Prevalence of Listeria spp. in ready to eat foods (RTE) from Algiers (Algeria)

The aim of this study was to establish the occurrence of Listeria spp., especially Listeria monocytogenes in ready to eat RTE food marketed in Algiers (Algeria).A total of 227 samples were collected from different producers and retailers.All samples were analyzed using a conventional cultivation method AFNOR V08-055.Out of 227 samples tested, 21 (9.3%) tested positive for Listeria spp. among them, 6 (2.6%) tested positive for L. monocytogenes. L. innocua was the most common Listeria species found being detected in 11 samples (4.8%), although both Listeria ivanovii and Listeria welshimeri were detected in 3 (1.3%) and 1 (0.4%) food samples respectively.The study of the antimicrobial sensitivity of Listeria monocytogenes strains showed no resistance.The study has enabled us to detect these contaminants in a wide range of RTE foods, to suggest that contamination likely occurs after heat treatment, and to assess the danger represented by this category of food for populations at risk.     

Influence of sunflower oil based nanoemulsion (AUSN-4) on the shelf life and quality of Indo-Pacific king mackerel (Scomberomorus guttatus) steaks stored at 20 °C

The influence of nanoemulsion (AUSN-4) on the microbiological, proximal, chemical, and sensory qualities of Indo-Pacific king mackerel (Scomberomorus guttatus) steaks stored at 20 °C was studied for a time period of 72 h. AUSN-4 treatment showed initial reduction (P > 0.05) in the heterotrophic, H₂S and lactic acid bacterial populations in 12 h, followed by a gradual increase in their respective populations. Irrespective of treatments, reduction in total carbohydrate, protein, and fat contents were observed in all samples with an increase in storage time (h), with AUSN-4 treated steaks having the lowest reduction. AUSN-4 treatment significantly (P < 0.05) decreased the values of chemical indicators of spoilage throughout the storage period. Organoleptic evaluation revealed that AUSN-4 treated steaks showed an extension of shelf life of 48 h, when compared with control and antibiotic treated samples, respectively. Based on the results obtained in our present study we conclude that sunflower oil based nanoemulsion preservative technique is able to extend the shelf life and maintain the quality of S. guttatus steaks during storage.     

Potential of Escherichia coli as a surrogate indicator for postchill broiler carcasses with high Campylobacter counts

Surrogating Campylobacter contamination level in broiler carcasses with other bacterial indicators, used to evaluate the hygienic status of the slaughterline operations, might be stimulation to the broiler meat industry to improve control of Campylobacter during slaughter. Theoretically, Escherichia coli might have some practical merits as a potential indicator for carcasses contaminated with Campylobacter. This study investigates the correlation between the counts of E. coli and Campylobacter in 231 postchill broiler carcasses. The impact of setting a process hygiene target based on E. coli counts on reducing the frequency of carcasses contaminated with Campylobacter at level of ≥3 log₁₀ CFU/g was also investigated. Almost half (48.9% (46/94)) of the carcasses with enumerable Campylobacter (≥1 log₁₀ CFU/g) had E. coli counts between 3 and 4 log₁₀ CFU/g. In addition, 54.8% (17/31) of the carcasses contaminated with Campylobacter of ≥3 log₁₀ CFU/g were correlated with E. coli count range of ≥3 & <4 log₁₀ CFU/g. A theoretical scenario assuming that hygiene and processing measures could allow achieving a target for E. coli that not exceeding 3 log₁₀ CFU/g showed a parallel impact on Campylobacter contamination in broiler carcasses. In such scenario, the overall number of Campylobacter-positive carcasses could be dropped from 40.6% to 12.5%; in addition, 80.6% (25/31) of the carcasses contaminated with Campylobacter of ≥3 log₁₀ CFU/g could be eliminated. Findings from this study reveal that a hygiene target based on E. coli count could be used as an indirect sanitary tool for reducing the level of Campylobacter contamination in postchill broiler carcasses.     

Organic thyme oil emulsion as an alternative washing solution to enhance the microbial safety of organic cantaloupes

Emulsions of organic essential oils may be used as postharvest alternative washing solutions in fresh produce production. In the present study, organic thyme oil was emulsified with whey protein concentrate, gum arabic, lecithin, or their equal mass mixtures without using specialized equipment. The stability of these emulsions was monitored by measuring hydrodynamic diameter during ambient storage up to 7 days. The antimicrobial activity of these emulsions against Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). All emulsions had lower MIC and MBC than free thyme oil pre-dissolved in ethanol. Thyme oil emulsified with gum arabic had the smallest and most stable hydrodynamic diameter (156–239 nm) and was chosen as the washing solution to evaluate its efficacy in reducing pathogens on organic cantaloupes. Cantaloupes inoculated with pathogens were immersed in 0.1%, 0.2%, or 0.5% emulsified or free thyme oil for 2 min. The counts of the three cultures inoculated on cantaloupes were reduced by either 0.2% or 0.5% thyme oil and the emulsions were more effective than free thyme oil (P < 0.05). Organic load (2% or 5%) had no effect on their antimicrobial efficacy (P > 0.05). During ambient storage (21 °C) up to 10 days, the counts for all three bacteria gradually declined for all treatments and the emulsion treatment had consistently lower populations than unwashed and water-washed treatments. Therefore, emulsions of organic essential oils have potential applications as postharvest washing solutions to improve the microbiological safety of organic fresh produce.     

Characterization and control of microbial black spot spoilage in dry-cured Iberian ham

The presence of black spots on dry-cured Iberian ham surface is an alteration related to microbial population. Although it provokes important economic losses for the meat industry, the microorganisms responsible for this spoilage still remain unclear. The aim of this study was to identify the microorganisms involved in dry-cured Iberian ham black spot spoilage and to study the conditions affecting their growth. Several microbial strains were isolated from Iberian hams spoiled with black spots at the beginning of post-salting stage. However just one strain produced black coloration in both culture media and fat from Iberian ham. This strain was tentatively identified as Pseudomonas fluorescens by using the API 20 NE system and phylogenetic analyses based on the 16S rRNA and carA genes. It was able to grow and produce blackening at 5, 25, and 30 °C and with salt concentrations of 2 and 5% at 25 °C and with 2% NaCl at 5 °C in culture media. When it was inoculated in sterile pork fat tissues, grow and blackening were only detected in samples without added salt at water activity of 0.94 and 0.97. In order to control the incidence of the alteration temperature throughout salting and post-salting stages should be under 5 °C until salt reaches at least a concentration of 2% or water activity is reduced on the surface of dry-cured Iberian ham below 0.92.     

Effect of Hibiscus sabdariffa extract and Nigella sativa oil on the growth and aflatoxin B1 production of Aspergillus flavus and Aspergillus parasiticus strains

This study was undertaken to evaluate the inhibitory effect of Hibiscus sabdariffa calyx extract at concentrations of 5, 7.5, 10 and 12.5 g/100 ml and Nigella sativa oil at concentrations of 1, 2 and 3 ml/100 ml on the growth and aflatoxin B₁ production by Aspergillus parasiticus (CBS 921.7) and Aspergillus flavus (SQU 21) strains. The inhibition of aflatoxin B₁ production by the different concentrations of H. sabdariffa calyx ranged between 91.5-97.9% and 87.1-93.3% for A. flavus and A. parasiticus strains, respectively, whereas the inhibition by different concentrations of N. sativa oil ranged between 47.9 and 58.3% for A. flavus and 32-48% for A. parasiticus strains. The different concentrations of H. sabdariffa calyx and N. sativa oil had no significant effect on the growth of either Aspergillus species. Neither H. sabdariffa calyx nor N. sativa oil detoxified pure aqueous aflatoxin. Our results suggest that H. sabdariffa calyx and N. sativa oil extracted from seeds had metabolic effects on aflatoxin biosynthesis pathway of both Aspergillus species and can be used as an effective biocontrol and non-toxic biopreservatives in food industry against aflatoxin contamination.     

Detection of Norovirus and Feline Calicivirus in spiked molluscs subjected to heat treatments

Aim of the study was the evaluation of the effect of heat treatment in shellfish experimentally contaminated with human Norovirus (NoV). Feline Calicivirus (FCV), often used as surrogate for human NoV, was examined in parallel to test for virus infectivity after treatment. The experiments were performed subjecting suspensions and spiked mussels to heat treatment (60 °C and 80 °C) for various times. Analysis by rRT-PCR showed a limited reduction of NoV (less than 1 log RT-PCR units ml⁻¹) and FCV (0.3-1.2 log TCID₅₀ ml⁻¹) both in virus suspensions and in spiked mussels, with the exception of NoV suspensions treated at 80 °C (reduction of 3 log RT-PCR units ml⁻¹). Cell culture assay showed that the infectious FCV in suspensions decreased of 4 log after 3 min of treatment while a lower reduction (2 log) was obtained in spiked samples treated for 15 min. Data showed that mussel matrix plays a protective role against heat inactivation of viruses. Despite the efficacy of rRT-PCR for viral nucleic acid detection, its inability to provide information on virus infectivity, represents a limit to the evaluation of product safety. Caution should therefore be used in the evaluation of efficacy of heat treatments on virus-contaminated foods and when the judgment of food product safety is based exclusively on rRT-PCR results.     

Adherence kinetics, resistance to benzalkonium chloride and microscopic analysis of mixed biofilms formed by Listeria monocytogenes and Pseudomonas putida

Comparison between the resistance to BAC and the microscopic structure between mixed-species biofilms formed by different strains of Listeria monocytogenes and Pseudomonas putida CECT 845 under different scenarios and that obtained by the corresponding monospecies L. monocytogenes biofilm was carried out. The association of P. putida with L. monocytogenes quickens biofilm formation and increases significantly (p < 0.05) the BAC-resistance of the biofilm after 4 days of incubation at 25 °C respecting to that formed by monospecies biofilms. According with the adherence profiles of P. putida, two different patterns of association between both species (A and B) were identified, being type A pattern found in the mixed biofilms much more resistant to BAC. After 11 days of incubation, a destructuration of mixed biofilms occurred in all experimental assays, being in 2 out of 5 experimental cases (4032 and BAC-adapted 5873 on polypropylene) accompanied by a sharp decrease in the number of adhered cells. Microscopic analyses demonstrated that complex three-dimensional microscopic structure showed the highest resistance to BAC (4032-SS). Obtained results clearly highlight that to improve disinfection protocols for assuring food safety, it is necessary to mimick those bacterial association that occur in nature.     

Safety aspects, genetic diversity and technological characterisation of wild-type Streptococcus thermophilus strains isolated from north Italian traditional cheeses

Antibiotic susceptibility, antimicrobial activity, genotypic and technological properties of 52 Streptococcus thermophilus isolates, collected from four north Italian traditional cheeses, was investigated. RAPD-PCR, was used to study genetic variability and distinguish closely related strains; the results showed a high degree of heterogeneity among isolates. With regard to technological properties, after 6 h of incubation in milk 25% of the streptococcal strains could be defined as fast acid producers, while after 24 h the majority of isolates (79%) displayed only weak acidification activity. Reduction activity was generally low; in fact, none of these S. thermophilus strains showed a Eh < −102 mV). All the studied S. thermophilus were susceptible to ciprofloxacin, levofloxacin, penicillin G, ampicillin, mupirocin, nitrofurantoin, quinupristin/dalfopristin and rifampicin. Nine isolates were classified as resistant to tetracycline, 6 to streptomycin, 5 to oxacillin, 3 to erytromycin, 3 to vancomycin and only one to chloramphenicol. PCR-based detection did not identify any of the common genetic determinants for vancomycin (vanA, vanB, vanC1, vanC2, vanC3, vanD, vanE, vanG) or erythromycin (ermB and ermC). The genetic basis of the tetracycline resistance phenotype in these strains was linked to tetS-tetL genes (8 isolates) or the tetM gene (1 isolate), and the integrase element int of the Tn916/Tn1545 family of transposons was negative. Four strains were able to produce antimicrobial compounds against Clostridium tyrobutyricum. The study provides new evidence of the resistance of S. thermophilus to antimicrobial agents, confirming the importance of including an accurate safety assessment of phenotypic/biotechnological data when carrying out strain selection for dairy applications.     

Effect of weak acids on Listeria monocytogenes survival: Evidence for a viable but nonculturable state in response to low pH

Listeria monocytogenes is capable of surviving environmental conditions that are normally fatal to many other bacteria, enabling this pathogen to remain on foods and eventually establish infection after consumption. This study investigated the extent to which L. monocytogenes is killed by several weak acids, with particular emphasis on the commonly used food preservative, potassium sorbate. The effects of potassium sorbate were found to be highly pH dependant, with significant killing only observed at a pH of 4.0. At this pH, the order of effectiveness of the acids tested was Na benzoate > Na propionate > acetic acid > K sorbate. Temperature had a significant impact on sensitivity, with K sorbate being effective at 50 mM and elevated (37 °C) temperature, but failing to affect survival when cells were at 4 °C or 21 °C. Finally, we found that cells grown in the presence of K sorbate at pH 4.0 entered a viable but nonculturable (“VBNC”) state, but became nonviable by 24 h. In contrast, cells incubated under these conditions in the absence of K sorbate entered the VBNC state and maintained viability throughout the 24 h study period.     

Anti-listeria activity of Pediococcus pentosaceus BCC 3772 and application as starter culture for Nham, a traditional fermented pork sausage

Lactic acid bacteria forming bacteriocin active against Listeria monocytogenes were screened for potential use in controlling growth of L. monocytogenes in Nham, a Thai traditional fermented pork sausage. Based on the spot-on-lawn assay, Pediococcus pentosaceus BCC 3772 was found to produce the highest anti-listeria activity. Through a series of chromatographic techniques, Edman N-terminal sequencing and mass spectrometry, this anti-listeria compound showed a complete homology to pediocin PA-1/AcH. Inoculation of P. pentosaceus BCC 3772 in Nham caused a significant decrease of 3.2 log in population of spiked L. monocytogenes within 18-24 h of fermentation, in comparison with the initial count with no significant changes in sensory properties and consumer acceptability on the overall characteristics of the final fermented Nham products. These results indicate that the P. pentosaceus BCC 3772 should be useful as a functional starter culture for control of L. monocytogenes without compromising the unique quality of Nham.     

Application of FINS and multiplex PCR for detecting genuine abalone products

Abalones rank first on the list of the four treasures from the sea in Chinese cuisine. It is regarded as a tonic for yin-enrichment and nourishing the lungs and liver. Hong Kong is a major entrepôt of abalones, importing 48% of the global production volume. Rapid detection of substitutes in abalone products is essential for safeguarding consumer interests and ensuring proper delivery of healthcare. Two molecular tools are applied to authenticate abalone products. Firstly, forensically informative nucleotide sequencing (FINS) analysis based on mitochondrial 16S rDNA region of abalone products retailed in Hong Kong revealed that all sliced and one canned products matched the sequences of other gastropods but not Haliotis species, suggesting the presence of substitutions. Secondly, multiplex PCR with a Haliotis-specific primer was used to facilitate rapid and reliable exclusion of substitutes from abalone products.     

Differential detection of small pelagic fish in Tunisian canned products by PCR-RFLP: An efficient tool to control the label information

Correct identification of fish species including their origin requires a compilation of recognized material to verify the conformity of the product with the labelling requirement. Sardine is among the internationally disputed example of species identification conflicts, with the lack of genetic reference material at the Southern part of the Mediterranean. Therefore a method was developed to discriminate between Tunisian small pelagic fish: Sardina pilchardus, Sardinella aurita and Engraulis encrasicolus. DNA extraction from fresh fish and from 12 canned sardine and 2 anchovy-type products, was followed by a PCR method that specifically amplifies a 252 bp fragment of the cytochrome b gene. The new sequences which were deposited in the NCBI GenBank, were searched against a nucleotide sequence database (GenBank) and phylogenetically analyzed with the MEGA software. Multiple alignments of 3 analyzed reference samples belonging to Clupeomorpha species was performed versus the canned samples. Low intraspecific variability was observed for canned anchovy and sardine (<0.05), whereas mean interspecific variability was 0.23. A phylogenetic tree was constructed, and the calculated bootstrap values (BP, 71-100%) were used as indicators of the correct assignment of unknown canned samples to reference species. Postamplification digestion with HaeIII and ALuI restriction enzymes, yielded specific profiles that enabled anchovy to be distinguished from either of the sardine species. This PCR-RFLP was found to be reliable for species identification of canned fish products.     

Application of disinfectant sprays after chilling to reduce the initial microbial load and extend the shelf-life of chilled chicken carcasses

The objective of this study was to evaluate the effects of various disinfectant sprays on initial microbial load and conduct a shelf-life study of chilled chicken carcasses. Chilled chicken carcass samples obtained after chilling were sprayed for 15 s in a purpose-built spray rig with sodium hypochlorite (SH, 50 & 100 mg/L), chlorine dioxide (CD, 50 & 100 mg/L), lactic acid (LA, 1 & 2%), acid electrolyzed oxidizing water (AEOW) and slightly acid electrolyzed oxidizing water (SAEOW). Untreated carcasses were used as the control. Back, leg and breast skin were removed from each carcass after treatment to determine the total viable counts (TVC) and total coliforms. Sprays of 2% LA, AEOW and SAEOW were the most effective treatments with reductions of 0.47–0.83 log CFU/cm² and 0.49–0.96 log MPN/cm² in TVC and total coliforms, respectively. Samples treated with AEOW and SAEOW had 2 days of microbial shelf-life extension compared to the controls, which exceeded the TVC limit of 7 log CFU/cm² at day 6. Even longer extension was obtained for the 2% LA treated samples. The total coliforms, pH and total volatile basic nitrogen (TVBN).TVBN values of the 2% LA, AEOW and SAEOW treated samples were significantly (P < 0.05) lower than those of the controls during storage. Predominantly, the TBARS values of the AEOW and SAEOW treated samples were not statistically different from those of the controls (P > 0.05); however, the 2% LA treatment accelerated lipid oxidization, manifested as the highest TBARS value (2.09 mg MDA/kg) at day 8. Conclusively, this study indicated that the application of AEOW and SAEOW sprays to chilled chicken carcasses after chilling can reduce initial microbial load and maintain quality attributes during refrigerated storage.     

Resistances to UV-C irradiation of Salmonella Typhimurium and Staphylococcus aureus in wet and dried suspensions on surface with egg residues

To clarify the effects of food sediments on ultraviolet-C (254 nm) sanitation in food-related environments, we examined the resistance of pathogenic bacteria (Salmonella Typhimurium and Staphylococcus aureus) cells, in wet and dried suspensions adhered with 1.5-15% w/v egg albumen, 1.5-15% yolk or 3.0-30% whole egg solutions, against UV-C irradiation. Bacterial suspensions (0.1 ml of 8 log CFU/ml) were put on 47 mmφ glass dishes and dried at room temperature (20-24 °C) for 180 min in a bio safety cabinet with ventilation. Viable S. Typhimurium and S. aureus cells in distilled water decreased during the drying period from 7.2 to 3.2 and from 8.0 to 6.5 log CFU/dish, respectively,. On the other hand, the bacteria cells were protected from drying by egg compounds, even by the lowest concentration. The UV-C treatment (0.16 mW/cm² for 10 min) showed a clear bactericidal effect in the absence of egg compounds. However, the bactericidal effect was inhibited by 15% yolk and 30% whole egg. Results in this study suggested that the small food sediment protect bacteria on the surfaces from dryness and UV-C irradiation and it might introduce cross contamination.     

Influence of temperature and surface kind on biofilm formation by Staphylococcus aureus from food-contact surfaces and sensitivity to sanitizers

This study aimed to assess the adhesion, detachment kinetic and biofilm formation of Staphylococcus aureus isolates from food services surfaces on stainless steel and polypropylene surfaces when cultivated in a vegetable-based broth at 7 and 28 °C, and the efficacy of peracetic acid (30 mg/L) and sodium hypochlorite (250 mg/L) in removing the bacterial cells from the matrix of the preformed biofilm. The isolates adhered over 4 Log cfu/cm² regardless the surface kind and incubation temperature. Cell detachment was around 3 Log cfu/cm² over the first six contacts with agar characterizing a high persistence of cells on the tested surfaces. Number of cells (5-7 Log cfu/cm²) needed for biofilm formation was noted at all experimental systems already after 3 days of incubation. A range of 2.0-3.3 and 1.5 to 2.1 Log cfu/cm² was observed in the reduction of cells in biofilm matrix caused by peracetic acid and sodium hypochlorite, respectively. The isolates of S. aureus revealed high capability to adhere and form biofilm on the tested surfaces in both assayed incubation temperature.     

Principal mycotoxins in wheat flour from the Serbian market: Levels and assessment of the exposure by wheat-based products

The presence of eleven principal mycotoxins from the wheat flour bought in supermarkets in Novi Sad, the capitol of the northern Serbian province of Vojvodina, was determined. The samples were prepared by simple one-step method and analyzed by ultra-high performance liquid chromatography with heated-electrospray ionization triple quadrupole mass spectrometry (UHPLC/HESI-MS/MS). Deoxynivalenol (DON) was the predominant mycotoxin for all analyzed samples followed by zearalenone (ZON) and T-2 toxin, with frequency of occurrence: 88.7%, 33.3% and 26.7%, respectively. Aflatoxins (AFs), ochratoxin A (OTA), HT-2 toxin, fumonisins B1 (FB1) as well as B2 (FB2) were below the limit of detection. All the samples complied with current European/Serbian legislation, except one sample that exceeded the DON maximum level of 750 μg/kg. In addition, mycotoxin intakes through consumption of wheat-based products were estimated for average adult consumers based on Serbian market basket and then compared with the tolerable daily intake (TDI) proposed by Scientific Committee on Food of the European Union. The calculated intakes of ZON and T-2 were lower than the respective TDIs. However, intakes of DON were assessed to be close to the level of TDI for adults. This is the first study on the intake assessment for mycotoxins present in the wheat flour through the consumption of wheat-based products on the Serbian market.     

Efficacy of pulsed electric fields for the inactivation of indicator microorganisms and foodborne pathogens in liquids and raw chicken

This study investigated the ability of pulsed electric fields (PEF) to inactivate a range of microorganisms in liquid media and on raw chicken meat. The susceptibilities of ten Campylobacter isolates (seven Campylobacter jejuni isolates and three Campylobacter coli isolates), Escherichia coli (ATCC 25922) and Salmonella Enteritidis (ATCC 13076) to PEF in liquid media were investigated. All Campylobacter isolates tested in liquid were susceptible to PEF treatment (65 kV/cm, 5 μs, 500 Hz) with reductions of between 4.33 and 7.22 log₁₀ CFU/mL observed for the longest treatment (30 s). Significant differences in susceptibility were observed between Campylobacter isolates subjected to equivalent PEF treatments ranging from 2.41 to 5.19 log₁₀ CFU/mL. Campylobacter isolates processed in liquid media were found to be more sensitive to PEF than E. coli and S. Enteritidis (P < 0.05). The application of PEF (3.75 and 15 kV/cm, 10 μs, 5 Hz) to inoculated samples of raw chicken did not result in any significant reductions in total viable counts, Enterobacteriaceae, C. jejuni, E. coli or S. Enteritidis. Therefore, under the conditions used in this study, PEF technology may not be suitable as a food safety intervention measure for the control of microbial contaminants on broilers during processing although it may have potential to reduce contamination of process water (e.g. scald or spin chill water).     

Identification of the wedge clam Donax trunculus by a simple PCR technique

The wedge clam Donax trunculus is an important bivalve commercial species in Portugal which can be easily mistaken with other three morphologically similar species (Donax semistriatus, Donax vittatus and Donax variegatus) that have a lower market price. This may lead fish sellers to make false claims about the authenticity of their products in order to get higher profits. To overcome this problem it is important to develop analytical techniques that can be used to test the authenticity of the species that is being sold. In this study we present two DNA extraction methodologies and a simple PCR method for the accurate identification of D. trunculus based on the amplification of the nuclear marker 5S rDNA. The PCR amplification results showed that this method is reliable to differentiate D. trunculus and D. variegatus from the remaining Donax species, since fragments of D. trunculus were about 275-300bp while D. variegatus were about ∼450 bp a little lower molecular weight than DNA fragments of the other two species (∼500 bp).