Microbiological Characteristics and Pathogenesis of Klebsiella pneumoniae Isolated from Hainan Black Goat
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Isolation and Identification of Bacteria
2.2. Drug Sensitive Test
2.3. Experimental Animals
2.4. Pathogenic Test of K. pneumoniae
2.5. Bacterial Colonization in Mice
2.6. Histopathology Examination of Lungs
2.7. RNA Preparation and Sequencing
2.8. RNA-seq Analysis and DEGs Identification
2.9. qRT-PCR Validation of DEGs
2.10. Statistical Analysis
3. Result
3.1. Colonies and Morphological Characteristics of Isolated Bacteria
3.2. 16S rRNA Identification of Bacteria
3.3. Biochemical Identification of Bacteria
3.4. Drug Sensitive Test of KPHN001
3.5. Mouse Pathogenicity Test
3.6. Bacteria Load in Viscera of Mice
3.7. Histopathology Examination of Mouse Lungs
3.8. RNA-seq Data Analysis
3.9. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes Analysis
3.10. qRT-PCR Validation of DEGs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Name of Genes | Primer Sequence (5′-3′) | Product Length (bp) |
---|---|---|
Cxcl2 | F: CACTCTCAAGGGCGGTCAA | 96 |
R: AGTTAGCCTTGCCTTTGTTCAG | ||
Ccl3 | F: TCCCAGCCAGGTGTCATTTTC | 105 |
R: GGCATTCAGTTCCAGGTCAGT | ||
Il1β | F: AATGCCACCTTTTGACAGTGATG | 139 |
R: AGCTTCTCCACAGCCACAAT | ||
Il6 | F: TGATGGATGCTACCAAACTGGA | 199 |
R: CTGTGACTCCAGCTTATCTCTTG | ||
C3 | F: ACTTCTTCATTGACCTGCGGC | 197 |
R: CGAGGACTTGGGAGGGATTT | ||
Tnf | F: CCCTCACACTCACAAACCAC | 134 |
R: ACAAGGTACAACCCATCGGC | ||
Ccl2 | F: CACTCACCTGCTGCTACTCA | 117 |
R: GCTTGGTGACAAAAACTACAGC | ||
Cxcl9 | F: GTGTGGAGTTCGAGGAACCCT | 173 |
R: GGCAGGTTTGATCTCCGTTC | ||
Myd88 | F: AAGCAGCAGAACCAGGAGTC | 150 |
R: GCAGTAGCAGATAAAGGCATCG | ||
Junb | F: CAGCCTTTCTATCACGACGAC | 96 |
R: GGTGGGTTTCAGGAGTTTGTAG | ||
Gng11 | F: CAAGTTGCAGAGACAACAGGTATC | 130 |
R: GCTGCCCTTTTCTTTGAAGGG | ||
Mapk3 | F: CAACACCACCTGCGACCTTA | 154 |
R: GGATTTGGTGTAGCCCTTGGAA | ||
β-actin | F: CCTCTATGCCAACACAGT | 146 |
R: TAGGAGCCAGAGCAGTAA |
Biochemical Project | Result | Biochemical Project | Result |
---|---|---|---|
Glucose | + | Sorbitol | + |
Sucrose | + | Urea | + |
Maltose | + | Malonate | + |
Mannose | + | Citrate | + |
Arabinose | + | Ornithine | − |
Raffinose | + | Lysine | + |
Rhamnose | + | Indole | − |
Xylose | + | Methyl red test (MR) | − |
Mannitol | + | Voges–Proskauer test | + |
Salicin | + | Hydrogen sulfide | − |
Inositol | + | Nitrate reduction | + |
Antimicrobial Agents | Inhibitory Zone Diameter/mm | Drug Resistance | Antimicrobial Agents | Inhibitory Zone Diameter/mm | Drug Resistance |
---|---|---|---|---|---|
Penicillin | 0 | R | Ciprofloxacin | 14 | R |
Oxacillin | 0 | R | Vancomycin | 0 | R |
Ampicillinum | 0 | R | Polymyxin B | 17 | S |
Carbenicillin | 0 | R | Trimethoprim | 0 | R |
Piperacillin | 10 | R | Furazolidone | 19 | S |
Cefalexin | 0 | R | Chloramphenicol | 0 | R |
Cefazolin | 0 | R | Amikacin | 0 | R |
Cefradine | 0 | R | Gentamicin | 0 | R |
Cefuroxime | 0 | R | Kanamycin | 0 | R |
Ceftazidime | 15 | R | Neomycin | 0 | R |
Ceftriaxone | 9 | R | Tetracycline | 7 | R |
Cefoperazone | 12 | R | Doxycycline | 0 | R |
Midecamycin | 0 | R | Minocycline | 10 | R |
Norfloxacin | 10 | R | Erythromycin | 0 | R |
Ofloxacin | 12 | R | Clindamycin | 0 | R |
Serial Number | Heart Bacteria Load (CFU/g) | Liver Bacteria Load (CFU/g) | Spleen Bacteria Load (CFU/g) | Lung Bacteria Load (CFU/g) | Kidney Bacteria Load (CFU/g) |
---|---|---|---|---|---|
E1 | 7.34 × 107 | 3.07 × 108 | 6.16 × 108 | 1.79 × 108 | 1.59 × 108 |
E2 | 2.25 × 107 | 1.27 × 108 | 9.88 × 108 | 3.27 × 107 | 7.60 × 107 |
E3 | 4.58 × 107 | 2.41 × 109 | 5.04 × 109 | 1.15 × 109 | 5.25 × 108 |
Serial Number | Total Reads | Q10 Bases Ratio(%) | Q20 Bases Ratio(%) | Q30 Bases Ratio(%) | GC Bases Ratio(%) | N Bases Ratio(%) |
---|---|---|---|---|---|---|
C1 | 71,187,134 | 100.00% | 98.98% | 96.12% | 51.62% | 0.00% |
C2 | 55,851,916 | 100.00% | 99.01% | 96.21% | 51.69% | 0.00% |
C3 | 55,792,564 | 100.00% | 98.99% | 96.17% | 51.72% | 0.00% |
E1 | 63,352,576 | 100.00% | 99.03% | 96.27% | 51.38% | 0.00% |
E2 | 61,343,784 | 100.00% | 98.98% | 96.1% | 51.69% | 0.00% |
E3 | 58,422,366 | 100.00% | 98.98% | 96.1% | 51.11% | 0.00% |
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He, M.; Li, H.; Zhang, Z.; Jiang, J.; Li, H.; Yang, W.; Cheng, Y.; Gao, H.; Chen, Q.; Du, L.; et al. Microbiological Characteristics and Pathogenesis of Klebsiella pneumoniae Isolated from Hainan Black Goat. Vet. Sci. 2022, 9, 471. https://doi.org/10.3390/vetsci9090471
He M, Li H, Zhang Z, Jiang J, Li H, Yang W, Cheng Y, Gao H, Chen Q, Du L, et al. Microbiological Characteristics and Pathogenesis of Klebsiella pneumoniae Isolated from Hainan Black Goat. Veterinary Sciences. 2022; 9(9):471. https://doi.org/10.3390/vetsci9090471
Chicago/Turabian StyleHe, Meirong, Haoyang Li, Zhenxing Zhang, Junming Jiang, Hong Li, Weijie Yang, Yiwen Cheng, Hongyan Gao, Qiaoling Chen, Li Du, and et al. 2022. "Microbiological Characteristics and Pathogenesis of Klebsiella pneumoniae Isolated from Hainan Black Goat" Veterinary Sciences 9, no. 9: 471. https://doi.org/10.3390/vetsci9090471