Comparison of four culture media for isolation of Mycobacterium avium complex from porcine tissues

The efficiency of four culture media was compared for the isolation of Mycobacterium avium complex from 197 procine tissues. In 82 tissues with microscopic granulomas and acid-fast bacilli, a significantly greater number of isolates were obtained on Middlebrook 7H10 medium with sodium pyruvate than on Stonebrink medium, Herrold egg yolk agar medium, or Lowenstein-Jensen medium (P=0.01). In 46 tissues in which no microscopic granulomas or acid-fast bacilli were observed, a significantly greater number of isolates were made on Middlebrook 7H10 medium or Herrold egg yolk agar medium than on Stonebrink medium or on Lowenstein-Jensen medium (P=0.01). The time required to grow M. avium complex on Lowenstein-Jensen medium was significantly greater than the time required to observe growth on Stonebrink, Middlebrook 7H10, or Herrold egg yolk agar medium (p=0.001).

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Comparison of Four Different Culture Media for Isolation and Growth of Type II and Type I/III Mycobacterium avium subsp. paratuberculosis Strains Isolated from Cattle and Goats

Applied and Environmental Microbiology ◽

10.1128/aem.00451-06 ◽

2006 ◽

Vol 72 (9) ◽

pp. 5927-5932 ◽

Cited By ~ 45

Author(s):

Lucï¿½a de Juan ◽

Julio ï¿½lvarez ◽

Beatriz Romero ◽

Javier Bezos ◽

Elena Castellanos ◽

...

Keyword(s):

Incubation Period ◽

Mycobacterium Avium ◽

Culture Media ◽

Egg Yolk ◽

Disease Diagnosis ◽

Type I ◽

Type Ii ◽

Sodium Pyruvate ◽

Tissue Samples ◽

Solid Media

ABSTRACT Culture is considered the definitive technique for Johne's disease diagnosis, and it is essential for later applications of certain molecular typing techniques. In this study, we have tested four solid media (Herrold's egg yolk medium [HEYM] with sodium pyruvate and mycobactin [HEYMm-SP], HEYM with mycobactin and without sodium pyruvate [HEYMm], Middlebrook 7H11 with mycobactin [Mm], and Lï¿½wenstein-Jensen with mycobactin [LJm]) for isolation of Mycobacterium avium subsp. paratuberculosis strains in 319 tissue samples from cattle herds and goat flocks. We have shown that each of the two main groups of M. avium subsp. paratuberculosis (type II and type I/III) has different requirements for growth in the culture media studied. The recommended solid media for isolation of type I/III strains are LJm and Mm, since the combination of both media allowed the recovery of all these strains. The most widespread culture medium, HEYM, is not suitable for the isolation of this group of M. avium subsp. paratuberculosis strains. Regarding the type II strains, HEYMm-SP was the medium where more strains were isolated, but the other three media are also needed in order to recover all type II strains. The incubation period is also related to the strain type. In conclusion, because the type of strain cannot be known in advance of culture, coupled with the fact that cattle and goats can be infected with both groups of strains, we recommend the use of the four solid media and the prolongation of the incubation period to more than 6 months to detect paratuberculous herds/flocks and to determine the true prevalence of the infection.

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Performance of Different Solid and Liquid Culture Media for the Improvement of Tuberculin Production

International Journal of Veterinary Science ◽

10.37422/ijvs/20.013 ◽

2020 ◽

Keyword(s):

Culture Media ◽

Synthetic Medium ◽

Chemical Properties ◽

Economic Loss ◽

Delayed Type Hypersensitivity ◽

Basic Medium ◽

Primary Methods ◽

Lowenstein Jensen ◽

Time Required ◽

Modified Media

Tuberculosis (TB) is one of the most important zoonotic bacterial diseases. A huge economic loss which could be direct or indirect are associated with the disease. Currently, the primary methods used for detection of TB in humans and cattle include the measurement of a delayed type hypersensitivity to purified protein derivative (PPD). So, the need for preparation of purified PPD with adequate properties and increasing the final PPD yield with decreasing the time of tuberculin production has stimulated the interest in the development of its preparation. Our study was performed to compare between the standard and modified media for improving tuberculin production. Middle brook 7H10 agar medium was used as a modified basic medium for mycobacterial growth, followed by cultivation of mycobacteria on Middle brook 7H9 broth medium. For the production, strains were inoculated onto the culture medium (Dorest Henly synthetic medium). Other steps for tuberculin production was done according to standard Weighbridge protocol. The results demonstrated that the using of both Middle brook 7H10 agar and Middle brook 7H9 broth instead of Lowenstein-Jensen (LJ) and glycerin broth media which used in currently produced tuberculin, have better physical and chemical properties. In addition, reducing the time required for production by accelerating the time of microbial growth. Also, it was found that the tuberculin produced using modified media was slightly more potent or the same as currently tuberculin produced. So, both Middle brook 7H10 agar and Middle brook 7H9 broth media are recommended for production of tuberculin saving time and increasing potency of the product but more investigation was recommended for estimation types of protein present in both locally prepared and modified tuberculin.

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Comparison of Methods for Processing Drinking Water Samples for the Isolation of Mycobacterium avium and Mycobacterium intracellulare

Applied and Environmental Microbiology ◽

10.1128/aem.02009-07 ◽

2008 ◽

Vol 74 (10) ◽

pp. 3094-3098 ◽

Cited By ~ 28

Author(s):

Rachel Thomson ◽

Robyn Carter ◽

Chris Gilpin ◽

Chris Coulter ◽

Megan Hargreaves

Keyword(s):

Mycobacterium Avium ◽

Water Samples ◽

Culture Media ◽

Tap Water ◽

Cetylpyridinium Chloride ◽

Sodium Thiosulfate ◽

Mycobacterium Intracellulare ◽

Solid Media ◽

Lowenstein Jensen ◽

Mycobacterial Growth

ABSTRACT Several protocols for isolation of mycobacteria from water exist, but there is no established standard method. This study compared methods of processing potable water samples for the isolation of Mycobacterium avium and Mycobacterium intracellulare using spiked sterilized water and tap water decontaminated using 0.005% cetylpyridinium chloride (CPC). Samples were concentrated by centrifugation or filtration and inoculated onto Middlebrook 7H10 and 7H11 plates and Lowenstein-Jensen slants and into mycobacterial growth indicator tubes with or without polymyxin, azlocillin, nalidixic acid, trimethoprim, and amphotericin B. The solid media were incubated at 32°C, at 35°C, and at 35°C with CO2 and read weekly. The results suggest that filtration of water for the isolation of mycobacteria is a more sensitive method for concentration than centrifugation. The addition of sodium thiosulfate may not be necessary and may reduce the yield. Middlebrook M7H10 and 7H11 were equally sensitive culture media. CPC decontamination, while effective for reducing growth of contaminants, also significantly reduces mycobacterial numbers. There was no difference at 3 weeks between the different incubation temperatures.

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Conjunctival Mycobacteriosis in Two Emus (Dromaius novaehollandiae)

Veterinary Pathology ◽

10.1177/030098589603300314 ◽

1996 ◽

Vol 33 (3) ◽

pp. 346-348 ◽

Cited By ~ 17

Author(s):

A. M. Pocknell ◽

B. J. Miller ◽

J. L. Neufeld ◽

B. H. Grahn

Keyword(s):

Young Adult ◽

Adult Female ◽

Mycobacterium Avium ◽

Mycobacterium Avium Complex ◽

Plasma Cells ◽

Giant Cells ◽

Broad Ring ◽

Acid Fast Bacilli ◽

Dromaius Novaehollandiae ◽

Multinucleate Giant Cells

Avian tuberculosis was diagnosed in two young adult female commercial emus ( Dromaius novaehollandiae) with granulomatous conjunctivitis. Histologically, the granulomas appeared typical of avian tuberculosis. Caseonecrotic cores were surrounded by a broad ring of palisading epithelioid macrophages and multinucleate giant cells with a moderate admixture of heterophils, lymphocytes, and plasma cells. One conjunctival granuloma had multifocal mineralization. At necropsy, granulomas were also found in visceral organs of both birds. Acid-fast bacilli were demonstrated in all lesions using Ziehl-Neelsen or Fite's stains. Culture confirmed the bacilli to be Mycobacterium avium (complex).

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