Journal: Brazilian Journal of Microbiology

Article Title: Inflammatory response of Haemophilus influenzae biotype aegyptius causing Brazilian Purpuric Fever

doi:

Figure Lengend Snippet: Bacterial Strains used in this work.

Article Snippet: βlac , Haemophilus influenzae strain serotype b ATCC 33533 , INCQS – FIOCRUZ.

Techniques: Amplification, Transformation Assay

Journal: Journal of Virology

Article Title: Stability of influenza A virus in droplets and aerosols is heightened by the presence of commensal respiratory bacteria

doi: 10.1128/jvi.00409-24

Figure Lengend Snippet: Decay of A/WSN/33 IAV in 1 µL PBS droplets at 40% RH and 22°C–25°C in a humidity-controlled chamber when mixed with microbiota bacteria. ( A ) A/WSN/33 virus was added to PBS alone (IAV Only), or added to PBS containing live commensal respiratory bacteria (IAV + Microbiota), for 5 × 10 7 plaque forming units (PFU)/mL final virus concentration. Equal colony forming units (CFU) of S. pneumoniae, S. aureus, Haemophilus influenzae, Moraxella catarrhalis , and P. aeruginosa were added for 10 8 CFU/mL total in the +microbiota case. One microliter droplets of each virus suspension were deposited on a non-binding 96-well plate and exposed to indoor air conditions for a total of 60 min. Triplicate droplets of each mixture were individually recovered in 300 µL of PBSi (PBS for infections) at time-points 0, 15, 30, 45, and 60 min post-deposition, and infectious viral titers were quantified by plaque assay (clear symbols indicate samples that were below plaque assay limit of quantification (LOQ), and were set at LOQ/√2 as described in Materials and Methods). Infectious viral titers were corrected for physical recovery (determined by genome quantification of IAV in each recovered droplet by digital PCR) relative to samples collected immediately after deposition (time 0, where no physical loss has occurred due to drying). Individual data points of triplicate droplets from two independent experimental repeats are shown ( n = 6 droplets total per group), with geometric mean ± geometric SD. Significant differences between IAV titers at time 60 were determined by Mann-Whitney U-test (**, P ≤ 0.01). ( B ) RH and temperature were monitored by a portable hygrometer across the 60-min period, with readings taken every minute. Solid and dotted lines distinguish readings from individual experimental repeats, with confidence intervals (±0.5°C, ±3% RH as provided by manufacturer) indicated by shaded regions. Bold text indicates the target RH.

Article Snippet: Gram-negative strains Moraxella catarrhalis (Strain Nell, DSM 9143), Pseudomonas aeruginosa (Strain CCEB 481, DSM 50071), and Haemophilus influenzae (Strain 572, serotype B, DSM 11969) were obtained from DSMZ (Germany).

Techniques: Bacteria, Virus, Concentration Assay, Suspension, Binding Assay, Plaque Assay, Digital PCR, MANN-WHITNEY

Journal: Journal of Virology

Article Title: Stability of influenza A virus in droplets and aerosols is heightened by the presence of commensal respiratory bacteria

doi: 10.1128/jvi.00409-24

Figure Lengend Snippet: Decay of A/WSN/33 in 1 µL PBS droplets at 22°C–25°C in a humidity-controlled chamber when mixed with Gram-negative bacteria. ( A ) A/WSN/33 virus was added to PBS alone (IAV Only), or added to PBS containing live Haemophilus influenzae (+ H.inf ), Moraxella catarrhalis (+ M.cat ), or Pseudomonas aeruginosa (+ P.aer ) bacteria at 10 8 CFU/mL final concentration. In all cases, virus was added for 5 × 10 7 plaque forming units (PFU)/mL final concentration. One microliter droplets of each virus suspension were deposited on a non-binding 96-well plate and exposed to indoor air conditions for a total of 60 min at 40% RH. Triplicate droplets of each mixture were individually recovered in 300 µL of PBSi (PBS for infections) at time-points 0, 15, 30, 45, and 60 min post-deposition, and infectious viral titers were quantified by plaque assay (clear symbols indicate samples that were below limit of quantification (LOQ) and were set at LOQ/√2). Infectious viral titers were corrected for physical recovery (determined by genome quantification of IAV in each recovered droplet by digital PCR) relative to samples collected immediately after deposition (time 0, where no physical loss has occurred due to drying). Data show individual data points of triplicate droplets, from two independent experimental repeats ( n = 6 droplets total per group), with geometric mean ± geometric SD. Significant differences in infectious titers at time 60 relative to IAV-only control were determined by Kruskal-Wallis test (*, P ≤ 0.05; ns, not significant). ( B ) One microliter droplets containing A/WSN/33 alone or A/WSN/33 mixed with H. influenzae, M. catarrhalis, or P. aeruginosa in PBS [as in ( A )] were recorded during drying at 40% RH. Images are representative of three individual droplets per group. Colored circles highlight the time-point where crystallization was first visible by eye for each sample. Scale bar = 2 mm. ( C ) Decay of A/WSN/33 in 1 µL droplets at 75% RH, when virus was added to PBS alone, or to PBS containing H. influenzae, M. catarrhalis, or P. aeruginosa [as in ( A )]. One microliter droplets were exposed to air at 75% RH for a total of 120 min. Triplicate droplets of each mixture were individually recovered in 300 µL of PBSi at time-points 0, 60, and 120 min post-deposition, and quantified and corrected as for ( A ). Clear symbols indicate samples that were below LOQ and were set at LOQ/√2. Individual data points of triplicate droplets from two independent experimental repeats ( n = 6 droplets total per group) are presented, with geometric mean ± geometric SD. Significant differences in infectious titers at time 120 relative to virus-only control were determined by Kruskal-Wallis test (*, P ≤ 0.05; ***, P ≤ 0.001; ns, not significant).

Article Snippet: Gram-negative strains Moraxella catarrhalis (Strain Nell, DSM 9143), Pseudomonas aeruginosa (Strain CCEB 481, DSM 50071), and Haemophilus influenzae (Strain 572, serotype B, DSM 11969) were obtained from DSMZ (Germany).

Techniques: Bacteria, Virus, Concentration Assay, Suspension, Binding Assay, Plaque Assay, Digital PCR, Control, Crystallization Assay

Journal:

Article Title: Evolution of the Major Pilus Gene Cluster of Haemophilus influenzae

doi:

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: Growth of E. coli and H. influenzae and purification of E. coli plasmids were as previously described ( 25 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Source a Relevant properties Reference(s) E. coli DH5α GIBCO BRL Host for E. coli plasmids E. coli plasmids Bluescript pSK − Stratagene pHic Cloned 468-bp fragment of R3001 purE-pepN region, obtained by PCR amplification with primers purE-F and pepN-R followed by subcloning of Alu I partial digestion fragment into Sma I site of pSK− This study pMCC1 Pst I (5.2-kb) fragment of the Hib Eagan hif operon in pUC19, subcloned from a cosmid library; includes the purE-hifA junction, hifA , hifB , and the 5′ end of hifC This study; 13 H. influenzae strains Eagan (E1A) CSF from meningitis patient Type b, streptomycin resistant, virulent in infant rat model 38 Rd (R906) = Goodgal Nonencapsulated derivative of type d strain; avirulent in rat model 8 INT1 Blood from meningitis patient Nonencapsulated; biotype V; virulent in rat model 31 R539 (ATCC 9006) ATCC Reference strain of serotype a R538 (ATCC 9795) ATCC Reference strain of serotype b R540 (ATCC 9007) ATCC Reference strain of serotype c R541 (ATCC 9008) ATCC Reference strain of serotype d R542 (ATCC 8142) ATCC Reference strain of serotype e R543 (ATCC 9796) ATCC Reference strain of serotype f U11 CSF Streptomycin-resistant derivative of U1 (Ramirez); nonencapsulated; avirulent in infant rat assay 38 R1965 (NCTC 8143) NCTC HK389; type strain of H. influenzae ; nonencapsulated, biotype II 22 R1967 (ATCC 11116) ATCC Type strain of H. influenzae biotype aegyptius R2140 Blood H. influenzae biotype aegyptius isolate from BPF; hemagglutinating (CDC F3031) 6 R2141 Blood H. influenzae biotype aegyptius isolate from BPF; nonhemagglutinating (CDC F3035) 6 R2777 CSF, from invasive disease Nonencapsulated This study R3001 Bronchial lavage of cystic fibrosis patient Nonencapsulated 25 C2836 Otitis media Nonencapsulated This study C2840 Tracheal infection in newborn Nonencapsulated This study C2843 Otitis media Type b This study C2853 Sputum from cystic fibrosis patient Nonencapsulated 25 C2859 CSF Nonencapsulated 25 C2861 CSF Nonencapsulated 25 Open in a separate window a ATCC, American Type Culture Collection; NCTC, National Collection of Type Cultures.

Techniques: Plasmid Preparation, Clone Assay, Amplification, Subcloning, Infection