Selection and evaluation of lactic acid bacteria from chicken feces in Thailand as potential probiotics.

Background: Lactic acid bacteria (LAB) are widely used as probiotics in poultry production due to their resilience to low pH and high bile salt concentrations, as well as their beneficial effects on growth performance and antagonistic activity against enteric pathogens. However, the efficacy of probiotics depends on strain selection and their ability to colonize the host's intestine. This study aimed to select, identify, and evaluate LAB strains isolated from chicken feces in Thailand for potential use as probiotics in the chicken industry. Methods: LAB strains were isolated from 58 pooled fresh fecal samples collected from chicken farms in various regions of Thailand, including commercial and backyard farms. Gram-positive rods or cocci with catalase-negative characteristics from colonies showing a clear zone on MRS agar supplemented with 0.5% CaCO3 were identified using MALDI-TOF mass spectrometry. The LAB isolates were evaluated for acid (pH 2.5 and pH 4.5) and bile salt (0.3% and 0.7%) tolerance. Additionally, their cell surface properties, resistance to phenol, antimicrobial activity, hemolytic activity, and presence of antimicrobial resistance genes were determined. Results: A total of 91 LAB isolates belonging to the Pediococcus, Ligilactobacillus, Limosilactobacillus, and Lactobacillus genera were obtained from chicken feces samples. Backyard farm feces exhibited a greater LAB diversity compared to commercial chickens. Five strains, including Ligilactobacillus salivarius BF12 and Pediococcus acidilactici BF9, BF14, BYF20, and BYF26, were selected based on their high tolerance to acid, bile salts, and phenol. L. salivarius BF12 and P. acidilactici BF14 demonstrated strong adhesion ability. The five LAB isolates exhibited significant cell-cell interactions (auto-aggregation) and co-aggregation with Salmonella. All five LAB isolates showed varying degrees of antimicrobial activity against Salmonella strains, with P. acidilactici BYF20 displaying the highest activity. None of the LAB isolates exhibited beta-hemolytic activity. Whole genome analysis showed that L. salivarius BF12 contained ermC, tetL, and tetM, whereas P. acidilactici strains BF9 and BF14 carried ermB, lnuA, and tetM. Conclusion: The selected LAB isolates exhibited basic probiotic characteristics, although some limitations were observed in terms of adhesion ability and the presence of antibiotic resistance genes, requiring further investigation into their genetic location. Future studies will focus on developing a probiotic prototype encapsulation for application in the chicken industry, followed by in vivo evaluations of probiotic efficacy.

The occurrence and genomic characteristics of the blaIMI-1 carbapenemase-producing Enterobacter cloacae complex retrieved from natural water sources in central Thailand.

AIM: Carbapenem resistance among Enterobacteriaceae is a serious threat to humans worldwide. This study aims to evaluate the phenotypic and genotypic characterization of carbapenemase-producing Enterobacter cloacae complex (ECC) retrieved from water sources in the central part of Thailand. METHODS AND RESULTS: Samples were collected from water bodies surrounding farms and communities in central Thailand. The species were identified by using MALDI-TOF MS. The minimum inhibitory concentration (MIC) and antibiotic susceptibility were determined. The carbapenemase-producing genes were detected by PCR and whole genome sequencing (WGS). ECC with chromosome-encoded blaIMI-1 carbapenemase were detected. These isolates were resistant to last-resort antibiotics such as carbapenems and colistin as well as penicillin. In addition, all blaIMI-1 genes isolated from this study were found to be associated with chromosomally integrated Xer-dependent integrative mobile elements (IMEXs). CONCLUSION: These findings highlight the diversity and dissemination of carbapenemases-producing Enterobacterales in environmental sources. With the increasing detection of carbapenemase genes worldwide, we should be aware of the blaIMI-producing E. cloacae complex with a high resistance profile and the ability to mobilize within the environment.

Practice, confidence and continuity of breast self-examination among women in Thailand during COVID-19 pandemic: a cross-sectional study.

OBJECTIVE: Breast self-examination (BSE) is the most feasible screening tool compared with clinical breast examination and mammography. It is crucial to address the associated factors of practising BSE to develop a targeted BSE promotion programme and improve the BSE quality in Thai women, particularly during the COVID-19 pandemic. DESIGN AND SETTING: We conducted a cross-sectional study in Thailand's north and northeast region from March 2020 to November 2022. PARTICIPANTS: This study involved 405 women aged 30-70 years old. VARIABLES AND OUTCOMES: Demographic information, health status and BSE were collected using a modified questionnaire based on the Champion Health Belief Model. The outcomes were ever-practising BSE, BSE practice within the last 6 months, continuity of BSE and confidence in doing BSE. Logistic regression and decision tree analysis identified the associated factors. RESULTS: 75.55% of participants ever performed BSE. Around 74.18% did BSE within the last 6 months. Diploma graduates (adjusted OR (aOR) 25.48, 95% CI 2.04 to 318.07), 21-40 reproductive years (aOR 4.29, 95% CI 1.22 to 15.08), ever pregnant (aOR 3.31, 95% CI 1.05 to 10.49), not drinking alcohol (aOR 2.1, 95% CI 1.04 to 4.55), not receiving hormone replacement (aOR 5.51, 95% CI 2.04 to 14.89), higher knowledge (aOR 1.29, 95% CI 1.09 to 1.52), attitude (aOR 1.15, 95% CI 1.05 to 1.26) and practice/cues of action towards BSE were associated with ever-practising BSE. Frequent high-fat diet, high awareness of breast cancer, lower knowledge of BSE and lower attitude toward BSE were associated with not practising BSE within 6 months and BSE discontinuation. Only high knowledge of BSE was associated with absolute confidence in BSE (p<0.05). CONCLUSION: Despite having a higher percentage than other studies in different countries prior to the pandemic, it is still crucial to improve knowledge of BSE to encourage BSE practice, confidence and continuity of BSE in Thai women. Moreover, the BSE campaign should target women with prolonged exposure to oestrogen and sedentary lifestyle.

The Cytokine, Chemokine, and Growth Factor Network of Prenatal Depression.

BACKGROUND: Neuro-immune pathways are engaged in antenatal and postpartum depression. AIMS: To determine if immune profiles influence the severity of prenatal depression above and beyond the effects of adverse childhood experiences (ACE), premenstrual syndrome (PMS), and current psychological stressors. METHODS: Using the Bio-Plex Pro human cytokine 27-plex test kit, we assayed M1 macrophage, T helper (Th)-1, Th-2, Th-17, growth factor, chemokine, and T cell growth immune profiles as well as indicators of the immune inflammatory response system (IRS) and compensatory immunoregulatory system (CIRS) in 120 pregnant females in the early (<16 weeks) and late (>24 weeks) pregnancy. The Edinburgh Postnatal Depression Scale (EPDS) was used to assess severity of antenatal depression. RESULTS: Cluster analyses showed that the combined effects of ACE, relationship dissatisfaction, unwanted pregnancy, PMS, and upregulated M1, Th-1, Th-2, and IRS immune profiles and the ensuing early depressive symptoms shape a stress-immune-depression phenotypic class. Elevated IL-4, IL-6, IL-8, IL-12p70, IL-15, IL-17, and GM-CSF are the cytokines associated with this phenotypic class. All immune profiles (except CIRS) were significantly associated with the early EPDS score, independent of the effects of psychological variables and PMS. There was a shift in immune profiles from early to late pregnancy, with an increase in the IRS/CIRS ratio. The late EPDS score was predicted by the early EPDS score, adverse experiences, and immune profiles, mainly the Th-2 and Th-17 phenotypes. CONCLUSIONS: Activated immune phenotypes contribute to early and late perinatal depressive symptoms above and beyond the effects of psychological stressors and PMS.

Age, comorbidities, c-reactive protein and procalcitonin as predictors of severity in confirmed COVID-19 patients in the Philippines.

Background: The Coronavirus Disease 2019 (COVID-19) pandemic has been affecting people globally, and the Philippines is one of the countries greatly struck by the virus. The continued rise of new positive cases has drawn attention to the urgent need for healthcare management to cope with this challenge. Severity prediction could help improve medical decision-making and optimise the patient's treatment plan with a good clinical outcome. This study aimed to identify the determinants of COVID-19 disease severity. Methods: Demographic characteristics and laboratory findings were collected from electronic medical records and paper forms of all confirmed COVID-19 cases reported by the University of Perpetual Help DALTA Medical Center between the September 1, 2020 and the October 31, 2021. We performed statistical analyses and interpretation of data to compare severe and non-severe groups. Results: 5,396 confirmed cases were examined. Most of the severe cases were elderly, male, had blood type A, and with comorbidities. Cycle threshold (Ct) values were lower in the severe group. Most patients had higher-than-normal levels of all blood parameters except platelet, white blood cell (WBC), neutrophil, and lymphocyte counts. Age, sex, ABO blood groups, comorbidities, open reading frame 1 ab (ORF1ab) and nucleocapsid (N) gene Ct values, ferritin, C-reactive protein (CRP), procalcitonin (PCT), D-dimer, white blood cell (WBC) count, neutrophil count, and lymphocyte count were significantly associated with disease severity. In multivariate analysis, age groups >60 and 30-59 years, presence of comorbidities, CRP level >5 ng/mL, and PCT >0.05 ng/mL were identified as disease severity predictors. Conclusions: Based on our results, age, comorbidities, CRP, and PCT level may be utilised as primary assessment factors for possible hospital admission and close monitoring upon testing. Early detection of these risk factors may provide strategic interventions that help reduce mortality, hospital admissions, and more expensive and extensive treatments.

Antenatal depressive symptoms are strongly predicted by the severity of pre-menstrual syndrome: results of partial least squares analysis

Background/Objective: Antenatal depression (AD) is the commonest morbidity during pregnancy. There is evidence that premenstrual syndrome (PMS) and AD share common immune-inflammatory and sex hormonal pathways. This study aims to evaluate the association between the severity of depressive PMS and AD in early and late pregnancy. Method: Participants were followed from early (<=16 weeks) to late pregnancy (>=20 weeks). The Premenstrual Symptoms Screening Tool (PSST) was used to assess PMS and AD symptoms were assessed using the Edinburgh Postnatal Depression Scale (EPDS). Results: Up to 57.6% of the variance in the early EPDS score was explained by the regression on the first factor extracted from 10 depression and anxiety PSST items (dubbed the DepAnx PSST), insomnia PSST, relation dissatisfaction, and partner abuse. There were specific indirect effects of DepAnx PSST (p < 0.001), insomnia PSST (p = 0.041), relation dissatisfaction (p = 0.023) and partner abuse (p = 0.007) on the late EPDS which were mediated by the early EPDS score. Conclusion: The affective, but not psychosomatic, symptoms of PMS strongly predict AD symptoms suggesting that the pathophysiology of affective PMS symptoms overlap with those of AD. (AU)

Antenatal depressive symptoms are strongly predicted by the severity of pre-menstrual syndrome: results of partial least squares analysis.

BACKGROUND/OBJECTIVE: Antenatal depression (AD) is the commonest morbidity during pregnancy. There is evidence that premenstrual syndrome (PMS) and AD share common immune-inflammatory and sex hormonal pathways. This study aims to evaluate the association between the severity of depressive PMS and AD in early and late pregnancy. METHOD: Participants were followed from early (<=16 weeks) to late pregnancy (>=20 weeks). The Premenstrual Symptoms Screening Tool (PSST) was used to assess PMS and AD symptoms were assessed using the Edinburgh Postnatal Depression Scale (EPDS). RESULTS: Up to 57.6% of the variance in the early EPDS score was explained by the regression on the first factor extracted from 10 depression and anxiety PSST items (dubbed the DepAnx PSST), insomnia PSST, relation dissatisfaction, and partner abuse. There were specific indirect effects of DepAnx PSST (p < 0.001), insomnia PSST (p = 0.041), relation dissatisfaction (p = 0.023) and partner abuse (p = 0.007) on the late EPDS which were mediated by the early EPDS score. CONCLUSION: The affective, but not psychosomatic, symptoms of PMS strongly predict AD symptoms suggesting that the pathophysiology of affective PMS symptoms overlap with those of AD.

Metagenomic analysis of the gut microbiota in piglets either challenged or not with enterotoxigenic Escherichia coli reveals beneficial effects of probiotics on microbiome composition, resistome, digestive function and oxidative stress responses.

This study used metagenomic analysis to investigate the gut microbiota and resistome in piglets that were or were not challenged with enterotoxigenic Escherichia coli (ETEC) and had or had not received dietary supplementation with microencapsulated probiotics. The 72 piglets belonged to six groups that were either non-ETEC challenged (groups 1-3) or ETEC challenged (receiving 5ml of 109 CFU/ml pathogenic ETEC strain L3.2 one week following weaning at three weeks of age: groups 4-6). On five occasions at 2, 5, 8, 11, and 14 days of piglet age, groups 2 and 5 were supplemented with 109 CFU/ml of multi-strain probiotics (Lactiplantibacillus plantarum strains 22F and 25F, and Pediococcus acidilactici 72N) while group 4 received 109 CFU/ml of P. acidilactici 72N. Group 3 received 300mg/kg chlortetracycline in the weaner diet to mimic commercial conditions. Rectal faecal samples were obtained for metagenomic and resistome analysis at 2 days of age, and at 12 hours and 14 days after the timing of post-weaning challenge with ETEC. The piglets were all euthanized at 42 days of age. The piglets in groups 2 and 5 were enriched with several desirable microbial families, including Lactobacillaceae, Lachnospiraceae and Ruminococcaceae, while piglets in group 3 had increases in members of the Bacteroidaceae family and exhibited an increase in tetW and tetQ genes. Group 5 had less copper and multi-biocide resistance. Mobile genetic elements IncQ1 and IncX4 were the most prevalent replicons in antibiotic-fed piglets. Only groups 6 and 3 had the integrase gene (intl) class 2 and 3 detected, respectively. The insertion sequence (IS) 1380 was prevalent in group 3. IS3 and IS30, which are connected to dietary intake, were overrepresented in group 5. Furthermore, only group 5 showed genes associated with detoxification, with enrichment of genes associated with oxidative stress, glucose metabolism, and amino acid metabolism compared to the other groups. Overall, metagenomic analysis showed that employing a multi-strain probiotic could transform the gut microbiota, reduce the resistome, and boost genes associated with food metabolism.

Microencapsulated probiotic Lactiplantibacillus plantarum and/or Pediococcus acidilactici strains ameliorate diarrhoea in piglets challenged with enterotoxigenic Escherichia coli.

Lactiplantibacillus plantarum (strains 22F and 25F) and Pediococcus acidilactici (strain 72N) have displayed antibacterial activity in vitro, suggesting that they could be used to support intestinal health in pigs. The aim of this study was to determine if microencapsulated probiotics could reduce the severity of infection with enterotoxigenic Escherichia coli (ETEC) in weaned pigs. Sixty healthy neonatal piglets were cross-fostered and separated into five groups. Piglets to be given the microencapsulated probiotics received these orally on days 0, 3, 6, 9, and 12. Only piglets in groups 1 and 5 did not receive probiotics: those in groups 2 and 4 received the three microencapsulated probiotic strains (multi-strain probiotic), and piglets in group 3 received microencapsulated P. acidilactici strain 72N. After weaning, the pigs in groups 3-5 were challenged with 5 mL (at 109 CFU/mL) of pathogenic ETEC strain L3.2 carrying the k88, staP, and stb virulence genes. The multi-strain probiotic enhanced the average daily gain (ADG) and feed conversion ratio (FCR) of weaned piglets after the ETEC challenge (group 4), whilst supplementing with the single-strain probiotic increased FCR (group 3). Piglets in groups 3 and 4 developed mild to moderate diarrhoea and fever. In the probiotic-fed piglets there was an increase in lactic acid bacteria count and a decrease in E. coli count in the faeces. By using real-time PCR, virulence genes were detected at lower levels in the faeces of pigs that had received the probiotic strains. Using the MILLIPLEX MAP assay, probiotic supplementation was shown to reduce pro-inflammatory cytokines (IL-1α, IL-6, IL-8, and TNFα), while group 4 had high levels of anti-inflammatory cytokine (IL-10). Challenged piglets receiving probiotics had milder intestinal lesions with better morphology, including greater villous heights and villous height per crypt depth ratios, than pigs just receiving ETEC. In conclusion, prophylactic administration of microencapsulated probiotic strains may improve outcomes in weaned pigs with colibacillosis.

Adverse childhood experiences combined with emotional and physical abuse by the partner predict antenatal depression.

BACKGROUND: Few studies examined the contributions of childhood adversities, intimate partner violence and social support to antenatal depression (AD). This study aims to 1) evaluate association of these psychosocial factors with AD symptoms in pregnancy; and 2) examine the mediating effect of social support on the relationship between psychosocial stressors and AD symptoms. METHODS: Participants were 120 pregnant women aged from 18 to 49 in less than 16 gestational weeks and attending at Antenatal Care Center at Khon Kaen hospital, Thailand. AD symptoms were assessed by the Edinburgh Postnatal Depression Scale (EPDS). Childhood adversities, intimate partner violence and social support were measured using the Adverse Childhood Experiences Questionnaire (ACE questionnaire), Abuse Assessment Screen (AAS), and Multidimensional Scale of Perceived Social Support (MSPSS). RESULTS: We found that the EPDS score was significantly and positively associated with adverse childhood experiences (ACEs) and negatively with social support. Partial Least Square analysis showed that 49.1% of the variance in the depressive subdomain of the EPDS score was predicted by ACEs, namely psychological and physical abuse and neglect, emotional or physical abuse by the partner, unplanned pregnancy, and no satisfaction with their relationship. The effects of adverse childhood experience due to neglect on the EDPS score was mediated by social support by friends. LIMITATIONS: ACEs were assessed retrospectively and, therefore, may be susceptible to recall bias. CONCLUSION: Prenatal depression scores are to a large extent predicted by psychological distress as indicated by early lifetime trauma, abuse by partner, relation satisfaction, and implications of unintended pregnancy.

Reducing the Risk of Transmission of Critical Antimicrobial Resistance Determinants From Contaminated Pork Products to Humans in South-East Asia.

Antimicrobial resistance (AMR) is a critical challenge worldwide as it impacts public health, especially via contamination in the food chain and in healthcare-associated infections. In relation to farming, the systems used, waste management on farms, and the production line process are all determinants reflecting the risk of AMR emergence and rate of contamination of foodstuffs. This review focuses on South East Asia (SEA), which contains diverse regions covering 11 countries, each having different levels of development, customs, laws, and regulations. Routinely, here as elsewhere antimicrobials are still used for three indications: therapy, prevention, and growth promotion, and these are the fundamental drivers of AMR development and persistence. The accuracy of detection of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARG) depends on the laboratory standards applicable in the various institutes and countries, and this affects the consistency of regional data. Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the standard proxy species used for indicating AMR-associated nosocomial infections and healthcare-associated infections. Pig feces and wastewater have been suspected as one of the hotspots for spread and circulation of ARB and ARG. As part of AMR surveillance in a One Health approach, clonal typing is used to identify bacterial clonal transmission from the production process to consumers and patients - although to date there have been few published definitive studies about this in SEA. Various alternatives to antibiotics are available to reduce antibiotic use on farms. Certain of these alternatives together with improved disease prevention methods are essential tools to reduce antimicrobial usage in swine farms and to support global policy. This review highlights evidence for potential transfer of resistant bacteria from food animals to humans, and awareness and understanding of AMR through a description of the occurrence of AMR in pig farm food chains under SEA management systems. The latter includes a description of standard pig farming practices, detection of AMR and clonal analysis of bacteria, and AMR in the food chain and associated environments. Finally, the possibility of using alternatives to antibiotics and improving policies for future strategies in combating AMR in a SEA context are outlined.

The efficacy of three double-microencapsulation methods for preservation of probiotic bacteria.

Lactic acid bacteria (LAB) are used as a probiotic alternative to antibiotics in livestock production. Microencapsulation technology is widely used for probiotic preservation. A variety of microencapsulation protocols have been proposed and compared based on chemicals and mechanical procedures. This study aimed to develop a double-encapsulated coating from alginate (1.5%) and chitosan (0.5%) by extrusion, emulsion, and spray drying methods using the LAB strains Lactobacillus plantarum strains 31F, 25F, 22F, Pediococcus pentosaceus 77F, and P. acidilactici 72N, and to monitor the basic probiotic properties of the encapsulated prototypes. The final products from each microencapsulation protocol were analysed for their appearance, probiotic properties and viable cell count. Using the spray drying method, particles smaller than 15 µm in diameter with a regular spherical shape were obtained, whereas the other methods produced larger (1.4-52 mm) and irregularly shaped microcapsules. After storage for 6 months at room temperature, the LAB viability of the spray-dried particles was the highest among the three methods. In all the LAB strains examined, the encapsulated LAB retained their probiotic properties in relation to acid-bile tolerance and antibacterial activity. This study highlights the efficacy of double-coating microencapsulation for preserving LAB properties and survival rate, and demonstrates its potential for probiotic application in livestock farms.

Use of Lactobacillus plantarum (strains 22F and 25F) and Pediococcus acidilactici (strain 72N) as replacements for antibiotic-growth promotants in pigs.

The lactic acid bacteria (LAB) Lactobacillus plantarum (strains 22F and 25F) and Pediococcus acidilactici (strain 72N) have appeared promising as replacements for antibiotics in in vitro studies. Microencapsulation, especially by the spray-drying method, has been used to preserve their numbers and characteristics during storage and digestion. This study compared the efficacy of these strains and their microencapsulated form with antibiotic usage on growth performance, faecal microbial counts, and intestinal morphology in nursing-finishing pigs. A total of 240 healthy neonatal pigs were treated on days 0, 3, 6, 9, and 12 after cross-fostering. Sterile peptone water was delivered orally to the control and antibiotic groups. Spray-dried Lactobacillus plantarum strain 22F stored for 6-months was administered to piglets in the spraydry group. Three ml of each the three fresh strains (109 CFU/mL) were orally administered to piglets in each group. All pigs received the basal diets, but these were supplemented with routine antibiotic for the antibiotic group. Pigs in all the probiotic supplemented groups exhibited a better average daily gain and feed conversion ratio than those of the controls in the nursery and grower phases. Probiotic supplementation increased viable lactobacilli and decreased enterobacterial counts. Antibiotic additives reduced both enterobacterial and lactobacilli counts. Villous height and villous height:crypt depth ratio were greater in probiotic and antibiotic supplemented pigs comparing to the controls, especially in the jejunum. The results demonstrated the feasibility of using these strains as a substitute for antibiotics and the practicality of the microencapsulation protocol for use in swine farms.

Anticonjugation and Antibiofilm Evaluation of Probiotic Strains Lactobacillus plantarum 22F, 25F, and Pediococcus acidilactici 72N Against Escherichia coli Harboring mcr-1 Gene.

Several species of lactic acid bacteria (LAB) are commonly used as probiotics and as an alternative to antibiotics in various industries, especially in the livestock industry. This study aimed to investigate the anticonjugation and antibiofilm activity of cell-free supernatant (CFS) of Thai LAB strains (Lactobacillus plantarum 22F, 25F, and Pediococcus acidilactici 72N) against colistin-resistant Escherichia coli isolates. A total of six colistin-resistant E. coli strains were isolated from different sources, including pigs, farmers, and farmhouse environments. The E. coli were characterized by plasmid profiling, PCR detection of mcr-1 gene, and antibiotic susceptibility patterns. The CFS at dilutions ≥1:16 was chosen as the proper dilution for anticonjugation assay. Besides, it could significantly reduce the transfer frequencies of resistance gene mcr-1 up to 100 times compared to the neutralizing CFS (pH 6.5). The biofilm production in the planktonic stage was reduced by non-neutralizing and neutralizing CFS determining with crystal violet staining assay up to 82 and 60%, respectively. Moreover, the non-neutralizing CFS also inhibited the biofilm formation in the sessile stage up to 52%. The biofilm illustration was confirmed by scanning electron microscopy (SEM). These results agreed with the findings of the crystal violet technique, which showed a significant reduction in cell density, aggregation, and extracellular polysaccharide (EPS) matrix. The application of Thai LAB may serve as an attractive alternative to antibiotics for reducing biofilm formation and limiting the proliferation of antibiotic-resistant genes.

Development and performance of CUHAS-ROBUST application for pulmonary rifampicin-resistance tuberculosis screening in Indonesia.

BACKGROUND AND OBJECTIVES: Diagnosis of Pulmonary Rifampicin Resistant Tuberculosis (RR-TB) with the Drug-Susceptibility Test (DST) is costly and time-consuming. Furthermore, GeneXpert for rapid diagnosis is not widely available in Indonesia. This study aims to develop and evaluate the CUHAS-ROBUST model performance, an artificial-intelligence-based RR-TB screening tool. METHODS: A cross-sectional study involved suspected all type of RR-TB patients with complete sputum Lowenstein Jensen DST (reference) and 19 clinical, laboratory, and radiology parameter results, retrieved from medical records in hospitals under the Faculty of Medicine, Hasanuddin University Indonesia, from January 2015-December 2019. The Artificial Neural Network (ANN) models were built along with other classifiers. The model was tested on participants recruited from January 2020-October 2020 and deployed into CUHAS-ROBUST (index test) application. Sensitivity, specificity, and accuracy were obtained for assessment. RESULTS: A total of 487 participants (32 Multidrug-Resistant/MDR 57 RR-TB, 398 drug-sensitive) were recruited for model building and 157 participants (23 MDR and 21 RR) in prospective testing. The ANN full model yields the highest values of accuracy (88% (95% CI 85-91)), and sensitivity (84% (95% CI 76-89)) compare to other models that show sensitivity below 80% (Logistic Regression 32%, Decision Tree 44%, Random Forest 25%, Extreme Gradient Boost 25%). However, this ANN has lower specificity among other models (90% (95% CI 86-93)) where Logistic Regression demonstrates the highest (99% (95% CI 97-99)). This ANN model was selected for the CUHAS-ROBUST application, although still lower than the sensitivity of global GeneXpert results (87.5%). CONCLUSION: The ANN-CUHAS ROBUST outperforms other AI classifiers model in detecting all type of RR-TB, and by deploying into the application, the health staff can utilize the tool for screening purposes particularly at the primary care level where the GeneXpert examination is not available. TRIAL REGISTRATION: NCT04208789.

Autochthonous lactic acid bacteria isolated from pig faeces in Thailand show probiotic properties and antibacterial activity against enteric pathogenic bacteria.

Lactic acid bacteria (LAB) play an important role in pig health and performance that arises from their beneficial impacts on the balance of gastrointestinal microbes, ability to fight enteric pathogens, and capacity to support the immune system. The aim of this study was to evaluate the functional and safety aspects of five previously isolated autochthonous LAB strains, (Lactobacillus plantarum 22F, 25F and 31F, Pediococcus acidilactici 72N and Pediococcus pentosaceus 77F) from pig faeces as potential probiotics for a pig feed supplement. The functional and safety properties of the strains were assessed by in vitro tests. The functional properties tested were their abilities in tolerating low pH values under simulated gastric conditions, their cell surface properties (hydrophobicity, auto- and co-aggregation), antibacterial activity against the common enteric pathogenic bacteria in pigs (such as pathogenic Escherichia coli, Salmonella Choleraesuis and Streptococcus suis), and diacetyl production. The safety of the strains was analyzed based on the absent of haemolysis on blood and bile salt hydrolase activity. Although all strains demonstrated diacetyl production, good survivability and antibacterial activities, L. plantarum 22F and 25F showed the best performance with the strongest antibacterial actions against the indicator pathogens. Of the strains, only P. pentosaceus 77F exhibited haemolysis or bile salt hydrolase activity. Furthermore, a principal component analysis revealed that L. plantarum 22F possessed superior functional and safety aspects compared to the other four autochthonous strains and to reference strains L. plantarum JCM 1149 and P. acidilactici DSM 20284. Further in vivo studies using oral administration of the strains are justified to assess their effectiveness as feed supplements for pigs.

Protective Effects of Cell-Free Supernatant and Live Lactic Acid Bacteria Isolated from Thai Pigs Against a Pandemic Strain of Porcine Epidemic Diarrhea Virus.

Porcine epidemic diarrhea virus (PEDV) is a coronavirus which causes severe diarrhea and fatal dehydration in piglets. In general, probiotic supplements could enhance recovery and protect piglets against enteric pathogens. Seven local lactic acid bacteria (LAB), (Ent. faecium 79N and 40N, Lact. plantarum 22F, 25F and 31F, Ped. acidilactici 72N and Ped. pentosaceus 77F) from pig feces were well-characterized as high potential probiotics. Cell-free supernatants (CFS) and live LAB were evaluated for antiviral activities by co-incubation on Vero cells and challenged with a pandemic strain of PEDV isolated from pigs in Thailand. Cell survival and viral inhibition were determined by cytopathic effect (CPE) reduction assay and confirmed by immunofluorescence. At 1:16, CFS dilution (pH 6.3-6.8) showed no cytotoxicity in Vero cells and was therefore used as the dilution for antiviral assays. The diluted CFS of all Lact. plantarum showed the antiviral effect against PEDV; however, the same antiviral effect could not be observed in Ent. faecium and Pediococcus strains. In competitive experiment, only live Lact. plantarum 25F and Ped. pentosaceus 77F showed CPE reduction in the viral infected cells to <50% observed field area. This study concluded that the CFS of all tested lactobacilli, and live Lact. plantarum (22F and 25F) and Pediococcus strains 72N and 77F could reduce infectivity of the pandemic strain of PEDV from pigs in Thailand on the target Vero cells.