Rothia nasimurium as a Cause of Disease: First Isolation from Farmed Chickens
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Isolation and Purification
2.2. Virus Detection
2.3. Bacterial Identification
2.3.1. Observation of Gram Staining
2.3.2. Bacterial Biochemical Test
2.3.3. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS)
2.4. Sequence Analysis of 16S rRNA
2.5. Antibiotic Susceptibility Testing
2.6. Detection of Antibiotic Resistance Genes
2.7. Animal Pathogenicity Test
3. Results
3.1. Virus Detection Results
3.2. Isolation and Purification
3.3. Animal Regression Experiment
3.3.1. Gram Staining
3.3.2. Bacterial Biochemical Test
3.3.3. MALDI-TOF MS Identification
3.4. Alignment of Bacterial 16S rRNA Sequences and Construction of Phylogenetic Tree
3.5. Antibiotic Susceptibility Testing
3.6. Detection of Antibiotic Resistance Genes
3.7. Animal Pathogenicity Experiment
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antimicrobial Types | Genes | Primer Sequences (5′→3′) | Annealing Temperature | Product Size/bp |
---|---|---|---|---|
β-lactams | bla TEM | F:CAGAAACGCTGGTGAAAGTA R:ACTCCCCGTCGTGTAGATAA | 55 | 719 |
bla SHV | F:ATGCGTATATTCGCCTGTG R:CCTCATTCAGTTCCGTTTCC | 55 | 502 | |
bla CTX-M | F:AGTGAAAGCGAACCGAATC R:CTGTCACCAATGCTTTACC | 55 | 365 | |
mecA | F:GTTGTAGTTGTCGGGTTTGG R:GTTGTAGTTGTCGGGTTTGG | 56 | 336 | |
TEM | F:AGGAAGAGTATGATTCAACA R:CTCGTCGTTTGGTATGGC | 51 | 535 | |
Sulfonamides | sul1 | F:CATTGCCTGGTTGCTTCAT F:CATTGCCTGGTTGCTTCAT | 54 | 238 |
sul2 | F:CATCATTTTCGGCATCGTC R:TCTTGCGGTTTCTTTCAGC | 54 | 793 | |
sul3 | F:AGATGTGATTGATTTGGGAGC R:TCTTGCGGTTTCTTTCAGC | 54 | 443 | |
Quinolones | aac(6′)-Ib | F:TTGCGATGCTCTATGAGTGGCTA R:CTCGAATGCCTGGCGTGTTT | 55 | 482 |
oqxAB | F:GATCAGTCAGTGGGATAGTTT R:TACTCGGCGTTAACTGATTA | 55 | 671 | |
qnrA | F:TCAGCAAGAGGATTTCTCA R:GGCAGCACTATTACTCCCA | 54 | 627 | |
gyrA | F:GGTGACGTAATCGGTAAATA R:ACCATGGTGCAATGCCACCA | 53 | 810 | |
gyrB | F:CTCCTCCCAGACCAAAGACA R:TCACGACCGATACCACAGCC | 59 | 448 | |
parC | F:GCGAATAAGTTGAGGAAT R:AGCTCGGAATATTTCGAC | 55 | 417 | |
parE | F:CTGAACTGCTGGCGGAGATG R:GCGGTGGCAGTGCGACGTAA | 59 | 483 | |
Aminoglycosides | aadB, | F:GAGGAGTTGGACTATGGATT R:CTTCATCGGCATAGTAAAA | 53 | 208 |
aacC2 | F:GCAATAACGGAGGCAATTCGA R:CTCGATGGCGACCGAGCTTCA | 56 | 697 | |
aph(3′)-Ia, | F:ATGGGCTCGCGATAATGTC R:CTCACCGAGGCAGTTCCAT | 56 | 600 | |
aac(6′)/aph(2″) | F:CCAAGAGCAATAAGGGCATA R:CACTATCATAACCACTACCG | 56 | 220 | |
ant(6)-I | F:ACTGGCTTAATCAATTTGGG R:GCCTTTCCGCCACCTCACCG | 56 | 597 | |
Tetracyclines | Tet(A) | F:GCTACATCCTGCTTGCCTTC R:CATAGATCGCCGTGAAGAGG | 59.5 | 210 |
tet(B) | F:TTGGTTAGGGGCAAGTTTTG R:GTAATGGGCCAATAACACCG | 59.5 | 659 | |
tet(C) | F:CTTGAGAGCCTTCAACCCAG R:ATGGTCGTCATCTACCTGCC | 59.5 | 418 | |
tetM | F:GTGTGACGAACTTTACCGAA R:GCTTTGTATCTCCAAGAACAC | 52 | 510 | |
Macrolides | ermB | F:GAAAAGGTACTCAACCAAATA R:AGTAACGGTACTTAAATTGTTTAC | 48 | 636 |
mefA | F:AACTATCATTAATCACTAGTGC R:TTCTTCTGGTACTAAAAGTGG | 50 | 346 | |
Lincosamides | LinA | F:GGTGGCTGGGGGGTAGATGTATTAACTGG R:GCTTCTTTTGAAATACATGGTATTTTTCGA | 57 | 323 |
Chloramphenicols | fexA | F:TTGGGAAGAATGGTTCAGGG R:ATCGGCTCAGTAGCATCACG | 50 | 977 |
Group | Quantity/pcs | Concentration/(CFU·mL−1) | Dosage/mL | Infection Method | Number of Deaths/pcs | Mortality Rate/% |
---|---|---|---|---|---|---|
1 | 3 | 3 × 105 | 0.2 | orally | 0 | 0 |
2 | 3 | 3 × 107 | 0.2 | orally | 0 | 0 |
3 | 3 | 3 × 108 | 0.2 | orally | 0 | 0 |
4 | 3 | 3 × 105 | 0.2 | injected intraperitoneally | 0 | 0 |
5 | 3 | 3 × 107 | 0.2 | injected intraperitoneally | 0 | 0 |
6 | 3 | 3 × 108 | 0.2 | injected intraperitoneally | 3 | 100 |
7 | 3 | 0.9% normal saline | 0.2 | injected intraperitoneally | 0 | 0 |
Substrate | Result |
---|---|
Maltose | + |
Sucrose | + |
Glucose | + |
Lactose | − |
Urea | − |
Mannitol | − |
Sodium citrate | − |
Hydrogen sulfide | − |
Drug Name | Judging Standard | Actual Result | Result | Drug Name | Judging Standard | Actual Result | Result |
---|---|---|---|---|---|---|---|
Penicillin | ≥15, ≤14 | 21 | susceptible | Ciprofloxacin | ≥21, ≤15 | 12 | resistant |
Ceftazidime | ≥18, ≤14 | 0 | resistant | Norfloxacin | ≥17, ≤12 | 0 | resistant |
Ampicillin/sulbactam | ≥15, ≤11 | 0 | resistant | Erythromycin | ≥23, ≤13 | 0 | resistant |
Levofloxacin | ≥17, ≤13 | 0 | resistant | Meropenem | ≥20, ≤15 | 13 | resistant |
Cefoxitin | ≥18, ≤14 | 0 | resistant | Imipenem | ≥23, ≤19 | 17 | resistant |
Tobramycin | ≥15, ≤12 | 0 | resistant | Azithromycin | ≥13, ≤12 | 0 | resistant |
Gentamicin | ≥15, ≤12 | 0 | resistant | Chloramphenicol | ≥18, ≤12 | 10 | resistant |
Amikacin | ≥17, ≤14 | 20 | susceptible | Moxifloxacin | ≥24, ≤20 | 0 | resistant |
Tetracycline | ≥15, ≤11 | 0 | resistant | Sulfamethoxazole-trimethoprim | ≥16, ≤10 | 0 | resistant |
Tigecycline | ≥15, ≤13 | 22 | susceptible | Clindamycin | ≥21, ≤14 | 0 | resistant |
Drug Name | Mic (mg/L) | Actual Result | Drug Name | Mic (mg/L) | Result |
---|---|---|---|---|---|
Penicillin | <0.12 | susceptible | Ciprofloxacin | 64 | resistant |
Ceftazidime | >128 | resistant | Norfloxacin | >16 | resistant |
Ampicillin/sulbactam | 32/16 | resistant | Erythromycin | >128 | resistant |
Levofloxacin | >64 | resistant | Meropenem | 128 | resistant |
Cefoxitin | >32 | resistant | Imipenem | 8 | resistant |
Tobramycin | >16 | resistant | Azithromycin | >8 | resistant |
Gentamicin | >16 | resistant | Chloramphenicol | >32 | resistant |
Amikacin | <16 | susceptible | Moxifloxacin | >8 | resistant |
Tetracycline | >16 | resistant | Sulfamethoxazole-trimethoprim | >320 | resistant |
Tigecycline | <0.5 | susceptible | Clindamycin | >128 | resistant |
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Zhang, J.; Mo, S.; Li, H.; Yang, R.; Liu, X.; Xing, X.; Hu, Y.; Li, L. Rothia nasimurium as a Cause of Disease: First Isolation from Farmed Chickens. Vet. Sci. 2022, 9, 653. https://doi.org/10.3390/vetsci9120653
Zhang J, Mo S, Li H, Yang R, Liu X, Xing X, Hu Y, Li L. Rothia nasimurium as a Cause of Disease: First Isolation from Farmed Chickens. Veterinary Sciences. 2022; 9(12):653. https://doi.org/10.3390/vetsci9120653
Chicago/Turabian StyleZhang, Jiahao, Shaojiang Mo, Hu Li, Ruizhi Yang, Xiangjie Liu, Xiaoyue Xing, Yahui Hu, and Lianrui Li. 2022. "Rothia nasimurium as a Cause of Disease: First Isolation from Farmed Chickens" Veterinary Sciences 9, no. 12: 653. https://doi.org/10.3390/vetsci9120653