A comparative review on methods of detection and quantification of mycotoxins in solid food and feed: a focus on cereals and nuts.

Many emerging factors and circumstances urge the need to develop and optimize the detection and quantification techniques of mycotoxins in solid food and feed. The diversity of mycotoxins, which have different properties and affinities, makes the standardization of the analytical procedures and the adoption of a single protocol that covers the attributes of all mycotoxins a tedious or even an impossible mission. Several modifications and improvements have been undergone in order to optimize the performance of these methods including the extraction solvents, the extraction methods, the clean-up procedures, and the analytical techniques. The techniques range from the rapid screening methods, which lack sensitivity and specificity such as TLC, to a spectrum of more advanced protocols, namely, ELISA, HPLC, and GC-MS and LC-MS/MS. This review aims at assessing the current studies related to these analytical techniques of mycotoxins in solid food and feed. It discusses and evaluates, through a critical approach, various sample treatment techniques, and provides an in-depth examination of different mycotoxin detection methods. Furthermore, it includes a comparison of their actual accuracy and a thorough analysis of the observed benefits and drawbacks.

Prevalence and Antibiotic Resistance of Staphylococcus aureus and Escherichia coli Isolated from Bovine Raw Milk in Lebanon: A study on Antibiotic Usage, Antibiotic Residues, and Assessment of Human Health Risk Using the One Health Approach.

The emergence, persistence, and spread of antibiotic-resistant microbes is a tremendous public health threat that is considered nowadays a critical One Health issue. In Lebanon, the consumption of raw bovine milk has been recently reported as a result of the financial crisis. The objectives of the current study were (1) to evaluate raw bovine milk samples in a comprehensive manner for the types of antibiotics used and their residues, (2) to determine the presence of mesophilic bacteria, extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA), and (3) to determine the associated human health risk caused by drinking raw milk with antibiotic residues among all age categories. LC-MS-MS was used to carry out the analysis. From 200 milk samples, 30 (15%) were found contaminated with four major antibiotics. The highest average concentration detected was for oxytetracyline 31.51 ± 13.23 µg/kg, followed by 5.5 ± 0.55 µg/kg for gentamicin, 4.56 ± 0.73 µg/kg for colistin, and 4.44 ± 0.89 µg/kg for tylosin. The mean contamination among most samples was below the maximum residue limits (MRLs). Upon comparison with the acceptable daily intake (ADI), the estimated daily intake (EDI) across all age groups was acceptable. The hazard quotient (HQ) was also below 1 across all age groups, signifying the absence of associated health risks for the Lebanese consumers. On the other hand, all milk samples were found exceeding the maximum tolerable value of mesophilic flora. Antibiotic-resistant bacteria (ARB) were detected and represented by ESBL-producing E. coli and MRSA isolates. Thus, the greatest threat of antibiotic use in Lebanon does not fall under antibiotic residues but rather the proliferation of antibiotic resistance in potentially pathogenic bacteria. In this study, the virulence profile of detected bacteria was not investigated; thus their pathogenicity remains unknown. Therefore, to mitigate this health threat in Lebanon, a "One Health" action plan against ABR is required. It will provide a framework for continued, more extensive action to reduce the emergence and spread of ABR in Lebanon.

Lactobacillus rhamnosus and Staphylococcus epidermidis in gut microbiota: in vitro antimicrobial resistance.

The gastrointestinal tract is one of the most complex microbiological niches containing beneficial and non-pathogenic bacterial strains of which some may evolve into virulent under specific conditions. Lactobacillus rhamnosus GG is of the most known beneficial species with an ability to protect the intestine as opposed to Staphylococcus epidermidis 444 which causes serious health risks due to its high antimicrobial resistance. This study investigates first the survival and coexistence ability of L. rhamnosus GG, and S. epidermidis 444 at different pH levels. Subsequently, lysozyme's antimicrobial and antibiofilm effect on these two strains was elucidated before adding different concentrations of oxytetracycline hydrochloride antibiotic. Results showed that 50% inhibition of L. rhamnosus GG, S. epidermidis 444, and a co-culture of these planktonic strains were obtained respectively at a lysozyme concentration of 30, 18, and 26 mg/mL after the addition of ethylenediamine tetra-acetic acid (EDTA). At a pH of 7.5, mixing lysozyme (at IC50) and EDTA with oxytetracycline hydrochloride (700 µg/mL) showed an additional bactericidal effect as compared to its known bacteriostatic effect. Similarly, the addition of lysozyme to the antibiotic further increased the biofilm eradication of S. epidermidis 444 and L. rhamnosus GG where a maximal eradication of 70% was reached. Therefore, the potential development of new drugs based on adding a lysozyme-EDTA mixture to different types of antibiotics may be highly promising.

Single, Subsequent, or Simultaneous Treatments to Mitigate Mycotoxins in Solid Foods and Feeds: A Critical Review.

Mycotoxins in solid foods and feeds jeopardize the public health of humans and animals and cause food security issues. The inefficacy of most preventive measures to control the production of fungi in foods and feeds during the pre-harvest and post-harvest stages incited interest in the mitigation of these mycotoxins that can be conducted by the application of various chemical, physical, and/or biological treatments. These treatments are implemented separately or through a combination of two or more treatments simultaneously or subsequently. The reduction rates of the methods differ greatly, as do their effect on the organoleptic attributes, nutritional quality, and the environment. This critical review aims at summarizing the latest studies related to the mitigation of mycotoxins in solid foods and feeds. It discusses and evaluates the single and combined mycotoxin reduction treatments, compares their efficiency, elaborates on their advantages and disadvantages, and sheds light on the treated foods or feeds, as well as on their environmental impact.

Assorted Methods for Decontamination of Aflatoxin M1 in Milk Using Microbial Adsorbents.

Aflatoxins (AF) are carcinogenic metabolites produced by different species of Aspergillus which readily colonize crops. AFM1 is secreted in the milk of lactating mammals through the ingestion of feedstuffs contaminated by aflatoxin B1 (AFB1). Therefore, its presence in milk, even in small amounts, presents a real concern for dairy industries and consumers of dairy products. Different strategies can lead to the reduction of AFM1 contamination levels in milk. They include adopting good agricultural practices, decreasing the AFB1 contamination of animal feeds, or using diverse types of adsorbent materials. One of the most effective types of adsorbents used for AFM1 decontamination are those of microbial origin. This review discusses current issues about AFM1 decontamination methods. These methods are based on the use of different bio-adsorbent agents such as bacteria and yeasts to complex AFM1 in milk. Moreover, this review answers some of the raised concerns about the binding stability of the formed AFM1-microbial complex. Thus, the efficiency of the decontamination methods was addressed, and plausible experimental variants were discussed.

A novel technique for aflatoxin M1 detoxification using chitin or treated shrimp shells: in vitro effect of physical and kinetic parameters on the binding stability.

This study aimed to investigate the ability of chitin and heat-treated shrimp shells to bind aflatoxin M1 (AFM1) in liquid matrix. Several concentrations of chitin or shrimp shells (grinded and ungrinded) were incubated in AFM1-contaminated phosphate-buffered saline (PBS) at different incubation times. The stability of the formed adsorbent-AFM1 complex was also tested in milk at different incubation times and temperatures. The unbound AFM1 was quantified by HPLC. Thereby, the percentages of the initial bounded AFM1 varied between 14.29 and 94.74%. Interestingly, in milk, an increase in incubation time coupled with a decrease in temperature affected positively the amount of bounded AFM1 to chitin and negatively those bounded to ungrinded shells. Results also revealed a partial reversibility in the binding of AFM1 to these adsorbents. These findings provided strong evidence on ability of chitin or shrimp shells by-product to bind AFM1 in milk and in PBS.

A comparative study of procedures for binding of aflatoxin M1 to Lactobacillus rhamnosus GG

ABSTRACT Several strains of lactic acid bacteria (LAB), frequently used in food fermentation and preservation, have been reported to bind different types of toxins in liquid media. This study was carried out to investigate the effect of different concentrations of Lactobacillus rhamnosus GG (ATCC 53103) to bind aflatoxin M1 (AFM1) in liquid media. AFM1 binding was tested following repetitive washes or filtration procedures in combination with additional treatments such as heating, pipetting, and centrifugation. The mixture of L. rhamnosus GG and AFM1 was incubated for 18 h at 37 °C and the binding efficiency was determined by quantifying the unbound AFM1 using HPLC. The stability of the complexes viable bacteria-AFM1 and heat treated bacteria-AFM1 was tested. Depending on the bacterial concentration and procedure used, the percentages of bound AFM1 by L. rhamnosus GG varied from as low as undetectable to as high as 63%. The highest reduction in the level of unbound AFM1 was recorded for the five washes procedure that involved heating and pipetting. Results also showed that binding was partially reversible and AFM1 was released after repeated washes. These findings highlight the effect of different treatments on the binding of AFM1 to L. rhamnosus GG in liquid matrix.

A comparative study of procedures for binding of aflatoxin M1 to Lactobacillus rhamnosus GG.

Several strains of lactic acid bacteria (LAB), frequently used in food fermentation and preservation, have been reported to bind different types of toxins in liquid media. This study was carried out to investigate the effect of different concentrations of Lactobacillus rhamnosus GG (ATCC 53103) to bind aflatoxin M1 (AFM1) in liquid media. AFM1 binding was tested following repetitive washes or filtration procedures in combination with additional treatments such as heating, pipetting, and centrifugation. The mixture of L. rhamnosus GG and AFM1 was incubated for 18h at 37°C and the binding efficiency was determined by quantifying the unbound AFM1 using HPLC. The stability of the complexes viable bacteria-AFM1 and heat treated bacteria-AFM1 was tested. Depending on the bacterial concentration and procedure used, the percentages of bound AFM1 by L. rhamnosus GG varied from as low as undetectable to as high as 63%. The highest reduction in the level of unbound AFM1 was recorded for the five washes procedure that involved heating and pipetting. Results also showed that binding was partially reversible and AFM1 was released after repeated washes. These findings highlight the effect of different treatments on the binding of AFM1 to L. rhamnosus GG in liquid matrix.