Indian Journal of Animal Research

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Research Article

Characterization of Staphylococcus pseudintermedius Isolates from Skin Infections of Dogs

Hridya Susan Varughese1, Murugesan Ananda Chitra1,*
1Department of Veterinary Microbiology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Vepery, Chennai-600 007, Tamil Nadu, India.

ABSTRACT

Background: Staphylococcus pseudintermedius is a part of the canine skin microflora and an opportunistic pathogen. It plays a central role in canine pyoderma, otitis and surgical wound infections. These conditions correlate with virulence genes distributed in the bacterial genome. These genes determine strain variability on typing, in turn aiding epidemiological surveillance. The aim of this study was to isolate, identify and characterize Staphylococcus pseudintermedius (SP) and Methicillin resistant Staphylococcus pseudintermedius (MRSP) from dogs with skin infections in Chennai, India.

Methods: SP and MRSP positive isolates were identified by multiplex PCR for nuc and mecA genes respectively. Characterization of the isolates for virulence genes responsible for biofilm formation (icaA, icaD), cell wall adherence (SpsO, SpsK, SpsP, SpsQ, SpsF), toxins (ExpA, ExpB, SIET, Sel, Se-int, LukS, LukF) and gene regulation (Agr, SarA) was performed.

Result: Out of 275 samples, 120 SP and 8 MRSP positive isolates were identified. Only one isolate could be typed as SCCmec Type V whereas other MRSP isolates were non typeable. Agr typing of MRSP isolates revealed type II in 7 isolates and type III in one isolate. Our study revealed that there was no significant difference in the detection of virulence genes between MSSP and MRSP.

INTRODUCTION

Staphylococcus pseudintermedius (SP) is a commensal bacterium of dogs and cats skin and mucous membranes. It is usually present in pharynx, rectum, nares, conjunctiva, perineum, perioral and axilla region. SP is responsible for pyoderma, otitis externa and post-operative infections in dogs. S. pseudintermedius can also colonize the nasal cavity of pet owners, veterinarians and animal handlers (Bannoehr and Guardabassi, 2012) and it is generally harmless in healthy individuals. However, individuals with compromised immune systems (e.g. HIV/AIDS, transplant and cancer patients) are more susceptible to SP infection and there have been reports of dermatitis and septicemia in human patients (Somayaji et al., 2016). Consequently S. pseudintermedius and especially methicillin resistant (MRSP) strains, are of public concern as potentially emerging zoonotic bacteria (Paul et al., 2011). Contamination with S. pseudintermedius in the environment in animal hospitals and households has been reported (Laarhoven et al., 2011; van Duijkeren et al., 2011) enabling re-infection and transfer of antimicrobial resistance genes (Frank et al., 2009).

The pathogenic potential of the organism can be explored through study of virulence factors such as cell adhering factors, toxins and enzymes. Particular genomic regions like the OriC in SP, controls virulence genes for adherence like SpsK, SpsP, SpsQ, SpsL, SpsM, SpsG and SpsJ genes. After adherence, epithelial colonization can be attributed to biofilm formation coded by the ica operon (Gerke et al., 1998). Following colonization, bacterial cell survival is dependent on toxin production for defense. Hence, this study was aimed to isolate, identify and determine the distribution and frequencies of various virulence factors in SP and MRSP isolates from skin infections of dogs in Chennai, India.

MATERIALS AND METHODS

Isolation and Identification
 
Skin swabs from pyoderma, otitis, allergy, demodicosis, dermatitis and other cases (hypothyroidism etc.) were collected during September 2017 to February 2018 from dogs presented to Dermatology ward of Teaching Veterinary Clinical Complex of Madras Veterinary College, Chennai, India. SP and MRSP isolation and identification were carried out as described previously (Ananda Chitra et al., 2016).

Table 1: Details of the primers used for identification and characterization of SP isolates.


 
Virulence genes detection
 
Molecular characterization for the presence of different virulence genes was performed by conventional PCR and primer details along with reference for these genes are given in Table 1. PCR was performed in a reaction volume of 10 µl containing approximately 100-150 ng of genomic DNA, 5 pmol of each primer in 2X master mix (Ampliqon, Denmark). Cycling conditions were 95°C for 3 min, followed by 30 cycles of denaturation at 95°C for 30 s, annealing at appropriate temperature for 30 s, extension at 72°C for 30/45 s and a final extension cycle of 5 min at 72°C. PCR products were loaded on a 2% agarose gel for electrophoresis, visualized with ethidium bromide and documented.
 
Typing of MRSP isolates
 
The SCCmec cassette of MRSP isolates was typed using previously described multiplex PCR methods developed by Zhang et al., (2012) and Perreten et al., (2010) for type II-III and VII. Agr typing was carried out using published primers (Ananda Chitra et al., 2015) and sequencing by Sanger’s sequencing technique.

RESULTS AND DISCUSSION

Isolation and Identification
 
A total of 275 swab samples were collected from various skin infections of dogs. Predominant samples (n=164) were collected from superficial bacterial folliculitis (pyoderma), followed by otitis (n=38), equal number of demodicosis and dermatitis cases (n=23 each). Out of 275 samples, 128 (46.5%) samples were found to be positive for SP by amplifying 780 bp amplicon of SP specific thermonuclease gene in PCR as shown in Fig 1. The prevalence of SP infection in various skin infections is given in Fig 2. SP has also been identified at varying rates with 26.6% incidence in Lithuanian dogs (Ruzauskas et al., 2016), 59% Chennai, India (Ananda Chitra et al., 2016) and 59.6% in Brazil (Scherer et al., 2018).

Fig 1: Agarose gel electrophoresis showing the results of PCR amplified products of nuc and mecA genes of SP isolates.



​

Fig 2: Case wise prevalence of MSSP and MRSP isolates



Among the 128 SP, eight isolates (6.25%) were identified as MRSP by detecting mecA gene (Fig 1). Six out of eight isolates were from pyoderma cases and two were from otitis cases. Higher prevalence of MRSP infection with 48% in China (Wang et al., 2012), 40.5% in Canada (Beck et al., 2012) and 28% in Chennai, India (Ananda Chitra et al., 2016). However, low MRSP prevalence rates have also been recorded such as 2% in Sweden (SWEDRES-SVARM, 2016) and 13.4% in Canada (Saab et al., 2018).
 
Detection of genes encoding cell wall associated proteins
 
The ability of bacteria to colonize and to cause infection is initiated by attachment to the host cells using surface proteins or cell wall anchored proteins. SpsK gene was present in all the SP isolates in this study and the same was observed in various studies (Bannoehr et al., 2011; Phumthanakorn et al., 2017). Prevalence of various virulence genes in MSSP and MRSP isolates are given in Fig 3 and 4 respectively.

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Fig 3: Prevalence of virulence genes in MSSP isolates.



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Fig 4: Prevalence of virulence genes in MRSP isolates.



SpsF gene was detected in 38/128 (29.68%) isolates which varied from 17.7% (Phumthanakorn et al., 2017) to 71.4% (Latronica et al., 2014) worldwide. SpsO was the one which was present in the least number of isolates (11/128=8.6%) in this study. It has been reported that the SpsO gene was detected in 28.6% of isolates by Latronica et al., (2014) and 40% by Phumthanakorn et al., (2017).

SP organism has two orthologues (SpsP and SpsQ) of staphylococcal protein A. SpsQ was more frequently present in 47/128 (36.71%) isolates than SpsP gene (36/128=28.12%). Both the orthologues (SpsP and SpsQ) were identified in 23 strains of SP and 69 isolates were negative for both the genes. In general, incidence of detection of cell wall protein genes was in a greater number of MRSP isolates than MSSP isolates. Phumthanakorn et al., (2017) and Latronica et al., (2014) reported the concomitant presence of SpsP and SpsQ genes in SP isolates whereas, Bannoehr et al., (2011) reported the differential presence of genes with more prevalence of SpsQ genes (60%) than SpsP (40%) as seen in the present study.
 
Detection of Biofilm forming genes
 
Staphylococci species have ica operon which contains icaADBC genes for biofilm formation and regulatory function. The icaA gene was present in 69/128 isolate (53.9%) and icaD gene was present in 86/128 (67%) SP isolates. Higher prevalence of icaD than icaA gene observed in the present study is contrary to the presence of 75.7% icaD and 77.9% icaA genes in SP isolates of Canada and USA reported by Singh et al., (2013). However, transcriptome analysis of SP isolates showed that MSSP isolates had an increased ability to form biofilm under acidic circumstances through up-regulation of the entire arc operon (Couto et al., 2016).
 
Detection of virulence regulatory genes
 
All staphylococcal species examined to date have been shown to encode AIP (Auto-inducible protein) peptides that are unique to each species based on AgrD gene. Accessory gene regulator genes were detected in all SP isolates in this study. Agr typing of 5 MSSP isolates were previously reported in India and type I and III AIP were produced by two strains of each and one isolate produced type II AIP (Ananda Chitra et al., 2015). Type IV agr AIP was the majority type found in MRSP isolates of USA (Black et al., 2009) whilst type III was predominantly seen in MRSP than MSSP isolates in Portugal (Couto et al., 2016). The present study identified Agr II in seven MRSP isolates and Agr III type in one MRSP isolate.

SarA is a DNA-binding protein, which binds to the agr promoter region affecting control of several virulence genes in S. aureus. SarA like gene is also present in SP and it was detected in 5/8 (62.5%) and 43/120 (35.8%) of MRSP and MSSP strains respectively. To the best of our knowledge, there is no available literature either to support or contradict this report. However, Couto et al., (2016) reported that higher expression of transcription of regulatory genes in MRSP isolate than MSSP isolate and, in MSSP isolates agrD regulatory genes had higher transcriptional expression.
 
Detection of exfoliative toxin genes
 
Exfoliative toxins (ET) - serine proteases, produced by staphylococci are involved in cutaneous infections of mammals. All the isolates of this study were detected to have SIET gene which concurs with previous studies (Ananda Chitra et al., 2016; Couto et al., 2016; Melter et al., 2017). In the present study, ExpA isoform of ET was detected in more number (14.84%) of isolates than ExpB isoforms (6.25%) as opposed to 4.7% ExpA and 9.52% ExpB in another study (Walther et al., 2012). The occurrence of ExpB was found to be 23.2% of isolates from dogs with superficial pyoderma and 6.1% of SP isolates from healthy dogs while ExpA gene was detected in 23.3% of SP isolates from canine pyoderma (Iyori et al., 2010).
 
Detection of Panton-Valentine Leucocidin (PVL) Like Toxin (Luk-I) in SP isolates
 
Panton-valentine Leucocidin (PVL) found in certain strains of S. aureus is a bi component-LukS-PV and LukF-PV, pore forming leukotoxin that causes leukocyte damage and tissue necrosis (Gillet et al., 2002). A similar bi component leukotoxin Luk-I, encoded by two genes, LukS/F, was detected in SP. All the SP isolates characterized in the present study as well as in the previous study (Ananda Chitra et al., 2016) possessed Luk-I genes. In other studies, 96.2% and 29.4% prevalence were reported (Melter et al., 2017; Ruzauskas et al., 2016). Luk-I gene was highly expressed in the MRSP isolate than MSSP isolate under transcriptome analysis (Couto et al., 2016).
 
Detection of enterotoxin se-int and superantigen like toxin (sel) genes in SP isolates
 
Staphylococcal enterotoxins (SE) are pyrogenic proteins associated with food poisoning and toxic shock syndrome. They are considered as superantigens as they bind to class II MHC molecules on antigen presenting cells and stimulate large populations of T cells releasing a cytokine bolus leading to an acute toxic shock. S. pseudintermedius produces two unique SEs - SECcanine which is an SEC variant (Cardona et al., 2006) and the other one is SE-int (Futagawa-Saito et al., 2004). In the present study, se-int gene was detected in 37.5% SP isolates where as 100% se-int gene prevalence was reported by Couto et al., (2016), Melter et al., (2017) and Futagawa-Saito et al., (2004). In another study, 37.5% and 75.9% of SP isolates from pyoderma and healthy dogs were found to possess se-int gene respectively (Tanabe et al., 2013).

In the present study, superantigen like protein (sel) gene was detected in all the SP isolates. 73.4% of the SP isolates from skin infection of Czech Republic was identified with sel gene (Melter et al., 2017).
 
SCCmec typing of MRSP isolates
 
In the present study, out of eight MRSP strains, only one isolate was identified as having SCCmec Type V by using multiplex PCR. Other seven strains were not typeable by the multiplex PCR methods. A 31% of the MRSP isolates from Italy were reported to be non-typeable (Gronthal et al., 2017).

SCCmec V has gained prominence in the UK (Maluping et al., 2014), Europe and North America (Perreten et al., 2010). In Asia, type V is dominant in Thailand, South China and Korea (Chanchaithong et al., 2014; Feng et al., 2012) and type II–III in Japan and North China (Ishihara et al., 2016; Wang et al., 2012). Therefore, SCCmec dissemination in Thailand and South China display a closer genetic relationship with Korea than Japan and North China.

CONCLUSION

This study showed the prevalence of various virulence genes in SP and MRSP in Chennai, India. Our study also revealed that there was no significant difference in the detection of virulence genes between MSSP and MRSP. However, it is possible that all the studied virulence factors may work in concert with other virulence factors to worsen skin infections of dog. Further studies are required to identify prevalent clonal types in this region to aid in molecular epidemiology.

ACKNOWLEDGEMENT

The authors are thankful to DST-SERB, Government of India for funding this study (EMR/2016/006141) and Tamil Nadu Veterinary and Animal Sciences University Chennai, India for providing necessary infrastructure to carry out the research work.

ETHICAL APPROVAL

Ethical approval was not required for this study as there were no invasive procedures. However, oral consent from the owners and complete confidentiality of the results were maintained.
 

CONFLICT OF INTEREST STATEMENT

The authors declare that there are no conflicts of interest.

REFERENCES


  1. Ananda Chitra, M., Jayanthy, C., Nagarajan, B. (2015). Detection and sequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates. Veterinary World. 8(7): 902 -907.

  2. Ananda Chitra, M., Jayanthy, C., Nagarajan, B. (2016). Virulence genes detection and antimicrobial susceptibility of Staphylococcus pseudintermedius isolates from canine skin infection in Chennai, India. Proceedings of National Academy of Sciences India Section B: Biological Sciences. https://doi.org/10.1007/s40011-016-0760-9.

  3. Bannoehr, J., Ben Zakour, N.L., Reglinski, M. (2011). Genomic and surface proteomic analysis of the canine pathogen Staphylococcus pseudintermedius reveals proteins that mediate adherence to the extracellular matrix. Infection and Immunity. 79: 3074-3086.

  4. Bannoehr, J., Brown, J.K., Shaw, D.J., Fitzgerald, R.J., Van Den Broek, A.H.M., Thoday, K.L. (2012). Staphylococccus pseudintermedius surface proteins SpsD and SpsO mediate adherence to ex-vivo canine corneocytes. Veterinary Dermatology. 23(2): 119-24.

  5. Beck, K.M., Waisglass, S.E., Dick, H.L., Weese, J.S. (2012). Prevalence of methicillin resistant Staphylococcus pseudintermedius (MRSP) from skin and carriage sites of dogs after treatment of their methicillin resistant or methicillin sensitive Staphylococcal pyoderma. Veterinary Dermatology. 23(4): 369-367.

  6. Black, C.C., Solyman, S.M., Eberlein, L.C., Bemis, D.A., Woron, A.M., Kania, S.A. (2009). Identification of a predominant multilocus sequence type, pulsed-field gel electrophoresis clusterand novel staphylococcal chromosomal cassette in clinical isolates of mecA-containing, methicillin-resistant Staphylococcus pseudintermedius. Veterinary Microbiology. 139: 333-338.

  7. Cardona, I.D., Cho, S.H., Leung, D.Y.M. (2006). Role of bacterial superantigens in atopic dermatitis: implications for future therapeutic strategies. American Journal of Clinical Dermatology. 7: 273-279.

  8. Chanchaithong, P., Perreten, V., Schwendener, S., Tribuddharat, C., Chongthaleong, A., Niyomtham, W., Prapasarakul, N. (2014). Strain typing and antimicrobial susceptibility of methicillin-resistant coagulase-positive staphylococcal species in dogs and people associated with dogs in Thailand. Journal of Applied Microbiology. 117: 572-586.

  9. Couto, N., Belas, A., Oliveira, M., Almeida, P., Clemente, C., Pomba, C. (2016). Comparative RNA-seq-Based Transcriptome Analysis of the Virulence Characteristics of Methicillin- Resistant and -Susceptible Staphylococcus pseudintermedius Strains Isolated from Small Animals. Antimicrobial Agents and Chemotherapy. 60(2): 962-967.

  10. Feng, Y., Tian, W., Lin, D., Luo, Q., Zhou, Y., Yang, T., Deng, Y., Liu, Y.H., Liu, J.H. (2012). Prevalence and characterization of methicillin-resistant Staphylococcus pseudintermedius in pets from South China. Veterinary Microbiology. 160(3- 4): 517-524.

  11. Frank, L.A., Kania, S.A., Kirzeder, E.M., Eberlein, L.C., Bemis, D.A. (2009). Risk of colonization or gene transfer to owners of dogs with meticillin resistant Staphylococcus pseudintermedius. Veterinary Dermatology. 20(5 6): 496-501.

  12. Futagawa Saito, K., Suzuki, M., Ohsawa, M., Ohshima, S., Sakurai, N., Ba Thein, W., Fukuyasu T. (2004). Identification and prevalence of an enterotoxin related gene, se int, in Staphylococcus intermedius isolates from dogs and pigeons. Journal of Applied Microbiology. 96(6): 1361-1366.

  13. Gerke, C., Kraft, A., Süßmuth, R., Schweitzer, O., Götz, F. (1998). Characterization of the N-Acetylglucosaminyl transferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin. Journal of Biological Chemistry. 273(29): 18586-18593.

  14. Grönthal, T., Eklund, M., Thomson, K., Piiparinen, H., Sironen, T., Rantala, M. (2017). Antimicrobial resistance in Staphylococcus pseudintermedius and the molecular epidemiology of methicillin-resistant S. pseudintermedius in small animals in Finland. Journal of Antimicrobial Chemotherapy. 72(4): 1021-1030.

  15. Gillet, Y., Issartel, B., Vanhems, P., Fournet, J.C., Lina, G., Bes, M., Vandensch, F., Piemont, Y., Brousse, N., Floret, D., Etienne, J. (2002). Association between Staphylococcus aureus strains carrying gene for panton-valentine leukocidin and highly lethal necrotising pneumonia in young immuno competent patients. Lancet. 359: 753-759.

  16. Ishihara, K., Koizumi, A., Saito, M., Muramatsu, Y., Tamura, Y. (2016). Detection of methicillin-resistant Staphylococcus pseudintermedius ST169 and novel ST354 SCCmec II- III isolates related to the worldwide ST71 clone. Epidemiology and Infection. 144: 434-442.

  17. Iyori, K., Hisatsune, J., Kawakami, T., Shibata, S., Murayama, N., et al. (2010). Identification of a novel Staphylococcus pseudintermedius exfoliative toxin gene and its prevalence in isolates from canines with pyoderma and healthy dogs. FEMS Microbiology Letters. 312(2): 169-175.

  18. Laarhoven, L.M., De Heus, P., Van Luijn, J., Duim, B., Wagenaar, J.A., van Duijkeren, E. (2011). Longitudinal study on methicillin-resistant Staphylococcus pseudintermedius in households. PloS one. 6(11): 27788.

  19. Latronico, F., Moodley, A., Nielsen, S.S., Guardabassi, L. (2014). Enhanced adherence of methicillin-resistant Staphylococcus pseudintermedius sequence type 71 to canine and human corneocytes. BMC Veterinary Research. 45: 70.

  20. Maluping, R.R., Paul, N.C., Moodley, A. (2014). Antimicrobial susceptibility of methicillin-resistant Staphylococcus pseudintermedius isolated from veterinary clinical cases in the UK. British Journal of Biomedical Sciences. 71(2): 55-57.

  21. Melter, O., Svec, P., Tkadlec, J., Doskar, J., Kinska, H., Pantucek, R. (2017). Characterisation of methicillin-susceptible Staphylococcus pseudintermedius isolates from canine infections and determination of virulence factors using multiplex PCR. Veterinary Medicine Czech. 62(2): 81-89.

  22. Paul, N.C., Moodley, A., Ghibaudo, G., Guardabassi, L. (2011). Carriage of methicillin resistant Staphylococcus pseudinter -medius in small animal veterinarians: indirect evidence of zoonotic transmission. Zoonoses and Public Health. 58(8): 533-539.

  23. Perreten, V., Kadlec, K., Schwarz, S.andersson, U.G., Finn, M., Greko, C. (2010). Clonal spread of methicillin-resistant Staphylococcus pseudintermedius in Europe and North America: an international multicentre study. Journal of Antimicrobial Chemotherapy. 65: 1145-1154.

  24. Phumthanakorn, N., Chanchaithong, P., Prapasarakul, N. (2017). Development of a set of multiplex PCRs for detection of genes encoding cell wall-associated proteins in Staphylococcus pseudintermedius isolates from dogs, humans and the environment. Journal of Microbiological Methods. 142: 90-95.

  25. Ruzauskas, M., Couto, N., Pavilonis, A., Klimiene, I., Siugzdiniene, R., Virgailis, M., Vaskeviciute, L., Anskiene, L., Pomba, C. (2016). Characterization of Staphylococcus pseudintermedius isolated from diseased dogs in Lithuania. Polish Journal of Veterinary Sciences. 19(1): 7-14.

  26. Saab, M.E.,  Muckle, CA., Stryhn, H., McClure, J.T. (2018). Comparison of culture methodology for the detection of methicillin- resistant Staphylococcus pseudintermedius in clinical specimens collected from dogs. Journal of Veterinary Diagnostic Investigation. 30(1): 93-98.

  27. Scherer, C.B., Botoni, L.S., Coura, F.M., Silva, R.O., Santos, R.D.D., Heinemann, M.B., Costa-Val, A.P. (2018). Frequency and antimicrobial susceptibility of Staphylococcus pseudinter -medius in dogs with otitis externa. Ciência Rural. 48(4).

  28. Singh, A., Walker, M., Rousseau, J., Weese, J.S. (2013). Characterization of the biofilm forming ability of Staphylococcus pseudintermedius from dogs. BMC Veterinary Research. 9(1): 93.

  29. Somayaji, R., Priyantha, M.A.R., Rubin, J.E., Church, D. (2016). Human infections due to Staphylococcus pseudintermedius, an emerging zoonosis of canine origin: report of 24 cases. Diagnostic Microbiology and Infectious Diseases. 85(4): 471-476.

  30. Tanabe, T., Toyoguchi, M., Hirano, F., Chiba, M., Onuma, K., Sato, H. (2013). Prevalence of staphylococcal enterotoxins in Staphylococcus pseudintermedius isolates from dogs with pyoderma and healthy dogs. Microbiology and Immunology. 57(9): 651-654.

  31. Van Duijkeren, E., Kamphuis, M., Van Der Mije Laarhoven, L.M., Duim, B., Wagenaar, J.A., Houwers, D.J. (2011). Transmission of methicillin-resistant Staphylococcus pseudintermedius between infected dogs and cats and contact pets, humans and the environment in households and veterinary clinics. Veterinary Microbiology. 150(3-4): 338-343.

  32. Walther, B., Hermes, J., Cuny, C., Wieler, L.H., Vincze, S., Elnaga, Y.A., et al. (2012). Sharing more than friendship-nasal colonization with coagulase-positive staphylococci (CPS) and co-habitation aspects of dogs and their owners. PloS one. 7(4): 35197.

  33. Wang, Y., Yang, J., Logue, C.M., Liu, K., Cao, X., Zhang, W., Shen, J., Wu, C. (2012). Methicillin-resistant Staphylococcus pseudintermedius isolated from canine pyoderma in North China. Journal of Applied Microbiology. 112: 623-630.

  34. Zhang, K., McClure, J.A., Conly, J.M. (2012). Enhanced multiplex PCR assay for typing of staphylococcal cassette chromosome mec types I to V in methicillin-resistant Staphylococcus aureus. Molecular and Cellular Probes. 26(5): 218-221.
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