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To explore the potential storage and safety of drinking leftover bottled tea beverages from various manufacturers after direct drinking from bottles, we conducted a screening experiment on the growth of salivary bacteria in plastic bottles of tea. The diluted saliva samples from 10 participants were inoculated into the test bottled beverages, which resulted in bacteria, particularly former members of the genus Lactobacillus, growing in some green tea beverages with a neutral pH. In contrast, tea beverages with less bacterial growth contained Streptococcus spp., and the leftovers may be safe to store and drink again.
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BACKGROUND: Dialysis-related amyloidosis (DRA) is a severe complication in end-stage kidney disease (ESKD) patients undergoing long-term dialysis treatment, characterized by the deposition of ß2-microglobulin-related amyloids (Aß2M amyloid). To inhibit DRA progression, hexadecyl-immobilized cellulose bead (HICB) columns are employed to adsorb circulating ß2-microglobulin (ß2M). However, it is possible that the HICB also adsorbs other molecules involved in amyloidogenesis. METHODS: We enrolled 14 ESKD patients using HICB columns for DRA treatment; proteins were extracted from HICBs following treatment and identified using liquid chromatography-linked mass spectrometry. We measured the removal rate of these proteins and examined the effect of those molecules on Aß2M amyloid fibril formation in vitro. RESULTS: We identified 200 proteins adsorbed by HICBs. Of these, 21 were also detected in the amyloid deposits in the carpal tunnels of patients with DRA. After passing through the HICB column and hemodialyzer, the serum levels of proteins such as ß2M, lysozyme, angiogenin, complement factor D and matrix Gla protein were reduced. These proteins acted in the Aß2M amyloid fibril formation. CONCLUSIONS: HICBs adsorbed diverse proteins in ESKD patients with DRA, including those detected in amyloid lesions. Direct hemoperfusion utilizing HICBs may play a role in acting Aß2M amyloidogenesis by reducing the amyloid-related proteins.
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Amiloidosis , Celulosa , Fallo Renal Crónico , Proteómica , Diálisis Renal , Microglobulina beta-2 , Humanos , Amiloidosis/metabolismo , Amiloidosis/sangre , Amiloidosis/terapia , Diálisis Renal/efectos adversos , Masculino , Femenino , Microglobulina beta-2/metabolismo , Microglobulina beta-2/sangre , Proteómica/métodos , Anciano , Celulosa/química , Persona de Mediana Edad , Adsorción , Fallo Renal Crónico/terapia , Fallo Renal Crónico/metabolismo , Fallo Renal Crónico/sangre , Espectrometría de Masas/métodos , Amiloide/metabolismo , Cromatografía LiquidaRESUMEN
OBJECTIVES: Breast milk is a valuable and useful source of nutrition; however, surplus milk is routinely discarded for hygiene reasons despite an unclear scientific basis. Here, we profiled the microbiota of expressed breast milk before and after feeding with an artificial nipple and examined the bacterial survival in breast milk stored at 4 °C. METHODS: Eleven mother-baby pairs were included in the study. Samples of expressed breast milk were collected before and after feeding with an artificial nipple and examined both immediately (0 h) and after storage for 3 and 12 h at 4 °C. Each sample was inoculated onto a blood agar plate and incubated anaerobically and aerobically at 37 °C. Genomic DNA was extracted from individual bacterial colonies, which were identified by 16S rRNA gene sequencing. RESULTS: Before feeding, the bacterial counts at 0 and 12 h were (1.4 ± 1.6) × 105 colony-forming units (CFU)/mL and (1.4 ± 0.6) × 105 CFU/mL, respectively. Staphylococcus (47.7% and 41.9%, respectively), Cutibacterium (20.7% and 36.0%, respectively), and Streptococcus (16.1% and 6.6%, respectively) were identified among the samples. In contrast, after feeding, the bacterial counts at 0 and 12 h were (2.7 ± 1.7) × 105 CFU/mL and (2.1 ± 2.5) × 105 CFU/mL, respectively. Staphylococcus (30.1% and 37.4%, respectively), Cutibacterium (11.7% and 31.7%, respectively), and Streptococcus (41.5% and 25.2%, respectively), were identified among the samples. CONCLUSIONS: Bacteria were present in the breast milk before feeding. Although the main component of the microbiota shifted from Staphylococcus to Streptococcus species after feeding, these results suggest that surplus expressed breast milk may be preserved safely in a refrigerator for at least 12 h after feeding with an artificial nipple.
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Microbiota , Leche Humana , Humanos , Lactante , Femenino , Leche Humana/microbiología , ARN Ribosómico 16S/genética , Pezones , Microbiota/genética , Bacterias/genética , Streptococcus/genéticaRESUMEN
This study aimed to characterize commensal microbiota on the skin before and after wearing masks, and to characterize the microbiota on the surface of used masks after 1 week of drying. From the 13 human subjects (age range, 19-26 years), mean bacterial concentrations of (6.1 ± 11.0) × 105 and (1.0 ± 1.4) × 106 colony-forming units (CFU)/mL were recovered from the skin of the buccal areas wiped with a sterile cotton swab before and after wearing non-woven fabric masks for 8 h, respectively. Furthermore (3.4 ± 4.9) × 104 CFU/mL of bacteria were recovered from the mask surfaces. The bacteria contained in the masks, which consisted mainly of Cutibacterium acnes and Staphylococcus epidermidis/aureus, virtually disappeared after drying the masks indoors for 1 week.
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Máscaras , Microbiota , Textiles , Adulto , Humanos , Propionibacterium acnes , Staphylococcus aureus , Adulto JovenRESUMEN
OBJECTIVES: The survival of bacteria in the sports drink and orange juice remaining in and at the mouth of bottles after direct drinking was examined after immediately drinking and incubation at 37 °C for 24 h. METHODS: Nine healthy participants were asked to drink approximately 100 mL of a plastic bottled sports drink or orange juice. The samples were cultured anaerobically at 37 °C for 7 days. Genomic DNA was extracted from the resulting individual colonies, and bacterial species were identified using 16 S rRNA gene sequencing. RESULTS: The mean amount of bacteria in the remaining sports drink and orange juice, immediately after drinking, were (1.6 ± 2.3) × 103 colony-forming units (CFU)/mL and (2.9 ± 3.3) × 103 CFU/mL, respectively. Additionally, bacteria recovered from the mouths of the sports drink and orange juice bottles were (2.5 ± 5.5) × 104 CFU/mL and (5.8 ± 2.4) × 103 CFU/mL, respectively. Oral bacteria, such as Streptococcus, Actinomyces, Neisseria, and Rothia were found to be transferred in the sports drink and orange juice, and the bacteria were scarcely detected after incubation at 37 °C for 24 h. CONCLUSIONS: The bacterial levels differed significantly from the previously reported levels in bottled tea 24 h after drinking, suggesting that remaining drinks with low pH levels can be preserved for a longer period.
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Citrus sinensis , Microbiota , Humanos , Microbiota/genética , Jugos de Frutas y Vegetales , Boca , PlásticosRESUMEN
OBJECTIVES: Profiling of oral microbiota has traditionally been performed using conventional methods. These methods are relatively time-consuming and labor-intensive. Metagenomic analysis of oral microbiota using high-speed next-generation sequencing is a highly promising technology. However, it is expensive. This study sought to develop a simple and cost-effective profiling method for oral microbiota using 16S rRNA gene polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of PCR-amplified 16S ribosomal RNA genes. METHODS: Oral isolates of 59 bacterial species from human saliva, including Streptococcus, Actinomyces, and Veillonella, were cultured anaerobically on CDC Anaerobe 5% sheep blood agar plates. Genomic DNA was extracted from single colonies and 16S rRNA genes were PCR-amplified using the 27F and 1492R universal primers. The PCR products were purified and characterized by single digestion with HpaII restriction endonuclease. 16S rRNA gene sequences were obtained from the GenBank database, and the expected restriction profiles were compared with the RFLP patterns obtained from agarose gel electrophoresis. RESULTS: Sixty-five RFLP patterns were obtained from 27 genera and 59 species. The expected fragment sizes of these species were calculated based on GenBank 16S rRNA gene sequences. Fifty-nine patterns were obtained from the analysis of GenBank sequences. The RFLP patterns produced with HpaII distinguished many oral bacterial species. RFLP patterns enabling identification of oral bacteria were generated. The 16S rRNA gene PCR-RFLP analysis did not require expensive equipment and reagents and was cost-effective. CONCLUSION: PCR-RFLP analysis based on 16S rRNA genes could be an alternative method for oral microbiota analysis in smaller laboratories.
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Microbiota , Boca/microbiología , Cartilla de ADN , Humanos , Microbiota/genética , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , ARN Ribosómico 16S/genéticaRESUMEN
It has been speculated that oral bacteria can be transferred to tea in plastic bottles when it is drunk directly from the bottles, and that the bacteria can then multiply in the bottles. The transfer of oral bacteria to the mouth of bottles and bacterial survival in the remaining tea after drinking directly from bottles were examined immediately after drinking and after storage at 37 °C for 24 h. Twelve healthy subjects (19 to 23 years of age) were asked to drink approximately 50 mL of unsweetened tea from a plastic bottle. The mouths of the bottles were swabbed with sterile cotton, and the swabs and the remaining tea in the bottles were analyzed by anaerobic culture and 16S rRNA gene sequencing. Metagenomic analysis of the 16S rRNA gene was also performed. The mean amounts of bacteria were (1.8 ± 1.7) × 104 colony-forming units (CFU)/mL and (1.4 ± 1.5) × 104 CFU/mL at the mouth of the bottles immediately after and 24 h after drinking, respectively. In contrast, (0.8 ± 1.6) × 104 CFU/mL and (2.5 ± 2.6) × 106 CFU/mL were recovered from the remaining tea immediately after and 24 h after drinking, respectively. Streptococcus (59.9%) were predominant at the mouth of the bottles immediately after drinking, followed by Schaalia (5.5%), Gemella (5.5%), Actinomyces (4.9%), Cutibacterium (4.9%), and Veillonella (3.6%); the culture and metagenomic analyses showed similar findings for the major species of detected bacteria, including Streptococcus (59.9%, and 10.711%), Neisseria (1.6%, and 24.245%), Haemophilus (0.6%, and 15.658%), Gemella (5.5%, and 0.381%), Cutibacterium (4.9%, and 0.041%), Rothia (2.6%, and 4.170%), Veillonella (3.6%, and 1.130%), Actinomyces (4.9%, and 0.406%), Prevotella (1.6%, and 0.442%), Fusobacterium (1.0%, and 0.461%), Capnocytophaga (0.3%, and 0.028%), and Porphyromonas (1.0%, and 0.060%), respectively. Furthermore, Streptococcus were the most commonly detected bacteria 24 h after drinking. These findings demonstrated that oral bacteria were present at the mouth of the bottles and in the remaining tea after drinking.
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OBJECTIVES: To clarify the characteristics and growth of bacteria that may infiltrate liquid baby formula during feeding and after storage for more than 3 h, the transfer of oral bacteria through artificial nipples, and bacterial survival in liquid baby formula and a baby drink were examined immediately after drinking and after storage at 4 °C for 12 h and 24 h. METHODS: Thirteen human subjects (aged 19-24 years) were asked to drink approximately 50 mL of liquid baby formula and a baby drink, via the artificial nipple of a baby bottle. Samples of the remaining liquid after storage at 4 °C for 12 h and 24 h were inoculated onto blood agar plates and incubated anaerobically at 37 °C for 7 days. Genomic DNA was extracted from individual colonies, and the bacterial species were identified by 16S rRNA gene sequencing. RESULTS: The mean concentrations of bacteria in the liquid baby formula were (2.6 ± 2.8) × 104 and (4.1 ± 6.6) × 104 colony-forming unit/mL after storage at 4 °C for 12 h and 24 h, respectively. Streptococcus (43.2%), Veillonella (9.3%), and Schaalia (8.2%) species were recovered from the remaining liquid baby formula after storage at 4 °C for 12 h. In contrast, no bacteria were detected in the remaining baby drink after storage at 37 °C for 24 h. CONCLUSIONS: The levels of bacteria immediately after drinking and after storage at 4 °C for 12 h or 24 h were similar, suggesting that remaining liquid baby formula may be preserved safely in a refrigerator for more than 3 h.
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Microbiota , Pezones , Bacterias/genética , Humanos , Fórmulas Infantiles , ARN Ribosómico 16S/genéticaRESUMEN
It is suspected that oral bacteria are transferred to the liquid baby formula through the artificial nipple and multiply in the bottle after feeding. In the present study, in order to understand the influence of bacteria on liquid baby formula after feeding, the transfer of oral bacteria through artificial nipples and their survival in liquid baby formula were examined immediately after drinking as well as after storage at 4°C for 3 h. Four healthy human subjects (20-23 years old) were asked to drink liquid baby formula (Aptamil®, ca. 50 mL) from baby bottles using artificial nipples. Samples of the liquid baby formula (immediately after drinking and 3 h later) were inoculated onto blood agar plates and incubated anaerobically at 37°C for 7 days. Salivary samples from each subject and 6 newborn infants were also cultured. Genomic DNA was extracted from individual colonies, and bacterial species were identified by 16S rRNA gene sequencing. The mean amounts of bacteria (CFU/mL) were (3.2 ± 3.0) ×104 and (3.4 ± 3.3) ×104 immediately after drinking and 3 h later, respectively. Streptococcus (41.6 and 40.5%), Actinomyces (24.3 and 21.5%) and Veillonella (16.2 and 11.0%) were recovered from the samples immediately after drinking and 3 h later, respectively. On the other hand, Streptococcus (38.9%), Actinomyces (17.1%), Neisseria (9.1%), Prevotella (6.9%), Rothia (6.9%) and Gemella (5.1%) were predominant in the saliva of adult subjects, and Streptococcus (65.2%), Staphylococcus (18.5%), Gemella (8.2%) and Rothia (5.4%) were predominant in the saliva of infant subjects. From these findings, oral bacteria, e.g., Streptococcus, Gemella and Rothia, were found to transfer into the liquid baby formula through artificial nipples, and the bacterial composition in the remaining liquid baby formula was found to resemble that of human saliva. The bacterial levels were similar between immediately after drinking and when stored at 4°C for 3 h, suggesting that the remaining liquid baby formula may be preserved in a refrigerator for a specified amount of time.