Figure 1:
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.
Figure 7.
Figure 8.
Sampling type of the 14 non-insecticidal B_ thuringiensis isolates (lab code), their crystal shapes, and the qualitative and quantitative quenching effects of different AT protein concentrations on the expression of S_ aureus spa and hla strains_
| Bt. Isolate (Code) sample | Crystal-Shape | Protein Conc. ug/ml | Spa | hla | ||||
|---|---|---|---|---|---|---|---|---|
| Effect | Bleaching zone (mm) | β-galactosidase Activity (MU) | Effect | Bleaching zone (mm) | β-galactosidase Activity (MU) | |||
| (M11) soil | Oval-spherical Nonattached to spores | 100 | + | 20±0.5 | 90±2.3 | + | 20±1.2 | 92±5.5 |
| 50 | + | 16±1 | 163±7.2 | + | 19±0.7 | 108±4.3 | ||
| 10 | − | 0 | 452±0 | − | 0 | 338±0 | ||
| (M13m) soil | Small spherical, not attached | 100 | + | 18±1 | 126±7.1 | + | 19±1.2 | 108±7.4 |
| 50 | + | 15.3±1.2 | 176.13.9 | + | 15±0.5 | 155±5.2 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M67t) Soil | Small spherical | 100 | - | 0 | 452±0 | - | 0 | 388±0 |
| 50 | + | 25±1.2 | 0±0 | + | 20±.2 | 92±1.9 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M 78) soil | Irregular, bi-pyramidal/cubic like crystals | 100 | + | 19.6±.3 | 100±1.4 | + | 21±1.2 | 62±3.6 |
| 50 | + | 16±0.5 | 163±5.1 | + | 14.3±1.2 | 167±14.1 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M91) soil | Nearly cubic, attached to spore | 100 | + | 20±1.5 | 90±6.8 | + | 19.3±1.3 | 89±6.1 |
| 50 | − | 16.3±0.8 | 158±7.8 | − | 15.6±0.7 | 147±6.6 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M154t1) Leaves/soil | Small, spherical non-attached to spore | 100 | + | 21.6±0.8 | 63±2.3 | + | 18.6±1.4 | 101±7.6 |
| 50 | + | 17±1.5 | 145±12.8 | + | 16.3±1.8 | 136±15.1 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M157) Dried leaves | Bipyramidal, attached to spore | 100 | + | 19±1 | 109±5.8 | + | 19.6±0.8 | 85±3.5 |
| 50 | + | 15.3±0.8 | 176±9.2 | + | 17±1.2 | 124±8.7 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M160) sheep manure | Blunt end bipyramidal attached to spore | 100 | + | 20.6±0.8 | 81±3.2 | + | 21±1 | 62±30 |
| 50 | + | 19±0.5 | 109±2.9 | + | 20±1.2 | 92±5.5 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M224) soil | Big spherical and hexagonal with inclined spore | 100 | + | 18.3±1.2 | 122±8.0 | + | 20.6±1.5 | 70±5.1 |
| 50 | + | 15±1.2 | 181±14.5 | + | 16.6±1.2 | 132±9,6 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388 | ||
| (M242) soil | bipyramidal, with blunt edges | 100 | + | 21.3±0.8 | 68±2.6 | + | 22.3±0.8 | 43±1.5 |
| 50 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M268P) Sewage W | Hexagonal attached to spores in long chains) | 100 | − | 0 | 452±0 | − | 0 | 388±0 |
| 50 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M296) soil | Large and small bipyramidal | 100 | − | 0 | 452±0 | − | 0 | 388±0 |
| 50 | + | 20.3±0.8 | 68±3.4 | + | 18.6±0.8 | 101±4.4 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| (M300) Rain W | Dark and bright ovoid (attached to spore) | 100 | + | 24±0.5 | 18±0.4 | + | 23.3±0.8 | 27±0.9 |
| 50 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
| A11 soil | Spherical non-spore attached | 100 | + | 16.6±1.2 | 154±11.2 | + | 19±0.5 | 93±2.5 |
| 50 | + | 13±1.7 | 217±29.7 | + | 15.6±0.8 | 147±7.6 | ||
| 10 | − | 0 | 452±0 | − | 0 | 388±0 | ||
Hemolytic activity of native Bt isolates on sheep blood agar_
| Bt isolate | Type of hemolysis | Hemolytic activity |
|---|---|---|
| Positive Control (S. aureus ATCC 25923) | β | + |
| M11 | γ | − |
| M78 | γ | − |
| M91 | γ | − |
| M154t1 | γ | − |
| A11 | γ | − |
Oligonucleotide primers in the RT-qPCR assay (Al-Mebairik et al_ 2016)_
| Gene | Sequence (5′–3′) |
|---|---|
| spa | F: GCGCAACACGATGAAGCTCAACAA |
| R: ACGTTAGCACTTTGGCTTGGATCA | |
| hla | F: CTGAAGGCCAGGCTAAACCACTTT |
| R: GAACGAAAGGTACCATTGCTGGTCA | |
| 16S rRNA | F: CTGGTAGTCCACGCCGTAAAC |
| R: CAGGCGGAGTGCTTAATGC | |
| RNAIII | F: GCACTGAGTCCAAGGAAACTAACTCT |
| R: AGCCATCCCAACTTAATAACCATGT |