Human Breast Milk Contamination with Aflatoxins, Impact on Children's Health, and Possible Control Means: A Review.

Aflatoxins are natural toxicants produced mainly by species of the Aspergillus genus, which contaminate virtually all feeds and foods. Apart from their deleterious health effects on humans and animals, they can be secreted unmodified or carried over into the milk of lactating females, thereby posing health risks to suckling babies. Aflatoxin M1 (AFM1) is the major and most toxic aflatoxin type after aflatoxin B1 (AFB1). It contaminates human breast milk upon direct ingestion from dairy products or by carry-over from the parent molecule (AFB1), which is hydroxylated in the liver and possibly in the mammary glands by cytochrome oxidase enzymes and then excreted into breast milk as AFM1 during lactation via the mammary alveolar epithelial cells. This puts suckling infants and children fed on this milk at a high risk, especially that their detoxifying activities are still weak at this age essentially due to immature liver as the main organ responsible for the detoxification of xenobiotics. The occurrence of AFM1 at toxic levels in human breast milk and associated health conditions in nursing children is well documented, with developing countries being the most affected. Different studies have demonstrated that contamination of human breast milk with AFM1 represents a real public health issue, which should be promptly and properly addressed to reduce its incidence. To this end, different actions have been suggested, including a wider and proper implementation of regulatory measures, not only for breast milk but also for foods and feeds as the upstream sources for breast milk contamination with AFM1. The promotion of awareness of lactating mothers through the organization of training sessions and mass media disclosures before and after parturition is of a paramount importance for the success of any action. This is especially relevant that there are no possible control measures to ensure compliance of lactating mothers to specific regulatory measures, which can yet be appropriate for the expansion of breast milk banks in industrialized countries and emergence of breast milk sellers. This review attempted to revisit the public health issues raised by mother milk contamination with AFM1, which remains undermined despite the numerous relevant publications highlighting the needs to tackle its incidence as a protective measure for the children physical and mental health.

Prevalence of Aflatoxins in Selected Dry Fruits, Impact of Storage Conditions on Contamination Levels and Associated Health Risks on Pakistani Consumers.

Dry fruits and nuts are nutritious foods with several health-promoting properties. However, they are prone to contamination with aflatoxins at all stages of production and storage. The present study aimed to determine the natural occurrence of aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), and total aflatoxins (AFT) in dates, pistachios, and walnuts collected from four districts of South Punjab (Pakistan), and to assess the associated health risks as estimated by dietary exposure and the Margin of Exposure (MoE) determinations. The contents of AFB1 and AFT in these food products were monitored during storage under three different conditions (open-air, hermetically closed jars, and refrigeration at 4 °C) to determine the most efficient conditions in preventing aflatoxin accumulation. HPLC-fluorescence analysis of 60 samples of these products for aflatoxin contamination showed that 52 (86.7%) samples were contaminated at different levels, with a maximum of 24.2 ng/g. The overall (all samples) mean concentrations of AFB1, AFB2, AFG1, AFG2, and AFT were 3.39 ± 2.96, 1.39 ± 1.68, 1.63 ± 1.48. 1.12 ± 1.23, and 7.54 ± 6.68, respectively. The Estimated Daily Intake (EDI) and MoE of aflatoxins through the consumption of the products ranged from 0.06 ng/kg bw/day to 2.0 ng/kg bw/day and from 84.84 to 2857.13, respectively, indicating that consumers are at high health risk. Significant differences were recorded between aflatoxin levels in the samples stored under different storage conditions, with storage under refrigeration (4 °C) being the most effective in controlling aflatoxin accumulation, although storage in closed jars was also efficient and offers a more flexible alternative to retailers. The findings of the study urge official authorities of Pakistan to implement appropriate regulatory and control measures and surveillance program to alleviate the potential public health risks associated with the consumption of dry fruits and nuts in the scope of their increased consumption.

Occurrence and Health Risk Assessment of Aflatoxins through Intake of Eastern Herbal Medicines Collected from Four Districts of Southern Punjab-Pakistan.

Eastern herbal medicines (HMs) are plant-derived naturally occurring substances with minimum or no industrial processing that have long been used in traditional medicine. Aflatoxins are frequent contaminants of plants. Therefore, these mycotoxins are likely to contaminate HMs and pose a health risk to individuals using them on a regular basis as preventive or curative treatments of various diseases. The present study aimed to determine aflatoxin levels in the most popular Pakistani HM formulations and to assess the health risk associated with the intake of aflatoxins. A total of 400 samples of HM formulations collected from four districts of Punjab were analyzed for the quantification of aflatoxins, out of which 52.5% were found to be contaminated. The average daily dose (ADD) of AFB1 and AFs through the intake of HM formulations ranged between 0.00483 and 0.118 ng/kg bw/day and between 0.00579 and 1.714 ng/kg bw/day, respectively. The margin of exposure (MOE) and population cancer risk ranged from 99.49 to 29378.8 and from 0.00011 to 0.0325 liver cancer cases/105 individuals/year (0.0075-2.455 liver cancer cases/105 individuals/75 years), respectively. Despite the low exposure to aflatoxins from HM formulations in the four studied Punjab (Pakistan) districts, the frequent contamination of the analyzed samples suggests that official measures should be considered to manage the associated risk.

Aflatoxins: Producing-Molds, Structure, Health Issues and Incidence in Southeast Asian and Sub-Saharan African Countries.

This review aims to update the main aspects of aflatoxin production, occurrence and incidence in selected countries, and associated aflatoxicosis outbreaks. Means to reduce aflatoxin incidence in crops were also presented, with an emphasis on the environmentally-friendly technology using atoxigenic strains of Aspergillus flavus. Aflatoxins are unavoidable widespread natural contaminants of foods and feeds with serious impacts on health, agricultural and livestock productivity, and food safety. They are secondary metabolites produced by Aspergillus species distributed on three main sections of the genus (section Flavi, section Ochraceorosei, and section Nidulantes). Poor economic status of a country exacerbates the risk and the extent of crop contamination due to faulty storage conditions that are usually suitable for mold growth and mycotoxin production: temperature of 22 to 29 °C and water activity of 0.90 to 0.99. This situation paralleled the prevalence of high liver cancer and the occasional acute aflatoxicosis episodes that have been associated with these regions. Risk assessment studies revealed that Southeast Asian (SEA) and Sub-Saharan African (SSA) countries remain at high risk and that, apart from the regulatory standards revision to be more restrictive, other actions to prevent or decontaminate crops are to be taken for adequate public health protection. Indeed, a review of publications on the incidence of aflatoxins in selected foods and feeds from countries whose crops are classically known for their highest contamination with aflatoxins, reveals that despite the intensive efforts made to reduce such an incidence, there has been no clear tendency, with the possible exception of South Africa, towards sustained improvements. Nonetheless, a global risk assessment of the new situation regarding crop contamination with aflatoxins by international organizations with the required expertise is suggested to appraise where we stand presently.

Chronic and Acute Toxicities of Aflatoxins: Mechanisms of Action.

There are presently more than 18 known aflatoxins most of which have been insufficiently studied for their incidence, health-risk, and mechanisms of toxicity to allow effective intervention and control means that would significantly and sustainably reduce their incidence and adverse effects on health and economy. Among these, aflatoxin B1 (AFB1) has been by far the most studied; yet, many aspects of the range and mechanisms of the diseases it causes remain to be elucidated. Its mutagenicity, tumorigenicity, and carcinogenicity-which are the best known-still suffer from limitations regarding the relative contribution of the oxidative stress and the reactive epoxide derivative (Aflatoxin-exo 8,9-epoxide) in the induction of the diseases, as well as its metabolic and synthesis pathways. Additionally, despite the well-established additive effects for carcinogenicity between AFB1 and other risk factors, e.g., hepatitis viruses B and C, and the hepatotoxic algal microcystins, the mechanisms of this synergy remain unclear. This study reviews the most recent advances in the field of the mechanisms of toxicity of aflatoxins and the adverse health effects that they cause in humans and animals.

Retrospective and Prospective Look at Aflatoxin Research and Development from a Practical Standpoint.

Among the array of structurally and toxicologically diverse mycotoxins, aflatoxins have attracted the most interest of scientific research due to their high toxicity and incidence in foods and feeds. Despite the undeniable progress made in various aspects related to aflatoxins, the ultimate goal consisting of reducing the associated public health risks worldwide is far from being reached due to multiplicity of social, political, economic, geographic, climatic, and development factors. However, a reasonable degree of health protection is attained in industrialized countries owing to their scientific, administrative, and financial capacities allowing them to use high-tech agricultural management systems. Less fortunate situations exist in equatorial and sub-equatorial developing countries mainly practicing traditional agriculture managed by smallholders for subsistence, and where the climate is suitable for mould growth and aflatoxin production. This situation worsens due to climatic change producing conditions increasingly suitable for aflatoxigenic mould growth and toxin production. Accordingly, it is difficult to harmonize the regulatory standards of aflatoxins worldwide, which prevents agri-foods of developing countries from accessing the markets of industrialized countries. To tackle the multi-faceted aflatoxin problem, actions should be taken collectively by the international community involving scientific research, technological and social development, environment protection, awareness promotion, etc. International cooperation should foster technology transfer and exchange of pertinent technical information. This review presents the main historical discoveries leading to our present knowledge on aflatoxins and the challenges that should be addressed presently and in the future at various levels to ensure higher health protection for everybody. In short, it aims to elucidate where we come from and where we should go in terms of aflatoxin research/development.

Barley protein concentrates: Extraction, structural and functional properties.

Protein concentrates were prepared from defatted barley (Hordeum vulgare L.) flour using alkaline and enzymatic treatments. Milder enzymatic treatments included (i) a bi-enzymatic method involving the use of starch-hydrolyzing enzymes, and (ii) a tri-enzymatic method using the former bi-enzymatic treatment followed by digestion with glucanase. The concentrate obtained through alkaline extraction (AI-BP) was comprised mainly of low molecular weight fractions of proteins. Bi-enzymatic treatment produced a protein concentrate with the highest protein content (49.0%), while those obtained by the tri-enzymatic treatment followed by an isoelectric precipitation step (TEI-BP) gave the highest protein recovery yield (78.3%). In both of the latter concentrates, 35 kDa B-hordeins were the major protein fraction. Divergence in secondary/tertiary structure elements (AI-BP; TEI-BP) was obtained and attributed to the difference in the protein profiles. Further characterization of protein concentrates indicated that they exhibit pseudoplastic behavior. Emulsifying capacity of concentrates was comparable to that of whey protein isolate.

Staphylococcal enterotoxins and enterotoxin-like toxins with special reference to dairy products: An overview.

Staphylococcal Enterotoxins (SEs) have been raising health concerns for food safety due to their association with staphylococcal foodborne poisoning (SFP). As superantigens, they also cause the life threatening toxic shock syndrome (TSS), the transmission of which via food cannot be ruled out despite the lack of epidemiological evidence. To date, at least 23 of these exotoxins are known and separated into SEs and Staphylococcal Enterotoxin-like (SEl) depending on whether or not they invoke emesis. This work presents an up-to-date overview on the presently known SEs/SEls from the perspective of their classification, pathogenesis, and genetic organisation. The incidence of these toxins in dairy products, the risk this poses to the public health, and possible control means are also reviewed.

Production of exopolysaccharides by selected Bacillus strains: Optimization of media composition to maximize the yield and structural characterization.

Selected Bacillus strains were investigated for the production of exopolysaccharides (EPS) by fermentation in three different culture media: (i) mineral base-medium with added yeast extract (M1), (ii) succinate-containing mineral base-medium with added yeast extract (M2) and (iii) tryptone and yeast extract-containing base-medium (M3). Modest EPS yield of 6.7g/L was recorded for Bacillus amyloliquefaciens 23350 grown on M1, where the EPS produced was characterized by a low MW (<5kDa) and of being mainly glucans. The most significant yield (48.57g/L) was obtained with Bacillus licheniformis 14580 in M2; the EPS produced was 5-30kDa in size and characterized by an exceptionally heterogeneous monosaccharide profile with galactose, fructose and glucose as the predominating monomers. The use of M1 medium for the growth of B. licheniformis 14580 resulted in low yield; however, the obtained heteropolymers EPS were characterized by a higher MW 30-100kDa. The effect of the concentrations of yeast extract, sodium succinate and sucrose on the EPS production by B. licheniformis 14580 were studied using response surface methodology analysis. Greater EPS yields were achieved with increased concentrations of sodium succinate and sucrose and with decreased concentrations of yeast extract in the mineral media.

Biogenic Amines in Dairy Products: Origin, Incidence, and Control Means.

Biogenic amines (BAs) are toxic compounds produced by a number of microorganisms (bacteria, yeasts, and molds) as a result of the metabolism of some amino acid, usually decarboxylation reactions. BA-producing microorganisms are not necessarily pathogenic, such as lactic acid bacteria, which are, on the contrary, among the most beneficial microbiota to human beings and some of which even have probiotic properties. However, the incidence of BAs in dairy products and their possible implication in serious dairy-borne intoxications has long been overlooked. Consequently, the implementation of control measures to limit such an incidence has not been considered among the priorities of the food safety authorities. Nonetheless, there is a growing concern with regard to the presence of BAs in dairy products, because their toxicological status as toxins that may have serious acute and/or chronic adverse health effects is becoming increasingly evident and well-documented. The main BAs associated with dairy products are reviewed herein from the perspective of their incidence in these food products, and to draw the attention of readers to the shortage in data to perform pertinent risk assessment, which is considered to be a key action to provide efficient control means and to help decision makers issue appropriate legislative and regulatory measures.

Behaviour of Listeria monocytogenes in raw sausages (merguez) in presence of a bacteriocin-producing lactococcal strain as a protective culture.

The effectiveness of a bacteriocin produced by Lactococcus lactis subsp. lactis M in reducing population level and growth of Listeria monocytogenes ATCC 7644 in fermented merguez sausage was examined. Two different formulas (with or without added nitrites) were assayed and predetermined numbers of Listeria (ca 10(6) cfu g(-1)) were added to sausage mixture. The effect of in situ production of the bacteriocin by Lactococcus lactis M on Listeria monocytogenes ATCC 7644 during fermentation and storage of merguez sausages at room (ca 22 °C) or at refrigeration (ca 7 °C) temperature was tested. Results indicated that counts of Listeria monocytogenes were decreased during fermentation of merguez samples fermented with either the bacteriocin-producing Lactococcus lactis M (Bac(+)) or a nonbacteriocin-producing Lactococcus lactis J (Bac(-)). However, reduction in Listeria cfu's was greater in samples fermented with the Bac(+) than in those fermented with the Bac(-) starter. In merguez sausage made without nitrites addition, the Bac(+) starter induced further decrease in Listeria counts by 1.5 log cycles compared with that induced by the Bac(-) starter. While in merguez samples with added nitrites (0.4%), the effect of the bacteriocin produced in situ was less important than in those made without nitrites addition.

Behavior of Listeria monocytogenes and Staphylococcus aureus in yogurt fermented with a bacteriocin-producing thermophilic starter.

Streptococcus salivarius subsp. thermophilus B producing a bacteriocin active against Listeria monocytogenes ATCC 7644 and Staphylococcus aureus SAD 30 was isolated from bakery yeast. The bacteriocin was partially purified by an adsorption/desorption technique, and its spectrum of action was compared to that of a neutralized cell-free supernatant (CFS). Although the CFS inhibited a number of gram-positive and -negative bacteria of health and spoilage significance, the spectrum of action of the partially purified bacteriocin was limited to gram-positive bacteria. L. monocytogenes was the most sensitive to both preparations. The bacteriocin-producing streptococcal strain was used in combination with a Bac- Lactobacillus delbrueckii subsp. bulgaricus CY strain isolated from commercial yogurt to assess the effectiveness of the resulting thermophilic starter in controlling L. monocytogenes and S. aureus in yogurt during fermentation and storage at refrigeration (ca. 7 degrees C) or abuse (ca. 22 degrees C) temperature. Yogurt samples were contaminated with L. monocytogenes or S. aureus to the approximate levels of 10(3) and 10(6) CFU/ml of milk, respectively. The results showed that in situ bacteriocin production was more active against L. monocytogenes than against S. aureus in vitro and in contaminated samples. While L. monocytogenes leveled off below the detectable limit in a 1-ml sample of yogurt within 24 h of processing, S. aureus survived in Bac+ and Bac- samples during 10 days of storage at room temperature (ca. 22 degrees C). Use of a Bac+ starter resulted in a 5-day extension of the shelf life.