Tryptone Soya Broth Research Articles

Dietary carbohydrates modulate Streptococcus mutans adherence and bacterial proteome.

Streptococcus mutans adherence to the tooth surface and subsequent biofilm development is modulated by the carbohydrate source, but the corresponding effect on bacterial proteome has not been previously studied. This study aimed to assess the effect of different carbohydrates on S. mutans viability and bacterial proteome at two-time points, early attachment (8 h) and biofilm maturation (24 h). Hydroxyapatite (HAp) discs coated with parotid saliva proteins were inoculated with S. mutans UA159 in Tryptone Soy Broth without dextrose supplemented with one of the following carbohydrates (n=12/treatment/time point): 1% Sucrose (S); 0.525% Glucose + 0.525% Fructose (G+F); 10% Xylitol (X); 10% Xylitol + 1% Sucrose (X+S); or culture medium without supplementation as negative control (C). Once inoculated, HAp discs were incubated for 8 h or 24 h at 37 °C and 10% CO2. After each incubation period, adhered bacteria were quantified using the plate-counting method for 6 HAp discs/group, and the remaining 6 HAp discs/group were used to extract bacterial cell wall proteins. Extracted proteins were analyzed using liquid chromatography coupled with mass spectrometry and then classified by their biological process. The study was conducted in three independent assays and the number of bacteria adhered to the HAp discs was determined at each time point and analyzed by two-way ANOVA followed by Bonferroni test (=5%). The results suggest that xylitol significantly repressed bacterial adherence and metabolism at 8 h and 24 h; however, bacterial adherence and metabolism were significantly enhanced when xylitol was combined with sucrose, showing no negative effect on S. mutans at both time points. Bacterial proteome was modulated by the carbohydrate source. The cariogenicity of S. mutans biofilms may be reduced by the alternative sweetener xylitol; however, the combination with fermentable sugars may inhibit such a beneficial effect.

Study of Plant Growth Promoting Abilities of Rhizobium Strains Isolated from Mungbean Root Nodules

Background: Plant Growth Promoting Rhizobacteria (PGPR) are a type of bacteria that use biofertilizers and other plant growth-promoting agents to increase plant output and growth. In the last several decades, there has been a global surge in the usage of beneficial soil microorganisms like PGPR for safe and sustainable agriculture due to the detrimental effects of artificial fertilizers on the environment and their rising prices. Plant-beneficial rhizobacteria have the potential to reduce the world's reliance on dangerous agricultural chemicals that upset agro-ecosystems. Today a large part of yield is lost due to the various stress mechanisms of plants. It could be classified into biotic and abiotic stress. These are generally a group of microorganisms that is found either in the rhizosphere or in the root nodules of plants. PGPR has beneficial impact, they colonize in the plant roots and improve crop yields and agricultural productivity and also produce some useful growth promoting growth hormones viz., IAA, Auxins, Cytokinin and Gibberellins. Methods: The bacterial isolates were screened for plant growth promoting traits such as phosphate solubilization, IAA production and siderophore production was tested qualitatively by using pikovskayas media, TSB (Tryptone Soy Broth) agar and Chrome Azurol Sulphate (CAS) blue agar respectively. Results: In present study out of the eight rhizobial isolates, five exhibited the phosphate solubilization zones, three shows negative result. However out of eight isolates five isolates produced IAA and three showed negative result. Among all isolates seven isolates were able to produce siderophore, while one showed negative result.

Emergence of Aeromonas salmonicida subsp. masoucida MHJM250: unveiling pathological characteristics and antimicrobial susceptibility in golden mahseer, Tor putitora (Hamilton, 1822) in India.

Aeromonas salmonicida subsp. masoucida, designated as laboratory strain MHJM250, was characterized from a naturally infected farmed golden mahseer, Tor putitora. The infected fish exhibited clinical signs of erosion at the caudal fin and hemorrhage onx the ventral body surface. Molecular identification through 16S rDNA and phylogenetic analysis revealed 100% similarity with a known strain A. salmonicida subsp. masoucida (MT122821.1). MHJM250 exhibited positive reactions for oxidase, catalase, esculin, MR-VP, O/F and utilized arginine and lysine. It also demonstrated siderophore activity, thrived at various NaCl concentrations, hydrolyzed gelatinase, skimmed milk and casinase. In vitro studies exhibited its hemolytic nature, significant biofilm production in glucose-rich tryptone soya broth and beta-hemolysis. MHJM250 didn't produce slime and was non-precipitated upon boiling. It showed crystal violet binding characteristics and auto-agglutination with relatively weak hydrophobicity (25%). In the challenge assay, intraperitoneal administration of MHJM250 to T. pitutora fingerlings at 108 CFU mL-1 resulted in pathogenicity with 3% mortality and mild hemorrhagic symptoms. Histopathological analysis revealed degenerative changes in gill, kidney, liver, muscle, and intestine samples. The bacterium displayed resistance to several antibiotics (µg/disc); ampicillin (10µg), ampicillin/ sulbactam (10/10 µg), clindamycin (2µg), linezolid (30µg), penicillin G (10µg) and rifampicin (5µg) and varied minimum inhibitory concentrations against oxytetracycline, erythromycin and florfenicol. Transmission electron microscopy showed its rod-shaped structure with single polar flagellum and lophotrichous flagella. An investigation on the molecular basis for virulence factors of A. salmonicida subsp. masoucida MHJM250 may offer crucial understandings to formulate disease prevention and control strategies in aquaculture.

Bacillus subtilis as an effective tool for bioremediation of lead, copper and cadmium in water

Bioremediation is a promising technique for the removal and recovery of contaminated areas, which is based on the ability of organisms to convert toxic substances into less harmful or inert compounds. Here, we evaluate the capacity of the Bacillus subtilis (BS) bacteria to bioadsorved heavy metals such as lead (Pb), copper (Cu) and cadmium (Cd) in water samples. Water samples were laboratory contaminated Pb at 500 ppm, Cu and Cd at 100 ppm. First, the growth curve of BS was plotted using Tryptone Soy Broth (TSB) at 100% (TSB100) and 33% (TSB33). Later, BS was studied in water containing all three metals separately and simultaneously. All solutions were stirred at 150 rpm, 35 ℃ for periods that ranged from 1 to 144 h. The heavy metal analyses were performed by X-ray fluorescence and Inductively Coupled Plasma Mass Spectrometry. The results showed that BS was resistant to Cu, Cd and Pb in water, with active multiplication and reduction in the concentration of the metals. The highest removal efficiency, in the presence of each heavy metal, reached 100% with Pb, 92.3% with Cd and 89% with Cu. In the solution with the mixture of heavy metals, Cd and Pb levels reduced significantly. However, the mixture of metals negatively affected the removal of Cu. In conclusion, the implement using Bacillus-type bacteria can be efficient in the biosorption of heavy metals from aqueous solutions and can be employed as a cost-effective treatment for the bioremediation of industrial effluents and contaminated areas.

Magnitude and antimicrobial susceptibility profile of bacteria isolated from pediatric sepsis cases at University of Gondar Hospital, Northwest Ethiopia

BackgroundSepsis is one of the major causes of morbidity and mortality among pediatric patients throughout the world. The varying microbiological pattern of sepsis warrants the need for researches on the causative organisms and their antimicrobial susceptibility pattern. The epidemiology of neonatal and pediatric sepsis in Ethiopia is under-research. The objective of this study was to evaluate the burden of bacterial pathogens and their antimicrobial susceptibility patterns among children suspected of sepsis.MethodsAn institutional-based prospective cross-sectional study was conducted on 370 pediatric(age birth-15 years) patients suspected of sepsis at the University of Gondar Comprehensive Specialized hospital from December 2020 to November 2021. Blood samples were collected aseptically and inoculated into Tryptone Soya Broth for culture. The organisms grown were identified by standard microbiological methods and subjected to antibiotic susceptibility testing by modified Kirby-Bauer disk diffusion method recommended by Clinical laboratory and standard institute. Methicillin resistance was confirmed using Cefoxitin disk diffusion method. Data entry and analysis were done using Statistical Package for Social Sciences (SPSS) version 26 software. A p-value less than 0.05 at 95% confidence interval was considered statically significant.ResultsOut of the total 370 study subjects, 21.6% (80/370) of them were culture positive. Of these, 43 (53.8%) and 37 (46.3%) were Gram-positive and Gram-negative bacterial pathogens, respectively. The most prevalent Gram-positive bacterial isolate was Staphylococcus aureus (n = 24; 30%) and coagulase negative staphylococci (n = 7; 8.8%). Among the Gram-negative bacterial isolates, the leading bacteria was Klebsiella pneumoniae (n = 20; 25%) followed by Escherichia coli (n = 7; 8.8%). Clindamycin, Chloramphenicol, Gentamicin and Ciprofloxacin were the most effective antibiotics against Gram-positive bacterial isolates while Amikacin, Meropenem and Chloramphenicol were effective against Gram-negative pathogens. Methicillin resistance was detected in 45.8% of Staphylococcus aureus. Multi-drug resistance (MDR) antimicrobial susceptibility pattern was observed in 76% of the bacterial isolates.ConclusionGram positive bacteria were the predominant isolates among pediatric sepsis cases and most of the bacterial isolates showed MDR. Staphylococcus aureus and Klebsiella pneumoniae were frequently isolated bacteria. The high prevalence of drug resistance warrants rational use of antibiotics and the need for regular antibiotic susceptibility surveillance studies.

Rapid and Sensitive Detection of Shiga Toxin-Producing Escherichia coli (STEC) from Food Matrices Using the CANARY Biosensor Assay.

Shiga toxin-producing Escherichia coli (STEC) causes a wide spectrum of diseases including hemorrhagic colitis and hemolytic uremic syndrome (HUS). Previously, we developed a rapid, sensitive, and potentially portable assay that identified STEC by detecting Shiga toxin (Stx) using a B-cell based biosensor platform. We applied this assay to detect Stx2 present in food samples that have been implicated in previous STEC foodborne outbreaks (milk, lettuce, and beef). The STEC enrichment medium, modified Tryptone Soy Broth (mTSB), inhibited the biosensor assay, but dilution with the assay buffer relieved this effect. Results with Stx2a toxoid-spiked food samples indicated an estimated limit of detection (LOD) of ≈4 ng/mL. When this assay was applied to food samples inoculated with STEC, it was able to detect 0.4 CFU/g or 0.4 CFU/mL of STEC at 16 h post incubation (hpi) in an enrichment medium containing mitomycin C. Importantly, this assay was even able to detect STEC strains that were high expressors of Stx2 at 8 hpi. These results indicate that the STEC CANARY biosensor assay is a rapid and sensitive assay applicable for detection of STEC contamination in food with minimal sample processing that can complement the current Food Safety Inspection Service (US) methodologies for STEC.

Impact of Growth Conditions on High-Throughput Identification of Repurposing Drugs for Pseudomonas aeruginosa Cystic Fibrosis Lung Infections.

Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) patients represent a therapeutic challenge due to antibiotic resistance. Repurposing existing drugs is a promising approach for identifying new antimicrobials. A crucial factor in successful drug repurposing is using assay conditions that mirror the site of infection. Here, the impact of growth conditions on the anti-P. aeruginosa activity of a library of 3386 compounds was evaluated. To this, after 24 h exposure, the survival rate of CF P. aeruginosa RP73 planktonic cells was assessed spectrophotometrically under "CF-like" (artificial CF sputum, pH 6.8, 5% CO2) and enriched (Tryptone Soya Broth, pH 7.2, and aerobiosis) conditions. Among non-antibiotic compounds (n = 3127), 13.4% were active regardless of growth conditions, although only 3.2% had comparable activity; 4% and 6.2% were more active under CF-like or enriched conditions, respectively. Interestingly, 22.1% and 26.6% were active exclusively under CF-like and enriched conditions, respectively. Notably, 7 and 12 hits caused 100% killing under CF-like and enriched conditions, respectively. Among antibiotics (n = 234), 42.3% were active under both conditions, although only 18.4% showed comparable activity; 9.4% and 14.5% were more active under CF-like and enriched conditions, respectively. Interestingly, 23% and 16.6% were active exclusively under CF-like and enriched conditions, respectively. Sulphonamides showed higher activity under CF-like conditions, whereas tetracyclines, fluoroquinolones, and macrolides were more effective under enriched settings. Our findings indicated that growth conditions significantly affect the anti-P. aeruginosa activity of antibiotics and non-antibiotic drugs. Consequently, repurposing studies and susceptibility tests should be performed under physicochemical conditions that the pathogen tackles at the site of infection.

Stability Studies of the Dilution Series of Different Antibiotic Stock Solutions in Culture Medium Incubated at 37 °C.

The long-term stability of antibiotics in culture media remains underexplored in scientific literature. This study evaluated the stability of eight distinct antibiotic stock solutions-amoxicillin, cefotaxime, neomycin, oxytetracycline, florfenicol, enrofloxacin, colistin, and potentiated sulfonamide-and their 10-fold dilution series in tryptone soy broth (TSB) at 37 °C, over 12 days. Samples were collected immediately after preparation and on days 1, 2, 5, 7, 9, and 12, with active substance concentrations measured using ultra-high-performance liquid chromatography (UHPLC) coupled with mass spectrometry. The results indicated that among the ultrapure water stock solutions, neomycin, florfenicol, and potentiated sulfonamide maintained stability (>95%). Within the culture medium, florfenicol showed consistent stability (100%) throughout the study, potentiated sulfonamide experienced minor degradation (>85%), and neomycin underwent significant degradation. Amoxicillin, oxytetracycline, and colistin displayed considerable degradation in both solution types but were more stable in ultrapure water solutions. The stability of cefotaxime and enrofloxacin in ultrapure water solutions and in the medium was very similar when compared; however, 3.6% of the former and 88.7% of the latter remained detectable by day 12. These findings are crucial for minimum inhibitory concentration (MIC) assessments, especially in minimum bactericidal concentration (MBC) studies, and in experiments concerning long-term evolution and co-selection. This study underscores the necessity of stability assessments in culture media to validate future experimental outcomes.

Overcoming Microbial Inhibition of S. Sonnei Through the Exploitation of Genomically Predicted Antibiotic Resistance Profiles for the Development of Food Enrichment Media

Linking outbreaks of Shigella spp. to specific foods is challenging due to poor selectivity of current enrichment media. We have previously shown that enrichment media, tailored to the genomically-predicted antimicrobial resistance (AMR) of Shiga toxigenic E. coli strains, enhances their isolation from foods. This study investigates the application of this approach for Shigella isolation. The AMR gene profiles of 21,908 published S. sonnei genomes indicated a high prevalence of genes conferring resistance to streptomycin (aadA, aph(3″)-Ib, aph(6)-Id, 92.8%), sulfonamides (sul1, sul2, 74.8%), and/or trimethoprim (dfrA, 96.2%). Genomic analysis and antibiotic susceptibility testing conducted with a panel of 17 outbreak-associated S. sonnei strains confirmed the correlation of AMR gene detection with resistance phenotypes. Supplementation of Shigella Broth (SB) with up to 400 µg/mL of trimethoprim or sulfadiazine did not suppress the growth of sensitive strains, whereas 100 µg/mL of streptomycin increased the selectivity of this broth. All three antibiotics increased the selectivity of modified Tryptone Soya Broth (mTSB). Based on these results, supplemented media formulations were developed and assessed by measuring the relative growth of S. sonnei in cultures coinoculated with a strain of bacteriocin-producing E. coli that is inhibitory to Shigella growth. S. sonnei was not recovered from cocultures grown in SB or mTSB without antibiotics. In contrast, media supplemented with streptomycin at 50 and 100 µg/mL, trimethoprim at 25 and 50 µg/mL, and sulfadiazine at 100 µg/mL increased the relative proportion of S. sonnei in postenrichment cultures. The enhanced recovery of resistant S. sonnei strains achieved in this study indicates that, in cases where genomic data are available for clinical S. sonnei isolates, customization of selective enrichment media based on AMR gene detection could be a valuable tool for supporting the investigation of foodborne shigellosis outbreaks.

Bacterial micro-aggregates as inoculum in animal models of implant-associated infections

Is it time to rethink the inoculum of animal models of implant-associated infections (IAI)? Traditionally, animal models of IAI are based on inoculation with metabolically active overnight cultures of planktonic bacteria or pre-grown surface-attached biofilms. However, such inoculums do not mimic the clinical initiation of IAI. Therefore, the present study aimed to develop a clinically relevant inoculum of low metabolic micro-aggregated bacteria. The porcine Staphylococcus aureus strain S54F9 was cultured in Tryptone Soya Broth (TSB) for seven days to facilitate the formation of low metabolic micro-aggregates. Subsequently, the aggregated culture underwent filtration using cell strainers of different pore sizes to separate micro-aggregates. Light microscopy was used to evaluate the aggregate formation and size in the different fractions, while isothermal microcalorimetry was used to disclose a low metabolic activity. The micro-aggregate fraction obtained with filter size 5–15 μm (actual measured mean size 32 μm) was used as inoculum in a porcine implant-associated osteomyelitis (IAO) model and compared to a standard overnight planktonic inoculum and a sham inoculum of 0.9 % saline. The micro-aggregate and planktonic inoculums caused IAO with the re-isolation of S. aureus from soft tissues, bones, and implants. However, compared to their planktonic counterpart, neither of the micro-aggregate inoculated animals showed signs of osteomyelitis, i.e., sequester, osteolysis, and pus at gross inspection. Furthermore, inoculation with low metabolic micro-aggregates resulted in a strong healing response with pronounced osteoid formation, comparable to sham animals. In conclusion, the formation and separation of low metabolic bacterial micro-aggregates into various size fractions is possible, however, planktonic bacteria were still seen in all size fractions. Inoculation with micro-aggregates caused a less-aggressive osteomyelitis i.e. combination of infected tissue and strong healing response. Therefore, the use of low metabolic micro-aggregates could be a relevant inoculum for animal models of less-aggressive and thereby slower developing IAI and add in to our understanding of the host-implant-bacteria interactions in slow-onset low-grade infections.

Strain-specific Recovery of S. sonnei from Artificially Contaminated Baby Carrots: Enhancing Food-safety Investigations with a Customized Shigella Detection Method Based on Genomically predicted Antibiotic Resistance Traits

Shigella spp. are Gram-negative gastrointestinal bacterial pathogens that cause bacillary dysentery or shigellosis in humans. Isolation of Shigella from outbreak-associated foods is often problematic due to the lack of selectivity of cultural enrichment broths. To facilitate Shigella recovery from foods, we have developed strain-specific enrichment media based on the genomically-predicted antimicrobial resistance (AMR) features of an outbreak-associated Shigella sonnei strain harboring resistance genes for streptomycin (STR) and trimethoprim (TMP). To assess performance of the method, baby carrots were artificially contaminated with the S. sonnei strain at low (2.4 CFU), medium (23.5 CFU), and high levels (235 CFU) along with 10-fold higher levels of a Shigella-inhibiting Escherichia coli strain. The target S. sonnei strain was successfully recovered from artificially-contaminated baby carrots when enriched in modified Tryptone Soya Broth (mTSB) supplemented with TMP, whereas Shigella was not recovered from Shigella broth (SB) or SB supplemented with STR. Quantitative PCR analysis indicated that supplementation of the enrichment broths with TMP or STR increased the relative proportion of S. sonnei in enrichment cultures, except at the lowest inoculation level for STR. Microbiome profiling of the baby carrot enrichment cultures conducted by 16S rRNA gene sequencing indicated that both SB-STR and mTSB-TMP repressed the growth of competing Enterobacteriaceae in the enrichment cultures, relative to SB without supplementation. Overall, improved Shigella recovery was achieved with the addition of the appropriate custom selective agent during cultural enrichments demonstrating that genomically informed custom selective enrichment of Shigella could be a valuable tool for supporting future foodborne shigellosis outbreak investigations.

Detection of Extended Spectrum ß-Lactamase-Producing Escherichia coli with Biofilm Formation from Chicken Meat in Istanbul.

Antimicrobial resistance is one of the major public health problems worldwide. This study aimed to detect the presence of extended-spectrum β-lactamase-(ESBL-)producing Escherichia (E.) coli in chicken meat in Istanbul, Türkiye. Raw chicken meat samples (n = 208) were collected from different sale points and analyzed for ESBL-producing E. coli. In total, 101 (48.5%) isolates were confirmed as E. coli by PCR, of which 80/101 (79.2%) demonstrated multiple antibiotic resistance. Resistance against amoxicillin-clavulanic acid was most frequent (87.1%). Eighteen isolates (17.8%) demonstrated phenotypical ESBL resistance, as assessed by the double disc synergy test (DDST). Isolates were tested for the presence of β-lactamase genes and mobilized colistin-resistant genes. The blaTEM group was most frequently detected (97.02%), followed by blaCTX m (45.5%), blaSHV (9.9%), and blaOXA-2 (0.9%). However, mcr genes and blaNDM,blaKPC, blaVIM, and blaOXA-48 genes were not found in any isolate. E. coli strains were tested for biofilm formation in six different media [Nutrient broth, LB broth, Tryptone Soya broth (TSB), TSB containing 1% sucrose, TSB containing 0.6% yeast extract, and BHI]. Biofilm formation by E. coli isolates (44/101, 43.5%) was highest in TSB with 1% sucrose. It is worth noting that all biofilm-producing isolates were found to harbor the blaTEM-1 gene, which can indicate a high level of antibiotic resistance. This is the first report about ESBL-producing E. coli in poultry meat, the exposure of consumers in Istanbul metropolitan areas, and the ability of E. coli from this region to produce biofilms.

Microwave Oven Application for the Preparation and Sterilization of Microbiological Culture Media: A Feasible Method with An Adapted Water Bath and Perforable Cap

There is currently increased interest in the use of alternatives to autoclaved culture media, in order to maintain the properties of the media, while saving energy and time. In this study, we assess a new system for culture media preparation, using a conventional microwave with a water bath and a glass bottle with a rubber cap that allows depressurization. Sterilization, using the proposed system (1000 W, 3 to 20 min), was compared with autoclaving for the preparation of tryptone soy agar (TSA), tryptone soy broth (TSB), Sabouraud 4% dextrose agar (SDA), and violet red bile glucose agar (VRBG). Microwave exposure for 7 min yielded sterile TSA plates. The productivity of both sterilization methods was assessed using the pour plate method, and significant increases in the growth of certain micro-organisms after using a microwave were observed for every culture medium, especially those that were sterilized by boiling (VRBG). The kinetics of microbial destruction showed that Escherichia coli and Bacillus subtilis spores were destroyed after 3 and 7 min in a microwave, respectively, while three decimal reductions were obtained for Geobacillus stearothermophilus spores after 15 min in an autoclave. This new sterilization method could be a feasible, rapid, and economical method to prepare microbiological media, with a quality similar to that obtained through autoclaving.

Vibrio chanodichtyis sp. nov., isolated from the intestine of swordfish Chanodichthys dabryi.

A Gram-stain-negative, facultatively anaerobic, motile, curved-rod-shaped flagellated bacterium, designated DSL-7T, was isolated from the intestine of Chanodichthys dabryi in the Yangtze river, PR China. The strain grew optimally in tryptone soy broth medium at 37 °C, pH 7.0 and with 1 % (w/v) NaCl. Strain DSL-7T showed less than 96.2 % 16S rRNA gene sequence similarity to type strains of the genus Vibrio. Phylogenetic analysis based on genomes indicated that strain DSL-7T belonged to the genus Vibrio and formed a subclade with Vibrio mimicus NCTC 11435T, Vibrio metoecus OP3HT, Vibrio cholerae ATCC 14035T, Vibrio albensis ATCC14547T, Vibrio paracholerae OP3HEDC-792T and Vibrio tarriae 2521-89T. The average nucleotide identity (ANI) and in digital DNA-DNA hybridization (dDDH) values between DSL-7T and closely related type strains were below the accepted threshold to delineate a new species of 95 and 70 %, respectively. The major cellular fatty acids were summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C16 : 0, summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and C14 : 0. The genomic DNA G+C content was 47.6 mol%. Based on the phenotypic, chemotaxonomic, phylogenetic and genomic data, strain DSL-7T represents a novel species of the genus Vibrio, for which the name Vibrio chanodichtyis sp. nov. is proposed, with strain DSL-7T (=KCTC 92851T=CCTCC AB 2022396T) as the type strain.

Prevalence and antibiotic resistance pattern of bacteria from sepsis suspected neonates at St. Paul’s Hospital Millennium Medical College, Addis Ababa, Ethiopia

BackgroundNeonatal sepsis is the major cause of neonatal mortality and morbidity, especially in low and middle-income countries. Continuous monitoring of pathogens and their antibiotic resistance pattern is crucial for managing neonatal sepsis. This study aimed to determine neonatal sepsis due to bacteria, antibiotic resistance patterns, associated risk factors and patient outcomes at St. Paul’s Hospital Millennium Medical College.MethodAn institutional-based cross-sectional study was conducted on 400 neonates suspected of sepsis at St. Paul’s Hospital Millennium Medical College from March 2020 to July 2020. A questionnaire was used to collect socio-demographic information, clinical parameters and potential risk factors from study participants. About 2ml of blood was drawn aseptically and inoculated into Tryptone Soya Broth at the patient’s bedside. Bacterial identification was performed by using standard microbiological techniques. The disk diffusion method was used to determine the antibiotic susceptibility patterns of each isolated bacteria. Data entry and analysis were done using Statistical Package for Social Sciences (SPSS) version 20 software. Bivariate and multivariable logistic regressions were used to assess associated risk factors of neonatal sepsis. A p-value less than 0.05 was considered statically significant with a 95% confidence interval.ResultsThe overall prevalence of neonatal septicemia was 21% (84/400). Of these, 67 (79.8%) and 17 (20.2%) were gram-negative and gram-positive bacteria, respectively. Klebsiella spp, 37 (44%), E. coli 19 (21.6%) and Coagulase negative Staphylococci 13 (15.47%) were the leading cause of neonatal sepsis. Ciprofloxacin and amikacin were the most effective antibiotics for gram-negative and gram-positive bacteria. Multidrug resistance was observed in 84% of the bacterial isolates. Low birth weight and preterm were associated with neonatal septicemia (AOR = 49.90, 95% CI = 15.14-123.081, P = 0.002) and (AOR = 18.20, 95% CI = 6.835–27.541, P = 0.004) respectively.ConclusionKlebsiella spp and E. coli were frequently isolated bacteria in our study. The proportion of multidrug-resistance was significantly high. Most isolated bacteria were resistant to ampicillin, ceftazidime, cefotaxime and gentamycin, which indicates the necessity of continuous evaluation of antibiotic resistance rate.

Dynamic Thermal Treatments in Green Coconut Water Induce Dynamic Stress Adaptation of Listeria innocua That Increases Its Thermal Resistance.

The global coconut water market is projected to grow in the upcoming years, attributed to its numerous health benefits. However, due to its susceptibility to microbial contamination and the limitations of non-thermal decontamination methods, thermal treatments remain the primary approach to ensure the shelf-life stability and the microbiological safety of the product. In this study, the thermal inactivation of Listeria innocua, a Listeria monocytogenes surrogate, was evaluated in coconut water and in tryptone soy broth (TSB) under both isothermal (50-60 °C) and dynamic conditions (from 30 to 60 °C, with temperature increases of 0.5, 1 and 5 °C/min). Mathematical models were used to analyse the inactivation data. The Geeraerd model effectively described the thermal inactivation of L. innocua in both TSB and coconut water under isothermal conditions, with close agreement between experimental data and model fits. Parameter estimates and analysis revealed that acidified TSB is a suitable surrogate medium for studying the thermal inactivation of L. innocua in coconut water, despite minor differences observed in the shoulder length of inactivation curves, likely attributed to the media composition. The models fitted to the data obtained at isothermal conditions fail to predict L. innocua responses under dynamic conditions. This is attributed to the stress acclimation phenomenon that takes place under dynamic conditions, where bacterial cells adapt to initial sub-lethal treatment stages, leading to increased thermal resistance. Fitting the Bigelow model directly to dynamic data with fixed z-values reveals a three-fold increase in D-values with lower heating rates, supporting the role of stress acclimation. The findings of this study aid in designing pasteurization treatments targeting L. innocua in coconut water and enable the establishment of safe, mild heat treatments for refrigerated, high-quality coconut water.

Acid adaptation increased the resistance of Escherichia coli O157:H7 in bok choy (Brassica rapa subsp. chinensis) juice to high-pressure processing.

Based on the U.S. Food and Drug Administration regulations, E. coli O157:H7 is a pertinent pathogen in high acid juices that needs to be inactivated during the pasteurization process. The results of this study suggest that the effect of acid adaptation should be considered in the selection of HPP parameters for E. coli O157:H7 inactivation to ensure that pasteurization objectives are achieved.

The Effects of Pectin-Honey Hydrogel in a Contaminated Chronic Hernia Model in Rats.

Incisional hernia is a frequent complication after abdominal surgery. A previous study on rats evaluated the use of a Pectin-Honey Hydrogel (PHH)-coated polypropylene (PP) mesh for the healing of acute hernias. However, there are no studies investigating the use of PHH in association with PP mesh in chronic contaminated hernia. The aims of this study are to assess the effectiveness of PHH in promoting abdominal hernia repaired with PP mesh and in counteracting infection. Twenty Sprague Dawley male rats were enrolled and a full thickness defect was made in the abdominal wall. The defect was repaired after 28 days using a PP mesh, and a culture medium (Tryptone Soy Broth, Oxoid) was spread onto the mesh to contaminate wounds in both groups. The rats were randomly assigned to a treated or untreated group. In the treated group, a PHH was applied on the mesh before skin closure. At euthanasia-14 days after surgery-macroscopical, microbiological and histopathological evaluations were performed, with a score attributed for signs of inflammation. An immunohistochemical investigation against COX-2 was also performed. Adhesions were more severe (p = 0.0014) and extended (p = 0.0021) in the untreated group. Bacteriological results were not significantly different between groups. Both groups showed moderate to severe values (score > 2) in terms of reparative and inflammatory reactions at histopathological levels. The use of PHH in association with PP mesh could reduce adhesion formation, extension and severity compared to PP mesh alone. No differences in terms of wound healing, contamination and grade of inflammation were reported between groups.

Characterization of Salmonella enterica Biofilms and Antibiofilm Effect of Carvacrol and 2-Aminobenzimidazole.

Biofilm-associated foodborne Salmonella infections in poultry have become increasingly challenging for veterinarians, particularly in developing countries, and warrant thorough investigation. We assessed the biofilm-forming tendency of poultry isolates of Salmonella enterica, namely Salmonella Typhimurium (n = 23), Salmonella Infantis (n = 28), and Salmonella Heidelberg (n = 18), in nutrient-rich Rappaport-Vassiliadis Soya (RVS) peptone broth and nutrient-deficient diluted Tryptone Soya Broth (TSB). Seven of the tested isolates exhibited moderate biofilm formation in diluted TSB, whereas two showed such formation in RVS. In addition, the Congo red agar assay revealed curli and cellulose production in seven isolates. Fourteen specific biofilm-associated genes were analyzed identifying sdiA and seqA to be the most prevalent (100%), and glyA the least prevalent (69.5%). The prevalence of the genes bcsA and csgA was significantly lower in moderate and weak biofilm formers, respectively, as compared with nonbiofilm formers in RVS peptone broth. Furthermore, the compounds carvacrol and 2-aminobenzimidazole (2-ABI) effectively inhibited biofilm formation by Salmonella serovars in RVS peptone and TSB media, respectively. Whereas the antibiofilm activity of 2-ABI against Salmonella has not been reported previously, we determined its most effective concentration at 1.5 mM among tested antibiofilm treatments. These findings indicate that Salmonella strains prevalent in poultry farms have the potential to form biofilms, and the tested compounds should be further explored as supportive or alternative antimicrobials.

Characterization of Biofilm Formation and Bacterial Resistance to Benzalkonium Chloride under Contrasting Cultivation Conditions

Benzalkonium chloride (BAC) is one of the most commonly used quaternary ammonium compounds in the pharmaceutical, cosmetic, and food industries. The aim of our study was to compare the physiological responses of Escherichia coli MSCL 332, Pseudomonas putida MCCL 650, and Staphylococcus epidermidis MSCL 333 on 50 mg/L BAC in rich and poor medium (100% and 5% tryptone soya broth (TSB)) in the temperature range from 8 °C to 37 °C, under static and shaking conditions. A high-throughput, 96-well microplate method was used to compare a broad range of cultivation conditions. The effect of BAC on growth, biofilm formation activity, and dehydrogenase and fluorescein diacetate hydrolysis activity was evaluated. Addition of BAC to 100% TSB inhibited biofilm formation at 37 °C by 2.4, 1.8, and 1.6 times for E. coli, P. putida, and S. epidermidis, respectively. In turn, BAC stimulated biofilm formation in E. coli in 5% TSB at 37 °C and 100% TSB at 8 °C, i.e., 1.4 and 1.3 times, respectively. Statistical optimization of broth composition with emphasis on biofilm formation and further testing under experimental conditions was performed with P. putida.