Food Poisoning Research Articles

Isolation and identification of bacterial pathogens associated with foodborne illnesses in stool samples: A prospective study

Introduction: Foodborne illnesses significantly impact public health, necessitating the isolation and identification of bacterial pathogens such as Salmonella, Escherichia coli, and Staphylococcus aureus. This study aims to isolate and identify bacteria from stool samples of individuals exhibiting food poisoning symptoms, enhancing understanding and prevention of these infections. Methods: This prospective study, conducted at GSL Medical College from August to September 2021, involved individuals over 18 years with food poisoning symptoms. Stool samples were collected, cultured on selective media, and analyzed for bacterial identification using Gram staining and biochemical tests, followed by statistical analysis of prevalence and symptoms. Results: The study included 100 participants, with 52% male and a majority (40%) aged 31–45 years. Abdominal pain (80%) and diarrhea (75%) were the most common symptoms. The predominant isolates were E. coli (30%) and Salmonella spp. (25%), with symptom correlations revealing distinct clinical presentations among the pathogens. Conclusion: This study underscores the significant role of pathogens in foodborne illnesses, particularly E. coli and Salmonella spp. The correlation between these pathogens and symptoms highlights diverse clinical presentations, emphasizing the need for effective food safety measures and public health interventions to mitigate the impact of foodborne infections on community health.

The possible link between venison poisoning and Sarcocystis spp. infection in Lithuania: a case report.

There have been report of possible food poisoning in Lithuania following the consumption of raw steaks from a roe deer (Capreolus capreolus). Two people experienced symptoms of abdominal pain, watery diarrhea, loss of appetite, and nausea. The parasitological examination of the roe deer revealed macrocysts of Sarcocystis sp. to the naked eye. Microscopic sarcocysts of Sarcocystis spp. were detected in different muscles, and a high sarcocyst load was observed in the roe deer muscles by a light microscope. Based on molecular results, S. capreolicanis, S. gracilis, S. linearis, S. oviformis, and S. silva were identified in meat samples of the roe deer. This is the first report of a possible link between venison poisoning and Sarcocystis spp. infection in Lithuania.

In-vitro Evaluation of Trichoderma Species and Some Selected Botanical Extracts against Alternaria helianthi (Hansf) Causing Alternaria Blight of Sunflower

This study investigates the in vitro efficacy of bioagents and selected botanicals in managing Alternaria helianthi, the pathogen responsible for Alternaria blight in sunflower (Helianthus annuus L.). Two bioagents, Trichoderma harzianum and Trichoderma viride, and six botanicals—Azadirachta indica (neem), Lantana camara (lantana), Curcuma longa (turmeric), Aloe barbadensis (aloe vera), Withania somnifera (ashwagandha), and Mentha spicata (pudina)—were evaluated using the dual culture and poison food technique, respectively. The results showed that T. harzianum exhibited the highest inhibition of mycelial growth at 76.32%, while neem extract was the most effective among botanicals, achieving a maximum inhibition of 75.03% at 15% concentration. These findings suggest the potential of bioagents and botanicals in eco-friendly management strategies for Alternaria blight in sunflower cultivation.

Avian IgY Antibodies for the Immunoprophylaxis and Therapy of Experimentally Infected Chicken by Salmonella enterica, serovar Enteritidis

The possibility of preventing salmonellosis in chicken (pathogen agent — Salmonella enterica, serovar Enteritidis (SE)) by using specific yolk IgY was discussed in presented article. There has been raising the number of reports in the scientific periodicals about increased antibiotic resistance of Salmonella. SE is one of the most common Salmonella serovars worldwide that causes food poisoning in humans. At the same time, infected chicken eggs and meat serve as the main source of infection. For extraction of the IgY, 10 laying hens were used. All layers were hyperimmunized by a vaccine containing live attenuated strains of Salmonella enterica, ser. Enteritidis and Salmonella gallinarum Pullorum. The control group with the same number of layers was not vaccinated. A feed additive for newly hatched chicken and solution with purified and isolated IgY were prepared from egg yolks of immunized and non-immunized birds. During oral administration as a feed additive in per os and intraperitoneal experimental infection by a lethal dose of SE the examination of the efficiency of the obtained product on six-day-old chickens were conducted. Thereby, conducted examinations have shown the high efficiency of the feed additive and preparation with purified immunoglobulins prepared from the yolks of eggs of immunized chicken in experimental infection with a lethal dose of Salmonella Enteritidis.

Nutrition, hydration and supplementation considerations for mountaineers in high-altitude conditions: a narrative review

Staying and climbing in high mountains (>2,500 m) involves changes in diet due to poor access to fresh food, lack of appetite, food poisoning, environmental conditions and physiological changes. The purpose of this review is to summarize the current knowledge on the principles of nutrition, hydration and supplementation in high-altitude conditions and to propose practical recommendations/solutions based on scientific literature data. Databases such as Pubmed, Scopus, ScienceDirect and Google Scholar were searched to find studies published from 2000 to 2023 considering articles that were randomized, double-blind, placebo-controlled trials, narrative review articles, systematic reviews and meta-analyses. The manuscript provides recommendations for energy supply, dietary macronutrients and micronutrients, hydration, as well as supplementation recommendations and practical tips for mountaineers. In view of the difficulties of being in high mountains and practicing alpine climbing, as described in the review, it is important to increase athletes’ awareness of nutrition and supplementation in order to improve well-being, physical performance and increase the chance of achieving a mountain goal, and to provide the appropriate dietary care necessary to educate mountaineers and personalize recommendations to the needs of the individual.

Etiological characteristics and whole genome sequence analysis of Clostridium perfringens causing a food poisoning outbreak

This study investigated the etiological characteristics and whole genome sequencing of clostridium perfringens (CP) from a food poisoning outbreak. Multiplex real-time PCR was employed to screen pathogens in collected samples. Based on the preliminary screening results, the isolation, cultivation, and identification of suspected pathogenic CP bacteria were performed. The nucleic acid tests were conducted for CP-related virulence genes on CP isolates, anal swab specimens and food samples. The molecular typing of CP isolates was analyzed by using pulsed-field gel electrophoresis (PFGE). A phylogenetic tree was established to analyze nucleotide and amino acid homology after sequencing the PLC gene. The whole genome sequencing and gene annotation on the representative strain QD2022FB4-CP4 were performed to explore its drug resistance, toxin genes and biological characteristics. The analysis revealed that the food poisoning was triggered by F-type Clostridium perfringens, which infected the consumers by contaminating'roujiamo' sandwiches. The whole genome sequencing of the strain QD2022FB4-CP found that it had active metabolic processes, multiple virulence genes and multidrug resistance characteristics.

Application of a liposomal subunit vaccine in chickens for reduction of Campylobacter gut colonisation

Abstract Introduction Campylobacter are the most common cause of food poisoning, which manifests itself in diarrhoea of varying severity. Additionally, because of the increasing number of people with immune deficiencies, more frequent serious complications of Campylobacter infections are being observed. The main source of infection is the consumption of contaminated poultry meat, which is a consequence of the insufficiency of current hygiene and biosecurity to control Campylobacter or eliminate it from the poultry food chain. Material and Methods Two hybrid proteins, presenting selected epitopes of the Campylobacter antigens CjaD and EF-Tu, were developed based on the highly immunogenic proteins CjaA and CjaC. Four groups of chickens were vaccinated with different preparations (a mixture of both hybrid proteins encapsulated in anionic or neutral liposomes) and different doses (a single dose given on the day of hatching or two doses given on days 1 and 14 of life). The number of Campylobacter was assessed in the intestinal contents of vaccinated birds. Results No statistically significant differences in colonisation levels were observed between chickens immunised with neutral liposomes containing hybrid proteins and their non-immunised counterparts, regardless of dosage regimen. Conclusion Although immunisation of chickens did not produce the expected results, the approach used has great potential, which is worth further investigation and development.

Genomic perspectives on foodborne illness

Whole-genome sequencing of bacterial pathogens is used by public health agencies to link cases of food poisoning caused by the same source of contamination. The vast majority of these appear to be sporadic cases associated with small contamination episodes and do not trigger investigations. A "contamination episode" refers to one or more contamination events from a single source over a period of time. We examine clusters of sequenced clinical isolates of Salmonella, Escherichia coli, Campylobacter, and Listeria that differ by only a small number of mutations (SNPs) to identify features of the underlying contamination episodes. These analyses provide additional evidence that the youngest age groups have greater susceptibility to infection by Salmonella, E. coli, and Campylobacter than older age groups. This age bias is weaker for the common Salmonella serovar Enteritidis than Salmonella in general. A large fraction of the contamination episodes causing sickness appear to have a long duration. For example, 50% of the Salmonella cases are in clusters that persist for almost 3 y. For all four pathogen species, the majority of the cases were part of genetic clusters with illnesses in multiple states and likely to be caused by contaminated commercially distributed foods. Salmonella infections in infants under 3 mo are predominantly acquired from the same contaminated food, pet food, or environmental sources as older individuals, rather than infant formula contaminated during production.

Bio-efficacy of Biochar's against Fusarium oxysporum f.sp. radicis-cucumerinum inciting Root and Stem Rot Disease of Cucumber in In vitro Condition

To assess in vitro evaluation of bio-efficacy different three biochar's viz., Green house waste (GHW), Eucalyptus wood (EW) and Citrus wood (CW) were evaluated at different three concentrations (each @ 1, 2 and 3%) against F. oxysporum f.sp. radicis-cucumerinum causing root and stem rot of cucumber by using poison food technique. Among the three different biochars, Green house waste (GHW) was found most effective by showing minimum mycelial growth of 50.78, 40.10 and 29.88 mm and 43.58, 55.44 and 66.80% per cent growth inhibition at 1, 2 and 3% concentrations, Whereas, Citrus wood (CW) depicted highest mycelial growth 66.26, 55.88, 44.30 mm with 26.38, 37.91 and 50.78% lowest per cent growth inhibition of the pathogen at same above concentrations, respectively.

Bacillus Cereus Control Using Lactoferrin and/or Propolis Incorporated Carboxymethyl Cellulose Edible Coating in Chilled Beef Fillets

Background: Bacillus cereus (B. cereus) is a major food poisoning bacterium that resists high cooking temperatures. The aim of this study was to prepare Carboxymethyl Cellulose (CMC) edible coats incorporated with lactoferrin, propolis and mixtures of them and evaluate their effects on B. cereu) experimentally inoculated in beef fillets stored in the refrigerator till spoilage. Methods: Prepare Carboxymethyl Cellulose (CMC) edible coats incorporated with lactoferrin, propolis and mixtures of them and evaluate their effects on bacillus cereus that experimentally inoculated in beef fillets stored refrigerated till spoilage. Results: The results revealed a significant and gradual increase in B. cereus count from ~8 log CFU/g. In first day the count reached a population of 9.69±0.12 and 9.27±0.02 log CFU/gm. at 9th day of storage in un-coated group (BF/BC) and blank coated group (CMC/BF/BC), respectively. While coated groups showed a significant decrease in B. cereus count from ~8 log CFU/gm. in first day of inoculation to 5.80±0.1, 5.41±0.06, 5.11±0.02 in CMC/BF/BC/LF, CMC/BF/BC/PR and CMC/BF/BC/LF/PR groups, respectively. The groups with coated beef showed a significant decrease (P<0.05) in B. cereus count from approximately 8 log CFU/g. On the first day of inoculation to 5.80, 5.41, and 5.11 in the Lactoferrin fortified CMC edible coating beef fillet, propolis fortified CMC edible coating beef fillet, and Lactoferrin/propolis fortified CMC edible coating beef fillet groups, respectively, on the 21st day of refrigerated storage. A synergistic antimicrobial effect between Lactoferrin and propolis was shown against B. cereus. Conclusion: It was revealed that CMC fortified with LF/PR plus its anti-B. cereus effect increased the shelf life and enhanced sensory profile of beef fillets during the 21 days of storage.

Biocontrol of Salmonella Schwarzengrund and Escherichia coli O157:H7 planktonic and biofilm cells via combined treatment of polyvalent phage and sodium hexametaphosphate on foods and food contact surfaces

Salmonella Schwarzengrund and Escherichia coli O157:H7 are ones of foodborne pathogens that can produce biofilms and cause serious food poisoning. Bacteriophages are an emerging antibacterial strategy used to prevent foodborne pathogen contamination in the food industry. In this study, the combined antibacterial effects of the polyvalent phage PS5 and sodium hexametaphosphate (SHMP) against both pathogens were investigated to evaluate their effectiveness in food applications. The combined treatment with phage PS5 (multiplicity of infection, MOI = 10) and 1.0% SHMP inhibited the growth of S. Schwarzengrund and E. coli O157:H7, and the viable counts of both decreased by more than 2.45 log CFU/mL. In KAGOME vegetable and fruit mixed juice, the combined treatment with PS5 (MOI = 100) and 1.0% SHMP also resulted in significant pathogen inactivation at 4 °C after 24 h. PS5 (1010 PFU/mL) and 1.0% SHMP showed stronger synergistic effects on biofilm formation and the removal of established biofilms on polystyrene plates. Additionally, we evaluated their combined effects on reducing the biofilms of S. Schwarzengrund and E. coli O157:H7 on glass tubes and cabbage leaves at 4 °C. These findings indicate the utility of this approach in the biocontrol of the planktonic and biofilm cells of S. Schwarzengrund and E. coli O157:H7 on foods and food contact surfaces.

Diversity of Staphylococcus aureus enterotoxin genes and its potential impact on severity of mastitis in dairy cows.

Staphylococcus aureus enterotoxins (SEs) can lead to food poisoning and appear to play a pivotal role in the development and severity of mastitis. This study aimed to evaluate whether the presence and diversity of S. aureus Enterotoxin genes mastitis-causing are associated with the development and severity of mastitis in dairy cows. Comparative analysis of S. aureus isolates from cows with clinical and subclinical mastitis (103 isolates each, CM and SM respectively) was conducted, examining the occurrence of enterotoxins (sea-see, seg-sez, sel26, sel27, sel01 and se02). Clinical isolates exhibited greater diversity, with 14 genes compared to four in subclinical isolates, and higher gene frequency (100% vs. 86.4%). Among the classical SEs, only sec (1%) and sed (4.9%) were detected in clinical isolates, suggesting a potential insignificance in the development of mastitis. Conversely, the selw and selx genes were among the most prevalent in both clinical and subclinical mastitis isolates. While their role in human food poisoning has not been definitively established, they appear to influence mastitis development. Statistical analysis revealed significant differences in clinical isolates carrying seg, seh, sei, seo, seul1, seul2, selw, and selx genes compared to subclinical isolates, suggesting their potential involvement in the severity of mastitis. Determining the frequency of SE gene and its role in mastitis establishment and in clinical mastitis severity, as well as understanding how enterotoxins contribute to pathogenicity in clinical and subclinical mastitis, is crucial for guiding optimal medical therapy for animals and establishing effective veterinary treatments to mitigate economic losses in the dairy industry.

Designing boron-doped carbon dot-functionalized COFs for fluorescence screening and liquid chromatography tandem mass spectrometry detection of toxins

In recent years, mushroom poisoning has been one of the most important factors of food poisoning in China, timely identification of the toxins contained in mushrooms is crucial for the treatment of patients. In this study, boric acid carbon dots (BA-CDs) can undergo specific boron affinity reactions with amatoxins toxins containing o-dihydroxyl groups by means of boric acid groups. Functional covalent organic framework (COF) and BA-CDs were combined to design a adsorbent with boric acid group (COF@VBC@BA-CDs) was designed to meet the requirements of both fluorescent and pretreated materials for amatoxins. The mushrooms and urine samples were rapid screening using fluorescence detection, and then, for positive samples, the target analytes on the COF@VBC@BA-CDs are collected and eluted for next liquid chromatography tandem mass spectrometry (LC-MS/MS) detection. According to the fluorescence characteristics of COF@VBC@BA-CDs, the fluorescence quenching intensity was linearly correlated with the concentration (2-200 μg/L) and the detection limit was 1.2 μg/L. Meanwhile, the detection limit of LC-MS/MS was 0.5 μg/kg for musroom and 0.2μg /L for urine, as well as the recovery rate was 72.7-110.1%. This noval method meets the methodological requirements and can be used for actual sample analysis.

Reports of tropane alkaloid poisonings and analytical techniques for their determination in food crops and products from 2013 to 2023.

Food safety is crucial to attaining food security and sustainability. Unsafe foods for human and animal consumption lead to product recalls and rejection, negatively impacting the global economy and trade. Similarly, climate change can adversely affect the availability of safe and nutritious food at the table. The changing climatic conditions and global food trade and transport can make the movement of toxic plants possible, resulting in food crops being increasingly invaded by some species of plants that produce toxic secondary metabolites, such as tropane alkaloids (TAs).Datura stramoniumfrom the Solanaceae plant family is an invasive and virulent plant that produces high amounts of two TAs, atropine and scopolamine. Various food poisoning events following accidental or deliberate ingestion of foods contaminated by atropine and scopolamine from seeds ofD. stramoniumhave been recorded in different locations globally. Due to these incidents, regulatory agencies require the development of plant toxin detection methods that can be used in the food chain as early as possible. This systematic review thus focuses on the TA determination techniques in food and feeds published between 2013 and 2023. A particular focus was given to the sample preparation methods, the improvements of each technique claimed, and data to support the performance of each method, especially the ability to measure at or below the maximum level. The review concludes with other technological advancements, including rapid spectroscopy, electrophoresis, and colorimetric methods, as well as the possibility of coupling with smartphones for use in on-farm detection and the challenges in applying them.

Phytochemical characterization, antioxidant activity and anthelmintic effect of Carica papaya L. (Caricaceae) seeds in Lohman Brown cockerels

Helminths are diseases which exist frequently with the poultry. To cure these parasitosis, breeders use very often synthetic products which are source of food poisoning for the consumers. However, past studies showed that some plants are known to their anthelmintic properties. For sanitary biosafety, the current study aimed to gauge helminthic capacity of papaya seed and established the relationship between phytochemical composition and anthelminthic activity. Qualitative and quantitative phytochemical analyses were carried out to identify phytoconstituents endowed with anthelmintic activity. Bioguided analyses were carried out in three months upon 260 Lohmann Brown cockerels which were divided in 5 batches: T-, T+, GP1, GP2, and GP. Cockerels of batches GP1, GP2 and GP3 received in their food papaya seed powder (5%) per month, respectively for one, two and three days. Batch T- did not receive any treatment but cockerels of batch T+ were treated by Levamisole (25%) on day per month. Phytochemical tests revealed the presence of alkaloids, reducing compounds, flavonoids, tannins, proteins and coumarins as chemical group in the seeds. Bioguided analyses proved that the reduction rates of number of EPG in batches T+, GP1, GP2, et GP3 are respectively: 90.08; 28.00; 53.00, and 90.35, but it was negative in batch T-. Batches T+ and GP3 presented higher average weekly gains, while average cumulative weight gains were identical for batches T+ and GP3. Nevertheless, it was batch GP3 that provided the lowest consumption index. In fact, cockerel treatment with papaya seed present almost the same anthelminthic efficacy as Levamisole.

Virulence and resistance genes in <i>Campylobacter spp</i> isolated from asymptomatic children at Kibera slum in Nairobi, Kenya

BACKGROUND Campylobacter spp. are among the global causes of food poisoning and harbour virulence and resistance genes which play a role in invasion and antibiotic resistance. This study aimed to detect selected resistance and virulence genes in archived Campylobacter spp. isolated from children in Kibera, Nairobi, Kenya. MATERIALS AND METHODS This was a retrospective laboratory-based cross-sectional study on archived samples of Campylobacter spp. at the Department of Public Health, Pharmacology and Toxicology, University of Nairobi. Campylobacter isolates were cultured on modified charcoal cefoperazone deoxycholate agar (mCCDA) media. The presence of selected virulence genes and resistance genes was detected by multiplex polymerase chain reaction (mPCR). STATA13 was used in data analysis. Binary logistic regression was used to ascertain the association between the genes and the Campylobacter spp. RESULTS The Campylobacter spp. under this study were Campylobacter jejuni (C. jejuni) (50%), Campylobacter coli (C. coli) (38%) and Campylobacter lari (C. lari) (12%). The virulence genes detected were cgtB 37 (25%), pldA (20%), cdtA (17%), cdtB (16%), cdtC (6%), wlaN (5%), iam (4%), ciaB (4%) and VirB11 (3%). The virulence gene waaC was not detected in any of the isolates. The resistance genes detected were TetA (42%), Tet B (26%), GyrB (18%) and GyrA (14%). CONCLUSION The study revealed the presence of various virulence and resistance genes in Campylobacter spp. isolated from children. This information could help guide the development and enhancement of guidelines for the management of Campylobacter infections.

UJI FORMALIN DAN BORAKS PADA IKAN ASIN, IKAN SEGAR, TAHU

Food safety is an effort that needs to be considered to prevent food from chemicals that can interfere, harm and endanger health. One of the causes of food poisoning is the presence of hazardous food additives in the food such as borax and formalin. Salted fish, fresh fish and tofu are foods that are widely favored by the public because of their taste and high nutritional content. The high water content of the three samples makes the samples not last long. The large number of foods sold on the market makes consumers have to be smart and careful in choosing food ingredients to consume. Formalin is a preservative that is sometimes misused as a preservative in food ingredients, the purpose of this study was to determine whether or not there was formalin and borax content in the three samples. One of the causes of food poisoning is the presence of food additives such as formalin, and borax, and textile dyes in a food. The purpose of this study was to test formalin and borax in the samples tested, namely salted fish, fresh fish and tofu. The method used was directly on the samples to be tested, namely salted fish, fresh fish, and tofu by testing the formalin and borax content in the samples. The conclusion of the results of this activity is that salted fish, fresh fish and tofu show positive for formalin and negative for borax.

Rapid and sensitive detection of staphylococcal enterotoxin E using a time-resolved fluorescence immunochromatography assay

Staphylococcal food poisoning is a significant foodborne illness caused by staphylococcal enterotoxins (SEs). Immunoassays have become the primary method for rapidly detecting harmful bacteria and toxins because of their excellent sensitivity and specificity. However, these assays have limitations in that they cannot differentiate between types of SEs and do not provide rapid, on-site, quantitative testing. In this study, a time-resolved fluorescence immunochromatography assay (TRFICA) was developed specifically for detecting staphylococcal enterotoxin E (SEE), which is commonly found in dairy products. Compared with a double antibody sandwich enzyme-linked immunosorbent assay, which had a detection limit of 0.028 ng/mL, TRFICA demonstrated comparable sensitivity, enabling SEE quantification with a detection limit as low as 0.081 ng/mL in infant formula. Validation by spiking infant formula samples confirmed no cross-reactivity with analogs (recoveries ranged from 93.17% to 128.77%). Furthermore, with an 8-min reaction time and interpretation delivered by a portable TRFICA strip reader, our method demonstrates potential for use in mobile and on-site detection. This study describes a rapid, easy, and reliable method for detecting trace levels of SEE in infant formula, which could serve as an early screening tool toward preventing food poisoning in infants and children.

Rapid, Sensitive, and Specific Microbial Whole-Cell Biosensor for the Detection of Histamine: A Potential Food Toxin.

Histamine is a biogenic amine; its level indicates food quality, as elevated levels cause food poisoning. Therefore, monitoring food at each step during processing until it reaches the consumer is crucial, but current techniques are complicated and time-consuming. Here, we designed a Pseudomonas putida whole-cell biosensor using a histamine-responsive genetic element expressing a fluorescent protein in the presence of the cognate target. We improved the performance of the proposed biosensor by optimizing the chassis, genetic regulatory element, and reporter gene. A sensitive and rapid biosensor variant was obtained with a limit of detection (LOD) of 0.39 ppm, manifesting a linear response (R2 = 0.98) from 0.28 to 18 ppm in 90 min. The biosensor showed minimal cross-reactivity with other biogenic amines and amino acids prevalent in food, making it highly specific. The biosensor effectively quantified histamine in spiked fish, prawn, and wine samples with a satisfactory recovery. Additionally, a colorimetric sensor variant PAlacZ was developed enabling histamine quantification in seafood via a smartphone application, with an LODgray of 0.23 ppm, exhibiting a linear response from 0 to 2.24 ppm. Overall, this study reports an efficient, specific, and highly sensitive biosensor with strong potential for the on-site detection of histamine, ensuring food safety.

In Vitro Antioxidant and Antifungal Activities of Extracts from Ocimum basilicum Leaves Validated by Molecular Docking and ADMET Analysis.

The aim of study is to investigate the various mechanism-based antioxidant and anti-fungal properties of a hydroalcoholic extract of Ocimum basilicum L leaves. Additionally, conduct molecular docking to simultaneously validate in vitro activities. Also, perform ADMET analysis to know pharmacokinetic properties and its toxicity for its safety. Prior extract's qualitative analysis has been performed to identify the bioactive compounds by phytochemical tests and GC-MS analysis. Different mechanism-based in vitro antioxidant methods are studied; in different methods, different IC50 values have come, which revealed the extract's antioxidant potentials. The antifungal potential of the extract has been observed by performing a modified poison food assay and a time-killing curve assay. In silico analysis with the human peroxiredoxin 5 enzyme (PDB ID: 1HD2) and secreted aspartic proteinase (PDB ID: 2QZX), which predict extract biological activity, has shown promising results of Ocimum basilicum L extract. In silico findings confirm the in vitro experimental outcome of the extract. The different IC50 values of extracts in different mechanisms indicate their therapeutic potential, and that encourages further research in formulation development. The time-killing assay method gives information about a dynamic interaction between extract and microbial strain. Concentration-dependent antifungal studies have significance in formulation development for dose determination.