Article Figures & Data
Tables
- Table 1.
Studies in which culture of M. avium subsp.paratuberculosis from sheep has been attempted
Facility and country of culturea Sample(s) Medium(a) Sensitivity Time Reference Difficulty United Kingdom Lymph nodes Modified Dubos 50 CFU/g NAb 6 United Kingdom Gut mucosa Modified Dubos-actidione-chloramphenicol 75 CFU/ft of intestine NA 6 New Zealand Feces Herrold’s 5 of 47 infected sheep; 1 to 15 colonies 6 mo 9 United States Tissues, feces Herrold’s, Lowenstein-Jensen 3 of 20 infected sheep 6 mo 32 United Kingdom Tissues Egg based Tiny colonies after 7 mo in 3 of 12 tubes 7 mo 15 Australiac Mucosa Herrold’s, Lowenstein-Jensen 1 of 3 infected ewes after 12 wk 5 mo 31 Australiac Mucosa, lymph node, feces Egg based 4 of 50 infected sheep 5 mo 7 Australiad Mucosa Herrold’s 0 of 1 infected sheep 9 mo 25 Morocco Feces Lowenstein-Jensen–mycobactin 2 of 10 infected ewes 16 wk 5 South Africa Feces, tissues Herrold’s 2% of 59 infected sheep NA 18 Iceland Mucosa, lymph nodes Finlayson’s, modified Dubos, Herrold’s 9 to 54% of 22 infected samples 8 mo 17 Spain Feces, tissues Herrold’s, Lowenstein-Jensen 7 and 93% of 41 infected samples, respectively but colonies minute 4 to 8 mo 20 Ease India Feces Serum agar 57% of 100 infected sheep NA 22 Iran Feces, tissues Finlayson-Taylor, Lowenstein-Jensen 75% (sheep and goats not differentiated) NA 3 China NA Potato broth 60% of 38 infected sheep NA 14 Spain Feces, tissues Lowenstein-Jensen, Middlebrook 7H11-OADC with or without mycobactin J 86% of 25 infected sheep 4 mo 1 - Table 2.
Composition of Middlebrook media used to culture strains of M. avium subsp. paratuberculosis from sheep in this study
Ingredient Amt/liter in: Modified BACTEC 12Ba Modified 7H10 agarb Modified 7H11 agarc For base medium Casein digest 667 mge 800 mgd 800 mg Ammonium sulfate 333 mg 400 mg 400 mg Monopotassium phosphate 667 mg 1.2 g 1.2 g Disodium phosphate 1.7 g 1.2 g 1.2 g Sodium citrate 67 mg 320 mg 320 mg Magnesium sulfate 33 mg 20 mg 40 mg Calcium chloride 0.33 mg 0.4 mg Zinc sulfate 0.67 mg 0.8 mg Copper sulfate 0.67 mg 0.8 mg l-Glutamic acid 333 mg 400 mg 400 mg Ferric ammonium citrate 27 mg 32 mg 32 mg Pyridoxine 0.67 mg 0.8 mg 0.8 mg Biotin 0.33 mg 0.4 mg 0.4 mg Malachite green 0.2 mg 0.8 mg Bacto Agar 12 g 12 g For enrichment Oleic acid 40 mg Albumin fraction V, bovine 3.3 ge 4.0 g 4.0 g Dextrose 1.6 g 1.6 g Catalase 32,000 Ue 2.4 mg 3.2 mg Sodium chloride 680 mg C14 palmitic acid 667 μCie As additive Egg yolk 167 ml 200 ml 200 ml Mycobactin J 0.83 mg 1.0 mg 1.0 mg PANTA PLUSf 33.3 ml 40 ml 40 ml ↵a Additives (and 700 μl of water) were added to the enriched BACTEC media at the following rates per 4-ml vial, resulting in a final volume of 6 ml/vial: 1 ml of egg yolk, 5 μg (100 μl of mycobactin J, and 200 μl of PANTA PLUS (4a, 13a).
↵b ADC was used for enrichment.
↵c OADC was used for enrichment.
↵d As added Casitone.
↵e Added by manufacturer to standard Middlebrook 7H9 broth to form BACTEC 12B medium.
↵f Consists of polymyxin B (1,000 U/ml), amphotericin B (100 μg/ml), nalidixic acid (400 μg/ml), trimethoprim (100 μg/ml), azlocillin (100 μg/ml), and polyoxyethylene stearate (4 mg/ml).
- Table 3.
Culture of ovine tissues and feces in modified BACTEC 12B radiometric mediuma
Histological grade Tissue samples Fecal samples n No. positive n No. positive No lesions 14 5 3 0 Paucibacillary 4 4 7 3 Multibacillary 18 18 19 19 Total 36 27 29 22 ↵a Group A. Sheep were selected from numerous farms based on clinical, pathological, and serological criteria. Tissues and feces were not from the same sheep. Data are the number of sheep with a positive radiometric GI and IS900 PCR result.
- Table 4.
Culture of ovine tissues and feces in modified BACTEC 12B radiometric medium and on modified Middlebrook agar slopesa
Histological grade n Modified BACTEC 12B Modified Middlebrook agar No. of positive tissue samples No. of positive fecal samples No. of tissue samples: No. of fecal samples: Positive on 7H10 Positive on 7H11 Contaminatedb Positive on 7H10 Positive on 7H11 Contaminatedb Paucibacillary 16 16 9 15 16 1 8 5 1 Multibacillary 14 14 13 14 12 12 11 0 Total 30 30 22 29 28 1 20 16 1 ↵a Group B. The sheep came from nine farms and were selected for culture based on histological findings alone. Tissues and feces were matched from the same sheep. Data are the number of sheep with a positive radiometric GI and IS900 PCR result or the number of sheep with growth on solid medium and a positive IS900 PCR result.
↵b Cultures on 7H11 that were overgrown by contaminants.
- Table 5.
Culture of ovine feces in modified BACTEC 12B radiometric mediuma
Histological grade n No. of positive fecal samples No lesions 11 3 Paucibacillary 6 2 Multibacillary 14 14 Total 31 19 ↵a Group C. Sheep were selected based on clinical signs and came from a single farm. Data are the number of sheep with a positive radiometric GI and IS900 PCR result.
- Table 6.
Culture of ovine tissues and feces in modified BACTEC 12B radiometric mediuma
Histological grade n No. of positive tissue samples No. of positive fecal samples No lesions 27 8 4 Paucibacillary 2 2 1 Multibacillary 11 11 11 Total 40 21 16 ↵a Group D. Twenty sheep were selected based on serology from each of two farms. Tissues and feces were matched from the same sheep. Data are the number of sheep with a positive radiometric GI and IS900 PCR result.
- Table 7.
Combined results of all trials with modified BACTEC 12B radiometric mediuma
Histological grade Tissue samples Fecal samples n No. (%) positive n No. (%) positive No lesions 41 13 (31.7) 41 7 (17.1) Paucibacillary 22 22 (100) 31 15 (48.4) Multibacillary 43 43 (100) 58 57 (98.3) ↵a Tissues and feces are not necessarily matched from the same sheep. Data are the number and percentage of sheep with a positive radiometric GI and IS900 PCR result.
