Probiotic Insights from the Genomic Exploration of Lacticaseibacillus paracasei Strains Isolated from Fermented Palm Sap.

This study focused on L. paracasei strains isolated from fermented palm sap in southern Thailand that exhibit potential probiotic characteristics, including antibiotic susceptibility, resistance to gastrointestinal stresses, and antimicrobial activity against various pathogens. However, a thorough investigation of the whole genome sequences of L. paracasei isolates is required to ensure their safety and probiotic properties for human applications. This study aimed to sequence the genome of L. paracasei isolated from fermented palm sap, to assess its safety profile, and to conduct a comprehensive comparative genomic analysis with other Lacticaseibacillus species. The genome sizes of the seven L. paracasei strains ranged from 3,070,747 bp to 3,131,129 bp, with a GC content between 46.11% and 46.17% supporting their classification as nomadic lactobacilli. In addition, the minimal presence of cloud genes and a significant number of core genes suggest a high degree of relatedness among the strains. Meanwhile, phylogenetic analysis of core genes revealed that the strains possessed distinct genes and were grouped into two distinct clades. Genomic analysis revealed key genes associated with probiotic functions, such as those involved in gastrointestinal, oxidative stress resistance, vitamin synthesis, and biofilm disruption. This study is consistent with previous studies that used whole-genome sequencing and bioinformatics to assess the safety and potential benefits of probiotics in various food fermentation processes. Our findings provide valuable insights into the potential use of seven L. paracasei strains isolated from fermented palm sap as probiotic and postbiotic candidates in functional foods and pharmaceuticals.

Lyophilized cell-free supernatants of Limosilactobacillus fermentum T0701 exhibited antibacterial activity against Helicobacter pylori.

Helicobacter pylori is a prominent gastrointestinal pathogen associated with various gastrointestinal illnesses. It presents substantial health risks due to its antibiotic resistance. Therefore, it is crucial to identify alternative treatments for H. pylori infections. Limosilactobacillus spp exhibit probiotic properties with beneficial effects in humans; however, the mechanisms by which it counteracts H. pylori infection are unknown. This study aimed to evaluate the potential of Limosilactobacillus fermentum T0701 lyophilized cell-free supernatants (LCFS) against H. pylori. The LCFS has varying antimicrobial activities, with inhibition zones of up to 10.67 mm. The minimum inhibitory concentration and minimum bacterial concentration of LCFS are 6.25-25.00 mg/mL and 6.25 mg/mL to > 50.00 mg/mL, respectively, indicating its capability to inhibit H. pylori. There is morphological damage observed in H. pylori treated with LCFS. Additionally, H. pylori adhesion to AGS cells (human gastric adenocarcinoma epithelial cells) reduces by 74.23%, highlighting the LCFS role in preventing bacterial colonization. Moreover, LCFS exhibits no cytotoxicity or morphological changes in AGS cells, and with no detected virulence or antimicrobial resistance genes, further supporting its safety profile. L. fermentum T0701 LCFS shows promise as a safe and effective non-toxic agent against H. pylori, with the potential to prevent gastric colonization.

Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae: insights from a tertiary hospital in Southern Thailand.

Broad-spectrum ampicillin-resistant and third-generation cephalosporin-resistant Enterobacteriaceae, particularly Escherichia coli and Klebsiella pneumoniae that have pathological features in humans, have become a global concern. This study aimed to investigate the prevalence, antimicrobial susceptibility, and molecular genetic features of extended-spectrum beta-lactamase (ESBL)-producing E. coli and K. pneumoniae isolates in Southern Thailand. Between January and August 2021, samples (n = 199) were collected from a tertiary care hospital in Southern Thailand. ESBL and AmpC-lactamase genes were identified using multiplex polymerase chain reaction (PCR). The genetic relationship between ESBL-producing E. coli and K. pneumoniae was determined using the enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction. ESBL-producing E. coli and K. pneumoniae isolates were mostly collected from catheter urine samples of infected female patients. The ESBL production prevalence was highest in the medical wards (n = 75, 37.7%), followed by that in surgical wards (n = 64, 32.2%) and operating rooms (n = 19, 9.5%). Antimicrobial susceptibility analysis revealed that all isolates were resistant to ampicillin, cefotaxime, ceftazidime, ceftriaxone, and cefuroxime; 79.4% were resistant to ciprofloxacin; and 64.3% were resistant to trimethoprim-sulfamethoxazole. In ESBL-producing K. pneumoniae and E. coli, blaTEM (n = 57, 72.2%) and blaCTX-M (n = 61, 50.8%) genes were prominent; however, no blaVEB, blaGES, or blaPER were found in any of these isolates. Furthermore, only ESBL-producing K. pneumoniae had co-harbored blaTEM and blaSHV genes at 11.6%. The ERIC-PCR pattern of multidrug-resistant ESBL-producing strains demonstrated that the isolates were clonally related (95%). Notably, the presence of multidrug-resistant and extremely resistant ESBL producers was 83.4% and 16.6%, respectively. This study highlights the presence of blaTEM, blaCTX-M, and co-harbored genes in ESBL-producing bacterial isolates from hospitalized patients, which are associated with considerable resistance to beta-lactamase and third-generation cephalosporins. IMPORTANCE: We advocate for evidence-based guidelines and antimicrobial stewardship programs to encourage rational and appropriate antibiotic use, ultimately reducing the selection pressure for drug-resistant bacteria and lowering the likelihood of ESBL-producing bacterial infections.

Factors associated with the worsening of COVID-19 symptoms among cohorts in community- or home-isolation care in southern Thailand.

Introduction: This study aimed to investigate factors associated with time-to-referral due to worsening symptoms in patients with laboratory-confirmed COVID-19 in southern Thailand. While underlying diseases have been evaluated to assess COVID-19 severity, the influence of vaccinations and treatments is also crucial. Methods: A cohort of 8,638 patients quarantined in home or community isolation with laboratory-confirmed COVID-19 was analyzed. Survival analysis and the Cox proportional hazard ratio were employed to assess factors influencing time-toreferral. Results: Age ≥ 60 years, neurologic disorders, cardiovascular disease, and human immunodeficiency virus infection were identified as significant risk factors for severe COVID-19 referral. Patients who received full- or booster-dose vaccinations had a lower risk of experiencing severe symptoms compared to unvaccinated patients. Notably, individuals vaccinated during the Omicron-dominant period had a substantially lower time-to-referral than those unvaccinated during the Delta-dominant period. Moreover, patients vaccinated between 1 and 6 months prior to infection had a significantly lower risk of time-to-referral than the reference group. Discussion: These findings demonstrate early intervention in high-risk COVID-19 patients and the importance of vaccination efficacy to reduce symptom severity. The study provides valuable insights for guiding future epidemic management strategies and optimising patient care during infectious disease outbreaks.

Evaluation of Antibacterial, Antibiofilm, Antioxidant, and Anti-Inflammatory Activities of Kratom Leaves (Mitragyna speciosa) Fermentation Supernatant Containing Lactobacillus rhamnosus GG.

Kratom (Mitragyna speciosa) leaves are commonly used to enhance endurance and treat various diseases. This study evaluated the effect of kratom leaf fermentation with Lactobacillus rhamnosus. Antibacterial activity was investigated against Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Escherichia coli, and E. coli O157:H7. Biofilm inhibition and eradication assays were also performed. Antioxidant properties were determined by measuring the total phenolic and flavonoid content and DPPH and ABTS scavenging activities. Nitric oxide and TNF-α, IL-1ß, and IL-6 expressions in LPS-stimulated RAW 264.7 macrophage cells were also measured. Aqueous kratom extract exhibited promising effects against free radicals and pro-inflammatory cytokines. Notably, all fermented kratoms showed significant antibacterial activity against the tested pathogens and antibiofilm formation by S. aureus and MRSA. Furthermore, the eradication of established biofilms of fermented kratoms was observed in S. aureus (day 2, 50 mg/mL) and E. coli (day 2, 100 mg/mL and day 4, 50 mg/mL). To the best of our knowledge, this study is the first to report that fermented and non-fermented kratoms could be nutraceutical sources of antibacterial, antibiofilm, antioxidant, and anti-inflammatory substances against related diseases and can be applied further in dietary or cosmetic products with health-promoting effects.

Relation between Increased IL-10 Levels and Malaria Severity: A Systematic Review and Meta-Analysis.

The roles of anti-inflammatory cytokines in the pathogenesis of severe malaria have been widely studied, and the role of IL-10 in the pathogenesis of severe malaria remains unclear. Therefore, we performed a systematic review and meta-analysis to determine the difference in IL-10 levels between patients with severe malaria and those with non-severe malaria. The search for relevant studies was performed using PubMed, Scopus, and Embase from 1 February 2022 to 12 February 2022. The quality of the included studies was assessed according to the guidelines of the Strengthening the Reporting of Observational Studies in Epidemiology. The random-effects model was used to estimate the pooled effect. In all, 1215 studies were identified, and 19 were included in the quantitative syntheses. The results showed that patients with severe malaria had a higher IL-10 level than those with non-severe malaria (p = 0.03, pooled standardized mean difference: 0.74, 95% CI: 0.08-1.40, I2: 97.22%, 19 studies/21 sub studies). The meta-analysis results demonstrated increased IL-10 levels in patients with severe malaria compared with those with non-severe malaria. However, with the heterogeneity of the meta-analysis results, further studies are required to confirm the changes in the IL-10 levels according to the severity of malaria and to investigate whether a combination of other severity parameters with IL-10 levels could be an alternative marker for severe malaria.

Prevalence and Associated Factors of Delayed Diagnosis of Type 2 Diabetes Mellitus in a Tertiary Hospital: A Retrospective Cohort Study.

Objective: To determine the prevalence and associated factors of delayed diagnosis of type 2 diabetes mellitus (DM) among outpatients in a tertiary hospital. Methods: This retrospective cohort study was conducted among outpatients aged ≥35 years with twice fasting plasma glucose (FPG) levels ≥126 mg/dl between 1 January 2018, and 31 December 2020. The prevalence and pattern of delayed diagnosis of DM were defined using the Thai Clinical Practice Guideline (CPG) for Diabetes, 2017, and the American Diabetes Association (ADA) 2017. The cut-off time for FPG level confirmation of 3 months was used to evaluate delayed diagnoses and associated factors. Multiple logistic regression was used to identify variables associated with delayed diagnoses. Results: Of 260 participants, 96.9% and 85.4% had delayed diagnoses as defined by the Thai CPG and the ADA, respectively. Factors significantly associated with delayed diagnosis were hypertension, non-cash insurance, and >10 years of physician experience. Conclusion: Undiagnosed diabetes and diagnosis delay should be a concern in tertiary settings. Senior physicians should focus on patients with higher FPG levels, particularly those who have hypertension, and use non-cash insurance schemes.

Characterization of probiotic properties and development of banana powder enriched with freeze-dried Lacticaseibacillus paracasei probiotics.

Lacticaseibacillus paracasei is one of the probiotic bacteria widely identified from fermented foods. The application of L. paracasei is commonly used in dairy and non-dairy products. To investigate the probiotic properties of L. paracasei cells including their acid, pepsin, pancreatin, and bile salt tolerances; adhesion ability; antipathogen activity; and antibiotic susceptibility, L. paracasei cells were incorporated into skim milk and lyophilized by freeze drying. Freeze-dried probiotic cells were add to green banana powder and low moisture additive food matrices and a storage analysis of the product was performed. The result showed that L. paracasei cells possessed potentially beneficial probiotic properties to survive stress in the gastrointestinal tract (GIT) and functional abilities as an anti-enteropathogenic agent; they were also safe to use and displayed antibiotic properties. Furthermore, the probiotic freeze-drying technique preserved high probiotic cell survivability (1011 CFU/g). In term of prolonged storage (60 days), the powder product was stable and maintained probiotic survival (107 CFU/g) while excluding non-probiotic growth. In conclusion, L. paracasei displayed probiotic properties in the GIT and was judged to be a highly acceptable product as a probiotics-banana rehydrated beverage.

Factors Associated with Anxiety and Depression Among Micro, Small, and Medium Enterprise Restaurant Entrepreneurs Due to Thailand's COVID-19-Related Restrictions: A Cross-Sectional Study.

Purpose: COVID-19 has become a global pandemic. Since the Thai government announced its pandemic-related restrictions, economic indicators have inevitably been impacted. Micro, small, and medium enterprise (MSME) entrepreneurs have adapted to the unexpected changes, undoubtedly requiring government assistance. Therefore, this study assessed the prevalence of anxiety and depression in MSME restaurant entrepreneurs after the government announced its COVID-19-related restrictions. Patients and Methods: An online questionnaire was developed and distributed to MSME entrepreneurs' restaurants in Songkhla province, southern Thailand, in November 2021. Thai Hospital Anxiety and Depression Scales (Thai HADS) were used to assess the prevalence of anxiety and depression. Multiple logistic regression was used to analyze the factors associated with depression and anxiety, and the adjusted odds ratio (OR) for both variables was reported at a P-value < 0.05 for significance. Results: In total, 181 MSME restaurant entrepreneurs responded to our online questionnaire in November 2021. The prevalence of anxiety and depression was 12.2% and 11.6%, respectively. Participants with a family history of psychological disorders (OR 34.9, P-value 0.003), an underlying disease (OR 7.14, P-value 0.032), a high debt-to-income ratio (OR 73.75, P-value 0.007), or a low education level were more likely to have anxiety. Similarly, participants who had an underlying disease (OR 14.29, P-value 0.032) and lacked access to supportive measures from the government (OR 6.94, P-value 0.009) were more likely to suffer from depression. Conclusion: The period of restrictive policies during the COVID-19 pandemic had a negative impact on MSME restaurant entrepreneurs' mental health. The factors that were significantly associated with anxiety were debt-to-income ratio and concern about COVID-19 prevention and treatment. The factors that were significantly associated with depression included inadequate access to government aid. Hence, the government should focus on providing and allocating healthcare resources during the COVID-19 pandemic.

Lacticaseibacillus spp.; Probiotic candidates from Palmyra palm sugar possesses antimicrobial and anti-biofilm activities against methicillin-resistant Staphylococcus aureus.

Background and Aim: Probiotics are beneficial microorganisms that play important roles by adhering to the gut and producing antimicrobial substances to inhibit pathogens. The objective of this study was to isolate and characterize the probiotic lactic acid bacteria (LAB) from Palmyra palm sugar, which can produce antimicrobial compounds against methicillin-resistant Staphylococcus aureus (MRSA), a new zoonotic and food-borne pathogens. Materials and Methods: Twenty-six LAB isolates were isolated from 30 Palmyra palm sugar samples. Three selected LAB were further characterized as probiotics. In addition, the antibacterial and anti-biofilm-forming activities of the probiotics' culture supernatants against MRSA and food-borne pathogens were investigated. Finally, the selected probiotics were identified by aligning 16S rRNA sequences. Results: The three confirmed probiotics, WU 0904, WU 2302, and WU 2503, showed strong antibacterial activities against S. aureus, MRSA, Escherichia coli O157:H7, and Listeria monocytogenes, as measured by a broth microdilution assay. Among the LAB isolates, 82.22-86.58%, 91.83-96.06%, and 64.35-74.93% exhibited resistance to low pH, pancreatin treatment, and bile salts, respectively. It was found that 59.46% and 83.33% auto-aggregation was observed in 2 and 24 h, respectively. Moreover, 50.25-57.24% adhesion was detected after the incubation of the bacterial cells to Caco-2 cells.. Biofilm inhibition (82.81-87.24%) was detected after the treatment of MRSA with the culture supernatants, when compared with that to the control. By the alignment of 16S rRNA sequences, the isolate WU 2302 was identified as Lacticaseibacillus spp. with 98.82% homology when compared to the GenBank database. Conclusion: This study indicates that isolated probiotics can produce antimicrobial compounds against MRSA and food-borne pathogens. The obtained results strongly suggest that these probiotics are promising candidates for pharmaceutical products.

Occurrence, Antimicrobial Resistance Profile, and Characterization of Extended-Spectrum ß-Lactamase-Producing Escherichia coli Isolates from Minced Meat at Local Markets in Thailand.

Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli exhibits strong multidrug resistance (MDR) to ampicillin and third-generation cephalosporins. This study examined the occurrence, antimicrobial susceptibility, and molecular genetic features of ESBL-producing E. coli isolates from three commonly consumed minced meat varieties, namely pork, chicken, and beef. In total, 150 samples were collected from 10 local markets in Thailand. ESBL-producing E. coli was identified in 78 samples (52%), and minced chicken meat was most contaminated (79.17%). The isolates exhibited potential susceptibility to amikacin (96.16%) and carbapenems (91-95%). However, ESBL-producing E. coli displayed strong resistance to ampicillin and cefpodoxime (100%) and high MDR to 3-5 antibiotic classes (94.87%). Most presumptive ESBL producers harbored ESBL resistance genes (97.44%), most commonly blaTEM (78.21%). Indeed, our results demonstrated that raw minced meat has a high occurrence of ESBL-producing E. coli harboring ESBL resistance genes, highlighting the importance of implementation of sanitary handling practices to reduce microbial contamination in commercial meat as well as the need for consumer education on safe handling and cooking of meat products.

Lyophilized cell-free supernatants of Lactobacillus isolates exhibited antibiofilm, antioxidant, and reduces nitric oxide activity in lipopolysaccharide-stimulated RAW 264.7 cells.

BACKGROUND: Probiotics can release bioactive substances known as postbiotics, which can inhibit pathogenic microorganisms, improve immunomodulation, reduce antioxidant production, and modulate the gut microbiota. METHODS: In this study, we evaluated the in vitro antimicrobial effects, antioxidant activity, and anti-inflammatory potential of 10 lyophilized cell-free supernatants (LCFS) of Lactobacillus isolates. LCFS was obtained via centrifugation and subsequent lyophilization of the supernatant collected from the culture medium ofeach isolate. The antibacterial and antibiofilm activities of the LCFS were determined using broth microdilution. The antioxidant potential was evaluated by measuring the total phenolic and flavonoid contents and 2,2-Diphennyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+) radical scavenging activities. RESULTS: All the isolates were able to inhibit the four tested pathogens. The isolates exhibited strong antibiofilm activity and eradicated the biofilms formed by Acinetobacter buamannii and Escherichia coli. All the prepared Lactobacillus LCFS contained phenols and flavonoids and exhibited antioxidant activities in the DPPH and ABTS+ radical scavenging assays. The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that LCFS was not cytotoxic to RAW 264.7 cells. In addition, the ten Lactobacillus LCFS decreased the production of nitric oxide. CONCLUSIONS: All the isolates have beneficial properties. This research sheds light on the role of postbiotics in functional fermented foods and pharmaceutical products. Further research to elucidate the precise molecular mechanisms of action of probiotics is warranted.

Effect of the COVID-19 Pandemic and Other Predictors of True Therapeutic Inertia on Patients with Hypertension in a Primary Care Clinic in Thailand.

INTRODUCTION: Hypertension (HT) has a significant impact on health care worldwide. Therapeutic inertia (TI) is defined as the failure to intensify therapy in the absence of an optimal goal and is widely used as a quality of care parameter. The coronavirus disease 2019 (COVID-19) pandemic has affected many health-care systems, including HT care. Therefore, the present study assessed the impact of the COVID-19 pandemic on TI and its predictors in patients with HT. METHODS: The electronic medical records of patients with HT who attended a primary care clinic at a tertiary hospital during pre-COVID-19 (February 2019 to February 2020) and COVID-19 (March to August 2020) periods were reviewed. RESULTS: Our study included 6089 visits during the 12-month pre-COVID-19 period and 2852 visits during the 6-month COVID-19 period. Most of the baseline characteristics of the HT patients were not significantly different between the two time periods. During the COVID-19 period, the percentage of uncontrolled HT visits decreased from 43% to 31%. Similarly, the prevalence of TI decreased from 81% to 77%. False TI was predominantly due to physicians' concerns regarding the in-clinic blood pressure measurement being inaccurate during both the periods. CONCLUSION: After readjustment for the physicians 'reasons, the true TI was 64% and 60% in the pre-COVID-19 and COVID-19 period. For adjusted physician and patient-related factors, multilevel modeling was used. Senior medical staff visits, elderly patients, prior diabetes mellitus diagnosis, patients who used more than one type of anti-HT medication, and patients with systolic blood pressure >150 mmHg were all predictors of TI. The COVID-19 period, on the other hand had no effect on TI with an adjusted odds ratio of 0.82 (95% confidence interval, 0.67-1.01).

Prevalence and characterization of extended-spectrum ß-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolated from raw vegetables retailed in Southern Thailand.

BACKGROUND: The increasing prevalence of broad-spectrum ampicillin-resistant and third-generation cephalosporin-resistant Enterobacteriaceae, particularly Escherichia coli and Klebsiella pneumoniae, has become a global concern, with its clinical impacts on both human and veterinary medicine. This study examined the prevalence, antimicrobial susceptibility, and molecular genetic features of extended-spectrum ß-lactamase (ESBL)-producing E. coli and K. pneumoniae isolates from 10 types of raw vegetables. METHODS: In total, 305 samples were collected from 9 markets in Nakhon Si Thammarat, Thailand, in 2020. RESULTS: ESBL-producing E. coli and K. pneumoniae isolates were found in 14 of the 305 samples obtained from 7 out of 10 types of vegetables (4.6% of the total). Further, 14 ESBL-producing E. coli (n = 5/14) and K. pneumoniae isolates (n = 9/14) (1.6% and 3.0%, respectively) were highly sensitive to ß-lactam/carbapenem antibiotics (imipenem, 100%). ESBL-producing E. coli (n = 4) and K. pneumoniae isolates (n = 8) were also sensitive to non-ß-lactam aminoglycosides (amikacin, 80.00% and 88.89%, respectively). ESBL producers were most resistant to ß-lactam antibiotics, including ampicillin (85.71%) and the cephalosporins cefotaxime and ceftazidime (64.29%). The most frequently detected gene in ESBL-producing E. coli and K. pneumoniae was blaSHV . However, two ESBL-producing E. coli isolates also carried three other ESBL-encoding variants, blaTEM , blaCTX-M1 , blaGES and blaTEM, blaSHV, blaCTX-M9 , which may be due to their association with food chains and humans. DISCUSSION: Indeed, our results suggest that raw vegetables are an important source of ESBL-resistant E. coli and K. pneumoniae, which are potentially transmittable to humans via raw vegetable intake.

Probiotic Properties of Lactobacillus Species Isolated from Fermented Palm Sap in Thailand.

Lactic acid bacteria (LAB), which are the most frequently used probiotics in foods, confer health benefits such as antimicrobial activity, immune stimulation, and anticancer activity. Fermented palm sap is a potential source of LAB. This study aimed to evaluate in vitro antimicrobial and probiotic properties of LAB isolated from traditional fermented palm sap in Thailand. Among 40 isolated LAB species, 10 were preliminarily selected for their antimicrobial activity. These 10 isolates were identified and confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA sequencing as Lactobacillus paracasei (8/10), Lactobacillus fermentum (1/10), and Lactobacillus brevis (1/10). They were evaluated for probiotic characteristics and antimicrobial activities against pathogens. These isolates were tolerant toward simulated gastrointestinal tract conditions, including low pH, pepsin, pancreatin, and bile salts. The 10 isolates retained strong auto-aggregation and cell surface hydrophobicity, and they adhered tightly to human intestinal epithelial cells. The isolates were susceptible to ampicillin, erythromycin, clindamycin, tetracycline, and chloramphenicol but resistant to vancomycin, kanamycin, and streptomycin. Moreover, all isolates exhibited no hemolytic activity. All isolates exhibited good antibacterial activity against nine pathogenic bacteria. Thus, these 10 Lactobacillus isolates from fermented palm sap are promising potential candidates for use as probiotics in functional fermented foods and pharmaceutical products.