Pulmonary Mycoplasma bovis infection in necropsied cattle due to bad-sequel of Bovine Respiratory Disease (BRD) in Sohag Governorate, South Egypt
Ahmed M.E.Y. KOUNOUR1, Ahmed M. A. ZAITOUN2
1Infectious Diseases, Dept. Animal Med., Faculty of Vet. Med. Sohag Univ.
2Infectious Diseases, Dept. Animal Med., Faculty of Vet. Med. Assiut Univ
*Ahmed M.A. ZAITOUN, Prof. Emeritus of Infectious Diseases, Faculty of Vet. Med. Assiut Univ., Assiut, Egypt
Samples and laboratory procedures:
Tissues’ specimens of the pneumonic lungs of the necropsied cases were aseptically excised and immediately immersed in screw-capped bottles containing Mycoplasma broth culture supplemented with (growth enhancers for bovine Mycoplasmas, and bacterial inhibitors as prescribed previously by Zaitoun, 1990). The broths were incubated at 37 OC. Two days later, the incubated broths were repeatedly blindly subcultured in new broths and incubated. Three blind passages were carried out. Thereafter, the incubated broths were platted onto Mycoplasma agar medium and incubated in Gas-pack Jar with 10% co2 atmosphere for two days. Post incubation, the plats were regularly examined for one week. The characteristic colonies were picked-up and purified by further subculturing processes.
Biochemically, the purified colonies were subjected to Genus determination and biochemical characterizations (glucose fermentation, arginine deamination and Film and spots production tests) as approved by Stalheim (1990). The biochemically glucose negative and arginine negative strains with production of film and spots were molecularly identified using conventional PCR technique.
PCR technique using species-specific premiers for Mycoplasma bovis.(Table DNA extraction of the tested and control samples were carried out based on manufacturer of QIAamp® DNA Mini Kit (Qiagen, Hilden, Germany, catalog no.: 511304). Forward and reserved sequences of PCR’s primer of both Mycoplasma bovis were illustrated onto Table 2. Protocol of PCR technique of the tested samples was carried out based on Kounour (2022) in The Biotechnology Unit of Faculty of Veterinary Medicine, Sohag University, Egypt.
Figure 1: Demographic factors, descriptive of demographic information
As shown in Figure (1) Demographic factors, 475 people participated in this study including 382 female (80.4%) and 93 males (19.6%) (p<0.05). Their age ware ranged between less than 16 and more than 35 with advantage for age group more than 35 (42.90%).
Table 1: Drug reactions: Descriptive analysis (n=475)
Figure 1A&B: Lung sequestration with thickening and fibrosis of the interlobular septa (A), Cut section in the diaphragmatic lobe revealed a much of yellowish pussy material (B). N.B: the bluish coloration of the lobes
Figure 2: Cut surface of pneumonic lung-lobe, caseonecrotic bronchopneumonia with bronchiectasis and yellowish caseated purulent materials inside bronchioles. The cut surface was edematous, and purulent material was found in small airways.
Figure 3: pulmonary sequestration with obviously demarcation of interlobular septa Fig
Figure 4: Characteristic fried-egg colonies of Mycoplasma on agar medium (x 32)
Figure 5: Agarose gel electrophoresis of PCR amplification products of 16S rRNA gene of Mycoplasma bovis. Lane M: DNA Ladder (Marker) 100 pb. Lane 1: Control positive (Zaitoun, 2000). Lane 2: control negative. Lanes 3 10: positive tested samples with amplified products at 360 pb.
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