Bacterial Isolates Research Articles

Microbial profile of diabetic foot osteomyelitis from the northwest of England

BackgroundOsteomyelitis of the diabetic foot is a common and challenging complication affecting patients with diabetic foot ulcers and infections. The complexity of these infections lies in their polymicrobial nature, high rates of persistence and recurrence. This study examined the microbiological profile of diabetic foot osteomyelitis from a teaching hospital in Northwest England and their resistance patterns to understand its impact on infection persistence and to direct effective treatment.MethodsA retrospective review of 105 patients who underwent surgical management for diabetic foot osteomyelitis between 2019 and 2024. We analysed three consecutive culture samples for each patient to assess for the microbiological profile and resistance patterns of these infections and to monitor infection recurrence and persistence rates.ResultsA total of 105 patients were identified. Infection eradication was noted in 42% of the cohort, infection persistence in 18%, and late infection recurrence in 40%. Polymicrobial growth was evident in 72% of our study sample. Gram-positive bacteria made up the majority of the bacterial isolates in all 3 culture samples, 74.81% in sample 1, 69.31% in sample 2, and 55.1% in sample 3. Staphylococcus aureus was the most prevalent gram-positive bacteria, at 52.38% in sample 1, 36.19% in sample 2, and 18.09% in sample 3, followed by Haemolytic Streptococcus, Enterococcus and Corynebacterium. The frequently identified gram-negative bacteria were Pseudomonas in sample 1 (7.61%), E. coli and Proteus in sample 2 (5,71%), Pseudomonas and Proteus in sample 3 (2.85%). Gram-positive bacteria were resistant to penicillin and macrolides with resistance of staphylococcus aureus to clarithromycin identified among all 3 culture samples. Gram-negative bacteria were most resistant to amoxicillin. Staphylococcus aureus was responsible for infection persistence in most of our cohort (12/19) 63.15%. Among those patients, Staphylococcus was resistant to clarithromycin in 6 of the cases. The 5-year mortality rate for our study sample was 32.38%.ConclusionThis study highlights the prevalence of polymicrobial growth and multi-drug resistant pathogens in the scope of diabetic foot osteomyelitis. It highlights the predominance of Staphylococcus aureus and its resistant strains among patients affected by diabetic foot osteomyelitis in Greater Manchester.

Recovery of clinically relevant multidrug‐resistant Klebsiella pneumoniae lineages from wastewater in Kumasi Metropolis, Ghana

AbstractAntimicrobial resistance (AMR) is under‐monitored in Africa, with few reports characterizing resistant bacteria from the environment. This study examined physicochemical parameters, chemical contaminants and antibiotic‐resistant bacteria in waste stabilization pond effluents, hospital wastewater and domestic wastewater from four sewerage sites in Kumasi. The bacteria isolates were sequenced. Three sites exceeded national guidelines for total suspended solids, biochemical oxygen demand, chemical oxygen demand and electrical conductivity. Although sulfamethoxazole levels were low, the antibiotic was detected at all sites. Multi‐drug‐resistant Klebsiella pneumoniae and Pseudomonas aeruginosa were isolated with multi‐locus sequence typing identifying K. pneumoniae strains as ST18 and ST147, and P. aeruginosa as ST235, all of clinical relevance. A comparison of ST147 genomes with isolates from human infections in Africa showed remarkable similarity and shared AMR profiles. Thirteen of the twenty‐one plasmids from ST147 harbored at least one AMR gene, including blaCTX‐M‐15 linked to copper‐resistance genes. Our study demonstrated high bacterial counts and organic matter in the analysed wastewater. The recovery of clinically significant isolates with multiple antibiotic and heavy metal resistance genes from the wastewater samples raises public health concerns.

Characterization and Selection of Potential Starters of Bacillus Sp. Involved in the Fermentation of Néré Seeds (Parkia biglobosa) in Côte d'Ivoire

Aims: The present study highlights about Bacillus sp. starter strains to control fermentation of Parkia biglobosa (néré) seeds. Place and Duration of Study: Between February and August 2023, this work was performed at Laboratory of Biochemistry, Microbiology and Valorization of Agroresources, of the Agropastoral Management Institute, which is part of Peleforo Gon Coulibaly University (Korhogo, Côte d'ivoire). Methodology: Fermented néré seeds were collected from soumbara producers in the Korhogo region in northern Côte d'Ivoire. Néré seeds were subjected to a physicochemical analysis to determine the pH, titrable acidity, moisture, dry matter and ash content. These fermented seeds were used for the isolation and enumeration of Bacillus bacteria. Production of extracellular enzymes and the influence of stress conditions were carried out on these strains to select potential starters. Results: The study of the physicochemical analysis of fermented néré seeds showed that these seeds have a pH of 6.59 ± 0.04, close to neutrality, with a very low titrable acidity of 0.06 ± 0.00%. The moisture content of the néré seeds is 38.76 ± 0.13% with a high dry matter content of 61.24 ± 0.22% coupled with a low ash content of 3.18 ± 0.05%. Thirty-three (33) Gram-positive bacilli were isolated from néré seeds with a microbial load of 7.1 log10 cfu/g. The production of extracellular enzymes and the influence of stress conditions led to the selection of five Bacillus strains (BS 08, BS 14, BS 19, BS 22 and BS 31) as potential starters. Conclusion: The control of the fermentation of néré seeds involves selecting strains with the best potential. Thus, five Bacillus sp. Isolates (BS 08, BS 14, BS 19, BS 22 and BS 31) were selected as potential starters for obtaining a soumbara of exceptional nutritional and organoleptic quality.

Isolasi Staphylococcus aureus dari swab tangan penjamah makanan di kantin Universitas Mataram

One of the main qualities of food is seen from the biological safety aspect, such as being free from Pathogenic microorganisms. Staphylococcus aureus is a pathogenic bacterium that can contaminate food ingredients come from the skin of food handlers. S. aureus produces enterotoxins that have a poisoning in consumers.The preliminary observationstated that there were still many food handlers in the Mataram University canteen who did not pay attention to hand hygiene and personal sanitation. This study aims to determine the bacterial contamination of S. aureus swabbed from the hands of food handlers in the Mataram University cafetaria. The method was used in this research is descriptive observational. The research consisted of two stages, namely isolation and biochemical identification. The isolation stage resulted 10 culturable bacterial isolates. All bacteria were able to ferment mannitol on Mannitol Salt Then, the biochemical test identification resulted that all isolates had a positive catalase test, a positive slide coagulase test, Gram-positive, and staphylococci cell. The TSIA test resulted A/A or (acid)/(acid) which indicated that all isolates were able to use diverse carbon sources from sucrose, lactose, glucose. Moreover, other test as sulfur, indole and motility tests were negative. Therefore, all isolates in this study are classified as S. aureus. This research is also a promotive and preventive effort for the campus in implementing hygiene and sanitation for food handlers in the cafetaria.

Phenotypic, Molecular Identification and Virulence Assessment of Cronobacter spp Isolated from Clinical Samples of Children Under Two Years in Mosul City/Iraq

Cronobacter spp. is known to be a foodborne causative agent for a variety of diseases in both humans and animals. This study focused on isolating Cronobacter species from 150 clinical samples of children under two years (60 from blood and stool samples, 30 from CSF samples) collected from Ibn al Atheer and Al Khanssa Hospitals. The phenotypic identification of bacterial isolates were performed through culture, microscopic, and biochemical examinations and vitek-2 system. The results revealed a mixture of bacteria in 6.5% of the clinical specimens. The identity of isolates was 99% using the vitek-2 system for identifying Cronobacter sakazakii, Cronobacter malonaticus, Cronobacter muytjensii, and Cronobacter pulveris. Out of 150 specimens there were 8 (5.33 %) of specimens gave positive for Cronobacter spp which included: C. sakazaki 2/60 blood specimens (3.3%), C. sakazaki 2/60 stool specimens (3.3%), C. malonaticus 2/60 stool specimens (3.3%), 1/60 C. muytjensii and C. pulveris 1/60 (3.3%). The eight isolates phenotypically identified as Cronobacter were confirmed at the molecular level through 16S rRNA sequencing and submitted to NCBI under the accession numbers (OR825874, OR825875, PP126443, PP126444, PP126445, PP126455). The virulence profile of these isolates showed that 7/8 (87.5%) of Cronobacter strains exhibited haemolysin activity, 5/8 strains (62.5%) were able to produce the protease enzyme and 2/8 (25%) of Cronobacter strains were positive for lipase and lecithinase. All strains lacked the ability to produce a slime layer. The results also showed the ability of Cronobacter strains to produce biofilm by using the tube method and microtiter plate method but with different levels.

An Etiology of Sputum Culture Contamination and their Drugs Susceptibility Patterns among TB-suspected Patients at Epicentre Mbarara Research Centre, Uganda

Aims: Contamination in TB cultures is problematic as it allows the growth of non-target bacterial or fungal species in sputum samples, potentially concealing M. tuberculosis. This study aimed to determine the prevalence, characterization, and drug susceptibility patterns of contaminants in sputum cultures from TB-suspected patients. Methods: A laboratory-based cross-sectional study was conducted from October 2023 to June 2024 at the Epicentre Mbarara Research Centre, involving 81 sputum samples from TB-suspected patients. Conventional decontamination procedures were used, and both solid Lowenstein Jensen (LJ) and liquid Mycobacterium Growth Indicator Tube (MGIT) culture methods were applied. Standard culture techniques and physicochemical analyses were employed. Bacteria were incubated at 37°C in O2 and Co2 incubators, with growth observed within 24 to 48 hours and up to three days for yeast contaminants. Sabouraud Dextrose Agar cultures were monitored for two weeks for filamentous fungi. Antibiograms for bacterial isolates were determined using the Kirby-Bauer, while broth microdilution was used for fungal isolates. Results: Of 81 samples, 14(17.28%) were contaminated. The most common bacterial contaminants were Staphylococcus aureus, Bacillus subtilis, Streptococcus viridans, Staphylococcus epidermidis, and Escherichia coli. Fungal contaminants included Aspergillus flavus, Penicillium species, Candida albicans, Cryptococcus neoformans, and non-albican Candida species. High resistance to erythromycin (73%) and tetracycline (50%) was noted among bacterial isolates, with sensitivity to vancomycin (67%), gentamycin (60%), and chloramphenicol (58%). Yeast isolates showed (87.5%) susceptibility to itraconazole and (75%) to amphotericin B but (50%) resistance to 5-flucytosine. Mold isolates exhibited (100%) susceptibility to itraconazole, amphotericin B, and 5-flucytosine but complete resistance to caspofungin. Conclusion: The level of contamination in TB cultures was noted, and Staphylococcus aureus and Candida albicans were observed as the most common contaminants. Bacterial isolates resisted erythromycin and tetracycline, while fungal isolates were generally susceptible to itraconazole and amphotericin B. We recommend Strengthening decontamination and monitoring common resistant contaminants.

Isolation and characterization of non-rhizobial bacteria and arbuscular mycorrhizal fungi from legumes

This study investigates non-rhizobial endophytic bacteria in the root nodules of chickpea (Cicer arietinum L), faba bean (Vicia faba), and cowpea (Vigna unguiculata L. Walp), as well as arbuscular mycorrhizal fungi in the rhizospheric soil of chickpea and faba bean. Out of the 34 endophytic bacterial populations examined, 31 strains were identified as non-rhizobial based on nodulation tests. All strains were assessed for their plant growth-promoting (PGP) activities in vitro. The results revealed that most isolates exhibited multiple PGP activities, such as nitrogen fixation, indole-3-acetic acid (IAA) and ammonia (NH3) production, phosphate solubilization, and exopolysaccharide production. The most effective PGP bacteria were selected for 16S rRNA analysis. Additionally, a total of 36 species of native arbuscular mycorrhizal fungi (AMF) were identified. Acaulospora (100%) and Scutellospora (91.66%) were the most prevalent genera in Cicer arietinum L. and Vicia faba L. plants, respectively. Acaulospora also exhibited the highest spore density and relative abundance in both plants. Moreover, the root colonization of Cicer arietinum L. and Vicia faba L. plants by hyphae, vesicles, and arbuscules (HVA) was significant. The findings of this study provide valuable insights into non-rhizobial endophytic bacteria associated with legume root nodules and the diversity of AMF. These organisms have great potential for PGP and can be manipulated by co-inoculation with rhizobia to enhance their biofertilizer effectiveness. This manipulation is crucial for promoting sustainable agriculture, improving crop growth, and advancing biofertilizer technology.

Evaluation of a microfluidic-based point-of-care prototype with customized chip for detection of bacterial clusters.

Especially in medical facilities, where morbid people are nursed in close distance to each other, pathogenic bacteria can accumulate and spread. To contain such infection clusters, usually time- and labor-intensive large-scale screening assays are conducted, where patients and patient-side surfaces are sampled, and PCR or whole-genome sequencing analyses are conducted to confirm or deny cluster affiliation of cultivated bacteria. Hence, fast solutions with easy application are required to complement the current state-of-the-art technology for cluster surveillance. Here, we developed a fully automated microfluidic point-of-care prototype that identified bacterial cluster isolates within 70 min from bacterial solutions, including swab eluates. The system requires only low hands-on time and can be applied apart from laboratory infrastructures near the patient, which considerably reduces the time from sampling to result. This ensures fast implementation of hygiene measures and quick containment of the infection cluster, which would enhance patients' safety and outcome.

Primary screening of growth-stimulating properties of rhizosphere microorganisms in relation to tomato seeds

The ability of bacterial isolates isolated from the rhizosphere of cultivated and wild plants of arid ecosystems of the Astrakhan region, which have the ability to tryptophan-induced synthesis of indolyl-3-acetic acid, to stimulate the germination of tomato seeds of the varieties “Sanka”, “Volgogradsky”, “Cosmonaut Volkov”, was studied. “Newbie” Experimental studies were carried out through laboratory experiments by incubating seeds in humid chambers; laboratory germination and morphometric parameters of seedlings were assessed. It was revealed that most of the studied suspensions of isolates have a stimulating effect on tomato seeds. High germination (90% or more) of seeds of the “Sanka” variety was observed when 12 isolates were treated with suspensions; variety “Volgogradsky” – suspensions of 9 isolates; variety “Cosmonaut Volkov” – suspensions of 2 isolates; variety “Novichok” – suspensions of 3 isolates. Suspensions of isolates 16/19 and 31/20 contributed to high germination of seeds of all varieties: up to 86.6–100% and 90–100%, respectively, and isolate 9/19 significantly reduced the germination of seeds of the varieties “Volgogradsky”, “Cosmonaut Volkov” and “Novichok” by 66.6–56.7%. When assessing the morphometric parameters of seedlings in the experiment, it was noted that the suspension of isolate 16/19 helps to increase the biomass of the varieties “Sanka”, “Volgogradsky” and “Cosmonaut Volkov” up to 273–449%. Suspension of isolate 31/20 has a stimulating effect on the morphometric parameters of seedlings of all varieties: it stimulates root development by 73.3–183.3%, stem development by 46.8–100.5%. The isolate 9/19 suspension not only inhibits seed germination, but also causes a reduced accumulation of seedling biomass, inhibits the development of roots and stems of seedlings of the varieties “Volgogradsky”, “Cosmonaut Volkov” and “Novichok”. Thus, during experimental studies, isolates (16/19 and 31/20) of rhizosphere bacteria were identified that have a stimulating effect on both the germination of tomato seeds and the morphometric parameters of seedlings.

Fosfomycin—Overcoming Problematic In Vitro Susceptibility Testing and Tricky Result Interpretation: Comparison of Three Fosfomycin Susceptibility Testing Methods

Background: Fosfomycin (FOS) is an older antimicrobial agent newly rediscovered as a possible treatment for infections with limited therapeutic options (e.g., Gram-negative bacteria with difficult-to-treat resistance, DTR), especially in intravenous form. However, for correct usage of FOS, it is necessary to have a reliable susceptibility testing method suitable for routine practice and robust interpretation criteria. Results: The results were interpreted according to 2023 interpretation criteria provided by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). DTR Gram-negatives were more likely to be resistant to FOS (45% in Enterobacterales and 20% in P. aeruginosa) than non-DTR (10% and 6.7%, resp.). All isolates of S. aureus were susceptible to FOS. In Gram-negatives, all agreement values were unacceptable. Etest® performed better in the DTR cohort (categorical agreement, CA, 80%) than in the non-DTR cohort (CA 45.7%). There were no very major errors (VREs) observed in P. aeruginosa. S. aureus had surprisingly low essential agreement (EA) rates (53% for MRSA and 47% for MSSA) for Etest®, but categorical agreement was 100%. Methods: A total of 130 bacterial isolates were tested and compared using the disc diffusion method (DD) and gradient strip method (Etest®) with the reference method (agar dilution, AD). The spectrum of isolates tested was as follows: 40 Enterobacterales (20 DTR vs. 20 non-DTR), 30 Pseudomonas aeruginosa (15 DTR vs. 15 non-DTR), and 60 Staphylococcus aureus (30 methicillin-susceptible, MSSA, vs. 30 methicillin-resistant, MRSA). Conclusions: Neither one of the tested methods was identified as a suitable alternative to AD. It would be beneficial to define more interpretation criteria, at least in some instances.

Comprehensive evaluation of the antibacterial and antibiofilm activities of NiTi orthodontic wires coated with silver nanoparticles and nanocomposites: an in vitro study.

Fixed orthodontic appliances act as a niche for microbial growth and colonization. Coating orthodontic wires with antimicrobial silver nanoparticles (AgNPs) and nanocomposite was adopted in this study to augment the biological activity of these wires by increasing their antibacterial and antibiofilm properties and inhibiting bacterial infections that cause white spot lesions and lead to periodontal disease. Three concentrations of biologically synthesized AgNPs were used for coating NiTi wires. The shape, size, and charge of the AgNPs were determined. Six groups of 0.016 × 0.022-inch NiTi orthodontic wires, each with six wires, were used; and coated with AgNPs and nanocomposites. The antimicrobial and antibiofilm activities of these coated wires were tested against normal flora and multidrug-resistant bacteria (Gram-positive and Gram-negative bacterial isolates). The surface topography, roughness, elemental percentile, and ion release were characterized. AgNPs and nanocomposite coated NiTi wires showed significant antimicrobial and antibiofilm activities. The chitosan-silver nanocomposite (CS-Ag) coated wires had the greatest bacterial growth inhibition against both Gram-positive and Gram-negative bacteria. The surface roughness of the coated wires was significantly reduced, impacting the surface topography and with recorded low Ni and Ag ion release rates. NiTi orthodontic wires coated with AgNPs, and nanocomposites have shown increased antimicrobial and antibiofilm activities, with decreased surface roughness, friction resistance and limited- metal ion release.

Caracterización fenotípica y simbiótica de las bacterias que nodulan Genista saharae en la región árida de Argelia

Twenty bacterial strains had been isolated from root nodules of Genista saharae that grew wild in Biskra and El Oued city (Northeastern Algerian Sahara). This study focused on obtaining isolates of legume nodule bacteria (LNB) from the plant G. saharae and evaluated their effectiveness in forming a symbiotic relationship with the legume species Vigna unguiculata through cross-inoculation. Additionally, the study aimed to identify the successful cross-inoculation group of LNB strains based on their phenotypic characteristics. The growth capacity of isolates under varying salinity conditions [NaCl] and pH levels was investigated using a spectrophotometer (96-microplate reader). The API 20NE and API 20E systems were used to identify the biochemical characteristics of the isolates. In addition, the rhizospheric soil samples from the two study sites were analyzed using standard analytical techniques of soil. All isolates established effective symbioses with Vigna unguiculata, were Gram-negative rods, and were fast-growing. The optimal growth temperature was between 28 °C and 37 °C; some isolates were thermophiles and specifically withstood extreme heat between 45-50 °C. Furthermore, they demonstrated a wide tolerance range to pH (5–10) with salt tolerance ranging from 100 mM to 500 mM. Biochemical results revealed that the isolates assimilated various sources of carbon and nitrogen and displayed numerous enzyme activities. Physicochemical analysis revealed that all the soils were deficient in nutrients and had an alkaline pH. This study enabled us to identify the effective stress-tolerant strains, which could be used in the future to inoculate plants for environmental applications. Keywords: Rhizobia, morpho-physiological and biochemical characterization.

Culture-Dependent Characterization of Hydrocarbon Utilizing Bacteria and Fungi from an Artisanal Crude Oil Refining Site

Crude oil refining is a global insert responsible for the production of a wide array of petroleum based products crucial for various sectors including energy, transportation, and manufacturing. However, the artisanal form of crude oil refining, often characterized by its small scale and limited technological infrastructure, has emerged as a major environmental concern in several regions around the world. Illegal refining of crude oil has led to oil spillages which in turn lead to a disruption of the microbial population of the soil. Hence, the aim of this study was to isolate and characterize the heterotrophic and hydrocarbon utilizing bacteria and fungi from an artisanal crude oil refining site. A total of thirty six (36) soil samples were collected from three (3) artisanal crude oil refining sites at iwofe, ogbogoro and ibaa community all in Rivers State, Nigeria. The samples were collected using soil auger and subjected to standard microbiological procedures such as culturing, isolation and identification. The results revealed significantly elevated levels of heterotrophic and hydrocarbon utilizing bacteria and fungi in the soil samples, with notable variations (p≤0.05) across the different artisanal crude oil refining sites. Total Heterotrophic Bacteria (THB) counts ranged from 3.75x105CFU/g in Iwofe to 2.70x105CFU/g in Ibaa, Total heterotrophic Fungal (THF) counts ranged from 2.80x104CFU/g recorded in Ogbogoro to 1.45x104CFU/g recorded in Iwofe, Hydrocarbon Utilizing Bacteria (HUB) counts ranged from 2.25x103CFU/g in Ibaa to 1.30x103CFU/g in Iwofe and Hydrocarbon Utilizing Fungi (HUF) counts ranged from 2.50x102CFU/g recorded in Ogbogoro to 1.40x102CFU/g recorded in Ibaa community with significant differences (p≤0.05). Bacteria isolated and characterized from the soil include Enterobacter, Pseudomonas putida, Serratia and Bacillus amyloliquefaciens while fungi include Fusarium, Aspergillus, Penicillium and Mucor. The microorganisms identified in this study have been reported to play key roles in the utilization of crude oil. Pseudomonas and Bacillus were the most dominant bacterial isolates and their dominance could imply that the soil could support the growth of crops. Thus, while they utilize the hydrocarbons, they also serve as plant-growth promoting microorganisms. Although some of the microorganisms in the present study have been reported to promote the growth of plant, the dumping of crude oil products legal or illegal should be minimized to avoid the extinction of plant-promoting microorganisms.

Personalized CZA-ATM dosing against an XDR E. coli in liver transplant patients; the application of the in vitro hollow fiber system.

A patient with an extensively drug-resistant (XDR) New Delhi metallo-β-lactamase (NDM) and oxacillinase (OXA-48) producing Escherichia coli (E. coli) infection was awaiting orthotopic liver transplant. There is no standardized antibiotic prophylaxis regimen; however, in line with the Infectious Diseases Society of America guidance, an antibiotic prophylactic regimen of ceftazidime-avibactam 2.5g TDS with aztreonam 2g three times a day (TDS) IV was proposed. The hollow fiber system (HFS) was applied to inform the individualized pharmacodynamic outcome likelihood prior to prophylaxis. A 4-log reduction in CFU/mL in the first 10 h of the regimen exposure was observed; however, the killing dynamics were slow and six 8-hourly infusions were required to reduce bacterial cells to below the limit of quantification. Thus, the HFS supported the use of the regimen for infection clearance; however, it highlighted the need for several infusions. Standard local practice is to administer prophylaxis antibiotics at induction of orthotopic liver transplantation (OLT); however, the HFS provided data to rationalize earlier dosing. Therefore, the patient was dosed at 24 h prior to their OLT induction and subsequently discharged 8 days after surgery. The HFS provides a dynamic culture solution for informing individualized medicine by testing antibiotic combinations and exposures against the bacterial isolates cultured from the patient's infection. .

Bacterial isolates and antibiotic sensitivity in canine bacterial keratitis in Korea.

To analyze bacterial isolates associated with canine bacterial keratitis and their antibiotic susceptibility patterns in Korea, focusing on multidrug resistance (MDR) and identifying effective antibiotic combinations for clinical treatment. A total of 146 dogs diagnosed with suspected bacterial keratitis between October 2022 and October 2023 in Korea, with 157 eye samples collected for analysis. Eye samples were cultured to isolate bacteria, and antibiotic susceptibility testing was performed using the minimum inhibitory concentration (MIC) method. Bacterial identification was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF). The study assessed the efficacy of individual antibiotics and combination therapies. Bacteria were isolated in 55.4% of the samples. The most common genera were Staphylococcus species (48.5%, 48/99), Streptococcus species (13.1%, 13/99), Pseudomonas species (9.1%, 9/99), and Escherichia coli (9.1%, 9/99). Amikacin (84.8%) showed the highest antibiotic susceptibility, while doxycycline exhibited the lowest (17.2%). The most effective antibiotic combinations were amikacin-moxifloxacin (93%). MDR isolates accounted for 52.5% (52/99) of the total bacterial samples. Staphylococcus species were the most common isolates, with 52.5% showing MDR, underscoring the need to curb antibiotic misuse. While antibiotics like amikacin demonstrated high susceptibility rates, their use should be reserved for resistant infections to prevent further resistance development. Rather than focusing solely on finding effective combinations of antibiotics, it is crucial to consider alternative treatment strategies that offer more sustainable solutions. Rather than relying on antibiotic combinations, attention should shift to sustainable alternatives to treat bacterial keratitis and reduce antibiotic dependence in clinical practice.

Deciphering Phyllomicrobiome of Cauliflower Leaf: Revelation by Metagenomic and Microbiological Analysis of Tolerant and Susceptible Genotypes Against Black Rot Disease.

Understanding the phyllomicrobiome dynamics in cauliflower plants holds significant promise for enhancing crop resilience against black rot disease, caused by Xanthomonas campestris pv. campestris. In this study, the culturable microbiome and metagenomic profile of tolerant (BR-161) and susceptible (Pusa Sharad) cauliflower genotypes were investigated to elucidate microbial interactions associated with disease tolerance. Isolation of phyllospheric bacteria from asymptomatic and black rot disease symptomatic leaves of tolerant and susceptible cultivars yielded 46 diverse bacterial isolates. Molecular identification via 16S rRNA sequencing revealed differences in the diversity of microbial taxa between genotypes and health conditions. Metagenomic profiling using next-generation sequencing elucidated distinct microbial communities, with higher diversity observed in black rot disease symptomatic leaf of BR-161. Alpha and beta diversity indices highlighted differences in microbial community structure and composition between genotypes and health conditions. Taxonomic analysis revealed a core microbiome consisting of genera such as Xanthomonas, Psychrobacillus, Lactobacillus, and Pseudomonas across all the samples. Validation through microbiological methods confirmed the presence of these key genera. The findings provide novel insights into the phyllomicrobiome of black rot-tolerant and susceptible genotypes of cauliflower. Harnessing beneficial microbial communities identified in this study offers promising avenues for developing sustainable strategies to manage black rot disease and enhance cauliflower crop health and productivity.

Prospects of iron solubilizing Bacillus species for improving growth and iron in maize (Zea mays L.) under axenic conditions

Iron (Fe) deficiency in calcareous soils is a significant agricultural challenge, affecting crop productivity and nutritional quality. This study aimed to isolate, characterize, and evaluate Fe solubilizing rhizobacterial isolates from maize rhizosphere in calcareous soils as potential biofertilizers. Forty bacterial isolates coded as SG1, SG2, …, SG40 were isolated and screened for siderophore production, with ten showing significant Fe solubilizing capabilities. These isolates were further assessed for phosphate solubilization and exopolysaccharides production. The selected bacterial isolates were also screened under axenic conditions for their ability to improve maize growth. The isolates SG8, SG13, SG24, SG30 and SG33 significantly enhanced growth parameters of maize. Notably, SG30 showed highest increment in shoot length (58%), root length (54%), root fresh and dry biomass (67% and 76%), SPAD value (67%), relative water contents (69%), root surface area (61%), and Fe concentration in shoots (79%) as compared to control. The biochemical characterization of these strains showed that all these strains have capability to solubilize insoluble phosphorus, produce indole-3-acetic acid (IAA), and ammonia with catalase, urease and protease activity. Molecular identification through 16s rRNA gene sequencing confirmed high similarity (99.7–100%) of the selected isolates to various Bacillus species, including B. pyramidoids, B. firmicutes, and B. cereus. The study provides a strong base for developing eco-friendly, cost-effective biofertilizers to address Fe deficiency in crops and promote sustainable agriculture.

Molecular Identification, Histopathology and Antibiotic Susceptibility Profiling of Aeromonas veronii Isolated from Oreochromis niloticus in Bangladesh.

Tilapia (Oreochromis niloticus) is the most widely cultured freshwater fish species in Bangladesh and worldwide. However, commercial tilapia culture systems face increasing challenges from bacterial infections. The objective of this study was to identify the bacterial isolates from infected tilapia inan intensive cage culture farm located along the Shitalakshya River in Bangladesh. Infected fish samples were collected and underwentcomprehensive clinical and post-mortem investigations, followed by phenotypic, biochemical and molecular identification of the bacterial isolates, as well as histopathological and antibiotic susceptibility examinations. Phenotypic and biochemical characterization showed similarities of the -collected isolates with Aeromonas veronii. Moreover, molecular analysis of the bacterial conserved region 16S rRNA also confirmed these isolates as A. veronii. The analysed 16S rRNA sequence (GenBank accession no. PP832815) showed a close relationship (100% identity) with A. veronii from China (GenBank accession no. MT071624) in the NCBI BLAST search, and in the phylogenetic tree, they grouped in a single clade. This close genetic relationship is also supported by the low genetic distance between the isolates. Histopathological analysis revealed gross pathological changes like necrosis, hypertrophy and inflammation in muscle tissues. The isolates were found to be sensitive to multiple antibiotics but resistant to trimethoprim and sulphamethoxazole. This study investigated the presence of A. veronii infection in tilapia (O. niloticus) in an intensive cage culture farm in Bangladesh.

Coexistence of aerobic vaginitis caused by Enterococcus faecalis and human papilloma virus (HPV) infection as a risk factor for the development of cervical intraepithelial lesions

<p><strong>Aim</strong> To determine the prevalence of aerobic vaginitis (AV) caused by <em>Enterococcus faecalis (E. faecalis)</em> in human papilloma virus (HPV)-positive women with pathological Pap test and to determine the most prevalent HPV type associated with <em>E. faecalis</em> infection.<br /><strong>Methods</strong> This prospective study was conducted at the Gynaecology Centre "Dr. Mahira Jahić" Tuzla and Primary Health Care Centre Tešanj (Bosnia and Herzegovina) in the period between February 2023 and March 2024. The research included 200 women aged 25 to 50 years. The examined group consisted of 100 women with a pathological (examined group) and 100 with a normal (control group) Pap test result.<br /><strong>Results</strong> Pathological Pap smears were found in 60 (out of 100; 60 %) women in the examined group: cervical intraepithelial neoplasia (CIN) 1 and CIN 2  in two women, respectively, CIN 3 in seven, atypical squamous cells of undetermined significance (ASCUS) in 29 and atypical squamous cells-high-grade cannot be excluded (ASC-H) in two women. Overall (both groups) prevalence of <em>E. faecalis</em> was 25.5% (51women); in 45 (22.5%) women <em>E. faecalis</em> was the only bacterial isolate, of which 42 (21%) in the examined group and three (1.5%) in the control group. High-risk HPV types were found in 62 (out of  100; 62%) women with the pathological Pap smear test. The association of <em>E. faecalis</em> and high-risk HPV positive women was found in 35 (35%) cases (moderately positive correlation; r=0.198).<br /><strong>Conclusion</strong> E. faecalis is very common in HPV 16 and 18 positive women and may represent a risk factor in the development of cervical intraepithelial lesions.</p>

Detection of E. coli using bacteriophage T7 and analysis of excitation‑emission matrix fluorescence spectroscopy

Conventional detection methods require the isolation and enrichment of bacteria, followed by molecular, biochemical, or culture-based analysis. To address some of the limitations of conventional methods, this study develops a machine learning (ML) approach to analyze the excitation-emission matrix (EEM) fluorescence data generated based on bacteriophage T7 and E. coli interactions for in-situ detection of live bacteria in the presence of fresh produce homogenate. We trained classification models using various ML algorithms based on the 3-D EEM data generated with bacteria and their interactions with a T7 phage. These ML algorithms, including linear Support Vector Classifier and Random Forest, demonstrate high accuracy (>0.85) for detecting E. coli at 102 CFU/ml concentration within 6 hours. Additionally, these ML models can differentiate among different E. coli concentration levels. For example, the Gaussian Process model achieved an accuracy of 92% in detecting different concentration levels of live E. coli. Application of these ML methods to detect E. coli in spinach homogenate yielded an accuracy of 89% using the linear-SVC model. Furthermore, feature selection techniques were employed to reduce the dimensionality of the data, revealing that only six features were necessary for achieving classification accuracy (>0.85) of spinach homogenate samples containing 102 CFU/ml of E. coli. These findings highlight the potential of this novel bacterial detection methodologies, offering rapid, specific, and efficient solutions for applications in food safety and environmental monitoring.