Microbial food spoilage occurs as a result of bacteria, yeast and mold growth in foods. It is a source of concern for public health and safety officials (Aar-Abbas and Halkman, 2004), which is one of the greatest challenges in food preservation because of health risks and economic losses (Nasar-Abbas and Halkman 2004). To overcome concerns about the harmful effects of these microbes on health. Chemical agents were used as industrial preservatives and it resulted in the development of resistant microbial strains, accumulation of toxic chemical residues in food and adverse or allergic reactions in consumers Ashraf et al., (2018). So many studies have focused extracting antimicrobials from safe, healthy, biodegradable,and effective natural sources that extend food shelf life (Manso et al., 2022). including alkaloids, flavonoids, phenols, tannins, terpenes, saponins and polyamines, displaying strong antifungal and antibacterial activities (Veg et al., 2009). Alsoufi and Aziz, 2026) as studied by (Djahida and Houcin, 2021) antimicrobial properties extracted from Thapsia garganica.                                     

Antimicrobial refers to any naturally occurring or synthetic chemical substance or agent produced or derived from plants or microorganisms that can kill bacteria or inhibit their growth and reproduction. Bialonska et al., (2010), by preventing bacteria from building their cell walls or other cell components Nasar-Abbas and Halkman (2004) also stop microbial reproduction by inhibiting the production of  essential protein for the replication process Studies have also shown that kernels contain a variety of chemical compounds believed to possess antimicrobial, antioxidant, antifungal and antibacterial properties (Saleem et al., 2009).
       
This study adopted the use of date pit seeds as a primary source for extracting antimicrobials, which are a major source of environmental pollution when burned.
       
Therefore, the study aimed to achieve the following:
• Evaluating the effectiveness of plant extracts as a natural means of combating microbial growth.
• Studying the effect of the extraction solvent type on the effectiveness of the antimicrobial extracts.
• Evaluating the effect of soaking time and solution type on the quantity and effectiveness of microbial extracts against microbial growth.

Experimental design
                                                                                                                           
The experiment was designed to determine the effect of the type of extraction solvent and the soaking time of date kernel powder on inhibiting the growth of different strains of microorganisms. 48 samples were collected to measure the minimum concentration of antimicrobials that inhibits microbial growth and to measure the diameter of the inhibition zone using a special ruler for these measurementmicrobes were provided by the General Company for Pharmaceutical Industries and laboratory tests were performed on Klebsiella pneumonia (10031 ATCC) American Culture Collection). Staphylococcus aureus (29737 ATCC), Candida albicans (1023 ATCC) and Saccharomyces cerevisiae (9763 ATCC), Aspergillus niger. (1015 ATCC) and Fusarium spp (76721 ATCC). ATCC: American Type Culture Collection, the world’s largest biobank stores and distributes microbial reference strains to ensure consistency of results across laboratories.It is a reference strain, specifically requested by name in international protocols (Cooper and Denny 1997). (United States Department of Health and Human Services) Control neomycin  sulfate (10 mg /ml) for Bacteria Nystatin (100 IU) for Fungi using Dimethyl Sulfoxide (DMSO).
 
Preparation of date kernel powder
                                                                                                                             
The date kernel are removed from the fruit and then cleaned. Of dirt and dust by water then drying by oven for 3 date 37c then ground into a fine powder, The resulting powder is sealed in containers, often under refrigeration. According (Venkatesan et al., 2021; Imran et al., 2021).
 
Antimicrobial extraction
 
Fifty grams (50 g) of ground date kernel powder were mixed with 100 mL of cold water (5-7°C), hot water (80°C), ethanol (95%) as extraction solvents. The mixtures were placed on a mechanical shaker (Griffin, England) for 24 h and 48 h to assess extraction efficiency.
       
After shaking, the mixtures were filtered using a vacuum filtration unit. The solvent was evaporated under reduced pressure using a rotary evaporator at 40°C. The resulting crude extracts were weighed  calculate extraction efficiency (% yield) using the formula (Venkatesan et al., 2021):.

 
The dried extracts were stored in airtight containers at 4 °C until analysis.
       
Preparation of crude antimicrobials several concentrations of crude antimicrobials were prepared to determine the lowest concentration that inhibits microbial growth  as follows: (1.25, 2.5, 5, 10, 12.5 and 15 mg/mL) of each solvent extract were prepared using DMSO as a diluent the following concentrations were prepared (Gajic et al., 2022).

Antimicrobial activity assay
 
The effectiveness of extracted antimicrobials is affected by the type of extraction solvent, the concentration of the antimicrobials and the type of plant- (AL-Saadi, 2016). Effectiveness was assessed by determining the minimum inhibitory concentration (MIC) and the average diameter of the inhibition zone.  using the well diffusion method, Mueller-Hinton agar (for bacteria) or Sabouraud Dextrose agar (for fungi) were used to inoculate sterile Petri dishes with a 1 mL aliquot of saline suspension containing roughly 10 CFU/mL of each microorganism (Coorevits et al., 2015).

Following even spreading, 100 µL of each extract concentration was added to six wells (6 mm in diameter) that were cut into the agar using a sterile cork borer. DMSO and reference antibiotics (neomycin or nystatin) were present in the control wells. For bacteria, plates were incubated at 37 °C for 24 hours and for fungi, at 28 °C for 48 hours. A digital caliper was used to measure the inhibition zone diameters (IZD) in) (Ghuman et al., 2016; Mulla et al., 2016; Wikler et al., 2006).
 
Statistical analysis
 
Every experiment was carried out in triplicate and the results were presented as mean ± standard deviation). One-way Analysis of Variance (ANOVA) was used in the statistical analysis to identify significant differences between treatments.

The effectiveness of extracted antimicrobials influenced by the type of microbe, the type of plant, the type of organic solvent used in extraction and the duration for soaking the ground raw material in organic solvents (Kothari et al., 2019).
       
It expresses the effectiveness of extracted antimicrobials ted from plants by minimal inhibitory concentration (MIC) and the diameter of zone of inhibition.

In general, MIC is considered the most basic laboratory measurement for determining antimicrobial activity against microorganisms.
 
Effectiveness of extracted antimicrobials against molds growth
 
The effectiveness of antimicrobial of date kernel extracts against mold growth was evaluated by determining the minimum concentrations of date kernel powder extracts and estimating the average diameter exacted of inhibition formed by the molds. The lowest concentration of date kernel powder extracts that inhibits Aspergillus activity after 24 hours of soaking in cold and hot water was 15mg/ml, with inhibition diameters of 11.8 ± 0.00 mm and 13.00 ± 0.00 mm (Table 1). The results also showed that the minimum concentration of antimicrobial extract of ate  kernel powder extracts that inhibits Aspergillus activity is 2.5 mg/ml, with an inhibition diameter of 10.0 mm. As for the minimum concentration of alcohol-soaked date kernel powder extracts that inhibits Fusarium, it is 10 mg/ml. With an inhibitory diameter Regarding the increased soaking time of 48 hours Table (2) we observe that the lowest concentration  of date kernel powder extracts in cold and hot water and alcohol after 48 hours of soaking showed inhibitory indicators at a concentration of 2.5 mg/ml. inhibitory diameter of 0.92  mm was observed. The concentration increased to 15 mg/ml and the inhibitory diameter to 12.46 mm for cold water and 12 mm for the extract. The ethanol extracts were more effective in inhibiting Aspergillus rot. At a concentration of 15 mg/ml, the inhibitory diameter was 21.0 mm. As for inhibiting Fusarium rot, the lowest  concentration was 10 mg/ml.




 
Effectiveness of extracted antimicrobials against bacteria
 
Antimicrobial extracts were also shown to inhibit bacterial growth, among which were Staphylococcus aureus and Klebsiella pneumoniae bacteria (Table 3 and 4). On K. pneumoniae, ethanolextract had a maximum inhibition zone diameter (IZD) of 14.0 ± 0 mm at 15 mg/mL with a 24-hour soaking duration and with a MIC value set at 10 mg/mL, IZD measured 11.46 ± 0.057 mm. Likewise, ethanol extracts on S. aureus had an IZD value of 14.13 ± 0.057 mm at 15 mg/mL. However, at 10 mg/mL MIC value, IZD measured 10.3 ± 0.0 mm.Hot-water extracts were less efficient, with MICs of 12.5 mg/mL (K. pneumoniae: IZD 9.5 ± 0 mm), suggesting that ethanol is more efficient as a solvent for extracting antibacterial agents. As evidenced previously, that aqueous extracts remain active at biologically active temperatures as high as 100°C for 30 minutes, it can be suggested that high temperatures do not reduce these active components. The antibacterial property of ethanol extracts remained consistent and had equal efficacy or greater efficacy compared with antibiotic gentamycin sulfate. Even extending the soaking duration for 48 hours did not significantly differ IZD values (Table 4), suggesting that it has more importance to focus on solvent and amount effects compared with duration. These findings support previous research suggesting that ethanol-extracted phytochemicals have more efficacy and at times surpass gentamycin sulfate.




 
Effectiveness of extracted antimicrobials against yeasts
 
C. albicans and S. cerevisiae are inhibited by date kernel powder ethanol extracts (Tables 5 and 6). IZDs rose with extract concentration after 24 hours of soaking. For instance, ethanol extracts at 15 mg/ml generated an IZD of 14.7±0 mm against S. cerevisiae and 14.53±0.0 mm against C. albicans. Alcohol is a better solvent for extracting bioactive substances like phenolics and flavonoids, as evidenced by the lower effectiveness of water extracts (Table 5). The inhibitory effects against C. albicans gradually increased at 48 hours (Table 6) and at the highest concentration of 15 mg/ml ethanol extract, the zone of inhibition reached 0.0±16.7 mm. The inhibition zones for S. cerevisiae grew, reaching 0.057±15.86 mm at 15 mg/ml ethanol. Ethanol extracts consistently showed the highest inhibition, followed by hot water, while cold water extracts were least effective. Extended soaking enhanced antimicrobial activity due to increased extraction of bioactive compounds.




       
The effectiveness of antimicrobials extracted from plants, such as ground dates, is affected by the type of microbe, extraction solvent and temperature of the extraction solvent (Kothari et al., 2019).
       
Also published that the extracts’ antimicrobial activity with ethanol is more successful at stopping the microorganisms’ growth (Hussain, 2019). This may be attributed to the fact that the alcoholic extracts contain some chemicals that have biological effects and properties to inhibit microbial cell growth. Also due to the high solubility of phenolic and flavonoid compounds, ethanol extracts demonstrated superior inhibition across molds, bacteria, and yeasts in line with previous reports (Kothari et al., 2019).
       
In line with earlier research on aqueous plant extracts, the finding that hot water extracts were more successful than cold water extracts implies that thermal energy improves the release of active compounds.
       
This confirms that high temperatures do not destroy the raw compounds extracted from date pits and  is consistent with what was published by some studies that the active components of aqueous extracts were not damaged by high temperatures, even when heated to 100°C for 30 minutes. It was published by (Atikya et al., 2014). The diameter of the inhibition zone or bacterial sensitivity of the raw extracts of guava leaves in boiling water was 22 mm. It was also noted that the diameter of the inhibition zone was (17 mm) (Nasar-Abbas and Halkman, 2004).
       
Date kernel powder’s potential as a natural antimicrobial agent was confirmed by the MIC values for ethanol extracts, which were either better or equal to those of gentamycin sulfate. Aspergillus and Candida albicans had the highest IZDs, while molds and yeasts were generally more sensitive than bacteria. Consequently, the findings encourage the use of date kernel extracts in food and pharmaceutical applications as a sustainable and environmentally beneficial substitute for artificial properties.

The type of microorganism, the extraction solvent and the soaking time all had a significant impact on the antimicrobial activity of date kernel extracts. Fusarium oxysporum, Staphylococcus aureus and Klebsiella pneumoniae showed more resistance to the extracts. Ethanol and hot-water extracts demonstrated more potent antimicrobial effects than cold-water extracts. The efficacy of the extracts produced after soaking the date kernels for 24 and 48 hours did not differ significantly. The ethanol extracts were more effective in inhibiting Aspergillus rot. At a concentration of 15 mg/ml, the inhibitory diameter was 21.0 mm. As for inhibiting Fusarium rot, the lowest concentration was 10 mg/ml.

My appreciation to Mutah University/Jordan for their provided financial support this research. The Author received financial support to conduct the practical experiments of this research. The author did not receive any financial support for the publication of this article.
 
Disclaimers
                                                                                                                                      
The opinions and findings presented in this article are those of the authors alone and may not be representative of those of the organizations with which they are affiliated. Although the writers are in charge of the information’s correctness and comprehensiveness, they disclaim all obligation for any losses, whether direct or indirect, that may arise from using this content.

They declare no conflicts of interest regarding the publication of this work. No sponsorship or funding had an impact on the study’s design, data collection, analysis, publication decision, or manuscript preparation.