Nachweis und Charakterisierung von Clostridium perfringens mittels real-time-PCR

Clostridium perfringens is a Gram-positive, anaerobic bacterium. The strains are classified into five types (type A to E), depending on the ability to produce alpha-, beta-, epsiloon and iota-toxin, but food-poisonings are mostly caused by C. perfringens type A isolates. Ingestion of contaminated food is followed by gastrointestinal disease, when enzyme-resistant C. perfringens enterotoxins (CpE) are set free during sporulation. In most cases the bacterium has to grow up to more than 10⁶ cfu/g food to cause a gastrointestinal disease (BVET, 2005; BfR, 2005). Cultural methods, normally used for the detection of C. perfringens, are not able to differentiate isolates into the five different toxin types. This is the reason, why the detection of C. perfringens in food under the present legal regulations can only used as an indication for inadequate production hygiene or a toxin-infection with contaminated food as source. For the detection of the toxin genes to differentiate C. perfringens in five different toxin types, three duplex real-time-PCR assays for the routine diagnostic were developed and validated. The assays can be used for quick detection and easy classification of C. perfringens isolates and as a rapid screening-system in suspected cases of C. perfringens food-poisonings.     

Nachweis und Charakterisierung von Clostridium perfringens mittels real-time-PCR

Clostridium perfringens is a Gram-positive, anaerobic bacterium. The strains are classified into five types (type A to E), depending on the ability to produce alpha-, beta-, epsiloon and iota-toxin, but food-poisonings are mostly caused by C. perfringens type A isolates. Ingestion of contaminated food is followed by gastrointestinal disease, when enzyme-resistant C. perfringens enterotoxins (CpE) are set free during sporulation. In most cases the bacterium has to grow up to more than 10⁶ cfu/g food to cause a gastrointestinal disease (BVET, 2005; BfR, 2005). Cultural methods, normally used for the detection of C. perfringens, are not able to differentiate isolates into the five different toxin types. This is the reason, why the detection of C. perfringens in food under the present legal regulations can only used as an indication for inadequate production hygiene or a toxin-infection with contaminated food as source. For the detection of the toxin genes to differentiate C. perfringens in five different toxin types, three duplex real-time-PCR assays for the routine diagnostic were developed and validated. The assays can be used for quick detection and easy classification of C. perfringens isolates and as a rapid screening-system in suspected cases of C. perfringens food-poisonings.

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Zuszammenfassung:  Clostridium perfringens geh?rt zur Gruppe der gram-positiven, anaerob bzw. aerotolerant wachsenden Bakterien. Abh?ngig von der F?higkeit der einzelnen C. perfringens St?mme, alpha-, beta-, epsilon und iota-Toxin zu bilden, erfolgt eine Einteilung in die Typen A bis E, wobei C. perfringens Typ A für den überwiegenden Teil der lebensmittelassoziierten menschlichen Erkrankungen verantwortlich ist. Ursache solcher Toxin-Infektionen ist das erst im Darm im Rahmen der Sporulation der mit den Lebensmitteln aufgenommenen vegetativen Zellen gebildete Enterotoxin (CpE). In der Regel muss sich der Erreger im Lebensmittel auf mehr als 10⁶ KbE/g Lebensmittel vermehren, damit beim Konsumenten eine Erkrankung ausgel?st wird (BVET, 2005; BfR, 2005). Da die derzeit vorhandenen kulturellen Nachweisverfahren keine M?glichkeit bieten, die gewonnenen Isolate einem bestimmten Toxintyp zuzuordnen, kann der Nachweis von C. perfringens in einem Lebensmittel nach gültiger Rechtslage lediglich als ein Hinweis auf eine mangelhafte Produktions- oder Verarbeitungtechnologie oder auf das Vorliegen einer lebensmittelbedingten Toxin-Infektion gewertet werden. Zum Nachweis der Toxingene, die eine Einteilung von C. perfringens in die fünf verschiedene Toxintypen erm?glichen, wurden drei Duplex-real-time-PCR-Assays etabliert und die Einsatzm?glichkeiten in der Routinediagnostik getestet. Die Systeme sollen für einen schnellen Nachweis und eine rasche Differenzierung von C. perfringens sowie als Screening-System bei Verdacht auf eine lebensmittelbedingte Erkrankung genutzt werden.

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Eingegangen: 30. Januar 2007     

Recovery of Staphylococcus aureus in Gray Mugil cephalus Roe (Bottarga): Investigation by an Integrated Cultural/Molecular Approach

In the Mediterranean area, salted and dried roe from the gray Mugil cephalus “bottarga” represent a speciality food with great commercial value. Bottarga is currently produced by a traditional handmade process and, the risk of human bacterial contamination during its manufacturing is still unknown; in this perspective the foodborne pathogen Staphylococcus aureus could potentially contaminate this product due to poor sanitation or bad handling during processing. The aim of this work is: to evaluate the contamination level of foodborne pathogens at different product manufacturing stages and, in addition, to describe a fast and realizable method for the rapid detection of S. aureus in bottarga samples in the field. A cultural procedure was initially used to investigate the occurrence of S. aureus and the other main foodborne pathogens in bottarga samples at the different manufacturing stages (from roe to final product). In addition, a molecular approach was used to rapidly determine the presence of total bacteria, S. aureus, and its potential toxigenicity. Of the 194 specimens analyzed, we identified: Clostridium perfringens, Enterococcus spp. and Enterobacteriaceae. However, some samples resulted as being contaminated with S. aureus (4% in roe and 8.7% in the final product). During the bottarga manufacturing process, we observed an increase in pathogen levels (from 10² to 10⁵ CFU/g) in contaminated samples, and entA and entB genotypes were identified. Reconstruction experiments suggest that the fresh roe and the bottarga (not completely dried) could represent a risk for the contamination and growth of pathogen bacteria.     

Culture‐Independent Rapid Detection Methods for Bacterial Pathogens and Toxins in Food Matrices

Rapid detection of bacterial pathogens and toxins in foods is necessary to provide real‐time results to mitigate foodborne illness outbreaks. Cultural enrichment methods, although the most widely used, are time‐consuming and therefore inadequate for rapid pathogen detection from food samples. The development of novel “rapid” detection methods has decreased detection time dramatically. This review presents an overview of detection methods for various foodborne pathogens, including Listeria monocytogenes, Salmonella enterica, and shiga toxin‐producing Escherichia coli, and bacterial toxins in food matrices, with emphasis on those methods which do not require cultural enrichment. Discussed methods include nucleic acid‐, immunological‐, and biosensor‐based techniques. A summary of each type of detection method is given, including referenced methods from the literature. Since these discussed methods do not require cultural enrichment, there is a higher probability of interference from the food matrices. Therefore, the review also discusses the potential interference of food components on detection methods and addresses preprocessing strategies to overcome matrix associated inhibition and to concentrate low quantities of pathogens and toxins in food. Development of rapid and sensitive detection technologies advances and ensures public health safety and security.     

STUDY ON BACTERIAL LOAD DURING GELATIN MANUFACTURING PROCESS AND ITS EFFECT ON FOOD GRADE GELATIN

A total of 138 samples from different stages of gelatin manufacturing process were analyzed to detect bacterial contamination of food grade gelatin and to see its effect on the quality of gelatin. The isolated bacterial species included Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Clostridium perfringens, which exhibited gelatinase activity and caused depletion in the nutritional quality of food grade gelatin. Marked changes in the amino acid content with incubation days indicated degradation of food grade gelatin because of the secretion of gelatinases that were depicted by paper chromatography. Because these isolated species are known pathogens and have gelatinolytic properties, their occurrence in food grade gelatin can make it an unsafe and unsuitable for food applications.     

Clostridium perfringens: A Dynamic Foodborne Pathogen

Clostridium perfringens is a spore-forming bacterium and natural inhabitant of soil and the intestinal tracts of many warm-blooded animals, including humans. The ubiquitous nature of this bacterium and its spores makes it a frequent problem for the food industry and establishments where large amounts of food are prepared. C. perfringens causes potentially lethal foodborne diseases in humans, including food poisoning and necrotic enteritis. This bacterium could be controlled properly following safety rules such as adequate heating and cooling of food during processing. Unfortunately, large C. perfringens outbreaks, sometimes with fatal outcomes are still frequently reported. This paper describes the main characteristics of C. perfringens that allow the bacterium to survive and grow in foods, and cause human disease as well as discusses strategies to control this microorganism during food processing.     

Staphylokokken-Enterotoxine: Bildung, Eigenschaften und Nachweis

Under appropriate conditions, Staphylococcus aureus can grow in food and produce enterotoxins which cause vomiting and diarrhoea when ingested. In contrast to the staphylococci staphylococci these toxins are resistant especially to high temperatures and are not inactivated by usual kitchen practice. As the presence of staphylococci is not essential to cause illness, this condition is a typical food poisoning. The estimation of a potential health risk by targeting the organism is therefore only appropriate to raw food. For products which underwent a procedure detrimental to the bacteria, e. g. heat treatment, testing for thermonuclease (indirect detection of a contamination with high numbers of staphylococci, irrespective of the presence of living bacteria at the time of testing) or the detection of enterotoxins can be applied. In this context, the detection of enterotoxin genes using molecular biological tests provides only supplemental information, as positive results are not evidentiary for gene expression and thus for the presence of toxins in the food. In this review, the causative organism and its toxins as well as methods of detection are discussed with special emphasis on molecular biological methods.     

Bacillus cereus food intoxication and toxicoinfection

Bacillus cereus is one of the leading etiological agents of toxin-induced foodborne diseases. Its omnipresence in different environments, spore formation, and its ability to adapt to varying conditions and produce harmful toxins make this pathogen a health hazard that should not be underestimated. Food poisoning by B. cereus can manifest itself as an emetic or diarrheal syndrome. The former is caused by the release of the potent peptide toxin cereulide, whereas the latter is the result of proteinaceous enterotoxins (e.g., hemolysin BL, nonhemolytic enterotoxin, and cytotoxin K). The final harmful effect is not only toxin and strain dependent, but is also affected by the stress responses, accessory virulence factors, and phenotypic properties under extrinsic, intrinsic, and explicit food conditions and host-related environment. Infamous portrait of B. cereus as a foodborne pathogen, as well as a causative agent of nongastrointestinal infections and even nosocomial complications, has inspired vast volumes of multidisciplinary research in food and clinical domains. As a result, extensive original data became available asking for a new, both broad and deep, multifaceted look into the current state-of-the art regarding the role of B. cereus in food safety. In this review, we first provide an overview of the latest knowledge on B. cereus toxins and accessory virulence factors. Second, we describe the novel taxonomy and some of the most pertinent phenotypic characteristics of B. cereus related to food safety. We link these aspects to toxin production, overall pathogenesis, and interactions with its human host. Then we reflect on the prevalence of different toxinotypes in foods opening the scene for epidemiological aspects of B. cereus foodborne diseases and methods available to prevent food poisoning including overview of the different available methods to detect B. cereus and its toxins.     

Rapid whole protein quantification of staphylococcal enterotoxin B by liquid chromatography

Food poisoning caused by Staphylococcus aureus is one of the most important foodborne diseases in the world. The ability of these bacteria to produce one or more enterotoxins in milk and dairy products is linked to staphylococcal food poisoning. Enterotoxin B (SEB) is an exotoxin produced by S. aureus and is one of the compounds most frequently involved in staphylococcal food poisoning worldwide. In this work, 20 samples of milk collected from restaurants have been studied for the presence of S. aureus enterotoxigenic strains. All the isolates from milk samples have been analysed by liquid chromatography-coupled with diode array detector for the rapid identification and quantification of SEB as intact protein. Limit of detection and limit of quantification values were 0.5 and 1 μg/mL, respectively. S. aureus was found in 35% of analysed samples but only one of them was an enterotoxigenic strain, which produced staphylococcal enterotoxin B at levels of 3.6 μg/mL.     

Staphylokokken-Enterotoxine: Bildung, Eigenschaften und Nachweis

Under appropriate conditions, Staphylococcus aureus can grow in food and produce enterotoxins which cause vomiting and diarrhoea when ingested. In contrast to the staphylococci staphylococci these toxins are resistant especially to high temperatures and are not inactivated by usual kitchen practice. As the presence of staphylococci is not essential to cause illness, this condition is a typical food poisoning. The estimation of a potential health risk by targeting the organism is therefore only appropriate to raw food. For products which underwent a procedure detrimental to the bacteria, e. g. heat treatment, testing for thermonuclease (indirect detection of a contamination with high numbers of staphylococci, irrespective of the presence of living bacteria at the time of testing) or the detection of enterotoxins can be applied. In this context, the detection of enterotoxin genes using molecular biological tests provides only supplemental information, as positive results are not evidentiary for gene expression and thus for the presence of toxins in the food. In this review, the causative organism and its toxins as well as methods of detection are discussed with special emphasis on molecular biological methods. Eingegangen: 18. Januar 2007; nach überarbeitung angenommen: 20. M?rz 2007     

Prevalence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in food samples associated with foodborne illness in Alberta, Canada from 2007 to 2010

Consumption of foods containing Staphylococcus aureus can cause severe gastro-intestinal illness. Given the fact that over the past decade, Canada has seen increasing rates of methicillin-resistant S. aureus (MRSA) carriage and infection, the objective of this study was to investigate the impact of methicillin-susceptible S. aureus (MSSA) and MRSA on foodborne illness in Alberta, Canada. Between January 2007 and December 2010, there were 693 food samples associated with foodborne investigations submitted to the Alberta Provincial Laboratory for Public Health (ProvLab). These foods were screened for: Bacillus cereus, Clostridium perfringens, S. aureus, Aeromonas spp., Campylobacter spp., Escherichia coli O157:H7, Salmonella, Shigella spp., and Yersinia spp. S. aureus was identified in 10.5% (73/693) of samples, and of these, 59% (43/73) were co-contaminated with at least one other organism on the screening panel. The S. aureus positive samples included 29 meat, 20 prepared foods containing meat, 11 prepared foods not containing meat, 10 dairy, and three produce. Methicillin-resistance was not detected in any isolates tested. These findings indicate that the presence of S. aureus in food associated with foodborne investigations is a cause for concern, and although MRSA was not found, the potential for outbreaks exists, and ongoing surveillance should be sustained.     

State of the art: Lateral flow assays toward the point-of-care foodborne pathogenic bacteria detection in food samples

Diverse chemicals and some physical phenomena recently introduced in nanotechnology have enabled scientists to develop useful devices in the field of food sciences. Concerning such developments, detecting foodborne pathogenic bacteria is now an important issue. These kinds of bacteria species have demonstrated severe health effects after consuming foods and high mortality related to acute cases. The most leading path of intoxication and infection has been through food matrices. Hence, quick recognition of foodborne bacteria agents at low concentrations has been required in current diagnostics. Lateral flow assays (LFAs) are one of the urgent and prevalently applied quick recognition methods that have been settled for recognizing diverse types of analytes. Thus, the present review has stressed on latest developments in LFAs-based platforms to detect various foodborne pathogenic bacteria such as Salmonella, Listeria, Escherichia coli, Brucella, Shigella, Staphylococcus aureus, Clostridium botulinum, and Vibrio cholera. Proper prominence has been given on exactly how the labels, detection elements, or procedures have affected recent developments in the evaluation of diverse bacteria using LFAs. Additionally, the modifications in assays specificity and sensitivity consistent with applied food processing techniques have been discussed. Finally, a conclusion has been drawn for highlighting the main challenges confronted through this method and offered a view and insight of thoughts for its further development in the future.     

Real-time-PCR-System zum Nachweis von Bacillus cereus (emetischer Typ) in Lebensmitteln

Bacillus cereus is a Gram-positive, facultative anaerobic, spore-performing bacterium. Some B. cereus strains have the ability to produce two different types of toxins: (a) diarrhoeic toxin: the disease is similar to a C. perfringens toxin-infection; it is caused by a heat-labile protein, (b) emetic toxin: the disease is similar to Staphylococcus-aureus intoxikation; it is caused by a heat-stable protein. Statements about the frequency of B. cereusfood-poisonings are difficult because a reporting system for this disease is missing in Germany and there exists no valid methodology to diagnose this disease with conventional microbiological or biochemical methods. Nevertheless B. cereus, beside S. aureus, seems to be the most important bacterium to cause food-associated intoxications.     

COMPARISON OF REVERSE PASSIVE LATEX AGGLUTINATION TEST AND IMMUNOBLOTTING FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN A AND B

Staphylococcal foodborne diseases resulting from consumption of food contaminated with staphylococcal enterotoxins (SEs) produced by certain strains of Staphylococcus aureus are the second most common foodborne illnesses in the world. Analytical methods are essential for routine monitoring purposes and safeguard public health. Different methods for SE detection have been proposed although their use in a complex matrix is often limited by the presence of substances that interfere with tests. In this article reverse passive latex agglutination (RPLA) and immunoblotting methods based on specific antibodies and currently available for SE detection have been compared. Culture filtrates from enterotoxin S. aureus strains isolated from cheese samples were identified by SET‐RPLA. Then the culture filtrates identified as staphylococcal enterotoxin A and staphylococcal enterotoxin B by RPLA test were analyzed with immunoblotting. The results obtained suggest that either SET‐RPLA or immunoblotting may be applied to culture filtrates for the detection of SEs with good correspondence of results. Although SET‐RPLA represents a simple method for routine monitoring purposes, a positive result by a rapid method (RPLA) is only regarded as presumptive and must be confirmed by standard methods ( Feng 1996 ), such as immunoblotting method.     

Effect of Mentha spicata L. and Mentha aquatica L. essential oils on the microbiological properties of fermented dairy product,kashk

The antibacterial activity of Mentha spicata and Mentha aquatica essential oils (EO) was tested against Staphylococcus aureus, Lactobacillus reuteri, Bifidobacterium animalis and Clostridium perfringens using agar well and disc diffusion techniques. Results showed that M. spicata EO had the highest inhibition activity against the studied microorganisms. Then, the antibacterial activity of both EO at 1500 and 2500 ppm was examined in industrial liquid kashk during the storage at 4 °C for 20 days. Both EO reduced the S. aureus viable count below 5 log CFU g^?1 after 4 days; however, the population of C. perfringens, L. reuteri and B. animalis decreased <1 log CFU g^?1 during the storage time. The least deteriorative effect on the lactic acid bacteria was related to M. aquatica. As revealed by organoleptic studies, kashk samples containing M. aquatica EO at 1500 and 2500 ppm were the most preferred samples.     

Dual-Labeled PCR-Based Immunofluorescent Assay for the Rapid and Sensitive Detection of Enterotoxic <Emphasis Type="BoldItalic">Staphylococcus aureus</Emphasis> Using Cocktail-Sized Liposomal Nanovesicles as Signal Enhancer

Staphylococcus aureus is a major foodborne pathogen worldwide, and as little as 1 μg of staphylococcal enterotoxins (SEs) can cause food poisoning. Among SEs, enterotoxin A is the most common toxin that causes staphylococcal food poisoning. Hence, this work has developed a dual-labeled PCR-based immunofluorescent assay using anti-digoxigenin (DIG) antibody-tagged immunomagnetic beads (IMBs) as the capture reagent and cocktail-sized NeutrAvidin-tagged liposomal nanovesicles (NA-LNs) that encapsulate fluorescent dyes as the detection reagent. In this approach, the amplicon of sea gene was doubly labeled with DIG and biotin using modified primers and biotin-11-dUTP. The system depends on the immunocapture of IMB to pre-concentrate the labeled amplicons, which were further quantified using cocktail-sized NA-LNs, based on the release and subsequent measurement of encapsulated fluorescent dyes following the lysis of NA-LNs. After optimization, the developed assay could detect S. aureus and differentiate it from other common foodborne bacteria, such as Salmonella enterica and Escherichia coli, with a limit of detection (LOD) of 10¹ CFU mL^?1 without pre-enrichment. With a 2-h pre-enrichment, this developed assay could detect as little as 1 CFU in 25 mL of milk within a workday. Hence, this work established a rapid and sensitive PCR-based immunofluorescent assay using liposomal nanovesicles as an instant signal enhancer to detect the contamination of enterotoxic S. aureus in milk.     

Optimising chitosan–pectin hydrogel beads containing combined garlic and holy basil essential oils and their application as antimicrobial inhibitor

Chitosan–pectin hydrogel beads that trap and release the maximal amount of combined garlic and holy basil essential oils to inhibit food microorganisms were developed based on the central composite design, with chitosan (0.2–0.7% w/v), pectin (3.5–5.5% w/v) and calcium chloride (CaCl₂) (5.0–20.0% w/v) contents. The optimal bead consisted of 0.3–0.6% w/v chitosan, 3.9–5.1% w/v pectin and 8.0–17.0% w/v CaCl₂, which had a high encapsulation efficiency (62.16–79.06%) and high cumulative release efficiency (31.55–37.81%) after storage at 5 °C for 15 days. Optimal hydrogel beads were packed into a cellulose bag to evaluate antimicrobial activity by the disc volatilisation method. The beads inhibited Bacillus cereus, Clostridium perfringens, Escherichia coli, Pseudomonas fluorescens, Listeria monocytogenes and Staphylococcus aureus but did not affect Lactobacillus plantarum and Salmonella Typhimurium. The oil-containing beads could potentially be applied in food packaging to inhibit the mentioned microorganisms.     

PHILIPPINE FOODBORNE-DISEASE OUTBREAKS (1995–2004)

The study presented details about 60 reported Philippines foodborne outbreaks for the period of 1995–2004. It was established that meat‐containing dishes were the more common causes of the outbreaks evaluated, with spaghetti as the leading food vehicle. Common risk settings for the outbreaks were the food services of schools and workplaces. Salmonella and Vibrio spp. were cited as the primary causes of infections, while human intoxications involved staphylococcal enterotoxins, paralytic shellfish poisoning (PSP) toxins and histamine.     

Prevalence of staphylococcal enterotoxins, toxin genes and genetic-relatedness of foodborne Staphylococcus aureus strains isolated in the Marmara Region of Turkey

Staphylococcus aureus is a major foodborne pathogen and it has the ability to produce a number of extracellular toxins. We analyzed 1070 food samples obtained from retail markets and dairy farms in the Marmara Region of Turkey for the presence of S. aureus. Out of 147 isolates, 92 (62.6%) were enterotoxigenic. PCR was used to investigate the presence of staphylococcal enterotoxin genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu), exfoliative toxin genes (eta and etb) and the toxic − shock syndrome toxin gene (tst). The PCR results showed that 53.3% of the isolates contained staphylococcal enterotoxin-like (SEl) toxin genes (seg, seh, sei, sej, sek, sel, sem, sen, seo, sep, seq and seu) which were more frequent than classical enterotoxin genes (sea to see). Furthermore, seo, sei, sem, seg, seu and sec were found in 37.0, 32.7, 30.4, 29.3, 29.3 and 27.2% of the isolates, respectively. The tst gene was detected and confirmed by DNA sequencing in 9 isolates. The presence of eta and etb were not found in the isolates. Enterotoxigenic capabilities of isolates with SEA-SEE were investigated by ELISA. Enterotoxigenic S. aureus isolates produced one to three enterotoxins, with the most frequently produced types being enterotoxin A and C. There was a correlation of 72.1% between production of a specific toxin and the presence of the respective genes. PFGE analysis was used to identify genetic-relatedness of enterotoxigenic S. aureus isolates and the results revealed that 13 groups of isolates from different or the same origin that contained the same genes showed 100% homology with indistinguishable band patterns. The other enterotoxigenic isolates showed related band patterns with 72-86% homology in sea-, 61-90% homology in sec-, 80-96% homology in seh-, and 69-96% homology in sep-positive isolates. To our knowledge, this is the first study to examine enterotoxins and related gene contents of S. aureus food isolates in the Marmara Region of Turkey.     

Inactivation strategy for Clostridium perfringens spores adhered to food contact surfaces

The contamination of enterotoxigenic Clostridium perfringens spores on food contact surfaces posses a serious concern to food industry due to their high resistance to various preservation methods typically applied to control foodborne pathogens. In this study, we aimed to develop an strategy to inactivate C. perfringens spores on stainless steel (SS) surfaces by inducing spore germination and killing of germinated spores with commonly used disinfectants. The mixture of l-Asparagine and KCl (AK) induced maximum spore germination for all tested C. perfringens food poisoning (FP) and non-foodborne (NFB) isolates. Incubation temperature had a major impact on C. perfringens spore germination, with 40 °C induced higher germination than room temperature (RT) (20 ± 2 °C). In spore suspension, the implementation of AK-induced germination step prior to treatment with disinfectants significantly (p < 0.05) enhanced the inactivation of spores of FP strain SM101. However, under similar conditions, no significant spore inactivation was observed with NFB strain NB16. Interestingly, while the spores of FP isolates were able to germinate with AK upon their adhesion to SS chips, no significant germination was observed with spores of NFB isolates. Consequently, the incorporation of AK-induced germination step prior to decontamination of SS chips with disinfectants significantly (p < 0.05) inactivated the spores of FP isolates. Collectively, our current results showed that triggering spore germination considerably increased sporicidal activity of the commonly used disinfectants against C. perfringens FP spores attached to SS chips. These findings should help in developing an effective strategy to inactivate C. perfringens spores adhered to food contact surfaces.