Use of Matrix-Assisted and Laser Desorption/Ionization Time-of-Flight Technology in the Identification of Aeromonas Strains Isolated from Retail Sushi and Sashimi.

The genus Aeromonas includes well-known pathogenic species for fishes and humans that are widely distributed in the aquatic environment and foods. Nowadays, one of the main issues related to wild Aeromonas isolates is their identification at the species level, which is challenging using classical microbiological and biomolecular methods. This study aims to test MALDI-TOF MS technology in the identification of Aeromonas strains isolated from n. 60 retail sushi and sashimi boxes using an implemented version of the SARAMIS software V4.12. A total of 43 certified Aeromonas strains were used to implement the SARAMIS database by importing the spectra obtained from their identification. The original SARAMIS version (V4.12) failed to recognize 62.79% of the certified strains, while the herein-implemented version (V4.12plus) allowed the identification of all the certified strains at least to the genus level with a match of no less than 85%. Regarding the sushi and sashimi samples, Aeromonas spp. was detected in n. 18 (30%) boxes. A total of 127 colonies were identified at the species level, with A. salmonicida detected as the most prevalent species, followed by A. bestiarum and A. caviae. Based on the results of the present study, we could speculate that MALDI-TOF technology could be a useful tool both for the food industry to monitor product contamination and for clinical purposes to make diagnoses effectively and quickly.

Microsporidian heavy infection in a batch of salted and dried cod.

The aim of this work is the description and characterization of a severe microsporidian infection in a batch of salted and dried cod. Particularly, the case involves a batch of approximately 800 kg obtained from Gadus macrocephalus (Food and Agriculture Organization Zone 61 - Northwest Pacific Ocean), which, after rehydration and sectioning operations, underwent routine company checks before packaging. In about 20% of the samples, the presence of whitish nodules with a diameter ranging from 1 to 2 mm was observed on the surface of the fillets and in cross-section. The lesions ranged from a few units to 10 per cm2. Some samples were subjected to fresh microscopic observation with the stereomicroscope, confirming the nodular nature of the lesions, which were often confluent, alternating with empty spaces, giving the tissue a honeycombing aspect. The histological examination at low magnification allowed us to observe the heavy vacuolization of nodular lesions irregularly surrounded by a spongy-like wall. The observation at higher magnification of other sections allowed us to identify intra-myofibrillar cists containing presumptive microsporidian elements. The tissue damage derived from the technological processes and gravity of lesions did not allow a morphological characterization of presumptive protozoans. The molecular examination of the nodular lesions and the analysis of the sequence of an 897 bp fragment of the small subunit 16S rRNA revealed 100% identity with Microsporidium theragrae (GenBank accession number MT928885-89) first isolated from the skeletal muscles of Gadus chalcogrammus specimens from the Sea of Okhotsk. This finding confirms the importance of selecting suppliers and raw materials in the seafood industry, as well as the usefulness of an effective traceability system.

The Occurrence of Microplastics in Donax trunculus (Mollusca: Bivalvia) Collected along the Tuscany Coast (Mediterranean Sea).

Microplastics (MPs) (0.1 µm-5 mm particles) have been documented in oceans and seas. Bivalve molluscs (BMs) can accumulate MPs and transfer to humans through the food chain. BMs (especially mussels) are used to assess MPs' contamination, but the genus Donax has not been thoroughly investigated. The aim of this study was to detect and characterize MPs in D. trunculus specimens collected along the Tuscan coast (Italy), and to assess the potential risk for consumers. The samples (~10 g of tissue and intervalval liquid from 35 specimens) were digested using a solution of 10% KOH, subjected to NaCl density separation, and filtered through 5 µm pore-size filters. All items were morphologically classified and measured, and their mean abundance (MA) was calculated. Furthermore, 20% of them were analyzed by Raman spectroscopy and, based on the obtained results, the MA was recalculated (corrected MA) and the annual human exposure was estimated. In the 39 samples analyzed, 85 items fibers (n = 45; 52.94%) and fragments (n = 40; 47.06%) were found. The MA was 0.23 ± 0.17 items/grww. Additionally, 83.33% of the items were confirmed as MPs (polyethylene and polyethylene terephthalate). Based on the correct MA (0.18 MPs/grww), D. trunculus consumers could be exposed to 19.2 MPs/per capita/year. The health risk level of MPs was classified as level III (moderate).

A stochastic approach for modelling the in-vitro effect of osmotic stress on growth dynamics and persistent cell formation in Listeria monocytogenes.

Persistent bacteria (or persisters) can be defined as a microbial subpopulation that, exposed to bactericidal treatment, is killed more slowly than the rest of the population they are part of. They stochastically originate in response to environmental stressors or spontaneously without external signals. When transferred into a fresh medium, persisters can resume active replication although they spend more time adapting to the new conditions remaining in the lag phase longer. They were studied for decades for their ability to survive antibiotic treatments while studies on their formation in food and potential impact on their safety are lacking. The most common food preservation techniques may act as stressors that trigger the formation of persistent bacteria able to survive bactericidal treatments and grow later in foods during storage. This study aimed to investigate a possible relationship between exposure to different salt concentrations (osmotic stress) and the amount of persisters triggered in a strain of Listeria monocytogenes. Furthermore, we described this phenomenon from a mathematical perspective through predictive microbiology models commonly used in the food field. The lag time distribution of a L. monocytogenes ATCC 7644 strain grown in broth with additional 2 %, 4 % and 6 % NaCl was evaluated using the software ScanLag. It uses office scanners to automatically record the colony growth on agar plates and evaluate the frequency distribution of their appearance times (lag phase) by automated image analysis. The same broth cultures were diluted to equalize salt concentration and transferred into a fresh broth to evaluate how the previous salt exposure impacted their growth kinetics. The observed growth curves were reproduced using predictive models in which the mean duration of the lag phase of the whole population took into account the occurrence of persisters with a longer lag phase. The models were solved first using a deterministic approach and then a stochastic one introducing a stochastic term that mimics the variability of lag phase duration due to the persisters occurrence. Results showed that the growth of L. monocytogenes in broth with additional NaCl might trigger the formation of persistent cells whose number increased consistently with salt concentrations. The proposed predictive approach reproduced the observed real curves in strong agreement, especially through the stochastic resolution of the models. Persistence is currently a neglected bacterial defence strategy in the food sector but the persisters' formation during production cannot be excluded; therefore, further insights on the topic are certainly desirable.

Aerococcus viridans and Public Health: Oral Carriage and Antimicrobial Resistance in Stray Dogs and Cats in Algeria.

The current study aimed to determine the occurrence and antimicrobial resistance of oral Aerococcus viridans in stray dogs and cats in Algeria. Oral swabs from 200 stray animals (100 dogs and 100 cats) were collected and cultured on Columbia agar medium supplemented with 5% defibrinated sheep blood. Isolates were identified using analytical profile index Rapid 20 Strep commercial kits, and antibiotic susceptibility was determined using the disk diffusion method. Of the 200 animals sampled, 34 carried A. viridans in their oral cavities, with 26 isolates (76.47%) resistant to at least 2 drugs. Multidrug resistance profiles (to more than three different antimicrobials) were observed only in cats (26.08% of isolates). More isolates were resistant to erythromycin and tetracycline (71% and 65%, respectively) than to other antimicrobials. This is the first research study in Algeria detecting antimicrobial resistance in oral A. viridans isolated from dogs and cats and highlights potential public health concerns. Clinical trials registration number: 01/2018.

The effects of herbicide application on two soil phosphate solubilizing bacteria: Pantoea agglomerans and Serratia rubidaea.

The application of synthetic pesticides is one of the fastest acting tools at farmers' disposal to prevent and mitigate the threats posed by plant pests in agriculture. However, the effects of these above-ground applications of pesticides are known to be detrimental to some belowground, non-target soil biota. At present, the effects many pesticides have on key functional microbial groups associated with phosphate (P) solubilization in the soil are still largely unknown. The purpose of this study was to compare the effects of two herbicides, glyphosate, and paraquat, on phosphate solubilizing bacteria (PSB) with and without pH adjustment (after herbicide addition) since pH is a major indicator of P solubilization. In our assay, two PSB strains (Pantoea agglomerans and Serratia rubidaea) were chosen to assess their ability to solubilize tricalcium phosphate (TCP) by using the vanadate-molybdate method (to measure the amount of P solubilized) in the presence of glyphosate (5.4 g/L and 10.8 g/L) or paraquat (2 g/L and 4 g/L) separately. To assess the effect of PSB treated by the herbicides, a growth experiment using PSB inoculated wheat seedlings was performed under greenhouse conditions (25 °C, light 16 h/8 h dark). After four weeks, wheat above-ground growth parameters were measured. Our results showed that even under recommended doses of glyphosate (5.4 g/L) and paraquat (2 g/L), a decrease in P solubilization activity was observed in P. agglomerans and S. rubidaea. Whilst paraquat affected TCP solubilization more than glyphosate with and without pH adjustment, there was a significant decrease (p < 0.05) in TCP solubilization, up to 39% and 93% in the presence of glyphosate and paraquat, respectively, for S. rubidaea, and up to 45% and 95% in the presence of glyphosate and paraquat, respectively, for P. agglomerans. The effect of the herbicides on the PSB had the same results as in the greenhouse test on wheat seedling growth, confirming that these herbicides have both above and belowground negative effects, despite being used at recommended doses.

Chemical Composition and Comprehensive Antimicrobial Activity of an Ethanolic Extract of Propolis from Tunisia.

In the present study, the chemical composition and the in vitro antimicrobial and antibiofilm activity of an ethanolic extract of propolis (EEP) from Tunisia against different ATCC and wild bacterial strains were evaluated. In situ antimicrobial activity and sensory influence of different EEP concentrations (0.5% and 1%), also in combination with 1% vinegar, were evaluated in chilled vacuum-packed salmon tartare. Furthermore, a challenge test was performed on salmon tartare experimentally contaminated with Listeria monocytogenes and treated with the different EEP formulations. The in vitro antimicrobial and antibiofilm activity was observed only against Gram-positive bacteria, such as L. monocytogenes and S. aureus, both ATCC and wild. Results of the in situ analyses revealed significant antimicrobial activity against aerobic colonies, lactic acid bacteria, Enterobacteriaceae and Pseudomonas spp. only when the EEP was used at 1% and in combination with 1% vinegar. The 1% EEP in combination with 1% vinegar was the most effective treatment also against L. monocytogenes, although 0.5% and 1% EEP used alone also showed antilisterial effects. After 7 days of storage, the sensory influence on odor, taste and color of salmon tartare was negligible for all EEP formulations. In this background, results obtained confirmed the antimicrobial efficacy of propolis which could be proposed as a suitable biopreservative to ensure safety and improve the quality of food.

Effects of osmotic stress on Listeria monocytogenes ATCC 7644: persistent cells and heat resistance.

Persistent bacteria are a microbial subpopulation that, exposed to bactericidal treatment, is killed at a slower rate than the rest of the population they are part of. They can be triggered either following stress or stochastically without external signals. The hallmark of persistent bacteria is the biphasic killing curve, a sign that, within a microbial population, two subpopulations are inactivated at a different rate. Furthermore, when plated into a fresh medium and in the absence of stressors, persistent bacteria typically remain in the lag phase longer before resuming active replication. This study aims to evaluate in vitro whether the formation of persistent cells in a strain of Listeria monocytogenes can be triggered by exposure to osmotic stress and if this phenomenon can increase heat resistance in the bacterial population. In a first experiment, the lag time distribution of a L. monocytogenes strain grown in a 6% NaCl broth was evaluated using the software ScanLag. A stationary phase broth culture was inoculated on agar plates placed on an office scanner inside an incubator at 37°C. The plates were scanned every 20' for 4 days and the acquired images were automatically elaborated with the aid of MatLab software in order to evaluate the appearance times of every single colony. The experiment was also carried out on a control culture obtained by growing the strain in the broth without salt. In a second experiment, the same broth cultures, after proper dilutions to rebalance NaCl concentration, were subjected to a heat treatment at 51°C and the death curves obtained were parameterized using the GinaFit system. Results showed that the lag phase of 31.40% of the salt culture colonies was long enough to suppose the formation of persistent bacteria. Analyses of the thermal survival curves showed that the shoulder and tail model was the one that best represented the inactivation trend of the salt culture, unlike the control culture, whose trend was essentially linear. Results of the present study show how exposure to salt could induce the formation of persistent bacteria in a L. monocytogenes strain. The last raises concerns as persistent cells may not only be undetected with common analytical techniques but they even show a greater heat resistance.

Quality index method developed for raw anglerfish (Lophius piscatorius) stored in ice.

The quality index method (QIM) is a widely accepted solution to establish the state of fish freshness quickly and effectively. The present study aims to determine increasingly reliable freshness parameters for fresh and whole Lophius piscatorius stored in ice. Sensory and microbiological analyses were performed on 148 anglerfishes. Sensory evaluations were performed based on the QIM, updating a previously proposed quality index (QI) scheme. Total viable count and specific spoilage organisms were determined through microbiological analyses of the tail musculature, evaluating their correlations with the QI scores over time. The updated QI scheme included 3 characters, namely appearance, eye, and fin, for a total of 18 demerits points. A positive linear correlation between QI score and storage time was observed such that the sensory rejection time (8th day) can be predicted within ± 1 day with the developed scheme. At the sensory rejection point, loads of the spoilage microbial flora were not high enough to be relatable to the appreciated alterations probably due to the anglerfish morphology in which the tail musculature is isolated from the gills and viscera, the main sources of bacterial contamination. The proposed scheme offers a ready-to-use freshness assessment of the anglerfish although further validations are needed.

Ginger and parsley essential oils: chemical composition, antimicrobial activity, and evaluation of their application in cheese preservation.

Chemical composition and in vitro antimicrobial activity of parsley [Petroselinum crispum (Mill.) Fuss] and ginger (Zingiber officinale Roscoe) essential oils from store-bought parsley plants and ginger rhizomes were evaluated. A sensory analysis was performed on aged cheese slices treated with a mix of each essential oil and seed oil. Parsley essential oil was selected as the best compromise between sensory influence and antimicrobial activity and tested to evaluate its antifungal effects on cheese slices experimentally contaminated with Aspergillus flavus. Monoterpenes and α-zingiberene were the most abundant compounds in parsley and ginger essential oil, respectively. Both essential oils showed in vitro antimicrobial activity against different fungal species while only a bacterial strain (Enterococcus faecalis) was sensible to ginger essential oil. Only the cheese slices treated with parsley essential oil were appreciated by the panellists and its use has been effective in preventing the A. flavus growth in cheese.

Study of the Effectiveness of Alumina and HDTMA/Alumina Composite in the Removal of Para-Nitrophenol and the Deactivation of Bacterial Effect of Listeria monocytogenes and Salmonella spp.

Removal of para-nitrophenol (p-NP) from an aqueous solution was studied under various batch adsorption experiments, using alumina (Al2O3) and its composite hexadecyltrimethylammonium bromide (HDTMA+-Br-) as adsorbents. These were later characterized, before and after adsorption of p-NP, by thermal analysis (DSC-TG), X-ray diffraction (XRD), Fourier transform infrared (FTIR), and UV/Visible spectroscopies. The results show that HDTMA+/Al2O3 adsorbents have a greater affinity toward p-NP than Al2O3 alone. Linear and non-linear forms of kinetics and isotherms were used to analyze the experimental data obtained at different concentrations and temperatures. The results indicate that the pseudo-second order kinetic model provided the best fit to the experimental data for the adsorption of p-NP on both adsorbents, and that the intra-particle diffusion was not only the rate controlling step. Both the Langmuir and Redlich-Peterson (R-P) models were found to fit the sorption isotherm data well, but the Langmuir model was better. Physical adsorption of p-NP onto the adsorbents proved to be an endothermic and spontaneous process at high temperatures, which mainly involves a hydrogen bonding mechanism of interactions between p-NP and functional groups of adsorbents. The antibacterial activity of Al2O3, HDTMA+-Br- and HDTMA+/Al2O3 were evaluated against Listeria monocytogenes and Salmonella spp. strains using both disc diffusion and broth microdilution methods. The HDTMA+-Br- and HDTMA+/Al2O3 displayed a bacteriostatic effect against all tested strains of Listeria monocytogenes and Salmonella spp., while Al2O3 exhibited no bacterial effect against all bacterial strains tested.

Chemical profile, antibacterial, antioxidant, and anisakicidal activities of Thymus zygis subsp. gracilis essential oil and its effect against Listeria monocytogenes.

In this work, the chemical composition of Thymus zygis subsp. gracilis collected from Ifrane, Morocco, along with the evaluation of the antibacterial, anti-biofilm of Listeria monocytogenes activities, larvicidal effect against L3 larvae of Anisakis, and antioxidant properties of its essential oil (TZG-EO), are reported. GC-MS and GC-FID analyses highlighted the presence of 84 volatile components and strong bactericidal and anti-biofilm activities against L. monocytogenes at a concentration of 0.02 % were demonstrated. Also, larvicidal effect against Anisakis larvae at concentrations of 0.01 and 0.005 % was attained leading to the death of all tested larvae within 4 h. The in situ antibacterial activity of TZG-EO (0.01 and 0.005 %) in smoked fish showed high efficiency against L. monocytogenes growth. TZG-EO could be used as potential antibacterial and larvicidal agents for fighting against foodborne pathogens and extending shelf life of food products.

Anisakicidal Effects of R (+) Limonene: An Alternative to Freezing Treatment in the Industrial Anchovy Marinating Process.

Anisakiasis is a fish-borne zoonotic disease caused by the ingestion of raw/undercooked fishes or cephalopods parasitized by members of the genus Anisakis. Freezing ensures the inactivation of viable Anisakis larvae; however, since it affects the organoleptic properties of food, essential oils and their compounds were proposed as an alternative. In this study, fresh anchovy fillets were experimentally parasitized with L3 Anisakis larvae to test the anisakicidal efficacy of R (+) limonene (LMN) in marinated fishery products. The anisakicidal effectiveness and organoleptic influence of several LMN concentrations (0.5%, 1%, and 5%) were tested during the marinating process (MS) and storage in sunflower seed oil (SO) of marinated anchovy fillets. Double treatment (DT) with 1% LMN was also performed both during marination and subsequent storage in oil. MS treatment resulted only in a reduction in larvae viability after 48 h, while a complete inactivation was observed in SO after 8, 10, and 20 days of treatment with 5%, 1%, and 0.5% LMN, respectively. DT was the most effective with complete larval inactivation after 7 days. Only 5% LMN influenced the sensory characteristics of the fillets, resulting, however, in a pleasant lemon-like odor and taste. Considering the results obtained, LMN might be a suitable natural alternative to manage Anisakis risk in the fishery industry.

Temperature fluctuations along food supply chain: A dynamic and stochastic predictive approach to establish the best temperature value in challenge tests for Listeria monocytogenes.

This study aims to evaluate the behaviour of Listeria monocytogenes under fluctuating temperature comparing the efficacy of deterministic and stochastic methods for its prediction. In the first part of the study, a strain of L. monocytogenes was maintained at two different fluctuating temperature regimes both from 2 to 8°C and regularly sampled for the quantitative determination. The first temperature regime lasted 204 hours with a fluctuation length of 12 hours whereas the second lasted 167 hours with a fluctuation length of 24 hours. A dynamic predictive model was implemented for the reproduction of the observed data. Model resolution has been carried out by using values of the recorded temperature as well as the value of the mean temperature, the kinetic mean temperature, the 75th and 95th percentile of the temperature. A stochastic resolution was also performed considering the mean temperature and Standard Deviation as stochastic variable. In the second part of the study, a temperature mean curve was constructed by monitoring temperature of 8 refrigerated conveyances, 10 display cabinet and 15 domestic refrigerators. This curve was used to obtain predictive scenarios for L. monocytogenes based on the above and also considering temperature regime suggested by the EURL Lm TECHNICAL GUIDANCE DOCUMENT on challenge tests and durability studies for assessing shelf-life of ready-to-eat foods related to Listeria monocytogenes (Version 4 of 1 July 2021). All predicted behaviours were compared to the observed ones through the Root Mean Squared Error. Firstly, dynamic predictive model as well as the stochastic one, provided the best level of reproducibility of the observed data. The kinetic mean temperature reproduced the observed data better than the mean temperature for the 12 hoursregime while for the 24 hours-regime was the opposite. The 75th and 95th percentile overestimated the observed growths. Secondary, predictions obtained with the mean temperature, kinetic temperature and stochastic approach well fitted the observed data. The 75th and 95th percentile of Temperature and the "Eurl LM" temperature regimes overestimated the observed prediction. Dynamic approach as well as the stochastic one allowed to obtain the lowest values of Root Mean Squared Error. The mean temperature and kinetic mean temperature appeared the most representative values in a deterministic "single-point" approach.

Development of a predictive model for the shelf-life of Atlantic mackerel (Scomber scombrus).

Despite its commercial value, the shelflife of the Atlantic mackerel (Scomber scombrus) during refrigerated storage was poorly investigated. In this regard, the Quality Index Method (QIM) was proposed as a suitable scoring system for freshness and quality sensorial estimation of fishery products. This study aims to develop a deterministic mathematical model based on dynamic temperatures conditions and a successive statistical analysis of the results obtained. This model will be exploited to predict the shelf-life of the Atlantic mackerel based on specific storage temperatures. A total of 60 fresh fishes were subdivided into two groups and respectively stored in ice for 12 days at a constant temperature of 1±0.5°C (Group A) and a fluctuating temperature ranging between 1 and 7°C (Group B). Microbiological analysis and sensory evaluation through the QIM were performed on each fish at regular time intervals. A critical value of 6 Log cfu/g of spoilage bacteria (mainly psychotropic) associated with a significant decay of the sensorial characteristics was exceeded after 9 days of storage for Group A and 3 days for Group B. A reliable prediction of fish freshness was obtained by modelling the QIM as a function of the spoilage bacteria behaviour. A coefficient ß of correlation was determined to convert the spoilage bacteria load into a Quality Index score. The adoption of mathematical predictive models to assess microbial behaviour under different environmental conditions is an interesting tool for food industries to maximize production and reduce waste.

Mineral and Microbiological Analysis of Spices and Aromatic Herbs.

Spices and aromatic herbs have always had great historical importance in human nutrition. Their use has been documented for centuries as a rich source of bioactive compounds; they have been used for their health benefits and also for flavoring or coloring food. However, despite the many health properties linked to the use of spices and aromatic herbs, these can represent biological hazards and can contain chemical substances of concern. Certainly, monitoring potential health hazards in spices and aromatic herbs includes microbiological safety and also the content of inorganic substances: both represent a key step. This research aims at monitoring the compliance of various spices and aromatic herbs from a non-European country market (namely: black cumin seeds, Iranian Tokhme Sharbati, clove buds, Shahjeera, Abbaszadeh saffron, organic fenugreek, whole black pepper, cinnamon, Abthul Ahmar (Asario), Ajwan seeds, whole coriander seeds, black sesame seeds, Sabja seeds) with the current European Union (EU) and WHO regulations, when available, regarding mineral and microbiological parameters. In particular, microbiological assays using rapid and conventional methods, and trace mineral determination by inductively coupled plasma mass spectrometry (ICP-MS) were performed. Results show the safety of the tested spices, given that the microbiological parameters were within the legal microbiological criteria set by the European Commission Regulation (EC) No. 2073/2005 and its amendment Regulation (EC) No. 1441/2007. With reference to potentially toxic Cd, Pb, As, Hg, these were within the limits set by the European Commission Regulation (EC) No. 1881/2006 and its amendments, Regulation (EU) No. 1317/2021 and Regulation (EU) No. 1323/2021, and WHO. According to EU regulations, for Pb content, 2 samples out of 16 showed values different from the set limits.

Virulence, Antimicrobial Resistance and Biofilm Production of Escherichia coli Isolates from Healthy Broiler Chickens in Western Algeria.

The aim of this study was to assess the virulence, antimicrobial resistance and biofilm production of Escherichia coli strains isolated from healthy broiler chickens in Western Algeria. E. coli strains (n = 18) were identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Susceptibility to 10 antibiotics was determined by standard methods. Virulence and extended-spectrum ß-lactamase (ESBL) genes were detected by PCR. The biofilm production was evaluated by microplate assay. All the isolates were negative for the major virulence/toxin genes tested (rfbE, fliC, eaeA, stx1), except one was stx2-positive. However, all were resistant to at least three antibiotics. Ten strains were ESBL-positive. Seven carried the ß-lactamase blaTEM gene only and two co-harbored blaTEM and blaCTX-M-1 genes. One carried the blaSHV gene. Among the seven strains harboring blaTEM only, six had putative enteroaggregative genes. Two contained irp2, two contained both irp2 and astA, one contained astA and another contained aggR, astA and irp2 genes. All isolates carrying ESBL genes were non-biofilm producers, except one weak producer. The ESBL-negative isolates were moderate biofilm producers and, among them, two harbored astA, two irp2, and one aggR, astA and irp2 genes. This study highlights the spread of antimicrobial-resistant E. coli strains from healthy broiler chickens in Western Algeria.

Nile Red staining for detecting microplastics in biota: Preliminary evidence.

Nile Red is a lipophilic, metachromatic and solvatochromic dye used as an alternative or complementary method to aid identification of microplastics in routine analysis of biological samples. It was rarely used in biota since organic residues after the digestion step can be co-stained with possible overestimation of microplastics. The limits of using Nile Red in biota were investigated in marine mussels experimentally contaminated with low-density polyethylene (LDPE) microplastics. Stained particles were detected through magnified images obtained by stitching together thirty photographs of the filter surface of each sample. LDPE particles appeared yellowish and fluorescent and could be confused with certain organic residues. The smaller the fragments, the greater the difficulty in recognizing them. In particular, it was difficult to recognize LDPE particles based on their fluorescence if <180 µm in size. Regardless of the size, fluorescence of the items aids the operator in LDPE particles identification also in biota.