Zbl. Bakt. 275, 269-271 (1991) © Gustav Fischer Verlag, StuttgartlNew York

Short Communication

Conspecificity of Hanseniaspora nodinigri and Hanseniaspora vineae: Comparison by Immunodiffusion and Immunoelectrophoresis

NEVA <::iFT<::iOGLU and NAMIK AKSOYCAN

Department of Microbiology, Faculty of Medicine, Ankara University, Slhhiye, Ankara, Turkey

With 1 Figure· Received September 11, 1990 . Accepted January 24, 1991

Summary

By using the methods of immunodiffusion and immunoelectrophoresis, we have found that there are antigenic differences between Hanseniaspora nodinigri and Hanseniaspora vineae which are concluded to be identical and synonymous by means of DNA reassocia­tion.

Zusammenfassung

Obwohl gezeigt wurde, daB H. nodinigri und H. vineae in ihrem DNA-Aufbau identisch und synonym sind, wurden durch Immundiffusion und Immunelectrophorese Antigenun­terschiede festgestellt.

Introduction

Meyer et aI., by means of DNA reassociatjon experiments supplemented with the physiological and morphological data, reported that there were six species in the genus Hanseniaspora (6). Lachance stated that Hanseniaspora nodinigri is a new species which should be accommodated in the genus Hanseniaspora (5). By means of DNA reassociation experiments, Smith and Poot concluded that H. nodinigri was identical and synonymous with Hanseniaspora vineae (8). Aksoycan et aI. reported that there was an antigenic relationship(s) among H. nodinigri and Escherichia coli (0: 75,

270 N. <;ift90glu and N. Aksoycan

0: 163), Salmonella aberdeen 0 antigens (3). Beside that in former publication, Ak­soycan also indicated that there was no antigenic relationship between H. vineae and the bacteria mentioned above (2).

The purpose of this study was to obtain the antigenic relationship(s) between H. nodinigri and H. vineae by the methods of immunodiffusion and immunoelectro­phoresis.

Materials and Methods

The type strains of H. nodinigri CBS 8031 and H. vineae CBS 2171 used in this study were provided by "Centraalburaeu voor Schimmelcultures, Yeast Division, Delft, Nether­lands". The antigens and the immune sera for the Hanseniaspora strains were prepared according to the method described previously by Aksoycan et al. for Candida antigens and immune sera production (1). In this method, 0.5% formalin is added to physiological saline suspensions of Hanseniaspora strains that had been grown on Sabouraud's dextrose agar in an incubator at 28 ac. After washing the antigens twice with physiological saline, 2 % suspensions (v/v) of the yeasts were prepared. These antigens were injected intravenously into rabbits which had been checked previously for the absence of the agglutinins for the Hanseniaspora strains. Injections were given every other day in 0.5, t.O, 2.0 and 4.0 ml amounts during the first week and 1.0, 2.0, 4.0 ml during the second and third week~. The sera were prepared from the blood of the rabbits taken one week after the last injection.

2171 (VEl Antigen

8031 (VEl Antigen

2171 Antiserum

2171 Antiserum

2171 (VEl Antigen

o L ~

~ 8031 (VEl Antigen

A

B

2171 (VEl Antigen

8031 (VEl Antigen

~ o

8031 Antiserum

8031 Antiserum

2171 (VEl Antigen

~ ~ =---~.~ 8031 (VEl Antigen

Fig. 1. Schematic diagrams of double diffusions (A) and immunoelectrophoretic (B) analyses showing cross-reactivity of the Hanseniaspora nodinigri CBS 8031 and Hanseniaspora vineae CBS 2171.

Conspecificity of Hanseniaspora nodinigri and H. vineae 271

The suspensions of antigens of Hanseniaspora strains for immunodiffusion and im­munoelectrophoretic analyses were prepared with veronal buffer (VE) (4). Immunological analyses were performed with the microplate double diffusion in gel technique described by Wadsworth (10) and with the immunoelectrophoretic technique in the modification of Wadsworth and Hanson (11). The electrophoretic separation was performed at a voltage of 6 Vlcm during 80 min in 0.05 M veronal buffer, pH 8.2 after the immunodiffusion period which was 3-4 days; precipitates were stained with Amidoblack 10 B (7, 9).

Results and Discussion

Schematic diagrams of immunodiffusion and immunoelectrophoretic analyses of H. nodinigri and H. vineae are shown in Fig. 1. Aksoycan et al. have reported that there is an antigenic relationship(s) among H. nodinigri and E. coli (0: 75,0: 163), S. aber­deen 0 antigens which is not present in the H. vineae.

Double diffusion and immunoelectrophoretic studies demonstrated different pre­cipitinogens between the type strains of H. nodinigri and H. vineae. So, although it is concluded that H. nodinigri and H. vineae are identical and synonymous by means of DNA reassociation, there is an antigenic difference between these type strains as shown by the method of immunodiffusion and immunoelectrophoresis.

References

1. Aksoycan, N., L. Le Minor et S. Le Minor: Antigenes communs entre les Candida et les Salmonella-Arizona. Ann. Inst. Pasteur 99 (1960) 723-728

2. Aksoycan, N.: Antigenic relationhip of Hanseniaspora and Kloeckera with Escherichia coli (0: 75,0: 163), Salmonella choleraesuis (0: 6, 7), Salmonella aberdeen (0: 11) 0 antigens. Zbl. Bakt. 273 (1990) 184-185

3. Aksoycan, N., B. Erdem, and i. Saganak: The antigenic relationship among Hansenias­pora nodinigri, Escherichia coli (0: 75, 0: 163) and Salmonella aberdeen 0 antigens. Infeks. Derg. 4 (1990) 279-283

4. Holmgren, J., G. Eggertsen, L. Hanson, and K. Lincoln: Immunodiffusion studies on Escherichia coli. Acta path. microbiol. scand. 76 (1969) 304-318

5. Lachance, M. A.: Hanseniaspora nodinigri, a new yeast species found in black knots (Dibotryon morbosum) of Prunus virginiana. Can. J. Microbiol. 27 (1981) 651-653

6. Meyer, S. A., M. T. Smith, and F. P. Simione: Systematics of Hanseniaspora Zikes and Kloeckeri:l Janke. Antonie v. Leeuwenhoek 44 (1978) 79-96

7. Ouchterlony, 0.: Handbook of immunodiffusion and immunoelectrophoresis, Ann Ar­bor-Humphrey Science Publishers, Ltd., London W.C.lEngland (1971)

8. B. Smith, M. Th. and G. A. Poot: Conspecificity of Hanseniaspora nodinigri and Han­seniaspora vineae: Comparison by DNA reassociation. Antonie v. Leeuwenhoek 51 (1985) 151-153

9. Svendsen, J. P., B. Weeke, and G. B. Johanson: Chemicals, solutions, equipment and general procedures. Scand. J. Immunol. 17, Suppl. 10 (1983) 3-20

~O. Wadsworth, C. A.: A slide microtechnique for the analysis of immune precipitates in gel. : Int. Arch. Allergy 10 (1957) 355-360

11. Wadsworth, C. A. and L. A. Hanson: Comparative analysis of immunoelectrophoretic technique. Int. Arch. Allergy 17 (1960) 165-177

Dr. Neva <::ift'rioglu, Ankara University, Faculty of Medicine, Department of Microbio­logy, Slhhiye, Ankara, Tiirkei

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