Introduction

Food safety is a pertinent issue globally that affects international trade and human health. Foodborne infections pose serious risks to consumer health and place a financial strain on healthcare systems around the world, making them a major global public health concern (El-Hawary et al., 2025). A wide range of microbial pathogens that contaminate different kinds of meat products are responsible for the millions of foodborne illnesses that are reported each year. Among these pathogens, Bacillus cereus has maintained its significance as a result of its association with foodborne outbreaks and the ability to cause serious illness (Foxcroft et al., 2024). B. cereus is a rod-shaped, aerobic, or facultatively anaerobic, gram-positive, motile, spore-forming bacterium that is common in nature and can also be found in food. Vegetative cells of B. cereus can live and replicate in a pH range of 5 to 10. They can also develop in a moderately broad variety of temperatures and are highly resistant to salting. Alternatively, spores are exceptionally resilient to a variety of extreme conditions, including chilling, drying, high temperatures, and gamma- and UV-irradiation. This enables B. cereus to persist on a variety of surfaces and in the environment (Tirloni et al., 2022).

B. cereus is capable of producing a variety of virulence influences and can infiltrate the gastrointestinal mucosa through digestion, resulting in diarrhea and vomiting (Song et al., 2019). Diarrhea is related to four distinct enterotoxins: hemolysin BL (HBL, encoded by hblA, hblC, and hblD), cytotoxin K (CytK, encoded by cytK), enterotoxin FM (EntFM, encoded by entFM), and nonhemolytic enterotoxin (NHE, encoded by nheA, nheB, and nheC). However, emesis, or vomiting, is related to a tiny, acid- and heat-stable toxin produced by cesB genes (Ehling-Schulz et al., 2015). Other than food-related illness, B. cereus is linked to severe sicknesses like pneumonia, endocarditis, osteomyelitis, endophthalmitis, and necrotizing fasciitis (Ikeda et al., 2015).

The public health relevance of B. cereus has already been established worldwide, but its specific survival capabilities and possible environmental adaptations may become more significant for food safety authorities and the food industry when considering our changing climate. B. cereus presents a substantial public health concern because of the quantity of meat products consumed in Egypt and the nature of their processing. A rise in its abundance in these foods increases the danger of exposure and the propagation of antibiotic-resistant bacteria. The significance of B. cereus for public health is highlighted by theories on its environmental and temperature range adaptability (Foxcroft et al., 2024). Antibiotics are still the best way to treat microbial illness, involving those produced by B. cereus. Conversely, the prevalent application of antibacterials has caused the development of antibacterial-resistant strains, linking strains that are resistant to more than one antibiotic (Friedman et al., 2016). Thus, it is imperative to discover the profile of antibiotic resistance of B. cereus to select the appropriate medications for therapy schemes.

Numerous investigations have been carried out to find novel techniques to prolong the duration of protection of meat and its products without using chemical supplements, as there is growing concern about the present developments regarding applying different common substitutes to improve the duration of keeping meat and its products safe and improve its shelf life. This is particularly relevant given the substantial rise in the manufacturing of meat products and their role in supplying the desired flavor and taste (Nady et al., 2024).

Apple cider vinegar (ACV) is an organic by-product of apple fermentation, consisting of apple, sugar, and yeast. It shows antibacterial efficacy against gram-positive microbes. Chemicals in plants, such as organic acids, minerals, flavonoids, vitamins, and polyphenols, work together to fight bacteria and other harmful substances (Mahmoud et al., 2024). The long-term tracking and trending of B. cereus in meat products can provide information on any significant changes in their prevalence, including specific food types. Such information could provide an early indication of changing factors in the environment, food chain, and food handling, and potentially impact food safety practices and standards. This research looked into the incidence, enterotoxin genes, and antibacterial resistance patterns of B. cereus strains in some meat products in Aswan, Egypt, as well as the effectiveness of ACV as a natural preservative against B. cereus.

Materials and methods

Samples

Between March and May 2023, 180 meat product -samples—20 samples each of minced beef, beef shawarma, beef burger, beef kofta, beef sausage, chicken shawarma, chicken kofta, and chicken kabab—were -collected from retail markets in the Aswan Governorate, Egypt. Every sample was stored under 4°C after being transferred to the lab from separate sterile bags in an icebox.

Enumeration, isolation, and identification of B. cereus

Every sample (25 g) was mixed with peptone water 0.1% (225 mL) (Oxoid, CM0009B) to make sequential dilutions and homogenized in a stomacher (Seward®400) (ISO 21871, 2006). One milliliter of the initial dilution was then placed on Mannitol Egg Yolk-Polymyxin agar (MYP) (Oxoid, CM09) and incubated for 24 h at 30°C. The colony counter was used to enumerate the lecithinase activity of a typical B. cereus colony, which was pink in color and surrounded by a precipitation zone. Then, a single colony was spread on chromogenic B. cereus agar plates (Huankai). Based on Quinn et al. (2002), various apparent colonies on chromogenic B. cereus agar plates were selected and incubated for 24 h at 37°C for additional biochemical characterization (Gram stain, starch hydrolysis, catalase, nitrate reduction, Voges–Proskauer, citrate utilization, lysozyme resistance, and anaerobic fermentation of glucose). In addition, the recognition of parasporal protein toxin crystal and rhizoid proliferation were conducted (Tallent et al., 2012).

PCR for detection of B. cereus toxins profile

GeneJET™ Genomic DNA Purification Kit (Thermo Fisher, K0722) assisted in extracting genomic DNA from the positively identified B. cereus culture, and the DNA is preserved at −20°C. After using gyrB gene primers to identify the B. cereus genotype, multiplex PCR was employed to recognize eight virulence factors (hblA, hblC, hblD, nheA, nheB, nheC, cytK, and cesB). The 25 μL PCR reaction consists of 12.5 μL COSMO PCR RED Master Mix, 1 μL each of reverse and forward primers (20 pmol), 6 μL DNA, and 4.5 μL free nuclease water. The amplification was conducted by the methods previously described (Ehling-Schulz et al., 2005; Hansen and Hendriksen, 2001; Tewari et al., 2015). Table 1 displays the thorough sequence of data used. The PCR product was analyzed by gel electrophoresis in a 1% agarose stained with SYBR SAFE (0.6 g/100 mL) at 100 V for 30 m (Elbarbary et al., 2024) and captured using a UV LED (BioRad).

Examining the antibiotic sensitivity of B. cereus strains

The Kirby–Bauer disk diffusion susceptibility technique, as described by Khairy et al. (2024), was employed to assess sensitivity to antimicrobials of all B. cereus isolates. Picked fresh isolate colonies were placed in 2 mL of sterile saline, combined, and then incubated at 37°C for 24 h. After that, the turbidity of the suspension was set by matching it to the 0.5 McFarland standard solution. On Muller-Hinton agar (MH) (Oxoid, CM0337), an immersed swab (HiMedia, PW009) from an inoculum tube was streaked three times. The antibacterial disks were put and distributed superficially on the MH agar with sterile forceps. After 24 h of incubation at 37°C, the inhibition area was finally assessed. The data were interpreted under Clinical Laboratory Standards Institute (CLSI 2017), and the strains were grouped as susceptible (S), intermediate (I), or resistant (R) following Magiorakos et al. (2012). Twenty antibiotics (Oxoid, UK) were tested, including ampicillin (AMP, 10 μg), quinupristin (QD, 15 mg), cefoxitin (FOX, 30 mg), cephalothin (kF, 30 μg), ciprofloxacin (CIP, 5 μg), cefotaxime (CTX, 30 µg), imipenem (IPM, 10 μg), vancomycin (VA, 30 μg), trimethoprim-sulfamethoxazole (SXT, 1.25 μg/23.75 μg), chloramphenicol (C, 30 mg), nalidixic acid (NL, 30 μg), clindamycin (DA, 2 μg), doxycycline (DO, 30 μg), erythromycin (E, 15 μg), colistin (CT, 10 μg), tetracycline (TE, 30 mg), gentamicin (CN, 10 mg), nitrofurantoin (FD, 300 mg), rifampicin (RA, 30 μg), and kanamycin (K, 30 mg). The antimicrobial agents that were analyzed are frequently employed in the veterinary and health sectors of Egypt. For each antibiotic and isolate, the multiple antimicrobial resistance (MAR) indices were assessed. A MAR rate of < 0.2 shows that the isolates developed from a polluted source with a low risk. On the other hand, isolates with a MAR > 0.2 have been from high-risk sources of pollution (Lozano et al., 2020).

In vitro assessment of antibacterial action of ACV

The ACV utilized in this investigation was obtained from Bragg Co., USA, via Amazon. eg. It was organic raw ACV, unfiltered, 5% acidic, unpasteurized, unheated, and had the amazing mother of vinegar.

The antibacterial effect of ACV was assessed using the agar well-diffusion technique against B. cereus isolates, as earlier reported by Balouiri et al. (2016). Using 0.9% sterile saline solution, the suspension turbidity of purified bacterial culture (10⁶ CFU/mL) was under 0.5 McFarland. Subsequently, a sterile cotton swab was employed for spreading 100 µL of the sample onto Mueller-Hinton agar (Hi-Media) plates. Using a sterilized cork borer, holes (7 mm) were created in the plates. These were then filled with 100 μL of produced ACV of 10%, 30%, 70%, and 100%, and incubated at 37°C for 24 h. A negative control was recognized by sterile demineralized water, although a positive control was established via antibiotic discs (ampicillin, 10 μg). The inhibitory halo diameter was measured using a gauge (mm). Assessments were taken in triplicate to establish the mean and standard deviations of the inhibition zone, which were determined. The strains were classified as resistant (0) for diameters < 8 mm, moderately sensitive (+) for 8–20 mm, sensitive (++) for 20–30 mm, and very sensitive (+++) for diameters >30 mm.

Minimal inhibitory concentration (MIC) and minimal -bactericidal concentration (MBC) assessments

Following CLSI (2012) references, MIC and MBC were assessed via a standard broth microdilution procedure. Fresh Mueller-Hinton broth was employed to produce the bacterial suspensions for the experiment, with the concentration of bacteria adjusted to 10⁶ CFU/mL. Twofold serial dilutions of ACV from the standard solution (1,016 μg/mL) were made in sterile distilled water. A volume of each ACV dilution (100 μL) was poured on U-shaped bottom, sterile polystyrene, 96-well culture plates (Techno Plastic Products, Switzerland). Every well got 100 µL of every bacterial suspension, kept at 37°C for 24 h. The MIC was found to be the smallest amount of antibacterial agent that completely stopped visual growth (CLSI, 2012). This means that there was no growth in the well that was related to the positive and negative growth wells. The MBC was recognized as the lowest dose that produced no observable growth following the incubation period (Andrews, 2001). The MBC was identified by subculturing 10 μL of the suspension from every well on MHA. The plates were subsequently left at 37°C for 24 h, or until growth was detected in the positive growth control. Every test was approved in triplicate, and the mean ± standard error of the mean was used to present the findings.

Statistical analysis

All of the data were examined using GraphPad Prism 9.0 under one-way analysis of variance (ANOVA). Outcomes were presented as mean ± SEM with a significance value of p < 0.05.

Results

Occurrence of B. cereus

From Table 2, it is evident that the incidence of B. cereus counts in the studied samples was the highest in beef kofta samples (2.44×10³ ± 0.16×10² CFU/g), followed by beef burger (2.02×10³ ± 0.18×10² CFU/g) and beef sausage (1.88×10³ ± 0.12×10² CFU/g), with no significant variations between them while there are significant differences observed between other samples. Chicken Kabab (0.17×10² ± 0.02×10² CFU/g) and chicken shawarma reported the lowest count (0.24×10² ± 0.01×10² CFU/g).

Of the 180 RTE samples evaluated, 54 (30%) samples have B. cereus. According to the findings, the incidence of B. cereus was 45% in beef kofta and beef burgers; 35% in minced beef, beef sausage, and chicken burgers; 25% in beef shawarma; and 10% in chicken shawarma and chicken kabab (Figure 1). B. cereus was recognized on special agar plates by their distinct wavy colony shapes. Standard B. cereus colonies appear pink in color and have a surrounding area that shows precipitation, which indicates lecithinase action, and they do not ferment mannitol (this shows a positive Nagler response). Tests showed that B. cereus samples had mobility and were able to use citrate, produce Voges-Proskauer, have catalase activity, ferment glucose, and break down gelatin. Similarly, the isolates were negative for oxidase, H₂S generation, methyl red, and indole. The samples did not have any protein crystals from Bacillus thuringiensis after being stained with carbol fuchsin using the Ziehl–Neelsen method.

Figure 1. Prevalence of Bacillus cereus in the examined products. Data with different superscript letters (a–c) are significant at p < 0.05.

Toxins profile of B. cereus by multiplex PCR

Following phenotypically suspicious isolates of B. cereus, PCR analysis—relying on the identification of a specific species—gyrB—was then performed. On agarose gel, all isolates generated 475 bp PCR products that were specifically B. cereus (Figure 2). The toxins’ profile established in this research using multiplex PCR and its distribution was assessed and categorized in Figure 3. The virulence genes of B. cereus are categorized into two groups based on their pathogenic properties: enterotoxin genes (hblA, hblC, hblD, nheA, nheB, nheC, and cytK) and cereulide synthetase genes (cesB). In hemolysin enterotoxin genes, the hblA gene occurs in most of the strains (16/54) and was considerably higher than hblC (7/54) and hblD (5/54). Nonhemolytic enterotoxin, nheA, was detected in 10/54 and was significantly higher than nheB (5/54) and nheC (3/54). Cytotoxin K (cytK) was detected in 10/54 of the strains. However, only 6 out of 54 samples tested positive for the cereulide synthetase gene (cesB), suggesting that emetic bacteria are less abundant in meat products compared to diarrheal ones (Figures 3 and 4). The incidence of toxigenic factors in the obtained strains showed statistically a notable variation (p < 0.05).

Figure 2. Electrophoretic profile of amplification products of the confirmed gyrB B. cereus gene at 475 bp. Lanes 1–7: minced beef, lanes 8–16: beef kofta, lanes 17–25: beef burger, lanes 26–30: beef shawarma, lanes 31–37: beef sausage, lanes 38–39: chicken shawarma, lanes 40–45: chicken kofta, lanes 46–52: chicken burger, and lanes 53–54: chicken Kabab. M: marker (50 bp), C+: positive control, C–: negative control.

Figure 3. Prevalence of virulence enterotoxin and emetic genes of Bacillus cereus isolates. There is a significant variance at p < 0.0001.

Figure 4. Electrophoretic profile of amplification products of enterotoxin and emetic genes in Bacillus cereus: hblA at 320 bp, hblC at 750bp, hblD at 430bp, nheA at 500 bp, nheB at 770 bp, nheC at 583bp, cytK at 238bp, and cesB at 1271bp. Lanes 1–7: minced beef, lanes 8–16: beef kofta, lanes 17–25: beef burger, lanes 26–30: beef shawarma, lanes 31–37: beef sausage, lanes 38–39: chicken shawarma, lanes 40–45: chicken kofta, lanes 46–52: chicken burger, and lanes 53–54: chicken Kabab. M: marker (50 bp), C+: positive control, C–: negative control.

Antibiogram profile of B. cereus

Every B. cereus isolate underwent testing for antibacterial sensitivity to 20 chosen antibiotics. Tables 3 and 4 displayed that most of the isolates were resistant to ampicillin, cefoxitin, and colistin (100% each), while being completely susceptible to imipenem, nalidixic acid, and vancomycin, making them the most significant antibiotics. The B. cereus strain showed multidrug resistance (resistance to at least three types of antibiotics) between 0.15 and 0.85, with an average of 0.517. The investigated B. cereus strains showed statistically a notable variation in their sensitivity to different antibacterials (p < 0.05).

Antimicrobial action of ACV against B. cereus

By the agar well diffusion experiment, all ACV doses (10%, 30%, 70%, and 100%) confirmed substantial inhibitory influence against B. cereus isolates, as shown in Table 5. More research was done on the inhibition zone ranges (mm) of the several antibiotics employed in the current investigation at their concentrations. The inhibitory zone diameter was between 11.5±0.6 and 16.4±0.3 mm (10% ACV), 13.6±0.4 and 19.4±0.8 mm (30% ACV), 17.2±0.2 and 24.7±0.5 mm (70% ACV), and 19.8±0.6 and 31.6±0.8 mm (100% ACV). Using 15 B. cereus isolates, MIC and MBC were inevitably carried out to ascertain exactly the antibacterial qualities of ACV. The MIC data revealed that ACV had high antibacterial action contrary to the tested isolates, with MICs ranging from 0.14 to 1.25 mg/mL. In the context of the AVC as a bacteriocidal agent, certain isolates that were investigated exhibited an MBC rate similar to the MIC rate.

Discussion

B. cereus is one of the most prevalent foodborne bacteria, causing serious food poisoning. The condition is primarily characterized by vomiting, diarrhea, liver failure, necrotic enteritis, and abdominal discomfort. Furthermore, B. cereus is commonly described to be the most common bacteria present in a variety of meat products. It is considered a public health concern (Algammal et al., 2024) following ingestion of a contaminated meal that has more than 10⁴–10⁵ B. cereus spores or vegetative cells/g (Gao et al., 2018). The study found that there was a potential danger of B. cereus, with counts ranging from 0.17×10² ± 0.02×10² in chicken kabab to 2.44×10² ± 0.16×10² in beef kofta. Alarmingly, B. cereus counts in certain of the investigated samples (beef kofta, beef burger, and beef sausage) surpassed the allowed limits (<10³ CFU/g) indicated by the Health Protection Agency (2009) in England. Furthermore, Stenfors et al. (2008) verified that low B. cereus levels in food could cause major occurrences of food poisoning among consumers. In addition, the product is not appropriate for human consumption if the count of B. cereus exceeds 10⁴ CFU/g or mL (FSANZ, 2001). In Hong Kong, ready-to-eat foods are sorted into three groups based on the amount of B. cereus they contain: satisfactory (<10³ CFU/g), acceptable (10³–10⁵ CFU/g), and unsatisfactory (>10⁵ CFU/g). It is illegal to sell “unsatisfactory” ready-to-eat foods (Centre for Food Safety, 2014). Therefore, regulations, directives, and decisions represent the main regulatory acts applicable to veterinary, sanitary, and food safety for the protection of consumers. All regulations are centered on the protection of the agro-alimentary line, food safety, and the protection of consumer interests. This is one of the main reasons why the involvement of civil society and consumers in debating and passing veterinary, sanitary, and food safety legislation, particularly food-related legislation, is increasingly obvious (Bondoc, 2016a, b).

Consequently, it is imperative to maintain appropriate temperature control, even throughout food preparation. Cold foodstuffs must be preserved at a temperature below 4°C, while hot foods must be preserved at a temperature above 60°C to prevent food with B. cereus (Mostafa et al., 2022). All of the isolates (30%) that were obtained for this study showed the distinctive phenotypic characteristics of B. cereus and showed agreement in their biochemical reactions. The higher occurrence of beef kofta and beef burgers compared to chicken kabab and chicken shawarma may be related to the preparation methods used for each type of meat and the inclusion of intestinal parts. In addition, adding spices and vegetables to meat may enhance the risk of B. cereus contamination and serve as another cause of contamination (Shawish and Al-Humam, 2016). Our findings matched those informed by Bashir et al. (2017), Tewari et al. (2015), and Yu et al. (2020), with respective percentages of 29.3%, 30.9%, and 35%. Higher ratios of B. cereus from meat products were noted by Abd El Tawab et al. (2015), Hwang and Park (2015), and Owusu-Kwarteng et al. (2017), who reported 38.3%, 47%, and 50.5%, respectively. Low percentages were recorded by Algammal et al. (2024), Amin and Tawfick (2021), Mahmoud et al. (2024), and Mostafa et al. (2022), who found B. cereus in 21%, 24%, 22.7%, and 11.1%, respectively, of the examined samples. In addition, common risk factors that contribute to the spread of B. cereus foodborne poisoning include ambient pollution, improper food temperature processing, and improper cleaning of food production equipment and preparation surfaces (Yu et al., 2020).

B. cereus has been linked to meat additives such as rice and flour that are used in the production of meat products (Giffel et al., 1996). Meat products were most likely contaminated during handling and preparation or after they had been processed. Furthermore, leaving the items out of the refrigerator for many hours promotes B. cereus proliferation and thus enterotoxin release (Shawish and Tarabees, 2017). Moreover, incorrect management of meat products next to cooking permits B. cereus spores to produce vegetative cells that grow and cause food poisoning (Hassan et al., 2019). Furthermore, additives, seasonings, and spices are added, which are regarded as a potential hazard since they increase the quantity of Bacillus spores and thus increase the chance of food illness (Shawish and Tarabees, 2017).

Molecular approaches are more precise for making conclusive identifications. As shown in Figures 2 and 3, the housekeeping gene gyrB of B. cereus, a molecular diagnostic marker, was positive in all detected phenotypic isolates of B. cereus. The public health significance of B. cereus strains as a reason for severe food illness in humans is highlighted by the fact that all of the strains obtained in this research inherited one or more enterotoxigenic genes. This matches the findings of Algammal et al. (2024), Amin and Tawfick, (2021), Fraccalvieri et al. (2022), Mahmoud et al. (2024), Owusu-Kwarteng et al. (2017), and Tewari et al. (2015). The pathogenicity is primarily supported by numerous virulence factors and toxins expressed by the appropriate genes. The consumption of B. cereus-polluted food causes illness. B. cereus cells adhere to the human intestinal mucosa, colonize, and produce enterotoxins (Algammal et al., 2024). The primary virulence determinants associated with food poisoning produced by B. cereus are the nheABC, hblABCD, cytK, and cesB genes (Berthold-Pluta et al., 2019). Many foodborne B. cereus outbreaks have been identified globally, and the sickness manifests in emetic and diarrheal forms. Cytotoxin K, a powerful heat-labile enterotoxin, is regarded as the primary virulence factor implicated in severe diarrhea, while emetic sickness is credited primarily to the cereulide toxin (ECDC, 2019). Furthermore, B. cereus has been linked to serious human diseases such as pneumonia, neonatal bacteremia, gas gangrene, bacterial meningitis, and ocular infections (Algammal et al., 2024).

Antibiotic therapy is the principal management for B. cereus infection. However, the failure of antibacterial treatment occurs from the development of antibacterial-resistant B. cereus strains, mostly from drug misuse or the gaining of resistance genes by horizontal gene transfer (Gao et al., 2018). Thus, the detection of the antibacterial resistance outline of B. cereus is of paramount importance to public well-being. In this investigation, imipenem, nalidixic acid, and vancomycin showed significant antibacterial activity against B. cereus strains from the various items tested. Furthermore, the retrieved strains were the consequence of high-risk contamination, as shown by the concerning MAR value of 0.517 (>0.2) (Qenawy et al., 2024). These outcomes coincide with those validated by Algammal et al. (2024) and Ikeda et al. (2015), who found that every acquired B. cereus strain was quite sensitive to vancomycin. In addition, the B. cereus isolates that were obtained were entirely resistant to ampicillin, cefoxitin, and colistin, and they exhibited exceptional resistance to almost all of the antibiotics that underwent testing, involving erythromycin, trimethoprim-sulfamethoxazole, doxycycline, and cefotaxime. Our results complement those emphasized by Mahmoud et al. (2024), Savić et al. (2016), and Yu et al. (2020). According to these findings, multidrug-resistant (MDR) B. cereus has been occurring in a diversity of meat products, indicating that it may be a major way for human consumers to contract foodborne MDR B. cereus (Bhunia, 2018). According to Algammal et al. (2024), the careless application of antibiotics in the medical and agricultural fields and the general public, as well as the ability to obtain medicines without a prescription and use them recklessly, promotes the emergence of MDR strains.

Various food preservation methods, particularly chemical antimicrobial agents, have long been applied industrially to stop the growth of bacteria in food products (Tropea, 2022), thereby improving the safety and extending the shelf life of the products. The scientific community and food companies have been motivated to look for efficient substitutes for the chemical antibacterial agents frequently employed in food preservation in recent years because of the growing understanding of the impact of diet on human well-being. Consumers are indeed skeptical of the use of these compounds, despite their stringent regulation, as a result of their potential long-term health risks (Primavilla et al., 2023). ACV is a fermented product that is categorized as a functional food because of its constituents and nutrients, including vitamins and minerals, as well as its ability to improve its production characteristics and have an inhibitory impact on a variety of bacteria by stopping the movement of nutrients via their cell membrane (Nady et al., 2024). The finding of this research displayed that ACV showed significant inhibitory effects against B. cereus at varying concentrations, as evidenced by the inhibition zone diameter determined by the agar well diffusion test. The zone diameter was between 11.5±0.6 and 16.4±0.3 mm (10% ACV), 13.6±0.4 and 19.4±0.8 mm (30% ACV), 17.2±0.2 and 24.7±0.5 mm (70% ACV), and 19.8±0.6 and 31.6±0.8 mm (100% ACV). Herein, with MIC ranging from 0.14 to 1.25 mg/mL, the MIC data revealed that ACV showed high antibacterial activity against the investigated isolates. Considering the ACV to be a bacteriocidal agent, some of the isolates under examination displayed MBC values at the same MIC value. These results matched those of earlier investigations by Gaber et al. (2020), Mahmoud et al. (2024), and Yagnik et al. (2021). The present findings are indicative of the efficacy of ACV as a natural preservative, as it holds a variety of active components, including antibacterial antagonists, as well as organic acids like malic acid, acetic acid, and phenolic mixtures like cresols, phenol, and ketone constituents (Nady et al., 2024). Organic acids work to stop bacteria from growing in several ways, such as by destroying the bacteria’s outer membrane, consuming the energy of the microbes, and increasing osmotic pressure, which breaks down the cell membrane and encourages the manufacture of antibacterial peptides in the host cells. This force the host cells to discharge many vital nutrients, like glutamic and acid ions, to equilibrate the osmotic pressure inside the cells, which stops bacteria from growing normally (Al-Hadidy et al., 2023). Minimizing the application of extra ingredients in organic manufacturing is often a substantial technological challenge, but it is being promoted by the usage of ACV in organic meat handling.

Conclusion

According to this investigation, the presence of MDR B. cereus that harbored one or more enterotoxin genes in meat products poses a significant threat to public health. As a result, stricter sanitation regulations must be implemented at all production, handling, and storage stages. Furthermore, employing ACV as a natural antibacterial agent may be a useful way to reduce the risk of B. cereus illness and its occurrence in the food sector, whether in public places or at home, suggesting that it could be a useful natural substitute for traditional preservatives. Future research must assess potential uses in food production for a sustainable strategy to safeguard the health of customers. Testing these substances against bacteria resistant to several drugs is crucial for developing several approaches to handle the rising issue of drug resistance. Furthermore, ACV derivatives can be used as preservatives in several industries, including healthcare (e.g., cosmetics and medications).