Expert-level microbiology covering bacterial and viral biology, microbial physiology, pathogenesis, antimicrobial resistance, microbiome science, and industrial microbiology.
Cell structure: peptidoglycan wall (Gram+/Gram-), membrane, flagella, pili. Gram staining: crystal violet — Gram+ purple, Gram- pink after decolorization. Growth phases: lag, exponential, stationary, death. Metabolism: aerobic, anaerobic, fermentation, chemolithotrophy. Genetics: horizontal gene transfer (conjugation, transformation, transduction).
Structure: capsid, envelope, genome (DNA or RNA, ss or ds). Replication cycle: attachment, entry, replication, assembly, release. Lytic vs lysogenic: phage lambda as model. Reverse transcriptase: retroviruses, HIV replication. Viral evolution: mutation rates, reassortment, recombination.
Virulence factors: adhesins, toxins, capsules, immune evasion. Toxins: endotoxin (LPS), exotoxins (A-B type, pore-forming, superantigens). Biofilm formation: quorum sensing, matrix production, antibiotic tolerance. Intracellular pathogens: Mycobacterium, Listeria, Salmonella — escape mechanisms.
Mechanisms: enzymatic inactivation, target modification, efflux pumps, reduced uptake. Beta-lactamases: ESBL, carbapenemases (KPC, NDM, OXA). MRSA: mecA gene, altered PBP2a. Resistance spread: plasmids, transposons, integrons — horizontal transfer.
Gut microbiome: 38 trillion bacteria, Bacteroidetes/Firmicutes ratio. Dysbiosis: altered composition linked to IBD, obesity, mental health. 16S rRNA sequencing: V3-V4 region, OTU clustering, diversity metrics. Shotgun metagenomics: functional profiling, strain-level resolution.
| Pitfall | Fix |
|---|---|
| Contamination in cultures | Use sterile technique, include negative controls |
| Ignoring biofilm in AMR studies | Planktonic MIC underestimates biofilm tolerance |
| Misidentifying Gram reaction | Some organisms have atypical staining |
| Confusing bacteriostatic and bactericidal | Time-kill assays needed to distinguish |