Mannheimia haemolytica suşlarının farklı besiyerlerinde üreme ve lökotoksin oluşturma özelliklerinin incelenmesi

Year 2020, Volume: 5 Issue: 2, 33 - 42, 31.08.2020

Mehmet Çelik , Sevil Erdenliğ Gürbilek

https://doi.org/10.31797/vetbio.692422

Abstract

Mannheimia haemolytica hayvanlarda pnömoniye sebebiyet veren en etkili bakterilerden biridir. Etkenin en önemli virülans faktörü olan lökotoksine karşı gelişen antikorlar, hastalığa karşı dirençte önemli bir rol oynarlar. Bu nedenle aşılama çalışmalarında uygun miktarda lökotoksin üretecek suşun ve besiyerinin seçimi önemlidir. Bu amaçla, çalışmada 4 ayrı besiyeri ve 4 ayrı suş test edildi. Test edilen besiyerlerinde canlılık sayımı bakımından RPMI besiyeri diğer test edilen besiyerlerinden önemli derecede farklı bulundu (P<0.001). Öbür taraftan BHIB, BHIB+ %5 serum ve BHIB+%1 yeast extract besiyerleri canlılık sayımı açısından birbirlerinden istatistiki olarakfarklı bulunmadı (P>0.05). Ancak, test edilen suşların tümünün 24 saatlik kültürleri en fazla canlılık sayımını BHIB + %5 at serumu içeren besiyerinde gösterdiler. Suşlar arasında bakteriyel sayıma ilişkin istatistiki olarak önemli bir fark saptanmadı (P>0.05). RPMI besiyeri üretilen lökotoksin açısından diğer besiyerlerine göre istatistiki olarak önemli derecede farklı bulundu (P<0.001). Tüm test suşları RPMI besiyerinde diğer 3 besiyerine göre çok daha fazla miktarda lökotoksin üretti. Alınan sonuçlara göre, MH05 suşu en fazla miktarda lökotoksin üretti ve bunu MH06 suşu izledi. MH04 ve MH03 suşları arasında bu yönden anlamlı bir farklılık saptanmadı. Test edilen kültür besiyerlerinde üreyen mikroorganizma sayısı ile üretilen lökotoksin arasında negatif bir korelasyon olabileceği düşünüldü. Sonuç olarak, açlık ve kısıtlı demir varlığı gibi stress faktörlerinin Mannheimia haemolytica’nın daha fazla lökotoksin ve muhtemelen diğer virülans faktörleri üretmesine yol açtığı kanısına varıldı. Mannheimia haemolytica is one of the most effective bacteria causing pneumonia in animals. Since antibodies against leukotoxin, which is the most important virulence factor of the agent, play an important role in resistance to disease, selection of strain and medium that produce leukotoxin is most important in vaccination studies. For this purpose, 4 different strains were tested in 4 different culture media tested in the study. Based on the viability counts in tested media, RPMI media was found significantly different from the rest of media (P<0.001). On the other hand, BHIB, BHIB+5% serum and BHIB+%1 Yeast extract media were not found statistically different from each others (P>0.05) for the viability counts. However, the viability counts measured after 24 hours of culture of tested strains were found to be highest in medium containing BHIB + 5% horse serum. There were no significant differences among strains related to bacterial counts in tested media (P>0.05). RPMI media was significantly different from the rest of media regarding to amount of produced leukotoxin in direct ELISA (P<0.001). All the strains produced more leukotoxins in RPMI media compared to other 3 tested media. According to our results, the strain MH05 produced the highest amount of leukotoxin followed by MH06. There was no significant differences between MH03 and MH04 strains in this regard. It was thought that there could be negative correlation between viability count of the bacteria and the produced leukotoxin amount in tested media. As conclusion, it was assumed that stress factors like starvation or iron restriction might cause Mannheimia haemolytica to produce more leukotoxin and possibly other virulence factors.

Keywords

Mannheimia haemolytica, pnömoni, lökotoksin, virülans

Supporting Institution

Harran Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

16107

Thanks

Bu çalışma, Harran Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü tarafından 16107 proje numarası ile Yüksek lisans tezi olarak desteklenmiştir. Destekleri için kuruma teşekkür ederiz.

Investigations on growth and leukotoxin production of mannheimia haemolytica strains growing in different culture media

Abstract

Mannheimia haemolytica is one of the most effective bacteria causing pneumonia in animals. Since antibodies against leukotoxin, which is the most important virulence factor of the agent, play an important role in resistance to disease, selection of strain and medium that produce leukotoxin is most important in vaccination studies. For this purpose, 4 different strains were tested in 4 different culture media tested in the study. Based on the viability counts in tested media, RPMI media was found significantly different from the rest of media (P<0.001). On the other hand, BHIB, BHIB+5% serum and BHIB+%1 Yeast extract media were not found statistically different from each other’s (P>0.05) for the viability counts. However, the viability counts measured after 24 hours of culture of tested strains were found to be highest in medium containing BHIB + 5% horse serum. There were no significant differences among strains related to bacterial counts in tested media (P>0.05). RPMI media was significantly different from the rest of media regarding to amount of produced leukotoxin in direct ELISA (P<0.001). All the strains produced more leukotoxins in RPMI media compared to other 3 tested media.
According to our results, the strain MH05 produced the highest amount of leukotoxin followed by MH06. There was no significant difference between MH03 and MH04 strains in this regard. It was thought that there could be negative correlation between viability count of the bacteria and the produced leukotoxin amount in tested media. As conclusion, it was assumed that stress factors like starvation or iron restriction might cause Mannheimia haemolytica to produce more leukotoxin and possibly other virulence factors.

Keywords

virulence, Mannheimia haemolytica, lökotoksin, pneumonia

Project Number

16107

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