Category Archives: Antimicrobials

Research -Antimicrobial-Resistant Listeria monocytogenes in Ready-to-Eat Foods: Implications for Food Safety and Risk Assessment

MDPI

Abstract

Antimicrobial resistance is an existential threat to the health sector, with far-reaching consequences in managing microbial infections. In this study, one hundred and ninety-four Listeria monocytogenes isolates were profiled for susceptibility using disc diffusion techniques. Possible foodborne listeriosis risk associated with ready-to-eat (RTE) foods (RTEF) and the risk of empirical treatment (EMPT) of L. monocytogenes infections, using multiple antimicrobial resistance indices (MARI) and antimicrobial resistance indices (ARI), respectively, were investigated. Twelve European Committee on Antimicrobial Susceptibility Testing (EUCAST) prescribed/recommended antimicrobials (EPAS) for the treatment of listeriosis and ten non-prescribed antimicrobials (non-PAS)] were evaluated. Antimicrobial resistance > 50% against PAs including sulfamethoxazole (61.86%), trimethoprim (56.19%), amoxicillin (42.27%), penicillin (41.24%), and erythromycin (40.21%) was observed. Resistance > 50% against non-PAS, including oxytetracycline (60.89%), cefotetan (59.28%), ceftriaxone (53.09%), and streptomycin (40.21%) was also observed. About 55.67% and 65.46% of the isolates had MARI scores ranging from 0.25–0.92 and 0.30–0.70 for EPAs and non-PAs, respectively. There was a significant difference (p < 0.01) between the MARI scores of the isolates for EPAs and non-PAs (means of 0.27 ± 0.21 and 0.31 ± 0.14, respectively). MARI/ARI scores above the Krumperman permissible threshold (>0.2) suggested a high risk/level of antimicrobial-resistant L. monocytogenes. The MARI risks of the non-success of empirical treatment (EMPT) attributed to EPAs and non-PAs were generally high (55.67% and 65.463%, respectively) due to the antimicrobial resistance of the isolates. MARI-based estimated success and non-success of EMPT if EUCAST-prescribed antimicrobials were administered for the treatment of listeriosis were 44.329% and 55.67%, respectively. The EMPT if non-prescribed antimicrobials were administered for the treatment of listeriosis was 34.53% and 65.46%, respectively. This indicates a potentially high risk with PAs and non-PAs for the treatment of L. monocytogenes infection. Furthermore, ARI scores ≤ 0.2 for EPAs were observed in polony, potato chips, muffins, and assorted sandwiches, whereas ARI scores for non-PAs were >0.2 across all the RTE food types. The ARI-based estimate identified potential risks associated with some RTE foods, including fried fish, red Vienna sausage, Russian sausage, fruit salad, bread, meat pies, fried chicken, cupcakes, and vetkoek. This investigation identified a high risk of EMPT due to the presence of antimicrobial-resistant L. monocytogenes in RTE foods, which could result in severe health consequences.

Research – Mānuka Oil vs. Rosemary Oil: Antimicrobial Efficacies in Wagyu and Commercial Beef against Selected Pathogenic Microbes

MDPI

Abstract

Essential oils possessing antimicrobial characteristics have acquired considerable interest as an alternative to chemical preservatives in food products. This research hypothesizes that mānuka (MO) and kānuka (KO) oils may possess antimicrobial characteristics and have the potential to be used as natural preservatives for food applications. Initial experimentation was conducted to characterize MOs (with 5, 25, and 40% triketone contents), rosemary oil (RO) along with kanuka oil (KO) for their antibacterial efficacy against selected Gram-negative (Salmonella spp. and Escherichia coli), and Gram-positive (Listeria monocytogenes and Staphylococcus aureus) bacteria through disc diffusion and broth dilution assays. All MOs showed a higher antimicrobial effect against L. monocytogenes and S. aureus with a minimum inhibitory concentration below 0.04%, compared with KO (0.63%) and RO (2.5%). In chemical composition, α-pinene in KO, 1, 8 cineole in RO, calamenene, and leptospermone in MO were the major compounds, confirmed through Gas-chromatography-mass spectrometry analysis. Further, the antimicrobial effect of MO and RO in vacuum-packed beef pastes prepared from New Zealand commercial breed (3% fat) and wagyu (12% fat) beef tenderloins during 16 days of refrigerated storage was compared with sodium nitrate (SN) and control (without added oil). In both meat types, compared with the SN-treated and control samples, lower growth of L. monocytogenes and S. aureus in MO- and RO- treated samples was observed. However, for Salmonella and E. coli, RO treatment inhibited microbial growth most effectively. The results suggest the potential use of MO as a partial replacement for synthetic preservatives like sodium nitrate in meats, especially against L. monocytogenes and S. aureus.

Research – Using TRIS-Buffered Plasma-Activated Water to Reduce Pathogenic Microorganisms on Poultry Carcasses with Evaluation of Physicochemical and Sensory Parameters

MDPI

Abstract

Foodborne diseases are mainly caused by the contamination of meat or meat products with pathogenic microorganisms. In this study, we first investigated the in vitro application of TRIS-buffered plasma-activated water (Tb-PAW) on Campylobacter (C.) jejuni and Escherichia (E.) coli, with a reduction of approx. 4.20 ± 0.68 and 5.12 ± 0.46 log10 CFU/mL. Furthermore, chicken and duck thighs (inoculated with C. jejuni or E. coli) and breasts (with natural microflora) with skin were sprayed with Tb-PAW. Samples were packed under a modified atmosphere and stored at 4 °C for 0, 7, and 14 days. The Tb-PAW could reduce C. jejuni on days 7 and 14 (chicken) and E. coli on day 14 (duck) significantly. In chicken, there were no significant differences in sensory, pH-value, color, and antioxidant activity, but %OxyMb levels decreased, whereas %MetMb and %DeoMb increased. In duck, we observed slight differences in pH-value, color, and myoglobin redox forms for the Tb-PAW, which were not perceived by the sensory test persons. With only slight differences in product quality, its application as a spray treatment may be a useful method to reduce C. jejuni and E. coli on chicken and duck carcasses.

Research – The Role of Biofilms in the Pathogenesis of Animal Bacterial Infections

MDPI

Abstract

Biofilms are bacterial aggregates embedded in a self-produced, protective matrix. The biofilm lifestyle offers resilience to external threats such as the immune system, antimicrobials, and other treatments. It is therefore not surprising that biofilms have been observed to be present in a number of bacterial infections. This review describes biofilm-associated bacterial infections in most body systems of husbandry animals, including fish, as well as in sport and companion animals. The biofilms have been observed in the auditory, cardiovascular, central nervous, digestive, integumentary, reproductive, respiratory, urinary, and visual system. A number of potential roles that biofilms can play in disease pathogenesis are also described. Biofilms can induce or regulate local inflammation. For some bacterial species, biofilms appear to facilitate intracellular invasion. Biofilms can also obstruct the healing process by acting as a physical barrier. The long-term protection of bacteria in biofilms can contribute to chronic subclinical infections, Furthermore, a biofilm already present may be used by other pathogens to avoid elimination by the immune system. This review shows the importance of acknowledging the role of biofilms in animal bacterial infections, as this influences both diagnostic procedures and treatment.

Research – A Systematic Quantitative Determination of the Antimicrobial Efficacy of Grape Seed Extract against Foodborne Bacterial Pathogens

MDPI

Abstract

Concerns regarding the role of antimicrobial resistance (AMR) in disease outbreaks are growing due to the excessive use of antibiotics. Moreover, consumers are demanding food products that are minimally processed and produced in a sustainable way, without the use of chemical preservatives or antibiotics. Grape seed extract (GSE) is isolated from wine industry waste and is an interesting source of natural antimicrobials, especially when aiming to increase sustainable processing. The aim of this study was to obtain a systematic understanding of the microbial inactivation efficacy/potential of GSE against Listeria monocytogenes (Gram-positive), Escherichia coli and Salmonella Typhimurium (Gram-negative) in an in vitro model system. More specifically, for L. monocytogenes, the effects of the initial inoculum concentration, bacterial growth phase and absence of the environmental stress response regulon (SigB) on the GSE microbial inactivation potential were investigated. In general, GSE was found to be highly effective at inactivating L. monocytogenes, with higher inactivation achieved for higher GSE concentrations and lower initial inoculum levels. Generally, stationary phase cells were more resistant/tolerant to GSE as compared to exponential phase cells (for the same inoculum level). Additionally, SigB appears to play an important role in the resistance of L. monocytogenes to GSE. The Gram-negative bacteria under study (E. coli and S. Typhimurium) were less susceptible to GSE as compared to L. monocytogenes. Our findings provide a quantitative and mechanistic understanding of the impact of GSE on the microbial dynamics of foodborne pathogens, assisting in the more systematic design of natural antimicrobial-based strategies for sustainable food safety.

Research – Foodborne Pathogen Biofilms: Development, Detection, Control, and Antimicrobial Resistance

MDPI

Bacteria can grow either as planktonic cells or as communities within biofilms. The biofilm growth mode is the dominant lifestyle of most bacterial species and 40–80% of microorganisms are associated with biofilms [1]. Biofilm is a sessile community that is irreversibly attached to a substratum or interface or to other members of the community [2]. It is surrounded by extracellular polymeric substances (EPS) that include extracellular polysaccharides, extracellular DNA, lipids, proteins, and other elements [3]. Biofilm formation is a complex but well-regulated process that can be classified into five distinct stages [4]. In the first stage, planktonic bacteria attach to a surface. Salmonella species, Listeria monocytogenesCampylobacter jejuni, or Escherichia coli have specific structures on the surface of the bacteria, such as flagella, curli, fimbriae, and pili, which help the bacteria attach [5].
The second stage is the adhesion step, which includes an initial reversible adhesion resulting in loose adhesion and a subsequent irreversible adhesion resulting in more stable adhesion. The third stage is to secrete EPS and form microcolonies. This is followed by biofilm maturation, which produces large amounts of EPS to grow in size and build three-dimensional structures. The final stage is the stage in which the biofilm is dispersed, releasing the planktonic cells and initiating the formation of a new biofilm at another location.
Microbial cells living within biofilms are protected from various environmental stresses such as desiccation, osmotic changes, oxidative stress, metal toxicity, radiation, antibiotics, disinfectants, and the host immune system [6]. Biofilms are much less sensitive to antimicrobial agents than planktonic cells, and several mechanisms contribute to their resistance to antimicrobials [7]. The exopolysaccharide matrix prevents the entry of antimicrobial agents by reducing diffusion and acting as a primary barrier [8]. Most antimicrobial agents kill rapidly dividing cells more effectively, but slow growth of biofilms leads to resistance [9]. Changes in metabolic activity within biofilms, genetic changes of antimicrobial resistant determinants in target cells, extrusion of antimicrobial agents using efflux pumps, and the presence of persistent cells also contribute to antimicrobial resistance [10].

Research – Evaluation of antibiotic efficacy of Ocimum gratissimum L. essential oil against Staphylococcus aureus and Pseudomonas aeruginosa

Academic Journals

Abstract

Ocimum gratissimum essential oil (EOGT) has been evaluated for its antibacterial efficacy, and its combinational therapy with antibiotics may enhance the therapeutic efficiency against infection-causing bacteria. Herein, we evaluated the chemical composition of EOGT and its antibiotic efficacy against Staphylococcus aureus and Pseudomonas aeruginosa. GC-MS and GC-FID analyzed EOGT. The antibiotic efficacy was determined by the agar diffusion method, microdilution, minimum inhibitory concentration (MIC), and fractional inhibitory concentration index (FICI). Eugenol (74.2%) was the main component of OGT. Using the agar diffusion method, the action of rifampicin, ciprofloxacin, and tetracycline was evaluated against S. aureus, while the action of cefepime and ciprofloxacin was evaluated against P. aeruginosa. FICI showed a reduced MIC of ciprofloxacin and tetracycline associated with EOGT. In the presence of EOGT, MIC of ciprofloxacin reduced from 0.6 to 0.0006 μg/mL and of tetracycline decreased from 0.028 to 0.0018 μg/mL against S. aureus and from 4 to 0.12 μg/mL against P. aeruginosa. EOGT enhanced the antibacterial efficacy of the antibiotics suggesting a synergistic effect, thereby enhancing the efficacy in treating infection against S. aureus and P. aeruginosa.

Research – Antimicrobial Effect of Moringa oleifera Leaves Extract on Foodborne Pathogens in Ground Beef

MDPI

Abstract

Consumers nowadays are becoming more aware of the importance of using only meat products containing safe and natural additives. Hence, using natural food additives for extending the shelf life of meat along with delaying microbial growth has become an urgent issue. Given the increasingly popular view of Moringa oleifera leaves as a traditional remedy and also the scarcity of published data concerning its antimicrobial effect against foodborne pathogens in meat and meat products, we designed the present study to investigate the antimicrobial effect of Moringa oleifera leaves aqueous extract (0.5%, 1%, and 2%) on ground beef during refrigerated storage at 4 °C for 18 days. MLE revealed potent antimicrobial properties against spoilage bacteria, such as aerobic plate count and Enterobacteriaceae count. MLE 2% showed a significant (p < 0.01) reduction in the counts of E. coli O157:H7, Salmonella enterica serovar Typhimurium, and Staphylococcus aureus artificially inoculated to ground beef by 6.54, 5.35, and 5.40 log10 CFU/g, respectively, compared to control, by the 18th day of storage. Moringa leaves extract (MLE) had no adverse effect on the overall acceptability and other sensory attributes; moreover, it induced a slight improvement in the tenderness and juiciness of treated ground beef, compared to the control. Therefore, MLE can be used as a healthy, natural, and safe preservative to increase meat products’ safety, quality, and shelf stability during cold storage. A promising approach for using natural food additives rather than chemical preservatives could begin new frontiers in the food industry, as they are more safe and do not constitute health risks to consumers.

Research – Zoonoses, foodborne outbreaks and antimicrobial resistance guidance for reporting 2022 data

EFSA

Abstract

This technical report of the European Food Safety Authority (EFSA) presents the guidance to reporting European Union (EU) Member States and non‐Member States in data transmission using extensible markup language (XML) data transfer covering the reporting of isolate‐based quantitative antimicrobial resistance data, as well as reporting of prevalence data on zoonoses and microbiological agents and contaminants in food, foodborne outbreak data, animal population data and disease status data. For data collection purposes, EFSA has created the Data Collection Framework (DCF) application. The present report provides data dictionaries to guide the reporting of information deriving from 2022 under the framework of Directive 2003/99/EC, Regulation (EU) 2017/625, Commission Implementing Regulation (EU) 2019/627 and Commission Implementing Decision (EU) 2020/1729. The objective is to explain in detail the individual data elements that are included in the EFSA data models to be used for XML data transmission through the DCF. In particular, the data elements to be reported are explained, including information about the data type, a reference to the list of allowed terms and any additional business rule or requirement that may apply.

Research – Manual for reporting on zoonoses and zoonotic agents, within the framework of Directive 2003/99/EC, and on some other pathogenic microbiological agents for information derived from the year 2022

EFSA

Abstract

This reporting manual provides guidance to European Union (EU) Member States (MSs) for reporting on zoonoses and zoonotic agents in animals, food and feed under the framework of Directive 2003/99/EC, Regulation (EU) 2017/625, Commission Implementing Regulation (EU) 2019/627 and of Commission Delegated Regulation (EU) 2018/772 and also on the reporting of other pathogenic microbiological agents or contaminants in food. The objective of this manual is to harmonise and streamline reporting by MSs to ensure that the data collected are relevant and comparable for analysis at the EU level. This manual covers all the zoonoses and zoonotic agents included under the current data collection system run by the European Food Safety Authority (EFSA). Detailed instructions are provided on the reporting of data in tables and information in text forms. The instructions given relate to the description of the sampling and monitoring schemes applied by the MSs, as well as the monitoring results. Special reference is made to data elements which allow trend watching over time and the analysis of sources of zoonotic agents at the EU level. This manual is specifically aimed at guiding the reporting of information deriving from the year 2022.