Phenotypic and genotypic detection of multi drug resistant coagulase-positive Staphylococcus spp. isolates from canine pyoderma

Document Type : Full paper (Original article)

Authors

1 MVSc in Veterinary Science, Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Junagadh, India

2 Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Junagadh, India

3 Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Kamdhenu University, Navsari, India

Abstract

Background: Dermatological infections in dogs are challenging to treat due to antibiotic resistance, which leads to longer recovery time and the need for stronger antibiotics. Aims: This study was undertaken to determine the prevalence of antimicrobial resistance in coagulase-positive staphylococcal isolates from pyoderma infection in dogs. This study also aimed to identify isolates with methicillin-resistance and multidrug resistance. Methods: 73 coagulase-positive staphylococci isolated from varying degrees of canine pyoderma cases. The samples were analyzed for the presence of Staphylococcus spp. using polymerase chain reaction (PCR) and resistance against antibiotics was studied by antimicrobial profile, minimum inhibitory concentration (MIC), and PCR on isolated bacteria. Results: Out of 75 bacterial isolates identified, 73 isolates were confirmed as Staphylococcus species by PCR. A higher percentage of antibiotic resistance was observed against penicillin-G (46.27%), followed by amoxiclav (38.81%), enrofloxacin (32.84%), cefpodoxime, oxytetracycline (28.36% each), levofloxacin (26.86%), and co-trimoxazole (22.39%). 29 (49.15%) S. pseudintermedius, three (50.00%) S. schleiferi subsp. coagulans, and two (100%) S. aureus isolates exhibited multidrug resistance. However, one (1.49%) isolate (S. pseudintermedius) revealed low-level mupirocin resistance in the E-test. Also, 12 (20.34%) methicillin-resistant Staphylococcus pseudintermedius (MRSP), one (16.67%) methicillin-resistant S. schleiferi subsp. coagulans (MRSS) and one (50%) methicillin-resistant S. aureus (MRSA) were reported using PCR. Conclusion: This study helps to understand the increased level and pattern of resistance in coagulase-positive staphylococci isolated from different types of canine pyoderma cases.

Keywords

Main Subjects


Abdulgader, SM; Lentswe, T; Whitelaw, A and Newton-Foot, M (2020). The prevalence and molecular mechanisms of mupirocin resistance in Staphylococcus aureus isolates from a Hospital in Cape Town, South Africa. Antimicrob. Resist. Infect. Control., 9: 2-7. doi: 10.1186/s13756-020-00707-8.
Abusleme, F; Galarce, N; Quezada-Aguiluz, M; Iraguen, D and Gonzalez-Rocha, G (2022). Characterization and antimicrobial susceptibility of coagulase-positive Staphylococcus isolated in a veterinary teaching hospital in Chile. Rev. Argent. Microbiol., 54: 192-202. doi: 10.1016/j.ram.2021.12.001.
Ankita, and Gandge, RS (2018). Prevalence and antibiotic susceptibility pattern of Staphylococcus species in canine skin infection. IJCMAS., 7: 2305-2313. https://doi.org/ 10.20546/ijcmas.2018.706.276.
Bauer, AW; Kirby, WM; Sherris, JC and Turck, M (1966). Antibiotic susceptibility testing by a standardized single disk method. Ame. J. Clin. Pathol., 45: 493-496.
Bhat, UR and Bhagwat, VG (2010). Study to assess the beneficial effects of immunol liquid in the management of canine pyoderma. Vet. World. 3: 78-81.
Bhatt, AH (2021). Bacterial zoonoses transmitted by household pets and as reservoirs of antimicrobial resistant bacteria. Microb. Pathog., 155: 1-10. https://doi.org/ 10.1016/j.micpath.2021.104891.
Chaudhary, AK; Kumar, A and Shrivastva, M (2019). Study on prevalence and resistance patterns of bacterial pathogens isolated from canine pyoderma. IJCMAS., 1: 2305-2311. https://doi.org/10.20546/ijcmas.2019.801.241.
Clinical and Laboratory Standards Institute (CLSI) (2017). Performance standards for antimicrobial susceptibility testing. NCCLS document M100S. 37: 19087.
Couto, N; Belas, A; Couto, I; Perreten, V and Pomba, C (2014). Genetic relatedness, antimicrobial and biocide susceptibility comparative analysis of methicillin-resistant and -susceptible Staphylococcus pseudintermedius from Portugal. Microb. Drug Resist., 20: 364-371.
Duran, N; Ozer, B; Duran, GG; Onlen, Y and Demir, C (2012). Antibiotic resistance genes & susceptibility patterns in staphylococci. IJMR., 135: 389-396.
Dziva, F; Wint, C; Auguste, T; Heeraman, C; Dacon, C; Yu, P and Koma, LM (2015). First identification of methicillin-resistant Staphylococcus pseudintermedius strains among coagulase-positive staphylococci isolated from dogs with otitis externa in Trinidad, West Indies. Infect. Ecol. Epidemiol., 5: 1-6. https://doi.org/10.3402/ iee.v5.29170.
Freney, J; Brun, Y; Bes, M; Meugnier, H; Grimont, F; Grimont, PA and Fleurette, J (1988). Staphylococcus lugdunensis sp. Nov. and Staphylococcus schleiferi sp. nov., two species from human clinical specimens. IJSEM., 38: 168-172. https://doi.org/10.1099/00207713-38-2-168.
Godbeer, SM; Gold, RM and Lawhon, SD (2014). Prevalence of mupirocin resistance in Staphylococcus pseudintermedius. J. Clin. Microbiol., 52: 1250-1252. https://doi.org/10.1128/JCM.03618-13.
Gomez-Sanz, E; Torres, C; Lozano, C and Zarazaga, M (2013). High diversity of Staphylococcus aureus and Staphylococcus pseudintermedius lineages and toxigenic traits in healthy pet-owning household members. Underestimating normal household contact? Comp. Immunol. Microbiol. Infect. Dis., 36: 83-94.
Gonzalez-Dominguez, MS; Carvajal, HD; Calle-Echeverri, DA and Chinchilla-Cardenas, D (2020). Molecular detection and characterization of the mecA and nuc genes from Staphylococcus species (S. aureus, S. pseudintermedius, and S. schleiferi) isolated from dogs suffering superficial pyoderma and their antimicrobial resistance profiles. Front. Vet. Sci., 7: 1-11. https://doi.org/10.3389/fvets.2020.00376.
Guardabassi, L; Larsen, J; Weese, JS; Butaye, P; Batisti, A; Kluytmans, J; Lloyd, DH and Skov, RL (2013). Public health impact and antimicrobial selection of meticillin-resistant staphylococci in animals. JGAR., 1: 55-62. https://doi.org/10.1016/j.jgar.2013.03.011.
Hariharan, H; Gibson, K; Peterson, R; Frankie, M; Matthew, V; Daniels, J; Martin, NA; Andrews, L; Paterson, T and Sharma, RN (2014). Staphylococcus pseudintermedius and Staphylococcus schleiferi subspecies coagulans from canine pyoderma cases in grenada, West Indies, and their susceptibility to beta-lactam drugs. Vet. Med. Int., 2: 1-7. https://doi.org/10.1155/2014/850126.
Huerta, B; Maldonado, A; Ginel, PJ; Tarradas, C; Gomez-Gascon, L; Astorga, RJ and Luque, I (2011). Risk factors associated with the antimicrobial resistance of staphylococci in canine pyoderma. Vet. Microbiol., 150: 302-308. https://doi.org/10.1016/j.vetmic.2011.02.002.
Janardhan, L; Kumar, VA; Kumar, KS and Rani, MU (2022). Prevalence studies on canine pyoderma. Pharma Innov. J., 11: 1237-1239.
Jane, ES; Terry, MN and Stephen, DW (2014). Canine and feline infectious diseases. Jane, ES (Ed.), Canine and feline infectious diseases: Pyoderma, otitis externa, and otitis media. (1st Edn.), Elsevier Publication, W. B. Saunders. PP: 800-813. https://doi.org/10.1016/B978-1-4377-0795-3.00084-3.
Khinchi, RK (2019). Superficial pyoderma in canines in southern part of Rajasthan-A detailed epidemiological study. Int. J. Chem. Stud., 3: 2737-2740.
Kizerwetter-Swida, M; Chrobak-Chmiel, D; Kwiecien, E; Rzewuska, M and Binek, M (2021). Molecular characterization of high-level mupirocin resistance in methicillin-resistant staphylococci isolated from companion animals. Vet. Microbiol., 259: 1-8. https://doi.org/10.1016/ j.vetmic.2021.109160.
Kumar, D; Bisht, D and Faujdar, SS (2020). Incidence of mupirocin resistance in Staphylococcus aureus isolated from rural population: A new emerging challenge. IJCRR., 12: 82-85. http://dx.doi.org/10.31782/IJCRR.2020.12225.
Lai, C; Ma, YC; Shia, WY; Hsieh, YL and Wang, CM (2022). Risk factors for antimicrobial resistance of Staphylococcus species isolated from dogs with superficial pyoderma and their owners. Vet. Sci., 9: 1-13. https://doi.org/10.3390/vetsci9070306.
Lee, GY; Hang-Ho, L; Sun, YH; Joonbae, H; Kwang-Soo, L and Soo-Jin, Y (2019). Carriage of Staphylococcus schleiferi from canine otitis externa: antimicrobial resistance profiles and virulence factors associated with skin infection. J. Vet. Sci., 20: e6.
Loeffler, A and Lloyd, DH (2018). What has changed in canine pyoderma? A narrative review. Vet. J., 235: 73-82. https://doi.org/10.1016/j.tvjl.2018.04.002.
Lynch, SA and Helbig, KJ (2021). The complex diseases of Staphylococcus pseudintermedius in canines: where to next? Vet. Sci., 8: 1-19. https://doi.org/10.3390/ vetsci8010011.
Magiorakos, AP; Srinivasan, A; Carey, RB; Carmeli, Y; Falagas, ME; Giske, CG; Harbarth, S; Hindler, JF; Kahlmeter, G; Olsson-Liljequist, B; Paterson, DL; Rice, LB; Stelling, J; Struelens, MJ; Vatopoulos, A; Weber, JT and Monnet, DL (2012). Multidrug-resistant, extensively drug resistant and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. CMI., 18: 268-281. https://doi.org/10.1111/j.1469-0691.2011.03570.
Mahmood, HA and Flayyih, MT (2014). Detection of vanA gene of vancomycin-resistant Staphylococcus aureus by PCR technique. IJAR., 2: 209-216.
Mahmoudi, S; Mamishi, S; Mohammadi, M; Banar, M; Ashtiani, M; Mahzari, M; Bahador, A and Pourakbari, B (2019). Phenotypic and genotypic determinants of mupirocin resistance among Staphylococcus aureus isolates recovered from clinical samples of children: an Iranian hospital-based study. Infect. Drug Resist., 12: 137-143. https://doi.org/10.2147/IDR.S185610.
Martineau, F; Picard, FJ; Ke, D; Paradis, S; Roy, PH; Ouellette, M and Bergeron, MG (2001). Development of a PCR assay for identification of staphylococci at genus and species levels. J. Clin. Microbiol., 39: 2541-2547. https://doi.org/10.1128/JCM.39.7.2541-2547.2001.
Mostafa, MS and Awad, AR (2020). Localization and characterization of mupA gene in high and low-level mupirocin resistant methicillin resistant Staphylococcus aureus. Egypt. J. Med. Microbiol., 29: 171-177. https://doi.org/10.51429/Ejmm29322.
Park, JH; Kang, JH; Hyun, JE and Hwang, CY (2018). Low prevalence of mupirocin resistance in Staphylococcus pseudintermedius isolates from canine pyoderma in Korea. Vet. Dermatol., 29: e95-e37. https://doi.org/10.1111/ vde.12518.
Perez-Sancho, M; Sergio, A; Teresa, G; Marta, H; David, R; Lucas, D; Marta, EG and Jose, LB (2020). Antimicrobial resistance of coagulase-positive Staphylococcus isolates recovered in a veterinary university hospital. Antibiotics. 9: 1-12. doi: 10.3390/ antibiotics911075.
Prior, CD (2021). Prevalence of methicillin resistance in Staphylococcus pseudintermedius isolates from dogs with skin and ear infections in South Africa. J. S. Afr. Vet. Assoc., 93: 40a-40h. http://hdl.handle.net/2263/83288.
Priyantha, MA; Fernando, PS and De Alwis, PS (2021). Emerging antimicrobial resistance in coagulase-positive staphylococci and E. coli isolated from bovine clinical mastitis in Sri Lanka. AJAVA., 28: 29-35.
Quinn, PJ; Markey, BK; Leonard, FC; Hartigan, P; Fanning, S and Fitzpatrick, E (2011). Veterinary microbiology and microbial disease. 2nd Edn., Wiley-Blackwell Publication.
Rana, EA; Islam, MZ; Das, T; Dutta, A; Ahad, A; Biswas,
PK and Barua, H
(2022). Prevalence of coagulase-positive methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in dogs in Bangladesh. Vet. Med. Sci., 8: 498-508. https://doi.org/10.1002/ vms3.701.
Ravens, PA; Vogelnest, LJ; Ewen, E; Bosward, KI and Norris, JM (2014). Canine superficial bacterial pyoderma: evaluation of skin surface sampling methods and antimicrobial susceptibility of causal Staphylococcus isolates. Aust. Vet. J., 92: 149-155. https://doi.org/10.1111/ avj.12176.
Reddy, BS; Kumari, KN and Sivajothi, S (2016). Methicillin-resistant Staphylococcus aureus (MRSA) isolated from dogs with recurrent pyoderma. JDVAR., 3: 62-65. https://doi.org/10.15406/jdvar.2016.03.00073.
Sambrook, J and Russell, DW (2001). Molecular cloning: a laboratory manual. 3rd Edn., Vol. 1, New York, Cold Spring Harbor Laboratory Press.
Senapati, SK; Patra, RC and Panda, HK (2014). Prevalence and antibiogram of bacterial pathogens isolated from canine pyoderma. IJFV., 9: 41-45.
Shah, B; Mathakiya, R; Rao, N and Nauriyal, DS (2017). Organisms recovered from cases of canine pyoderma and their antibiogram pattern. J. Ani. Res., 7: 1067-1073. https://doi.org/10.5958/2277-940X.2017.00159.0.
Silva, V; Oliveira, A; Manageiro, V; Caniça, M; Contente, D; Capita, R; Alonso-Calleja, C; Carvalho, I; Capelo, JL; Igrejas, G and Poeta, P (2021). Clonal diversity and antimicrobial resistance of methicillin-resistant Staphylococcus pseudintermedius isolated from canine pyoderma. Microorganisms. 9: 1-10. https://doi.org/ 10.3390/microorganisms9030482.
Somayaji, R; Priyantha, MA; Rubin, JE and Church, D (2016). Human infections due to Staphylococcus pseudintermedius, an emerging zoonosis of canine origin: report of 24 cases. Diagn. Microbiol. Inf., 85: 471-476.
Strommenger, B; Layer, F and Werner, G (2018). Staphylococcus aureus and methicillin-resistant Staphylococcus aureus in workers in the food industry. 1st Edn., Robert Koch Institute, Wernigerode, Germany, Academic Press. PP: 163-188. https://doi.org/10.1016/ B978-0-12-809671-0.00009-7.
Sum, S; Park, HM and Oh, JY (2020). High-level mupirocin resistance in Gram-positive bacteria isolated from diseased companion animals. J. Vet. Sci., 21: e40. https://doi.org/ 10.4142/jvs.2020.21.e40.
Taha, S; Kamel, N and Metwally, L (2022). Detection of mupirocin resistance in methicillin-resistant Staphylococcus aureus isolates in an Egyptian Hospital. Egypt. J. Med. Microbiol., 31: 51-55. https://doi.org/ 10.21608/ejmm.2022.247201.
Tamakan, H and Gocmen, H (2022). Genetic characterization of methicillin resistant Staphylococcus pseudintermedius in dogs and cats in Cyprus: Comparison of MRSP and MRSA results. Pak. J. Zool., 54: 1-6. https://dx.doi.org/10.17582/ journal.pjz/20211101121137.
Wu, MT; Burnham, CA; Westblade, LF; Dien Bard, J; Lawhon, SD; Wallace, MA; Stanley, T; Burd, E; Hindler, J and Humphries, RM (2016). Evaluation of oxacillin and cefoxitin disk and MIC breakpoints for prediction of methicillin resistance in human and veterinary isolates of Staphylococcus intermedius group. J. Clin. Microbiol., 54: 535-542. https://doi.org/10.1128/ JCM.02864-15.