Page 48 - Гахай, тахианы микоплазмозын оношлогоо

Гахай, тахианы микоплазмозын оношлогоо

Materials and methods

A total of 674 samples including 456 blood serum, 218 swabs of both esophagus and

anus from randomly selected chickens and a total of 753 samples including 581

blood serum and 172 swabs of anal and nasal mucus from randomly selected pigs
were collected in some farms in Bulgan, Uvs, Orkhon, Darkhan-Uul, Tuv, Dornod,

Uvurkhangai, Bayankhongor, Govi-Altai, Zavkhan and Arkhangai aimags and peri-
urban areas around Ulaanbaatar city, where the country’s swine and chicken farms

are developed. As a whole 1427 samples were collected and used for the testing.

Serological testing: Blood serum of pigs involved in the study were tested by ELISA
kit (ID Screen® Mycoplasma hyopneumoniae indirect) according to the

manufacturer’s instruction. Chicken blood serum samples were tested by ELISA kit
(VDPro® MG AB ELISA, VDPro® MS AB ELISA) according to the manufacturer’s

instruction, and the test results were obtained by using software of micro plate
reader.

Bacteriological testing: Causative agents such as M. hyopneumoniae, M.

gallisepticum and M. synoviae were detected and cultivated by adding blood serum
of healthy horse (10%), glucose and newly prepared yeast extract (25%), DNA

sodium pyruvate of DNA Herring fish (25%), phenolic red (1%) and penicillin to
“PPLO (Pleuropneumonia-like organism) broth” and “PPLO agar”, incubated in 5-

10% carbon dioxide (CO2).

Molecular biological testing: DNA was extracted from culture of mycoplasma like
organisms isolated from swine and PCR testing was performed using primers Mh-F

5'- GAG CCT TCA AGC TTC ACC AAG A -3' and Mh-R 5'- TGT GTT AGT ACT TTT
GCC ACC -3' containing specific sequence of 16S rRNA gene of swine

mycoplasmosis causative agent M. hyopneumoniae.

Primers pair MG-14F 5'-GAG CTA ATC TGT AAA GTT GGT C -3' and MG-13R 5'-
GCT TCC TTG CGG TTA GCA AC -3' containing specific sequence of 16S rRNA

gene to identify causative agent species M. gallisepticum of chicken mycoplasmosis
and MS-F 5'- GAG AAG CAA AAT AGT GAT ATC A -3' and MS-R 5'- CAG TCG TCT

CCG AAG TTA ACA A -3' of 16S rRNA gene of M. synoviae were used for PCR

testing.

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