Bacterial vaginosis-associated bacteria G. vaginalis (ATCC 14018, ATCC 49145), candidiasis pathogen Candida albicans (ATCC 14053), trichomoniasis pathogen Trichomonas vaginalis (ATCC 30001), and human vagina-derived lactobacilli, Lactobacillus crispatus (ATCC 33820), Lactobacillus gasseri (ATCC 33323), Lactobacillus plantarum (ATCC 14917), and Lactobacillus jensenii (ATCC 25258), were purchased from the American Type Culture Collection (ATCC). Gardnerella vaginalis MR is a spontaneous metronidazole-resistant mutant of ATCC 14018. The clinical isolates of BV-associated bacteria and healthy human vagina lactobacilli were kindly provided by Dr. Chuang at National Yang Ming Chiao Tung University, Taiwan (all bacterial isolates are listed in Table 1 ). All BV-associated bacteria were cultured in NYCIII broth (ATCC medium 1685) and grown at 37°C in anaerobic conditions, using AnaeroPack®-Anaero (MGC, Japan). All Lactobacillus spp. were cultured in MRS broth (Difco BD) and grown at 37°C in facultatively anaerobic conditions, using AnaeroPack®-MicroAero (MGC, Japan), except L. gasseri was grown in aerobic conditions. Candida albicans was cultured in YM broth (Difco BD) and grown at 37°C in aerobic conditions.
Lactobacillus gasseri
Manufactured by American Type Culture Collection
Sourced in United States
Lactobacillus gasseri is a species of bacteria commonly found in the human gastrointestinal tract. It is an anaerobic, gram-positive, rod-shaped bacterium that is known for its probiotic properties. This strain is available for laboratory and research purposes.
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Lab products found in correlation
Lactobacillus plantarum, by American Type Culture Collection (2 mentions)
Bifidobacterium adolescentis, by American Type Culture Collection (2 mentions)
Staphylococcus epidermidis, by American Type Culture Collection (2 mentions)
Bacillus cereus, by American Type Culture Collection (2 mentions)
Streptococcus mutans, by American Type Culture Collection (2 mentions)
Deinococcus radiodurans, by American Type Culture Collection (2 mentions)
Rhodobacter sphaeroides, by American Type Culture Collection (2 mentions)
Escherichia coli, by American Type Culture Collection (2 mentions)
Enterococcus faecalis, by American Type Culture Collection (2 mentions)
Mrs broth, by BD (2 mentions)
Candida albicans, by American Type Culture Collection (1 mentions)
Ym broth, by BD (1 mentions)
Lactobacillus rhamnosus, by American Type Culture Collection (1 mentions)
Lactobacillus casei, by American Type Culture Collection (1 mentions)
Lactobacillus fermentum, by American Type Culture Collection (1 mentions)
Lactobacillus helveticus, by American Type Culture Collection (1 mentions)
Lactobacillus reuteri, by American Type Culture Collection (1 mentions)
Lactobacillus strains, by American Type Culture Collection (1 mentions)
Acinetobacter baumannii, by American Type Culture Collection (1 mentions)
Helicobacter pylori, by American Type Culture Collection (1 mentions)
5 protocols using lactobacillus gasseri
1
Cultivation of Vaginal Microbiome Pathogens
2
Heat-Killed Lactobacillus Strains Characterization
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The following bacterial strains were used in the study: Lactobacillus gasseri JCM 1131T and JCM 5814; Limosilactobacillus fermentum (previously Lactobacillus fermentum) ATCC 9338; Lactobacillus helveticus YIT 0049; L. plantarum TUA 5099L, NRIC 1067T, and YIT 0139; and Lacticaseibacillus rhamnosus (previously Lactobacillus rhamnosus) ATCC 7469T. These bacteria were initially obtained from the American Type Culture Collection (ATCC), the Japan Collection of Microorganisms (JCM), the Nodai Research Institute Culture Collection (NRIC), Tokyo University of Agriculture (TUA), and Yakult Central Institute (YIT). All strains were cultured at 37°C for 20 h in lactobacilli-MRS broth (Difco), washed with 20 mM Tris–HCl (pH 7.4), and then heated at 100°C for 20 min to obtain heat-killed bacteria.
3
Characterization of Bacterial DNA Mixture
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A mixture of bacterial DNA (10 Strain Even Mix Genomic Material, MSA-1000) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), comprising genomic DNA prepared from the following ten bacterial strains: Bacillus cereus (ATCC 10987), Bifidobacterium adolescentis (ATCC 15703), Clostridium beijerinckii (ATCC 35702), Deinococcus radiodurans (ATCC BAA816), Enterococcus faecalis (ATCC 47077), Escherichia coli (ATCC 700926), Lactobacillus gasseri (ATCC 33323), Rhodobacter sphaeroides (ATCC 17029), Staphylococcus epidermidis (ATCC 12228), and Streptococcus mutans (ATCC 700610).
4
Lactobacillus Strains Preparation Protocol
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The PP mixture comprised various Lactobacillus strains supplied by the American Type Culture Collection (ATCC) (Manassas, VA, USA). The strains included Lactobacillus gasseri (ATCC 33323), Lactobacillus plantarum (ATCC BAA-793), Lactobacillus reuteri (ATCC 23272), Lactobacillus helveticus (ATCC BAA-2840), Lactobacillus fermentum (ATCC 23271), Lactobacillus rhamnosus (ATCC BAA-2836), and Lactobacillus casei (ATCC BAA-2843). Following the supplier’s protocol, we cultured the strains in MRS broth (Beckton Dickinson, Sparks, MD), and preserved them as glycerol stocks at −80°C. To initiate culture growth, we prepared 5 mL of pre-warmed MRS broth with the glycerol stocks and then incubated these starter cultures at 37°C in a 5% CO2 environment for 2 h. The bacteria were cultured overnight in 1 L of MRS broth under identical conditions until they reached the logarithmic growth phase, confirmed by measuring optical density at 600 nm (OD600). After growth, the bacterial cells were collected through several centrifugation steps at 3000×g and 4°C, followed by a wash in cold PBS. We then resuspended the bacterial pellets in 10% glycerol in PBS and promptly froze them in 1 mL aliquots for future administration to mice. The viability and concentration of the bacteria were verified by serial dilution and colony-forming unit (CFU) enumeration on agar plates.
5
Multi-Strain Bacterial Reference Sample
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MSA-2002 was purchased from ATCC, Manassas, VA. The sample contains a mixture of 20 different bacterial strains distributed equally (5% ea.): Acinetobacter baumannii (ATCC 17978), Actinomyces odontolyticus (ATCC 17982), Bacillus cereus (ATCC 10987), Bacteroides vulgatus (ATCC 8482), Bifidobacterium adolescentis (ATCC 15703), Clostridium beijerinckii (ATCC 35702), Cutibacterium acnes (ATCC 11828), Deinococcus radiodurans (ATCC BAA-816), Enterococcus faecalis (ATCC 47077), Escherichia coli (ATCC 700926), Helicobacter pylori (ATCC 700392), Lactobacillus gasseri (ATCC 33323), Neisseria meningitidis (ATCC BAA-335), Porphyromonas gingivalis (ATCC 33277), Pseudomonas aeruginosa (ATCC 9027), Rhodobacter sphaeroides (ATCC 17029), Staphylococcus aureus (ATCC BAA-1556), Staphylococcus epidermidis (ATCC 12228), Streptococcus agalactiae (ATCC BAA-611), and Streptococcus mutans (ATCC 700610).
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