Carretto, Edoardo

Kosecka-Strojek, Maja

Marrollo, Roberta

Międzobrodzki, Jacek

Savini, Vincenzo

Polish Journal of Microbiology

English

Jagiellonian Univeristy Repository

Polish Journal of Microbiology 2013, Vol. 62, No 4, 465-466SHORT COMMUNICATIONCAMP test Detected Staphylococcus delphini ATCC 49172 P-Haemolysin ProductionVINCENZO SAVINI1, MAJA KOSECKA2, ROBERTA MARROLLO1, EDOARDO CARRETTO3and JACEK MI^DZOBRODZKI21 Clinical Microbiology and Virology, Spirito Santo Hospital, Pescara (PE), Italy 2 Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology Jagiellonian University, Krakow, Poland 3 Microbiology, IRCCS Arcispedale S. Maria Nuova, Reggio Emilia (RE), ItalySubmitted 10 October 2013, revised 26 October 2013, accepted 16 November 2013A b s t r a c tThrough a CAMP test, we first observed a Staphylococcus delphini strain (ATCC 49172) to release (J-haemolysin. Production of the lat­ter in this coagulase-positive species of the 'Staphylococcus intermedins Group) in fact, has been labeled to be undetermined, thus far. Of course, a wider number of strains have to be investigated in order to define whether this property is constitutive (like in Staphylococcus (pseud)intermedius), or strain-dependent (like in Staphylococcus aureus), and which clinical impact it has; nevertheless, we can state that S. delphini ATCC 49172 indeed produces this toxin.K ey  w o r d s ;  Staphylococcus delphini, (3-haemolysin, CAMP testIn 1944, Christie and Atkins observed that arrow- shaped haemolysis (called “candle flame-shaped” or “arrowhead”) occurred when Streptococcus agalactiae (group B streptococcus, GBS) was grown in a zone of Staphylococcus aureus (3-haemolysin activity (Christie etal, 1944; Darling 1975). This phenomenon was later called ‘CAMP test’, after the names of authors who first studied it (Christie, Atkins, and Munch-Petersen) and is currently used in microbiology laboratories as one among the most reliable methods to identify GBS, Lis­teria monocytogenes and Rhodococcus equi (Darling 1975; Munch-Petersen etal, 1945; Ramsey etal, 2010; Savini etal, 2011).To perform the assay, reference S. aureus strains that produce (3-haemolysin are streaked perpendicu­larly to the tested isolate, and arrowheads are observed after 24 h incubation. Indeed, any |3-haemolysin- producing S. aureus isolate may be used (Darling 1975); among coagulase-positive microorganisms other than S. aureus, nevertheless, (3-haemolysin is constitu- tively produced by Staphylococcus (pseud)intermedius (Ramsey etal, 2010; Devriese etal, 2005), while pro­duction by Staphylococcus delphini (that forms, together with S. (pseud)intermedius, the ‘Staphylococcus inter-medius Group’) is labeled as undetermined (Devriese etal, 2005; Savini etal, 2013; Van Hoovels etal, 2006; Varaldo etal, 1988).Therefore, we carried out a CAMP test by streak­ing S. delphini strain ATCC 49172 on a sheep blood plate perpendicularly to S. agalactiae (identification confirmed through latex agglutination) (Ramsey etal, 2010) and observed arrowhead (Fig. 1), surprisingly, meaning S. delphini (3-haemolysin production. As a confirmation, strain ATCC 49172 was cultivated on horse and rabbit blood media, where the dark, S. (pseud)intermedius-like, a-haemolytic band (which is p-haemolysin-related) was not formed; it was instead clearly visible on sheep blood (Darling 1975; Dinges etal, 2000; Savini etal, 2013).(3-haemolysin, in fact, has been known to be highly haemolytic for sheep but not rabbit and horse erythro­cytes, and is neither lethal in mice nor dermonecrotic in guinea pigs (Darling 1975; Dinges etal, 2000). It is secreted into the culture medium as an exotoxin by cer­tain S. aureus strains, particularly those from animal habitats, as well as from all S. (pseud)intermedius iso­lates (Dinges etal, 2000; Savini etal, 2013). Although its role in disease pathogenesis is not completely* Corresponding author: V. Savini, Clinical Microbiology and Virology, Spirito Santo Hospital, Pescara (PE), via Fonte Romana 8, CAP 65124, Italy; phone: +39-340-7379737; e-mail; vsavini2013@gmail.com466 Salvini V. et al. 4Fig. 1 - CAMP test with S. delphini (ATCC 49172). Horizontal inocula: GBS strains; vertical inoculum: S. delphini A (ATCC 49172) -  yellow arrows indicate zones of CAMP reaction; white arrows indicate edges of the (S-haemolysin-related incompletely haemolytic band (a-haemolysis).understood, thus far, high level expression in veteri­nary strains seem to indicate that producing organisms garner selective advantages from this toxin secretion (Dinges etal, 2000).We first showed that an S. delphini strain produces (3-haemolysin; however, this species is rarely isolated and further studies on a wider number of strains are needed, as soon as they are collected and identified, to define whether production is constitutive (like in S. (pseud)- intermedius) or strain-dependent (like in S. aureus). Nevertheless, we suggest, for the moment, that S. del­phini (3-haemolysin production be no more considered to be undetermined, but potential, and observed.AcknowledgmentsThis study was supported in part by the grant NCN 401 017740 from the Polish Ministry of Science and University Education.Conflict of interests statementThe authors have no conflict of interest to declare.LiteratureChristie R., N.E. Atkins and E. Munch-Petersen. 1944. A note on a lytic phenomenon shown by group B streptococci. Aust. J. Exp. Biol. Med. Sei. 22:197-200.Darling C.L. 1975. Standardization and evaluation of the CAMP reaction for the prompt, presumptive identification of Streptococcus agalactiae (Lancefield group B) in clinical material. J. Clin. Micro­biol. 1:171-174.Devriese L.A., M. Vancanneyt, M. Baele, M. Vaneechoutte, E. De Graef, C. Snauwaert, I. Cleenwerck, P. Dawyndt, J. Swings, A. Decostere and F. Haesebrouck. 2005. Staphylococcus pseudin- termedius sp. nov., a coagulase-positive species from animals. Int. J. Syst. Evol. Microbiol. 55: 1569-1573.Dinges M.M., P.M. Orwin, Pand and M. Schlievert. 2000. Exotox­ins of Staphylococcus aureus. Clin. Microbiol. Rev. 13:16-34. Munch-Petersen E., R. Christie, and R.T. Simmons. 1945. Further notes on a lytic phenomenon shown by group B streptococci. Aust. J. Exp. Biol. Med. Sei. 23: 193-195.Ramsey, K.J., E.C. Carter, M.L. McKee, and B.J. Beck. 2010. Reclassification of the Lisferia-CAMP test strain ATCC 49444 Staphylococcus aureus as Staphylococcus pseudintermedius. J. Food. Prot. 73:1525-1528.Savini V., C. Passeri, G. Mancini, O. Iuliani, R. Marrollo, A.V. Argentieri, P. Fazii, D. D’Antonio and E. Carretto. 2013. Coagu­lase-positive staphylococci: my pet’s two faces. Res. Microbiol. 164: 371-374.Savini, V., P. Salutari, M. Sborgia, I. Mancini, G. Masciarelli, C. Catavitello, D. Astolfi, C. D’Amario, G. Fioritoni, A. Spadano, and D. D’Antonio. 2011. Brief tale of a bacteraemia by Rhodococcus equi, with concomitant lung mass: what came first, the chicken or the egg? Mediterr. J. Hematol. Infect. Dis. 3: e2011006.Van Hoovels L., A. Vankeerberghen, A. Boel, K. Van Vaerenbergh, and H. De Beenhouwer. 2006. First case of Staphylococcus pseudin­termedius infection in a human. J. Clin. Microbiol. 44:4609-4612. Varaldo P.E., M. Vancanneyt, M. Baele, M. Vaneechoutte, E. De Graef, C. Snauwaert, I. Cleenwerck, P. Dawyndt, J. Swings, A. Decostere, F. Haesebrouck. 1988. Staphylococcus delphini sp. nov., a coagulase-positive species isolated from dophins. Int. J. Syst. Bacteriol. 38: 436-439.

CAMP test detected Staphylococcus delphini ATCC 49172 \beta-Haemolysin production

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CAMP test detected Staphylococcus delphini ATCC 49172 \beta-Haemolysin production

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