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The Effect of Long-Term Storage on Mycobacterium bovis

OLEXIY TKACHENKO
OLEXIY TKACHENKO
, 
NATALI KOZAK
NATALI KOZAK
, 
MARYNA BILAN
MARYNA BILAN
, 
VOLODYMYR HLEBENIUK
VOLODYMYR HLEBENIUK
, 
NATALIA ALEKSEEVA
NATALIA ALEKSEEVA
, 
LILIYA KOVALEVA
LILIYA KOVALEVA
, 
VITALII NEDOSEKOV
VITALII NEDOSEKOV
 et 
OLEXANDR GALATIUK
OLEXANDR GALATIUK
   | 17 sept. 2021
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Article Category: original-paper
Publié en ligne: 17 sept. 2021
Pages: 327 - 337
Reçu: 11 mars 2021
Accepté: 14 juil. 2021
DOI: https://doi.org/10.33073/pjm-2021-031
Mots clés
© 2021 Olexiy Tkachenko et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Cultures of Mycobacterium bovis: a) passage No. 54, b) passage No. 180.
Cultures of Mycobacterium bovis: a) passage No. 54, b) passage No. 180.

Fig. 2.

Morphology of initial cultures of Mycobacterium bovis: a) passage No. 54, b) passage No. 180 (stained by Ziehl-Neelsen). Bar = 10 µm.
Morphology of initial cultures of Mycobacterium bovis: a) passage No. 54, b) passage No. 180 (stained by Ziehl-Neelsen). Bar = 10 µm.

Fig. 3.

Subcultures of Mycobacterium bovis: a) passage No. 54, b) passage No. 115 (at 37°C), c) passage No. 115 (at 3°C), d) passage No. 135, e) passage No. 143, f) passage No. 171 (at 37°C), g) passage No. 171 (at 3°C), h) passage No. 180.
Subcultures of Mycobacterium bovis: a) passage No. 54, b) passage No. 115 (at 37°C), c) passage No. 115 (at 3°C), d) passage No. 135, e) passage No. 143, f) passage No. 171 (at 37°C), g) passage No. 171 (at 3°C), h) passage No. 180.

Fig. 4.

Morphology of Mycobacterium bovis subcultures: a) passage No. 54, b) passage No. 115 (at 37°C), c) passage No. 115 (at 3°C), d) passage No. 135, e) passage No. 143, f) passage No. 171 (at 37°C), g) passage No. 171 (at 3°C), h) passage No. 180; 1 – rod-shaped bacteria, 2 – thread-like forms, 3 – grains (elementary bodies), 4 – output of grains from thread-like forms, 5 – L-forms. Bar = 10 µm.
Morphology of Mycobacterium bovis subcultures: a) passage No. 54, b) passage No. 115 (at 37°C), c) passage No. 115 (at 3°C), d) passage No. 135, e) passage No. 143, f) passage No. 171 (at 37°C), g) passage No. 171 (at 3°C), h) passage No. 180; 1 – rod-shaped bacteria, 2 – thread-like forms, 3 – grains (elementary bodies), 4 – output of grains from thread-like forms, 5 – L-forms. Bar = 10 µm.

Fig. 5.

The spleen of the guinea pig: 1) normal, 2) with tuberculous foci.
The spleen of the guinea pig: 1) normal, 2) with tuberculous foci.

Fig. 6.

Growth of pathogenic Mycobacterium bovis strain and its subcultures (variants) on nutrient media. FPA – nutrient agar, FPB – nutrient broth.
Growth of pathogenic Mycobacterium bovis strain and its subcultures (variants) on nutrient media. FPA – nutrient agar, FPB – nutrient broth.

Fig. 7.

Catalase activity of Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs (repeatedness, %): “–” – negative reaction, “±” – doubtful reaction, “+”, “++”, “+++” – the magnitude of the positive reaction.
Catalase activity of Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs (repeatedness, %): “–” – negative reaction, “±” – doubtful reaction, “+”, “++”, “+++” – the magnitude of the positive reaction.

Fig. 8.

Dehydrogenase activity of Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs (repeatedness, %): “–” – negative reaction, “±” – doubtful reaction, “+”, “++”, “+++” – the magnitude of the positive reaction after 24 hours.
Dehydrogenase activity of Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs (repeatedness, %): “–” – negative reaction, “±” – doubtful reaction, “+”, “++”, “+++” – the magnitude of the positive reaction after 24 hours.

Fig. 9.

Reduction of nitrates by Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs (repeatedness, %): “–” – negative reaction, “±” – doubtful reaction, “+”, “++”, “+++” – the magnitude of the positive reaction.
Reduction of nitrates by Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs (repeatedness, %): “–” – negative reaction, “±” – doubtful reaction, “+”, “++”, “+++” – the magnitude of the positive reaction.

Fig. 10.

Tween-80 hydrolysis by Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs: A) after five days, B) after ten days.
Tween-80 hydrolysis by Mycobacterium bovis initial subcultures and the subcultures passaged through the guinea pigs: A) after five days, B) after ten days.

The results of DNA amplification of mycobacteria

Type of mycobacteriaStrain of mycobacteriaResult of detection of amplification products
Mycobacterium bovisfast-growing (original)+
after the long-term storage and reinoculation of cultures on the nutrient medium), passages 54, 115, 135, 135, 171, and 180–
Vallee strain+
BCG strain+
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