Duration Of Incubation Research Articles

BLOODSTREAM INFECTIONS AND ITS ANTIBIOGRAM OF INTENSIVE CARE UNITS IN A TERTIARY CARE HOSPITAL OF CENTRAL INDIA

Objective: To perform bacteriological analysis and to study drug susceptibility patterns of isolates from bloodstream infections. Methods: This retrospective study was conducted in the microbiology department of NSCB Medical College, Jabalpur, spanning from July 2023 to December 2023. The study included blood culture samples from all adult intensive care unit patients of the medical college. Blood samples were collected with aseptic guidelines and cultured for 7 days. Growths were identified using standard biochemical tests and subjected to sensitivity testing according to Modified Kirby-Bauer’s disk diffusion method. Data for the source of blood collection and duration of incubation were noted and compared. Results: A total of 100 (21.7%) pathogens were isolated from 460 bacteremia suspect patient blood specimens. Pseudomonas spp. Were predominant organisms recovered followed by Klebsiella spp. and Staphylococcus aureus, coagulase-negative staphylococci, and Acinetobacter spp. were the primary pathogens isolated. Carbapenems, glycopeptides, and aminoglycosides were the most effective drugs for treating bacteremia. Conclusions: Early diagnosis and proper antimicrobial therapy lead to successful treatment of sepsis and decreased morbidity and mortality. Antibiogram of a particular area helps in rationalizing verified treatment strategies.

Optimal incubation duration of liquid cultures for assessing culture negative conversion in patients with Mycobacterium avium complex and Mycobacterium abscessus pulmonary diseases.

Mycobacterial liquid culturing typically requires six weeks or longer, primarily because of the slow growth rate of Mycobacterium tuberculosis. This study aimed to evaluate the potential of shortening the duration of mycobacterial liquid culturing in healthcare settings with high prevalence rates of non-tuberculous mycobacteria. We retrospectively analyzed the relationship between mycobacterial species and time to positive testing of liquid cultures from sputum samples using the Mycobacteria Growth Indicator Tube system over a 3.5-year period beginning in July 2020 at a university hospital in Japan. We analyzed 15,147 sputum culture samples and found a 1.1% positivity rate for Mycobacterium tuberculosis complex, while the rates for Mycobacterium avium complex and Mycobacterium abscessus were 17.6% and 2.1%, respectively. The median time to positivity was 17 days for Mycobacterium tuberculosis complex, 9 days for Mycobacterium avium complex, and 4 days for Mycobacterium abscessus. Comparing a 4-week culture period with an eight-week period, the positivity rates for Mycobacterium avium complex and Mycobacterium abscessus were 97.0% and 99.4%, respectively. In settings with a high incidence of non-tuberculous mycobacteria, the basic liquid culturing period can be safely shortened to 4 weeks without significantly compromising detection sensitivity, except for the samples that are highly suspected to contain tuberculosis, extremely slow-growing mycobacteria, smear-positive, or nucleic acid amplification testing positive.

A molecular detection method for HBV pgRNA without RNA extraction based on nucleic acid hybridization

BackgroundEarly diagnosis is critical for patients with chronic hepatitis B. Here, we utilized a sandwich RNA hybridization assay to directly detect HBV pgRNA, thereby avoiding the need for RNA extraction and purification. MethodsWe designed a hybridization cascade reaction on a solid surface using a set of oligonucleotide probes that target several highly conserved regions in pgRNA. The detection performance was validated by concurrently testing serum samples from CHB patients and healthy individuals. ResultsThe optimal detection conditions were: a universal probe coating concentration of 0.003 µg/µl with a coating duration of 2 h; capture probe and detection probe concentrations of 0.1 nM; a hybridization capture duration of 4 h; and an antibody incubation duration of 60 minutes. The sensitivity and specificity were calculated to be 91.47 % and 90.63 %, respectively. ConclusionThis novel detection method is both simple and high-throughput, making it particularly suitable for active CHB infection screening

A cost-effective protocol for single-cell RNA sequencing of human skin.

Single-cell RNA sequencing (scRNAseq) and flow cytometry studies in skin are methodologically complex and costly, limiting their accessibility to researchers worldwide. Ideally, RNA and protein-based analyses should be performed on the same lesion to obtain more comprehensive data. However, current protocols generally focus on either scRNAseq or flow cytometry of healthy skin. We present a novel label-free sample multiplexing strategy, building on the souporcell algorithm, which enables scRNAseq analysis of paired blood and skin samples. Additionally, we provide detailed instructions for simultaneous flow cytometry analysis from the same sample, with necessary adaptations for both healthy and inflamed skin specimens. This tissue multiplexing strategy mitigates technical batch effects and reduces costs by 2-4 times compared to existing protocols. We also demonstrate the effects of varying enzymatic incubation durations (1, 3, and 16 hours, with and without enzyme P) on flow cytometry outcomes. Comprehensive explanations of bioinformatic demultiplexing steps and a detailed step-by-step protocol of the entire experimental procedure are included. The protocol outlined in this article will make scRNAseq and flow cytometry analysis of skin samples more accessible to researchers, especially those new to these techniques.

Recombinant MS087-based indirect ELISA for the diagnosis of Mycoplasma synoviae.

Accurate detection is a prerequisite for effective prevention and control of Mycoplasma synoviae infection. ELISA is the most popular method for the clinical detection of M. synoviae because of its convenience, low cost, and high detection rate. However, the cross-reactivity of commercially available ELISA kits with other avian pathogen-positive sera needs to be addressed. The aim of this study was to establish an ELISA method with high specificity for the detection of anti-M. synoviae antibodies in chicken serum to evaluate the M. synoviae infection status on poultry farms. The recombinant MS087 (rMS087) protein was expressed in Escherichia coli BL21 (DE3) and purified by Ni2+ affinity chromatography. An antibody against rMS087 was generated by immunizing BALB/c mice. Bioinformatic analysis revealed that MS087 was conserved among M. synoviae strains. Western blotting and indirect immunofluorescence results indicated that MS087 was not only localized in the cytoplasm and on the membrane but also secreted by the organism. For the established ELISA method based on rMS087, the optimal antigen concentration, blocking buffer, blocking duration, serum dilution, serum incubation duration, secondary antibody dilution, secondary antibody incubation duration and colorimetric reaction duration were 2 μg/mL, 1% BSA, 3 h, 1:500, 1.5 h, 1:20,000, 2 h and 5 min, respectively. Validation of the rMS087-based ELISA revealed a cut-off value of 0.5. The coefficients of variation of both the intra-batch and inter-batch methods were less than 9%. The assay was able to differentiate positive serum against M. synoviae from antisera against nine other avian pathogens and was able to recognize M. synoviae-positive sera at a dilution of 1:1,000. Compared with the commercial ELISA method, the rMS087-based ELISA has the potential to recognize more positive sera against M. synoviae. Collectively, the rMS087-based ELISA is a reproducible, specific, and sensitive serological method for detecting antibodies against M. synoviae in chicken serum and has robust potential for large-scale serological epidemiology of M. synoviae infection on poultry farms.

Influence of storage periods of quail eggs of Radonezh breed on the results of incubation and the development of young poultry

The storage period of eggs has a negative influence on the incubation period, embryo development, egg hatchability and hatching of young animals, which ultimately reduces the economic efficiency of the poultry farm. In breeding quail farms, the collection period of hatching eggs can reach 2–3 weeks. Our studies have studied the infl uence of the storage period of hatching eggs on the duration and results of incubation, the development of young quails. These issues have been studied not enough in meat-productive quails. In Russia, similar studies have been conducted on egg-productive quails of Japanese gray breed. However, poultry with meat productivity differs in both the duration of incubation and its results due to differences in metabolism in the body. Research has been carried out at the Siberian Research Institute of Poultry Farming on the eggs of Radonezh quail breed. It has been established that storing quail eggs for more than 7 days reduced the capacity of egg hatchability by 3.3–29.0 abs.%, and the hatching of young animals by 6.2–36.5 abs.%. Extending the shelf life of quail eggs to 14–21 days increased the average incubation hour by 2.5–9.7 hours, had a statistically signifi cant infl uence during embryogenesis on the live weight of day-old quails, the use of nutrients in the yolk sac and the development of the heart, liver and muscular stomach. It is advisable to store the hatching eggs of meat-productive quails of Radonezh breed before incubation for no more than 7 days. The results of these research can be used in breeding, industrial and farm quail farms for planning the sampling time, the number of eggs laid for incubation and places for planting day-old young animals, the population of future layers, and the volumes of necessary feed.

Mechanical Microvibration Device Enhancing Immunohistochemistry Efficiency.

Immunohistochemistry (IHC) is a widely used technique in diagnostic pathology and biomedical research, but there is still a need to shorten the operation process and reduce the cost of antibodies. This study aims to assess a novel IHC technique that incorporates mechanical microvibration (MMV) to expedite the process, reduce antibody consumption, and enhance staining quality. MMV was generated using coin vibration motors attached to glass slides mounted with consecutive tissue sections. Multiple antibodies targeting various antigens were used to stain cancerous and normal tissues, with and without microvibration. Various parameters were tested, including incubation durations, temperatures, and antibody dilutions. The novel method showed the potential to achieve comparable or superior outcomes in significantly less time, utilizing over 10 times less antibody than controls. MMV improved specific staining quality, yielding stronger, and better-defined positive reactions. This was validated through a multicenter double-blind assessment and quantitative image analysis. The possible mechanisms were also investigated. MMV shortens immunohistochemical staining duration, reduces antibody usage, and enhances staining specificity, likely by accelerating antibody movement and diffusion. These improvements translate to time and cost savings, offering clinical and financial value for diagnostic pathology and biomedical research.

Biosynthesis of antimicrobial nanoparticle from Swertia spp. (Chirayita) against bacterial pathogens of poultry- A way forward to green nanotechnology and nano-medicines

The increasing prevalence of multidrug-resistant microorganisms in poultry has led to a rise in bacterial infections, causing significant economic loss. Green nanotechnology, such as silver nanoparticles (AgNPs), has the potential to address this issue by providing potent antifungal, antiviral, and antibacterial properties. This study explored the combined potential of AgNPs and the local herb Swertia chirayita against established poultry pathogens, employing a non-factorial Central Composite Design (CCD) to evaluate the factors affecting the production of nanoparticles induced by silver nitrate from the selected herb. The optimal values for temperature, wavelength, silver nitrate concentration, incubation duration, and pH were found to produce the highest nanoparticles. The functional groups in Swertia chirayita stimulated nanoparticles were confirmed using FTIR spectroscopy, and the stability of ScNPs was elucidated using zeta potential. The crystalline structure of ScNPs was confirmed using diffraction intensity patterns. Silver nanoparticles demonstrated antibacterial activity against Salmonella spp. and Escherichia coli (E.coli), both known as significant poultry pathogens, using the agar well diffusion method, with inhibition zones of 25.0 mm and 35.0 mm, respectively.This study explored the green manufacturing of silver nanoparticles by using plants and microorganisms, focusing on their antibacterial properties. The exact mechanism of synthesis and action in AgNPs is still poorly understood. Researchers should prioritize the use of accessible, easy-to-extract plants or bacteria, especially non-pathogenic and fast-growing microorganisms for safe handling. Analyzing biomolecules in plant extract, microbial biomass, or culture supernatants, including probiotic bacteria, is crucial for creating and stabilizing AgNPs, which could be effective synthetic agents. It is crucial to optimize conditions for rapid, stable, and large-scale synthesis. Based on this research, Sc-NPs may be proposed as nanomedicine for treating infections in poultry caused by E. coli and Salmonella spp.

Antioxidant Activity of Gold Nanofilms Synthesized via Cyclic Voltammetry Technique

Abstract Gold nanoparticles (AuNPs) have attracted much attention as one of the most effective agents with high catalytic activities for radical scavenging reactions. Many studies have investigated its antioxidant activity using various in vitro methods and have shown promising scavenging action. In this research, we demonstrated the synthesis of Au nanofilms (AuNFs) by cyclic voltammetry technique and evaluated for their antioxidant activity. The application of a range of voltage levels was utilized in the synthesis of AuNFs, and their antioxidant potential was subsequently assessed through in vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. All AuNFs exhibited antioxidant activity which increased as the upper potential was reduced from 1,5 V to 0,75 V. The antioxidant activity of AuNFs was shown to increase gradually with longer incubation times before the DPPH assay, indicating a significant impact on their performance. The obtained inhibition percentage was ranging from 8,26% to 22,91%. The highest antioxidant activity could be achieved by synthesizing AuNFs using 0,75V of upper potential and incubating it for 90 minutes prior to DPPH assay. The variations in the applied potential influenced the morphological characteristics of AuNFs, leading to increased surface area, while the duration of incubation was found to enhance the efficacy of the scavenging reaction between the AuNFs and DPPH. In conclusion, this cyclic voltammetry technique was shown to successfully synthesize AuNFs with significant antioxidant activity.

Initial development of a rapid, portable, stall-side ELISA for the measurement of equine adrenocorticotropic hormone.

Pituitary pars intermedia dysfunction (PPID) is a neurodegenerative disease of senior horses. Loss of dopaminergic inhibition of the melanotropes of the pars intermedia leads to increased concentrations of pro-opiomelanocortin (POMC)-derived peptides. Diagnosis is challenging due to pre-analytical variables, such as sample storage, handling, and time to analysis. Our objective was to develop an ELISA for ACTH measurement, which could ultimately form the basis for a stall-side equine ACTH test. We selected 2 ACTH-specific monoclonal antibodies, CBL57 and EPR20361-248, based on the recognition of separate epitopes, strong and rapid color change, and minimal background interference, including no cross-reactivity with themselves, each other, and the test reagents. CBL57 was chosen as the detection antibody (or secondary antibody). EPR20361-248, functionalized on superparamagnetic iron oxide beads, was chosen as the capture antibody (or primary antibody) to bind ACTH in plasma. The incorporation of magnetic beads marks the initial stage in establishing a platform that could potentially be utilized in the field, similar to other stall-side tests. The concentrations of antibodies, magnetic beads, and incubation durations were optimized. Our immunoassay detected unglycosylated rat recombinant ACTH. Further studies are ongoing to optimize and validate our assay using equine plasma and serum samples.

Molecular phylogenetic characterization of L-asparaginase-producing endophytic fungi inhabiting Prunus africana and Periploca linearifolia: Effect of incubation time and pH on enzyme production

The therapeutic use of L-asparaginase derived from bacterial sources has been constrained by various challenges, including toxicity and repression. This has prompted the exploration of alternative sources, particularly eukaryotic microorganisms like fungi, to enhance the safety and effectiveness of the L-asparaginase enzyme. This study aims to investigate the fungal endophytes inhabiting Periploca linearifolia and Prunus africana as a potential source of novel L-asparaginase. Additionally, the study seeks to examine the impact of incubation time and pH on L-asparaginase production. Standard surface sterilization techniques were used for isolation of endophytic fungi. Based on the research findings, 24 % of the fungal endophytes demonstrated a positive reaction for L-ASNase activity and were identified as Colletotrichum sydowii, Fusarium sporotrichioides, Fusarium solani, Cercospora canescens, Penicillium crustosum, Penicillium pancosmium, Phoma sp, Penicillium ubiquetum, septoria sp and Penicillium commune. A significant variation in the production of L-asparaginase based on the time of incubation and pH was observed. Most endophytic fungal isolates exhibit optimal enzyme activity on the 6th day of incubation. However, both Septoria sp and Colletotrichum sydowii recorded the highest L-asparaginase activity on the 9th day of incubation. Fusarium solani demonstrated peak activity of 12.4 ± 1.12 UI/mL on the 12th day of incubation. The optimal pH for L-asparaginase production by fungal endophytes was found to be between 5 and 6. The fungal endophytes isolated from medicinal plants have the potential to serve as sources of novel L-asparaginase. Furthermore, it is evident that fungal endophytes display considerable variation in L-asparaginase production, influenced by the duration of incubation and pH conditions.

Magnetic nanoparticles and quantum dots coupled immuno nano fluorescence assay for visual detection of HPV16-induced cervical cancer cells from cytology/biopsy samples

Biopsy-based histopathology and immunohistochemistry for cervical cancer detection are costly, time-consuming, and require expert personnel for data interpretation. We developed a simple magnetic nanoparticle (MNPs) and quantum dots (QD) coupled immuno nano fluorescence assay (MNPQDCINFA) for visual detection of HPV16-induced cervical cancer cells under UV light from cytology/biopsy samples exploiting host cancer cells expressing viral E7 protein as a biomarker. The E7 domain-specific polyclonal antibodies were generated against the 1–44 amino acid N-terminal (anti-domainN antibody) and 48–98 amino acid C-terminal domain (anti-domainC antibody). These antibodies were bioconjugated with nonfluorescent MNPs (60 % efficiency) and fluorescent QDs (66 % efficiency) to generate capturing (MNPs-anti-domainN antibody) and detecting (QDs-anti-domainC antibody) nano-complex, respectively. Assay conditions, such as concentration of capturing (20 μM) and detecting (50 nM) antibody nano-complexes and incubation duration (30 min), were standardized. The analytical sensitivity using pure HPV16 E7 protein was recorded up to 200 ng with very high specificity to differentiate from other HPV strains E7 proteins. The diagnostic performance characteristics with cytology samples showed 100 % sensitivity and specificity compared to immunofluorescence and biopsy-based histopathology analysis. The present invention can be effectively used for a quick, disposable, rapid cervical cancer cell detection system as an alternate test for immunofluorescence and histopathology.

Stability of Epinephrine in a Normal Saline Solution.

Dilution of concentrated epinephrine prior to intravenous use during clinical emergencies can delay urgent interventions. The objective of this study was to determine whether diluted epinephrine remains stable and sterile over time in common hospital settings. Epinephrine samples were prepared by clinically relevant double dilution techniques. Samples were stored in 10-mL syringes and incubated under 1 of 4 simulated hospital environments with a mixture of lighting and temperature settings: 4°C/20°C, with or without fluorescent lighting. Samples were incubated for 0, 15, 30, 60, or 90 days. Capillary zonal electrophoresis was used to quantify the concentration of epinephrine and/or presence of any degradation products. All samples were tested for the presence of bacterial growth using blood agar cultures. Diluted epinephrine samples remained stable for up to 90 days in all 4 simulated clinical storage conditions. No bacterial colony-forming units were detected in any of the environmental samples regardless of incubation duration, light, or temperature conditions. Diluted epinephrine for anticipated clinical emergencies may remain clinically useful for up to 90 days, thus improving patient safety, access to medications, and overhead costs by reducing waste.

Factors Affecting Cell Viability during the Enzymatic Dissociation of Human Endocrine Tumor Tissues.

The enzymatic dissociation of human solid tissues is a critical process for disaggregating extracellular matrix and the isolation of individual cells for various applications, including the immortalizing primary cells, creating novel cell lines, and performing flow cytometry and its specialized type, FACS, as well as conducting scRNA-seq studies. Tissue dissociation procedures should yield intact, highly viable single cells that preserve morphology and cell surface markers. However, endocrine tissues, such as adrenal gland tumors, thyroid carcinomas, and pituitary neuroendocrine tumors, present unique challenges due to their complex tissue organization and morphological features. Our study conducted a morphological examination of these tissues, highlighting the intricate structures and secondary degenerative changes that complicate the dissociation process. We investigated the effects of various dissociation parameters, including the types of enzymes, incubation duration, and post-dissociation purification procedures, such as debris removal and nontarget blood cell lysis, on the viability of cells derived from different tumor types. The findings emphasize the importance of optimizing tissue digestion protocols to preserve cell viability and integrity, ensuring reliable outcomes for downstream analyses.

Evaluation of d 18 in ovo administration of soursop (Annona muricata) leaf extract into the air space on hatch performance and physiology of Noiler chicks

Efficient poultry production can be accomplished using combined technologies. A combination of in ovo and ethno-veterinary technologies can deliver significant benefits, including reduced labour and production costs. This study evaluated the effect of in ovo administration of soursop leaf extract (SLE) on the hatching performance and physiology of Noiler chicks. A total of 550 eggs were incubated, and 460 fertile eggs were randomly distributed into 4 groups with 3 replicates on the 18th d of incubation. The eggs were then injected with 0.75 µg SLE, 1.5 µg SLE, and saline solution (negative control) at a rate of 0.2 ml in the air space. The noninjected group served as the positive control. At the end of the hatching period, the various groups were evaluated for embryo mortality, hatchability, hatch duration, organ weight, serum biochemistry, and chick quality. The results showed no significant differences (P > 0.05) in embryonic mortality, hatchability, organ weight, total protein, albumin, globulin, aspartate aminotransferase, alanine aminotransferase, and glucose among the treatment groups. However, chick weight, chick quality, and serum triglyceride levels were significantly (P < 0.05) higher in the extract-injected group. Additionally, incubation and hatch times were significantly lower (P < 0.05) in the SLE group compared to the other groups. In ovo administration of soursop leaf extract resulted in reduced incubation duration, hatch time, and embryo mortality. In conclusion, the in ovo injection of SLE improved hatch performance and chick quality.

Update on methods used for mycological testing: wide diversity and opportunities for improvement persist

Past analysis of laboratory methods used for mycology specimens revealed significant variation in practices, many of which fell short of recommended procedures. In 2016 these findings led to a set of recommendations for laboratories to consider modification of their methods where appropriate, to analyse current laboratory methods used by participants in the Royal College of Pathologists of Australasia Quality Assurance Programs (RCPAQAP) Mycology module, and to compare these to the 2016 recommendations. Seven test items, with 105-107 participants each, were analysed. Several laboratories (7-12%) did not handle specimens as recommended in an appropriate biological safety cabinet. Direct microscopy was not performed on tissue specimens 23-25% of the time. The most used staining method was potassium hydroxide with an optical brightener for fluorescent microscopy (49%) followed by Gram stain (33%). While 17-25% of laboratories used three or more media, use of four or more was uncommon (<3%). Between 9-13% of participants used only a single non-inhibitory medium for cultures. Urine specimens were incubated longer than recommended with 57% of laboratories incubating for >7days and 24% >21 days. Duration of incubation was shorter than recommended for several specimen types with 36% of skin specimens and 37-48% of tissue specimens being kept ≤21 days. For cultures kept >7 days, 13% were inspected daily but for those incubating >14 days only 3%. The methods of several laboratories remain outside recommended practice. An updated set of recommendations are made.

Acute flaccid paralysis on 11 years old child in Bali: a case report

Introduction: As medical professionals, we need to be aware of cases of acute flaccid paralysis, known as the current outbreak of polio infections in Indonesia. Polio is a highly contagious illness that primarily affects children under five. It can result in death (2-10% of those who were paralyzed) or lifelong paralysis (about 1 in 200 infections). The virus spreads from person to person mostly by the fecal-oral pathway. Contaminated foods and water are also less common routes of transmission. After the virus grows in the gut, it can enter the nerve system and paralyze people. Though it may vary from 4 to 35 days, the duration of incubation is typically 7 to 10 days. Approximately ninety percent of infected individuals either show no symptoms at all or have minor ones, hence the illness is typically not diagnosed. The purpose of this case report is to describe a case of acute flaccid paralysis and increase our awareness about the current polio outbreak. The Case: Eleven-year-old Korean child weighing 41 kg arrived to the clinic complaint of weakness in both limbs for the past five days. It happened throughout the day, particularly during the morning, and was felt abruptly. He also had instability when walking and trembling in both legs. He also experienced three episodes of watery diarrhea that day. In addition, he reported having puked twice in the two days before his clinic appointment. His calves had a motoric grade of 3 and a grade of 5 on his soles, indicating that his calves were weaker than his soles. It was discovered that the muscular tone was weak. No evidence of muscular atrophy was present. Positive tripod indication was seen. There were no abnormalities in sensibility. He was diagnosed with acute flaccid paralysis et causa suspected poliomyelitis dd Guillain Barre Syndrome dd hypokalemia paralysis et causa diarrhea and vomiting. Conclusion: Facing the polio outbreak in Indonesia, this case would serve as an example to raise awareness of acute flaccid paralysis cases, enabling early detection, differential diagnosis exclusion, and effective surveillance.

Antioxidant and biofertilizing effect of compost extracts on horticultural crops to minimize the use of agrochemicals

Excessive chemical fertilizers harm the environment, economy, and health, while compost and its extracts provide a sustainable solution. Consequently, the development of liquid organic amendments with biofertilizing and antioxidant capabilities is of significant interest in intensive agriculture. To achieve this, four Compost Extraction Protocols (CEP1–4) were applied to different compost to obtain a range of aqueous extracts. These treatments varied in temperature, incubation duration, and agitation. The raw materials for the compost used were Agri-food Waste (AW), Sewage Sludge (SS), Vegetable Waste (VW), and Olive Mill Waste (OMW). The extracts were characterized in physicochemical terms and their potential to promote radicle germination in cucumber and lettuce seeds. Additionally, counts of microbial groups associated with biofertilizing capacity were conducted. Three extracts were chosen based on the germination index to conduct an in vivo bioassay on seedlings. Finally, oxidative stress in radicles and seedlings resulting from the preceding tests was evaluated by quantifying malondialdehyde (MDA), total phenolic compounds (TPC), and ascorbate-glutathione cycle enzymes. The results established that protocols with milder temperatures, such as CEP1 and CEP4, yielded aqueous compost extracts with good biofertilizing and antioxidant properties, although the effect was dependent on crop sensitivity. Specifically, the extracts selected for the seedling trial, OMW-A CEP4, AW-A CEP1, and especially AW-A CEP4, demonstrated a remarkable biofertilizing and antioxidant properties in lettuce, by increasing growth parameters and TPC while decreasing MDA. The results indicate that aqueous compost extracts are a suitable alternative to reduce the consumption of chemical fertilizers.

Anticandidal activity of a wild Bacillus subtilis NAM against clinical isolates of pathogenic Candida albicans

BackgroundResistance to antifungal medications poses a significant obstacle in combating fungal infections. The development of novel therapeutics for Candida albicans is necessary due to the increasing resistance of candidiasis to the existing medications. The utilization of biological control is seen as a more advantageous and less hazardous strategy therefore the objective of this study is to identify the antifungal properties of Bacillus subtilis against pathogenic C. albicans.ResultsWe conducted a study to evaluate the antifungal properties of three bacterial isolates against the human pathogen Candida albicans. One of the bacterial isolates exhibited a potent antifungal activity against this fungal pathogen. This bacterium was identified as Bacillus subtilis based on the 16Sr RNA gene sequence. It exhibited inhibitory efficacy ranging from 33.5 to 44.4% against 15 Candida isolates. The optimal incubation duration for achieving the maximum antifungal activity was determined to be 48 h, resulting in a mean inhibition zone diameter of 29 ± 0.39 mm. The Potato Dextrose agar (PDA) medium was the best medium for the most effective antifungal activity. Incubation temperature of 25oC and medium pH value of 8.0 were the most favorable conditions for maximum antagonistic activity that resulted fungal growth inhibition of 40 ± 0.16 and 36 ± 0.94 mm respectively. Furthermore, the addition of 10.5 mg/ml of bacterial filtrate to C. albicans colonies resulted in 86.51%. decrease in the number of germinated cells. The fungal cell ultrastructural responses due to exposure to B. subtilis filtrate after 48 h were investigated using transmission electron microscopy (TEM). It revealed primary a drastic abnormality that lead to cellular disintegration including folding and lysis of the cell wall, total collapse of the yeast cells, and malformed germ tube following the exposure to the filtrate. However, the control culture treatment had a characteristic morphology of the normal fungal cells featuring a consistently dense central region, a well-organized nucleus, and a cytoplasm containing several components of the endomembrane system. The cells were surrounded by a uniform and intact cell wall.ConclusionThe current study demonstrates a notable antifungal properties of B. subtilis against C. albicans as a result of production of bioactive components of the bacterial exudate. This finding could be a promising natural antifungal agent that could be utilized to combat C. albicans.

Ineffectiveness of phosphorus-containing amendments to reduce Pb bioaccessibility in an urban alkaline soil.

Urban soils contaminated by historical and current anthropogenic activities present an alarming human health risk requiring redress. Federal and state governments continue to lower residential soil lead (Pb) screening standards, which will likely require new risk-based approaches to address urban soil Pb contamination. Phosphorus (P) soil amendments have long been presented as a solution to sequester Pb, thereby reducing exposure risk. In this study, P-containing sources (biosolids incinerator ash, poultry litter, biosolids compost, and triple superphosphate) of varying solubilities were assessed as soil amendments to reduce Pb bioaccessibility and serve as an inexpensive remediation strategy for urban soil. Contaminated soil (1624mg kg-1Pb, pH 7.43) from Cleveland, OH, was treated with the four P-containing soil amendments at a 1:5Pb:P molar ratio and two combination treatments at 1:10Pb:P molar ratio and incubated for 3 months. A batch equilibration analysis was also conducted to assess reduction in in vitro bioaccessible Pb (IVBA Pb). Pb bioaccessibility was evaluated using US EPA Method 1340 at pH 1.5 and the Physiologically Based Extraction Test pH 2.5 at 1 and 3 months. In general, treatments were ineffective in reducing IVBA Pb regardless of IVBA extraction method, incubation duration, batch equilibration analyses, or P source. The results of this study suggest P-containing amendments are not suitable to address Pb exposure in the study soil. Site-specific efficacy testing to determine reductions in IVBA Pb from P-containing amendments should be performed before making recommendations for remediation of Pb-contaminated urban soil.