacterianos, canes.

INTRODUCTION



Bacterial infections have negative effects on the health of companion animals. Due to the short period of time between generations and the capacity of exchanging genetic material, the development of resistance to antibiotics for many animal bacteria has been inevitable (McKellar, 1998). The antibiotic resistance epidemic is an ecological disaster of unknown consequences and without any evident solution (Gould, 2009). To find microorganisms resistant to diverse antibiotics is not unusual (Arriola et al., 2011; Luján et al., 2013; Luján et al., 2014). This has increased in developing countries since the use of antibiotics to treat people and animals is not regulated (Hart and Kariuki, 1998). This problem has been observed in pathogenic bacteria recovered from dogs, although the reports are scarce (Cavalcanti and Coutinho, 2005). Appropriate knowledge of antibiotic resistance in these pathogenic bacteria is useful to establish recommendations regarding the wise use of these drugs. The objective of this communication was to identify resistance to antibiotics in bacteria isolated from dogs. MATERIALS AND METHODS This retrospective study was carried out at Clínica Veterinaria La Colonial, located in Bellavista district, Callao, Peru. The results of bacterial cultures and antibiograms from routine hospital patients submitted or not to previous treatment with antibiotics during the period from January 1 2003 to December 31 2012 were reviewed. Only one isolate for each patient was considered. The evaluation of the isolates was accomplished at the microbiology and parasitology laboratory of the Veterinary Medicine faculty at Universidad Nacional Mayor de San Marcos in Lima city, Peru. Microorganism identification was accomplished according to cultural, morphological, staining and biochemical characteristics. The analysis of susceptibility was carried out using the Bauer-Kirby disk diffusion technique (Bauer and Kirby, 1966). Strains with intermediate susceptibility were defined as sensitive. The following antibiotics were included: amoxicillin-clavulanic acid (20/10 µg), cephalexin (30 µg), ceftriaxone (30 µg), ciprofloxacin (5 µg), norfloxacin (10 µg), enrofloxacin (5 µg), gentamicin (10 µg), amikacin (30 µg), tobramycin (10 µg), clindamycin (2 µg) and trimethoprim/sulfamethoxazole (25 µg).



A commercial statistical software package (STATA 12.0) was used to determine simple proportions and confidence intervals (Wald test). RESULTS During the study period, a total of 92 bacteriological cultures provided 81 pathogenic bacteria (87%, CI95% = 80.08-93.84). The dogs included in the study were a heterogeneous population comprised of 43 (53.1%) males and 38 (46.9%) females. A median age was 6 years 18 days. Isolates were recovered from different affected sites: ear (36), skin (25), infected wounds (7), urine (6), nostrils (4) and rectum (3). The most frequent breeds were: mongrel (19), cocker spaniel (9) and schnauzer (7) (Table 1). The highest frequencies of the different pathogenic species were SIG and P. aeruginosa, both with 22 strains each one (Table 2). The resistance rates during the analysis showed that SIG was highly resistant to trimethoprim/sulfamethoxazole (31%), while ciprofloxacin resistance was found in 5%. In the case of P. aeruginosa, high resistance rates were found against cephalexin (86%), while 100% sensitivity to gentamicin was observed. Regarding the E. coli isolates, these showed high resistance to clindamycin (78%) and trimethoprim/sulfamethoxazole (75%); otherwise, 100% sensitivity to amikacin was observed (Table 3). Table 1.- The breeds of dogs, Clínica Veterinaria La Colonial, 2003-2012, Callao, Peru. _______________________________________________________________________________ Breed nº % _______________________________________________________________________________ Boxer Bulldog Chihuahua Cocker spaniel Collie Dachshund Fila brasileiro Fox terrier

1 4 1 9 1 1 1 4

5.6 3.8 1.2 14.4 1.2 1.2 0.6 1.8



German shepherd 4 3.8 Golden retriever 1 1.2 Labrador retriever 4 6.4 Mongrel 19 23.5 Pekingese 4 4.4 Pit Bull 1 2.4 Rottweiler 5 2.4 Samoyed 1 1.8 Schnauzer 7 8.6 Shar-Pei 5 1.8 Shih tzu 4 6.4 Siberian husky 4 1.2 _______________________________________________________________________________ Total

81

100%

Table 2 .- Frequency of pathogenic bacteria isolated from dogs, Clínica Veterinaria La Colonial, 2003-2012, Callao, Peru. Year ____________________________________________________________

Total _____________________

2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 nº (%) CI 95% ____________________________________________________________________________________________________________________ SIG Staphylococcus aureus Staphylococcus albus Pseudomonas spp. Pseudomonas aeruginosa Escherichia coli Proteus spp. Proteus mirabilis Proteus vulgaris Citrobacter spp. Enterobacter spp.

2 1 0 0 1 0 0 0 0 0 0

1 0 0 0 0 0 0 0 0 0 0

3 2 0 0 2 0 0 0 0 0 0

2 1 0 1 1 0 0 0 0 0 0

2 0 0 2 1 0 0 0 0 0 0

3 1 0 1 2 0 0 0 0 0 0

6 0 0 0 2 1 0 0 0 1 1

1 0 0 1 6 3 0 1 0 1 0


1 0 0 0 1 3 1 2 1 2 0

1 0 1 0 6 7 0 1 0 1 0

22 5 1 5 22 14 1 4 1 5 1

27.2 6.2 1.2 6.2 27.2 17.3 1.2 4.9 1.2 6.2 1.2

17.5-36.9 0.9-11.4 0.0-3.6 0.9-11.4 17.5-36.9 9.1-25.5 0.0-3.6 0.2-9.7 0.0-3.6 0.9-11.4 0.0-3.6


Table 3.- Antibiotic resistance in SIG, P. aeruginosa and E. coli isolated from dogs, Clínica Veterinaria La Colonial, 20032012, Callao, Peru. SIG

P. aeruginosa

E. coli

Antibiotic _______________________________________________ nº % nº % nº % _______________________________________________________________________________ Amoxicillin/clavulanic acid 2/14 14 13/22 59 6/13 46 Cephalexin 3/17 17 12/14 86 5/10 50 Ceftriaxone ± ± 2/12 17 ± ± Ciprofloxacin 1/20 5 0/16 0 6/10 60 Norfloxacin 1/17 6 ± ± ± ± Enrofloxacin 3/13 23 3/20 15 6/13 46 Gentamicin 2/14 14 0/11 0 2/6 33 Amikacin 1/13 7 1/12 8 0/7 0 Tobramycin ± ± 5/12 42 2/7 29 Clindamycin 2/17 12 13/17 76 7/9 78 Trimethoprim/sulfamethoxazole 4/13 31 17/21 81 9/12 75 _______________________________________________________________________________ ± = Non tested



DISCUSSION The amount of antibiotics used in veterinary practice is in general more than the levels used for human medicine. In veterinary medicine, the role of antibiotic use in the development of resistance has only been recently discussed; in this context, surveillance of antibiotic resistance is very important (da Silva, 2013). SIG is one of the commonly isolated Staphylococci of skin infections and ears in dogs. It is usually part of the flora of the dog, but it can also be an opportunist pathogen (Cristina and Degi, 2013). In our study, this microorganism presented resistance to trimethoprim/sulfamethoxazole (31%), a lower prevalence than has been reported locally (43.3%) (Antúnez et al., 2009), but greater than rates in South Africa (24%) (Blunt, 2013) and the United States (23%) (Petersen et al., 2002). P. aeruginosa can be isolated from pyoderma and chronic otitis. It represents a challenge to antibiotic therapy due to its intrinsic resistance to several antibiotics (Luján, 2014). In this case, resistance to cephalexin (86%) was observed at a lower rate than that reported in Italy (100%) (Ghidini, 2011) but higher than in Romania (62.5%) (Dégi, 2010). E. coli is usually encountered in intestinal tract of dogs and it can be a potential source of resistance genes for pathogenic bacteria (Harada, 2011). Considerable resistance to cephalexin (78%) was found, similar to the prevalence observed in Brazil (82.3%) (Ishii, 2011). It is noteworthy that, in general, the resistance found in SIG, E. coli and P. aeruginosa to trimethoprim/sulfamethoxazole was high (31%-81%), with the exception of P. aeruginosa (with intrinsic resistance) the resistance to this antibiotic is relatively low (Matanović et al., 2012). These resistance levels may be due to frequent use of this antibiotic and reveals a potentially agent which might select for resistance to other antibiotics. The problem of resistance to antibiotics in dogs is complex; according to available data, the susceptibility profiles reflect the use of antibiotics at a given time (Prescott et al., 2002). In Peru, several problems have resulted from the quality of prescription, dispensation and the use of antibiotics in veterinary medicine;



these include the use of antibiotics in viral illnesses, the use of the same antibiotics in veterinary medicine and in human medicine and the weakness of public and private services to surveil and control veterinary products (Falcón et al., 2010). It is advisable not to stop treatment early, since this provides bacteria the opportunity to develop resistance to the antibiotic, hindering the remission of the infection and making it necessary to use a more potent antibiotic (Arias et al., 2012). CONCLUSIONS Moderately high resistance profiles against the evaluated antibiotics were observed in P. aeruginosa and E. coli, which presented the highest levels of resistance. The lowest levels of resistance were found for amikacin. It is advisable to evaluate a greater number of samples and to determine the molecular patterns of resistance. ACKNOWLEDGMENTS To Maria G. F. Penido BA for her administrative support.

REFERENCES     

Antúnez O, Calle S, Morales S, Falcón N, Pinto C (2009) Frecuencia de patógenos aislados en casos clínicos de dermatitis bacteriana canina y su susceptibilidad antibiótica. Rev Inv Vet Perú 20:332-338. Arias MVB, Carrilho CMDM (2012) Resistência antimicrobiana nos animais e no ser humano. Há motivo para preocupação?. Semina: Ci Agrárias 33:775-790. Arriola CS, Güere ME, Larsen J, Skov RL, Gilman RH, Gonzales AE, Silbergeld EK (2011) Presence of methicillin-resistant Staphylococcus aureus in pigs in Peru. PLoS ONE 6(12):e28529. Bauer AW, Kirby EM (1966) Antibiotic susceptibility testing by standardized single disk method. Am J Clin Pathol 45:493-496. Blunt CA, van Vuuren M, Picard J (2013) Antimicrobial susceptibility profiles of Staphylococcus intermedius isolates from clinical cases from canine pyoderma in South Africa. J S Afr Vet Assoc 84:E1-6.



          

   

Cavalcanti S, Coutinho S (2005) Identificação e perfil de sensibilidade antibacteriana de Staphylococcus spp isolados da pele de cães sadios e com piodermite. Clín Vet 58:60-66. Cristina RT, Degi J (2013) Multiresistant Staphylococcus intermedius isolated from otitis externa in dogs and them human owners. A practical approach. Afr J Pharm Pharmacol 7:1351-1356. da Silva KC, Knöbl T, Moreno AM (2013) Antimicrobial resistance in veterinary medicine: mechanisms and bacterial agents with the greatest impact on human health. Braz J Vet Res Anim Sci 50:171-183. Dégi J, Cristina RT, Stancu A (2010) Otitis externa caused by bacteria of the genus Pseudomonas in dogs. Lucrări 43:143-147. Falcón N, Ortega C, Gorniak S, Villamil LC, Ríos C, Simón MC (2010) El problema de la resistencia a antibióticos en salud pública. Una Salud 1:75-88. Ghidini F, Piancastelli C, Taddei S, Gandolfo E, Cavirani S, Cabassi CS (2011) Antibiotic sensitivity of bacterial isolates from cases of canine dermatitis. New Microbiol 34:403-408. Gould IM (2009) Antibiotic resistance: the perfect storm. Int J Antimicrob Agents 34:S2-S5. Harada K, Morimoto E, Kataoka Y, Takahashi T (2011) Clonal spread of antimicrobial-resistant Escherichia coli isolates among pups in two kennels. Acta Vet Scand 53:11. Hart CA, Kariuki S (1998) Antimicrobial resistance in developing countries. Br Med J 317:647-650. Ishii JB, Freitas JC, Arias MVB (2011) Resistência de bactérias isoladas de cães e gatos no Hospital Veterinário da Universidade Estadual de Londrina (2008-2009). Pesq Vet Bras 31:533-537. Luján DA, Ibarra JO, Mamani E (2013) Resistencia a los antibióticos en cepas de Klebsiella pneumoniae, Serratia spp. y Acinetobacter spp. aisladas de pacientes con infección del tracto urinario - Lima, Perú. Electron J Biomed 1:25-30. Luján DA, Málaga EJ (2014) Resistencia a quinolonas en aislados clínicos de Staphylococcus aureus. Perspect Med 25:48-50. Luján DA (2014) Pseudomonas aeruginosa: un adversario peligroso. Acta Bioquim Clin Latinoam 48:465474. Matanović K, Mekić S, Šeol B (2012) Antimicrobial susceptibility of Staphylococcus pseudointermedius isolated from dogs and cats in Croatia during a sixth-month period. Vet Archiv 82:505-517. McKellar QA (1998) Antimicrobial resistance: a veterinary perspective. Br Med J 317:610-611.



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

Petersen AD, Walker RD, Bowman MM, Schot II HC, Rosser Jr EJ (2002) Frequency of isolation and antimicrobial susceptibility patterns of Staphylococcus intermedius and Pseudomonas aeruginosa patterns isolates from canine skin and ear samples over a 6-year period. J Am Anim Hosp Assoc 38:407413. Prescott JF, Brad Hanna WJ, Reid-Smith R, Drost K (2002) Antimicrobial drug use and resistance in dogs. Can Vet J 43:107-116.

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