Laboratorial diagnostics of anaerobic dysentery lambs
UDK 619.616-084 :616.9:636.22/28
Laboratorial diagnostics of anaerobic dysentery lambs.
E.M. Agayeva, S.H.Huseynova
Azerbaijan State Agrarian University. Az2000. Ganja Pr. Ataturk,262
The effectiveness of medical treatment of infectious illnesses of new born lambs with symptoms of acorn-bowels illness, especially anaerobic dysentery, which is one of more often met pathologies, depends on accurate and timely diagnosis. In this case laboratorial diagnostics may be necessary.
Laboratorial diagnosis of anaerobic dysentery in lambs is based on discovering the presence of toxins in bowels that damage the walls of the bowels and internal organs. [epizootic and clinic and anatomic pathology given excretory of clean plant and its type.]
For the research in the laboratory a fresh piece of bowel is sent in, in bandaged form which contains parenchymatic organs, mesentery lymphatic nodes, and bone in tube form.
In this work we have provided cultures and identified anaerobic micro- organisms from both environmental sources and internal organs of laboratory animals.
In the first stage we took cultures of mixed feces and soil from farms in the surrounding area. Samples were taken to a depth of 5cm (weight 1-2 gr).
Samples were ground and mixed in NaCI with following mixtures 1:10, 1:100, 1:1000, then 1:10000. The resulting solution was incubated in test tubes with Kitt-Tarossi boullion. One part was heated to 80°C for 20 minutes, the other was not heated. Both samples were incubated in anaerobic conditions at 37°C for 24 hours.
Resulting toxins were used to infect white mice in a standard neutralization reaction test. The reaction solutions were made with antitoxin whey Cl. perfringens A, B, C, D, E, F. Subsequent discovery in the bowels of the surviving mouse of B-type toxin Cl.perfringens gave an adequate basis for making microbiological diagnosis.
Smears were prepared from the parenchymatic organs and from the small intestines colored according to the Qramm method. Resulting analysis revealed many gramm-formed thick, short straws with hooked ends. Discovery of these bacteria confirmed our diagnosis.
For getting a clear culture, glucose-blood agar was made according Seyslery generally-accepted weights.
1.We can confirm the nature of the bacteria by noting the features of existing cells colored according to the Qramm method, 4-8 mkm in length, 0,1-1,5 mkm thick.
2.In glucose-blood agar bacterial columns in S and R forms were observed with two or one zone of strati, the first zone spontaneously touching the columns with a narrow contour, after it more bright zone, with a larger contour.
After contact with air, Cl.perfringens columns on glucose-blood aqar became greener.
3. After an incubating 4-6 hours, darkening of the surrounding Vilson-Blera area was seen.
4. When Cl.perfringens is introduced into milk it causes putrefaction in 4-6 hours; whey will precipitate, and milk solids to form a gassy curd,
5. A sample of liver placed in the Kitt-Tarossi bullion, also produced gas after 3-4 hours of isolation.
6. Cl.perfringens also caused fermentation in the following carbohydrates: glucoses, lactoses, maltoses, galaktoses, and sakharoses. During the fermentation process two gases were produced: mannita and dulcitta.
In order to differentiate between the characteristis of different types of Cl.perfringens, neutralization reaction tests and biological tests were used.
It was established that anti-toxin serum type A will only neutralize A-type bacteria; similarly B serum neutralizes B-type bacteria; C serum neutralizes C-type bacteria, etc. Therefore, since type A and B bacteria are always found together outside the laboratory, it is necessary in treating a diseased animal to inject both A and B serum together.
Identification procedures for Cl.perfringens types A and B were followed on white mice. It was found that injections of 0,1 ml of type A bacteria did not result in the death of the mice. However as little as 0,01 ml of type B resulted in death.
During the pathogenic process, blood was extracted from the dead mice and cultured in Kitt-Tarossi boullion for 24 hours. Then culture was mixed with a solution of 0,85%-m NaCI to produce bacterial mass 25-30 s. of optic standard turbidity. Suspension was injected into the muscle of white mice in doses of 0,5 ml per day for three days. In our experience the mice died 3 days after infection.
Biological tests carried out during this process produced positive identification of type B Cl.perfringens bacteria. We were also able to observe the general symptoms characteristic of this bacteria.
The final outcome of this research was that we were able to document the general symptomatic characteristics of type B Cl.perfringens bacteria; and we were able to isolate a clear culture of type B Cl.perfringens bacteria.
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Reference Research literature
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