Actinobacillus pleuropneumoniae Features and Culture

                       Actinobacillus pleuropneumoniae. Features and Culture.

                                                     Carlos Rodriguez G  BIOFARMA LABORATORIES      2014

Actinobacillus pleuropneumoniae is the etiologic agent of swine pleuropneumonia directly involved in the porcine respiratory disease complex, its distribution is worldwide and has become more important since the 80s of last century.
It occurs acutely, causing mortality and its chronically delayed growth or fattening.
Etiology, the causative agent is Actinobacillus pleuropneumoniae one gram microorganism negative hemolytic when grown on blood agar, form cocoidal with a capsule of lipopolysaccharide, very similar to that of other gram-negative bacteria, but by itself antigenic cacacterísticas, his only guest is the pig isolates vary in virulence and pathogenicity.
Actinobacillus pleuropneumoniae (Ap) is easily grown on blood agar, requiring nicotinamide adenine dinucleotide NAD for growth, there are reports showing strains that do not need it, but in a laboratory culture isolation is preferably made in s Blood Agar and NAD is provided

Stapphilococcus epidermidis, which is easily obtained from the skin.
To make planting, we have first samples of freshly killed animals in its acute form, the lesions are very characteristic, show hepatized in the lung area, which contains serous exudate.
For insulation, use a spatula laboratory, which is heated to red in a bunsen burner and surface Leson, then 10 ml syringe is used burns. With 18-gauge needle into the lesion indroduce and suction, the liquid obtained is planted on blood agar and when it has dried a strip of Staphylococcus (Nurse) is added, by placing the Petri plates incubated in a wide mouth jar has a lighted bela, closing the jar candle fire consumes the oxygen in the bottle and cultivation is done with a low oxygen tension.
The cultivation takes place in 18 hours, showing a few small colonies, which grow abundantly near the Mothership, we undertake planting Agar and Brain Heart Infusion supplemented with serum, colonies show a very noticeable iridescence due to capsule bacteria, tones ranging from light blue to green.
Taking some colonies with a straight strip of platinum, embryos may be inoculated 5 days of incubation, the embryo culture kills 18 hours incubation.
The liquid yolk sac harvested gives us very high titers.
Can also be grown in liquid media.
A crop can be made in large quantities using:
Primagen 22 grams (tissue culture medium)
5 g Sodium Chloride
2.5 g Sodium diphosphate.
2.5 g Dextrosa
Destilled Water c.b.p. 1000 ml.
This medium is adjusted to pH 7.2
It is autoclaved, 15 pounds pressure for 20 minutes.
To plant this means is necessary to add pig serum should be clarified that the blood plasma, is a liquid obtained from the blood of pigs after adding an anticoagulant and remove cellular components. Blood serum is the liquid obtained from the blood without added anticoagulant, after removal of fibrin and cellular components.
The serum was inactivated at 56 degrees Celsius for 30 minutes, cooled to 5 ° and centrifuged in a Westfalia centrifuge, obtaining a liquid of similar color to red wine, the liquid is "polished" in a filter press until well lens, then passed through a series of filters with the last capsule sterile 0.22 micron membrane. (Sartorius)
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NAD (B-nicotinamide adenine dinucleotide disodium salt N 6005.SIGMA CHEMICAL COMPANY) .Viene powder is diluted in double distilled water and filtered through a 0.22 membrane sterilized, a sample was also taken to check the sterilization.
Positive cultures were tested for agglutination serotypes and their characteristics are widely published article,
For rapid diagnosis in cases of acute infection and high mortality exist ELISAs.
If you want to be cultured in large volumes, a Bio-reactor is used at 600 rpm for 20 hours at 37 degrees Celsius

References ::
Experiences of the author:.carlosrodriguezg25@gmail.com
HAP Proceedins of the 1996 International Conference Mexico

The author is a veterinarian Zootecnista by the Faculty of Veterinary Medicine of the National Autonomous University of Mexico, an incomplete MS in Industrial Microbiology from the School of Chemical Sciences of the Autonomous University of Nuevo León. Former member of The American Association Of Avian Pathologists and a member of the Mexican Association of Microbiology, Biotechnology and Bioengineering. Mexico DF

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