Marmara Bölgesindeki sağlam koyunların kan serumlarında ELISA yöntemi ile Listeria monocytogenes'e karşı oluşan antikorların saptanması ve listeriozis üzerinde etiyolojik-epizootiolojik çalışmalar

Bu çalışmada Marmara Bölgesindeki sağlıklı koyunların kan serumlarında Enzyme-Linked Immunosorbent Assay (ELISA) ile Listeria monocytogenes'e karşı oluşan spesifik antikorlar araştırıldı. Antijen olarak dializ membran filtreleri ile elde edilen Listeriolysin O, L.monocytogenes serotip 1/2a ve serotip 4b'nin somatik O antijenleri kullanıldı.Toplanan 1000 koyun serumu 3 farklı antijene göre test edildi ve sonuçlar karşılaştırıldı. ELISA sonuçlarında, kan serumlarında Listeriolysin O antijenine %3.6, somatik l/2a'ya %11.5 ve somatik 4b'ye karşı %10.4 oranında pozitiflik saptandı.Ayrıca, çalışmada laktasyon dönemindeki koyunların süt ve dışkılarından etken izolasy-onu yapıldı. FDA yöntemi ile süt örneklerinden yapılan izolasyon çalışmalarında PALCAM Ağar ve Listeria Selektif Ağar (Oxoid), soğuk zenginleştirme yöntemi ile dışkılardan yapılan izolasyon çalışmalarında da Listeria Selektif Ağar (Oxoid) ve Listeria Selektif Ağar (Merck) besiyerleri kullanıldı.Sağım yolu ile koyunlardan sterirköşullarda alınan 100 süt örneğinde Listeria etkenleri saptanmadı. 180 koyunun rektumundan alınan dışkı örneklerinin yapılan araştırmasında iki L.monocytogenes, iki L.innocua, iki L.welshimeri, bir L.grayi ve bir L.murrayi suşu izole edildi.

Listeriosis in the Marmara Region and the detection of antibodies to Listeria monocytogenes in the sera of clinically healthy sheep by ELISA

In this study, the presence of antibodies to Listeria monocytogenes were investigated in sheep sera by Enzyme-linked Immunosorbent Assay (ELISA) in the Marmara region. Listeriolysin O was obtained from the supernatant of cultured L.monocytogenes using dialysing membran and used in ELISA. Whole cell antigens of L.monocytogenes serotypes l/2a and serotype 4b were also used as antigens.A thousand sheep sera tested by ELISA using three different antigens. Antibodies to liste-riolysin O were detected in 3.6% sheep sera while antibodies to serotype l/2a and serotype 4b have been detected 11.5% and 10.4% respectively.In addition to serology, isolation and identification studies were also performed in milk and feaces of sheep. The heat enrichment method was used for isolation from milk samples using PALCAM and LSA (Oxoid). The cold enrichment method was used from feaces using LSA (Oxoid) and LSA (Merck). No Listeria was isolated from 100 milk samples taken in sterile condition by hand milk-ing. Whereas, two L.monocytogenes, two L.innocua, two L.welshimeri, one L.grayi and one L.murrayi were isolated from the feaces of 180 sheep taken from rectum.

PDF

___

1- Art, D., Andre, P. (1991): Comparison of three selective isolation media for the detection of Listeria monocytogenes in foods. Zbl. Bakt. 275, 79-84.
2- Barlow, R.M., McGorum, B. (1985): Ovine listerial encephalitis: Analysis, hypothesis and synthesis. Vet. Rec.116, 233-236.
3- Berche, P., Reich, K.A., Bonnichon, M., Beretti, J-L., Geoffroy, C., Raveneau, J., Cossart, P., Gaillard, J-L., Geslin, P., Kreis, H., Veron, M. (1990): Detection of anti-Listeriolysin O for serodiagnosis of human listeriosis. Lancet. 335, 624-627.
4- Bocklisch, V.H., Wilhelms, D., Mirle, C., Lange, S., Kucken, U. (1991): Listerienabort des Rindes - bakteriologie, serologie und epizootiologie. Berl. Münch. Tierarztl. Wschr.104, 309-313.
5- Curtis, G.D.W., Mitchell, R.G., King, A.F., Griffin, E.J. (1989): A selective differential medium for the isolation of Listeria monocytogenes. Lett. App. Microbiol. 8, 95-98.
6- Çiftçioğlu, G. (1992): Kıyma, sucuk ve tavuk etlerinde Listeria monocytogenes 'in mevcudiyeti üzerine araştırmalar. İst.Üniv.Sağ.Bil.Enst. Doktora tezi.
7- Davies, J.W., Ewan, E.P., Varughese, P., Acres, S.E. (1984):Listeria monocytogenes infections in Canada. Clin. Invest. Med. 7 (4), 315-320.
8- Dhanda, M.R., Lall, J.M., Seth, R.N., Chandrasekariah, P. (1960): A case of listeric abortion in an ewe with a small scale survey of the incidence of agglutinings to listeria in the sera of sheep. Indian Vet. J. 36, 113-124.
9- Doğuer, M. (1961): Türkiye'de Listeriosis. Etlik Vet. Bak. Enst. Derg. 1 (4), 345-346.
10- Donker-Voet, J. (1972):Listeria monocytogenes: some biochemical and serological aspects. Acta. Microbiol. Acad. Sci. Hung. 19, 287-291.
11- Donnelly, C.W., Brackett, R.E., Doores, S., Lee, W.H., Lovett, J. (1992): Listeria. Compendium of methods for the microbiological examination of foods. (Third edition, Edited by Carl Vonderzant, Don F. Splittestoesser. American Public Health Associotion.). Chap. 38, 637-663.
12- Farber, J.M., Sanders, G.W., Johnston, M.A. (1989): A survey of various foods for the presence of listeria species. J. Food Protec. 52, 7, 456-458.
13- Fedio, W.M., Jackson, H. (1992): On the origin of Listeria monocytogenes in raw bulk-tank milk. Int. Dairy J. 2, 197-208.
14- Finci, E. (1968): Türkiye'de listeria durumu üzerinde araştırmalar. Ank. Üniv. Vet. Fak., Doktora tezi.
15- Gellın, B.G., Broome, C.V. (1989): Listeriosis. JAMA 261 (9), 1313-1320.
16- Genigeorgis, C.A., Dutuliscu, D., Garayzabal, J.F., (1989): Prevalence of Listeria spp. in Poultry meat at the supermarket and slaughterhouse level. J. Food Protec. 52 (9), 618-624.
17- Gitter, M., Stebbings, R.S., Morris, J.A., Hannam, D., Harris, C. (1986): Relationship between ovine listeriosis and silage feeding. Vet. Rec. 22, 207-208.
18- Gronstol, H. (1979):Listeria monocytogenes excretion and immunological state in healthy sheep. Acta Vet. Scand. 20, 168-179.
19- Gronstol, H. (1987): Listeriosis - a review. Norwegian Coll. Vet. Med. Pub. 5-14.
20- Gronstol, H. (1990): Listeria. Pathogeneses of Bacterial Infections in Animals. (Ed. by Carlton L. Gyles, Charles O. Thoen. Iowa State University Press / Ames). Chap. 6, 48-55.
21- Gunasinghe, C.P., Henderson, C., Rutter, M.A. (1994): Comparative study of two plating media (PALCAM and Oxford) for detection of listeria species in a range of meat products following a variety of enrichment procedures. Lett. App. Microbiol. 18, 156-158.
22- Gün, H. (1994):İstanbul ve yöresindeki sağlıklı ineklerin sütlerinde listeria türlerinin izolasyon, identifikasyon ve patojenitesi üzerine araştırmalar. İstanbul Üniv. Sağ. Bil. Enst., Doktora tezi.
23- Hitchins, A.D. (1992): Listeria monocytogenes. FDA Bacteriol. Anal. Man. 7th Ed. 10, 141-151.
24- Hudak, A.P., Lee, S.H., Issekutz, A.C., Bortolussi, R. (1984): Comparison of three serological methods ELISA, Complement fixation, and Microagglutination in the diagnosis of human perinatal Listeria monocytogenes infection. Clin. Invest. Med. 7 (4), 349-354.
25- Husu, J.R. (1990): Epidemiological Studies on the Occurrence of Listeria monocytogenes in the Feces of Dairy Cattle. J. Vet. Med. B. 37, 276-282.
26- İzgür, M., Esendal, Ö.M., Akay, Ö., Gülcü, B. (1990): Koyunların atık fötuslarından Listeria monocytogenes izolasyonu. KÜKEM Derg. 13 (2), 31-35.
27- Jones, D. (1990): Foodborne listeriosis. Lancet. 336 (10), 1171-1174.
28- Kutsal, A., Alpan, O., Arpacık, R. (1990):İstatistik uygulamaları. Bizim Büro Basımevi, Ankara.
29- Lovett, J. (1988): Isolation and enumeration of Listeria monocytogenes. Food Technol., 4, 172-175.30- Low, J.C., Davıes, R.C., Donachıe, W. (1992): Purification of Listeriolysin O and Development of an Immunoassay for diagnosis of listeric infections in sheep. J. Clin. Microbiol. 30,2705-2708.
31- Low, J.C., Donachıe, W. (1991): Clinical and serum antibody responses of lambs to infection by Listeria monocytogenes. Res. Vet. Sci. 51, 185-192.
32- Low, J.C., Renton, C.P. (1985): Septicaemia, encephalitis and abortions in a housed flock of sheep caused by Listeria monocytogenes type 1/2. Vet. Rec. 116, 147-150.
33- Low, J.C., Wright, F., Mclauchlin, J, Donachie, W. (1993): Serotyping and distribution of Listeria isolates from cases of ovine listeriosis. Vet. Rec. 133, 165-166.
34- McLauchlin, J. (1987):Listeria monocytogenes, recent advances in the taxonomy and epidemiology of listeriosis in humans. J. App. Bacteriol. 63, 1-11.
35- McLauchlın, J. (1990): Distribution of serovars of Listeria monocytogenes isolated from different categories of patients with listeriosis. Eur. J. Clin. Microbiol. Infect Dis. 9, 210-213.
36- Miettinen, A., Husu, J. (1991): Antibodies to listeriolysin O reflect the acquired resistance to Listeria monocytogenes in experimentally infected goats. FEMS Microbiol. Lett. 77, 181-186.
37- Miettinen, A., Husu, J., Tuomi, J. (1990): Serum antibody response to Listeria monocytogenes,listerial excration, and clinical characteristics in experimentally infected goats. J. Clin. Microbiol. (2), 340-343.
38- Murray, E.G.D., Webb, R.A., Swann, M.B.R. (1926): A disease of rabbits characterised by a large mononuclear leucocytosis, caused by a hitherto undescribed bacillus Bacterium monocytogenes. J. Pathol. Bacteriol. 29, 407-439.
39- Muller, H.E. (1988): Listerosis in animals. İnfek. Derg. 2 (4), 505-519.
40- Parlakgül, D. (1991): Brucella bakterilerinin peynirden ayrılması için balıklı besiyeri geliştirilmesi ve İstanbul piyasasında satılan peynirlerde Brucella ve Listeria bakterilerinin araştırılması. İst. Üniv. Cerrahpaşa Tıp Fak. Uzmanlık tezi.
41- Peel, M., Donachie, W., Shaw, A. (1988): Temperature-dependent expression of flagella of Listeria monocytogenes studied by electon microscopy, SDS-PAGE and western blotting. J. Gen. Microbiol. 2171-2178.
42- Prevot, A.R. (1961): Listeria. Traite Syst. Bacteriol. 2. 511-512.
43- Ralovich, B. (1993): Detection and Epidemiological typing of Listeria strains. Diagnostic methods for Listeria infections. Acta Microbiol. Hung. 40 (1), 3-38.
44- Rocourt, J. (1988): The recognition and identification of Listeria species by classical methods. İnfek. Derg. 2 (4), 471-485.
45- Rodriguez, L.D., Fernandez, G.S., Garayzabal, J.F.F., Ferri, E.R. (1984): New methodology for the isolation of Listeria microorganisms from heavily contaminated environments. App. Environ. Microbiol. 47 (5), 1188-1190.
46- Scotter, S.L., Wood, R., Williams, A.P. (1993): Methods for the detection of Listeria monocytogenes in foods. Results of an Inter-laboratory study. J. Assoc. Publ. Anal. 29, 221-251.
47- Seeliger, H.P.R. (1984): Modern taxonomy of the Listeria group relationship to its pathogenicity. Clin. Invest. Med. 7 (4), 217-221.
48- Seeliger, H.P.R., Jones, D. (1986): Listeria. (Ed. by Sneath,P.H.A., Mair,N., Sharpe,M.E., Holt,J.G. "Bergey's manual of Systematic Bacteriology". Williams and Wilkins, Baltimore, London, Los Angeles, Sydney. Vol. 2, 1235-1245)
49- Skovgaard,N., Norrung,B. (1989): The incidence of Listeria spp. in faeces of danish pigs and in minced pork meat. Inter. J. Food Microbiol. 8, 59-63.
50- Slade, P.J., Collins-Thompson, D.L. (1988): Comparison of two-stage and direct selective enrichment techniques for isolating listeria spp. from raw milk. J. Food Sci. 53 (6), 1694-1702.
51- Turilli, C., Marcolin, G., Prosperi, S. (1988): Evaluation of ELISA versus CF for the diagnosis of bovine listeriosis. Clin.Vet. 111 (3), 154-157.
52- Ünlü, G. (1990): Sivas yöresindeki çiğ sütlerde Listeria monocytogenes ve diğer türlerin aranması. Cumhuriyet. Üniv. Tıp Fak. Uzmanlık Tezi.
53- Van Netten, P., Perales, I., Van De Moosdijk, A., Curtis, G.D.W., Mossel, D.A.A. (1988):Liquid and solid selective differential media for the detection and enumeration of Listeria monocytogenes in more or less severely contaminaed food: An attempt to reach concensus at the international level. Proceedings 10. Inter.Congress on Listeriosis, Pesc, Hungary.1-15.
54- Vazquez-Boland, J.A., Dominguez, L., Blanco, M., Rocourt, J., Fernandez-Garayzabal, J.F., Gutierrez, C.B., Tascon, R.I., Rodriguez-Ferri, E.F. (1992): Epidemiologic investigation of a silage-associated epizootic of ovine listeric encephalitis, using a new Listeria selective enumeration medium and phage typing. Am. J. Vet. Res. 53 (3), 368-371.
55- Vazquez-Boland, J.A., Dominguez, L., Rodriguez-Ferri, E.F., Suarez, G. (1989): Purification and characterization of two Listeria ivanovii cytolysins, a sphingomyelinase C and a thiol-activated toxin (Ivanolysin O). Infect. Immun. 57 (12), 3928-3935.
56- Wehr, H.M. (1987):Listeria monocytogenes -Special reports. J. Assoc. Anal. Chem. 70 (5),769-772.
57- Wılesmıth, J.W.,Gıtter, M. (1986): Epidemiology of ovine listeriosis in Great Britain. Vet. Rec. 119, 467-470.