Food Security and Foodborne Mycotoxicoses-What Should Be the Adequate Risk Assessment and Regulation?

The purpose of this review is to elucidate the actual threat of the most prevalent mycotoxins in agricultural commodities and human/animal food/feed for the induction of foodborne diseases or ailments. The underestimated hazard of combined mycotoxin uptake by animals or humans is critically discussed with regard to synergistic or additive interaction between some target mycotoxins. The real toxicity of target mycotoxin combinations as it happens in practice is evaluated and possible lower limit values or control measures are suggested in such cases. Some critical points on adequate risk assessment, hygiene control, and regulation of mycotoxins are discussed. The efficiency of current mycotoxin regulations and control measures is evaluated in regard to human/animal health hazards. The risk assessment in the case of multiple mycotoxin exposure of humans/animals via food/feed or agricultural commodities is evaluated and some suggestions are proposed in such cases. Appropriate control measures and food safety issues throughout the food supply chain are proposed in order to prevent the target foodborne diseases. Some preventive measures and possible veterinary hygiene controls or risk evaluations are proposed in some natural cases of foodborne diseases for preventing mycotoxin contamination of animal products designed for human consumption and to avoid possible public health issues.

Natural feed additives and bioactive supplements versus chemical additives as a safe and practical approach to combat foodborne mycotoxicoses.

This review highlights the possible hazard of mycotoxins occurrence in foods and feeds in regards to foodborne diseases. The possible management of the risk of contamination of foods and feeds with mycotoxins by using natural feed additives, protecting against deleterious effects of mycotoxins or inhibiting the growth of fungi and mycotoxin production, is deeply investigated in the available literature and some effective measures for safe utilization of mycotoxin contaminated feed/food are proposed. The biological methods of decontamination, degradation or biotransformation of mycotoxins are deeply analyzed and discussed. Some natural antagonists against target fungi are also reviewed and a comparison is made with conventional fungicides for ensuring a safe prevention of mycotoxin contamination. The most common and useful chemical methods of mycotoxins decontamination of agricultural commodities or raw materials are also investigated, e.g., chemical additives inactivating or destroying and/or adsorbing mycotoxins as well as chemical additives inhibiting the growth of fungi and mycotoxin production. The practical use and safety of various kind of feed/food additives or herbal/biological supplements as possible approach for ameliorating the adverse effects of some dangerous mycotoxins is deeply investigated and some suggestions are given. Various possibilities for decreasing mycotoxins toxicity, e.g., by clarifying the mechanisms of their toxicity and using some target antidotes and vitamins as supplements to the diet, are also studied in the literature and appropriate discussions or suggestions are made in this regard. Some studies on animal diets such as low carbohydrate intake, increased protein content, calorie restriction or the importance of dietary fats are also investigated in the available literature for possible amelioration of the ailments associated with mycotoxins exposure. It could be concluded that natural feed additives and bioactive supplements would be more safe and practical approach to combat foodborne mycotoxicoses as compared to chemical additives.

Foodborne Diseases Due to Underestimated Hazard of Joint Mycotoxin Exposure at Low Levels and Possible Risk Assessment.

The subject of this review paper is to evaluate the underestimated hazard of multiple mycotoxin exposure of animals/humans for the appearance of foodborne ailments and diseases. The significance of joint mycotoxin interaction in the development of foodborne diseases is discussed, and appropriate conclusions are made. The importance of low feed/food levels of some target mycotoxins co-contaminations in food and feedstuffs for induction of target foodborne mycotoxicoses is also studied in the available literature. The appropriate hygiene control and the necessary risk assessment in regard to possible hazards for animals and humans are also discussed, and appropriate suggestions are made. Some internationally recognized prophylactic measures, management of the risk, and the necessity of elaboration of new international regulations in regard to the maximum permitted levels are also carefully discussed and analysed in the cases of multiple mycotoxin contaminations. The necessity of harmonization of mycotoxin regulations and control measures at international levels is also discussed in order to facilitate food trade between the countries and to ensure global food safety.

New Evidences about the Carcinogenic Effects of Ochratoxin A and Possible Prevention by Target Feed Additives.

A review of the carcinogenic effects of ochratoxin A (OTA) on various tissues and internal organs in laboratory and farm animals is made. Suggestions are made regarding how to recognize and differentiate the common spontaneous neoplastic changes characteristic for advanced age and the characteristic neoplasia in different tissues and organs in laboratory animals/poultry exposed to OTA. The synergistic effects of OTA together with its natural combination of penicillic acid are also investigated regarding possible carcinogenic effects. The malignancy and the target location of OTA-induced neoplasia is studied. The sex-differences of such neoplasia are investigated in the available literature. The time of appearance of the first neoplasia is investigated in long-term carcinogenic studies with OTA-treated animals. The possibility of target feed additives or herbs to counteract the toxic and carcinogenic effects of OTA is studied in the available literature. Some effective manners of prophylaxis and/or prevention against OTA contamination of feedstuffs/foods or animal production are suggested. The suitability of various laboratory animals to serve as experimental model for humans with regard to OTA-induced tumorigenesis is investigated.

Studies on teratogenic effect of ochratoxin A given via mouldy diet in mice in various sensitive periods of the pregnancy and the putative protection of phenylalanine.

Embryotoxic and teratogenic effects were found in OTA-treated pregnant Swiss albino mice, which were particularly strong at OTA contamination levels of 20 ppm (corresponding to 2.8 mg OTA/kg bw per day), when fed between the day 7 and day 12 of the pregnancy. Slighter embryotoxic and teratogenic effects were found when OTA was given up to day 7 or after day 12 of mice pregnancy. The feed levels of 10 ppm OTA (corresponding to 1.4 mg OTA/kg bw per day) have a significantly weaker embryotoxic and teratogenic effects on pregnant mice and no such effects were found at 5 ppm OTA (corresponding to 0.7 mg OTA/kg gt per day). The main OTA-induced malformations wre seen in the craniofacial structures, e.g. anophthalmia, monophthalmia, microphthalmia, astomia, microstomia, maxillary hypoplasia, microcephaly, macrocephaly, in newborn mice as compared to OTA-induced somatic malformations, e.g. peromelia, micromelia, spina bifida and facial cleft. Phenylalanine given at 20 ppm in the diet was found to have a protective effect against embryotoxic and teratogenic effects of OTA.

Follow up long term preliminary studies on carcinogenic and toxic effects of ochratoxin A in rats and the putative protection of phenylalanine.

Carcinogenic effects of ochratoxin A (OTA) on liver, kidneys, intestine, lung and eyes of Wistar rats exposed to 10 ppm or 5 ppm OTA in the diet and additionally supplemented or not with phenylalanine (PHE) were examined during 24-months experimental period. OTA was seen to provoke strong degenerative changes and slight pericapillary oedema in most internal organs, e.g. kidneys, liver, intestine, spleen and brain. Six of total nine neoplasms were identified as malignant and three as benign. Five of total six malignant neoplasms and two of total three benign neoplasms were seen in male rats. The pathological finding in rats after two weeks feeding with OTA-contaminated feed was dominated by degenerative changes in various internal organs, which were weaker in the group additionally supplemented with PHE. The protective effect of PHE was evident with respect to OTA-induced decrease of serum glucose and serum protein, but this protection was not singnificant with respect to serum enzymes activity. The number of neoplasms in PHE-supplemented group exposed to 10 ppm OTA was similar to that in the group exposed to twice lower feed levels of OTA alone, suggesting about a possible protective effect of PHE. The rats would not be able to serve as experimental model for humans with regard to OTA-induced tumorigenesis, because the target organ of OTA-toxicity in humans and pigs is mainly the kidney as opposed to the significant damages and carcinogenic effects seen in various organs in rats exposed to OTA.

Long term preliminary studies on toxic and carcinogenic effect of individual or simultaneous exposure to ochratoxin A and penicillic acid in mice.

Carcinogenic effects of ochratoxin A (OTA) on liver, kidneys, intestine, muscles and subcutaneous tissue of BALB/c albino mice divided in three experimental and one control groups (30 mice in each group - 15 males and 15 females) and exposed to 10 ppm OTA and/or 50-60 ppm penicillic acid (PA) in the diet were seen. A total 22 neoplasias were found to be induced in the mice during 20 months' experimental period. Among them 14 were malignant and 8 benign. The number of neoplasias was significantly higher in the mice treated simultaneously with OTA and PA (14) compared to those in mice treated with OTA only (8). The number of malignant neoplasias was also higher (14) compared to benign neoplasias (8). Nine of a total fourteen malignant neoplasis and five of a total eight benign neoplasias were seen in the male mice. Pathological changes in mice after two weeks' experimental period were characterized by degenerative changes in kidneys, liver and gastrointestinal tract, which were better expressed in the mice exposed simultaneously to OTA and PA. A strong synergistic effect was found between OTA and PA towards tumorogenesis. It seems that mice are not a good experimental model for humans with regard to OTA-induced tumourigenesis, because the target organ of OTA-toxicity in humans or pigs is mainly the kidney, but not the liver, intestines, subcutaneous tissue or muscles as seen in this study.

Balkan Endemic Nephropathy - Still continuing enigma, risk assessment and underestimated hazard of joint mycotoxin exposure of animals or humans.

The spreading of mycotoxic nephropathy in animals/humans was studied. The possible etiological causes provoking this nephropathy were carefully reviewed and analyzed. The natural content of the most frequent nephrotoxic mycotoxins in target feedstuffs/foods were investigated, in addition to their significance for development of renal damages in endemic areas. An estimation of the level of exposure of humans to the nephrotoxic mycotoxin, ochratoxin A (OTA), is made. The possible synergism or additive effects between some target mycotoxins in the development of nephropathy is also covered. The significance of joint mycotoxin interaction and masked mycotoxins, in addition to some newly isolated fungal toxic agents in the complicated etiology of mycotoxic nephropathy ranged in Balkan countries is discussed. The importance of some target fungal species which can induce kidney damages was evaluated. The morphological/ultrastructural, functional and toxicological similarities between human and animal nephropathy are studied. The possible hazard of low content of combinations of some target mycotoxins in food or feedstuff ingested by pigs, chickens or humans under natural conditions is evaluated and a risk assessment was made. Some different but more effective manners of prophylaxis and/or prevention against OTA contamination of feedstuffs/foods are suggested. A survey was made in regard to the best possible ways of veterinary hygiene control of OTA-exposed animals at slaughter time for preventing the entrance of OTA in commercial feedstuffs/food channels with a view to reduce the possible health hazard for humans. The economic efficacy and applicability of such preventive measures is additionally discussed and some practical suggestions are made.

Foodborne mycotoxicoses, risk assessment and underestimated hazard of masked mycotoxins and joint mycotoxin effects or interaction.

The existing hazard of joint mycotoxin exposure of animals/humans and the significance of masked mycotoxins in foods or feeds and their respective contributions to the development of some food born mycotoxicoses is briefly reviewed. The importance of joint mycotoxin interaction in the complex etiology of some foodborn mycotoxicoses is covered in depth. The toxicity of low contamination levels of some combinations of mycotoxins ingested often by farm animals was carefully studied. The appropriate hygiene control and the necessary risk assessment in regard to mycotoxin contamination of foods and feeds are briefly analyzed and some useful prophylactic measures and management of the risk of mycotoxin contamination, in addition to tolerable daily intakes are also described. A reference is also made to the most suitable methods of veterinary hygiene control in some practical situations in order to prevent mycotoxins contaminating commercial food commodities and endangering public health.

Porcine/chicken or human nephropathy as the result of joint mycotoxins interaction.

A survey was made of the literature concerning the occurrence and incidence of mycotoxic nephropathy in pigs and chicks in different countries. Various etiological factors contributing to the development of the disease were considered. The main nephrotoxic fungi as well as the specific conditions for their growth and toxins production were briefly described. A survey was made about the most frequent nephrotoxic fungal contaminants in various feedstuffs from plant origin. In addition, their natural quantities and importance for development of mycotoxic porcine/chick nephropathy (MPN/MCN) are also explored. In addition, a survey was made of the feedstuffs representing the most favorable environment for nephrotoxic fungal growth as well as the most favorable storehouse conditions for this fungal growth were shortly described. The significance of some underestimated fungal species, which can provoke kidney damage, was studied. The importance of joint mycotoxin interaction and newly identified fungal metabolites in the complex etiology of mycotoxic nephropathy ranged in some countries is deeply investigated. The toxicity of the low contamination levels of some combinations of mycotoxins often administered by pigs and chicks in the practice was carefully studied.

Food safety and increasing hazard of mycotoxin occurrence in foods and feeds.

The possible hazard of mycotoxin occurrence in foods and feeds and some food-borne mycotoxicoses is reviewed. Management of the risk of mycotoxin contamination using some useful preventive measures against mycotoxin contamination of foods/feeds during pre- and post-harvesting periods is considered. The physical and chemical methods of mycotoxin decontamination of foods/feeds are briefly described. The use of various feed additives as a method for prevention of the adverse effects of mycotoxins is reviewed. The processing of various foods and feeds is considered in a view to possible mycotoxin decontamination. The necessary hygiene control and risk assessment in regard to mycotoxin contamination of foods and feeds in addition to some useful prophylactic measures are briefly described. A short reference is made concerning the most successful methods of veterinary hygiene control in order to prevent a possible entering of some mycotoxins in commercial channels with a view to human health.

Experimental mycotoxic nephropathy in pigs provoked by a mouldy diet containing ochratoxin A and fumonisin B1.

Mycotoxic nephropathy was induced in eighteen young pigs by mouldy diets containing 0.5 ppm ochratoxin A (OTA) and/or 10 ppm fumonisin B1 (FB1) for three months. While the most obvious damages provoked by OTA were seen in the kidneys as expressed by the strong degenerative changes in proximal tubules and fibrosis in kidneys, FB1 was found to induce an increase in permeability of vessels mainly in lung, brain, cerebellum or kidneys and slight to moderate degenerative changes in kidneys. Pathomorphological damages in pigs exposed to both mycotoxins simultaneously present a combination of the main lesions provoked by each mycotoxin alone being stronger in their expression. Biochemical investigations as expressed by the increase of serum creatinine, urea and enzyme activity of ASAT/ALAT and by the decrease of serum cholesterol, total protein, albumin and glucose were strongest in pigs exposed to both mycotoxins simultaneously as can be anticipated form the strongest lesions in the kidneys. Both mycotoxins and their combination were found to disturb powerfully humoral immune response in all experimental pigs as expressed by the strong decrease in antibody titer against Morbus Aujesky at days 21 and 35 after vaccination. Having in mind that the feed levels of the both mycotoxins as well as the exposure time and the pathological findings corresponded to those in some spontaneous cases of porcine nephropathy in Bulgaria and South Africa, it can be concluded that the same mycotoxins are involved in the observed field cases of that nephropathy.

Studies on some feed additives and materials giving partial protection against the suppressive effect of ochratoxin A on egg production of laying hens.

The protective effects of various feed supplements against the harmful effect of ochratoxin A on egg production and sexual maturation of two-weeks old Plymouth Rock female chicks designed for laying hens were studied. A significant protective effect of the feed additives or materials: water extract of artichoke (WEA), sesame seed (SS), Roxazyme-G (RG) and l-beta phenylalanine (PHE) against the suppressive effect of ochratoxin A (OTA) on egg production of laying hens was found. A similar protection was also seen on the toxic effect of OTA on various internal organs of the same hens. A significant protection was found against the decrease of the weight or the quantity of eggs as well as against the delay of the beginning of the laying period of chicks, both of which were provoked by ochratoxin A. These protective effects were strongest in chicks treated with SS or WEA, but were slightest in chicks treated with l-beta PHE.

Complex etiology and pathology of mycotoxic nephropathy in South African pigs.

Spontaneous nephropathy in pigs seen in South Africa was found to have multi-mycotoxic etiology involving several mycotoxins such as ochratoxin A (OTA), penicillic acid (PA) and fumonisin B1 (FB1) in addition to a not yet identified mycotoxin. Contamination levels of OTA were comparatively low (67-75 µg/kg) in contrast to high contamination levels of FB1 (5,289-5,021 µg/kg) and PA (149-251 µg/kg). A heavy contamination with Gibberella fujikuroi var. moniliformis and Penicillium aurantiogriseum complex (mainly P. polonicum) was observed in the fed forages in contrast to the light contamination with Aspergillus ochraceus, P. verrucosum and P. citrinum. The pathomorphological picture of this nephropathy was found to differ from the classical description of mycotoxic porcine nephropathy as originally made in Scandinavia by the extensive vascular changes.

Simultaneous occurrence of mycotoxins in human food commodities from Cameroon.

Eighty-two samples of dried food commodities from Cameroon were screened and quantified for different mycotoxins, including fumonisin B1 (FB1), zearalenone (ZEA), deoxynivalenol (DON), aflatoxin (AF) and ochratoxin A (OTA), by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), respectively. The percentage of positive samples was as follows: FB1 41%, AF 51%, ZEA 57%, DON 65% and OTA 3%. High FB1 contents were found in maize, averaging 3,684 µg/kg (range: 37-24,225 µg/kg), whereas the highest average ZEA level was found in peanuts (70 µg/kg), followed by maize (69 µg/kg), rice (67 µg/kg) and beans (48 µg/kg) with no ZEA was detected in soybeans. DON contents were low, ranging from 13 to 273 µg/kg, and for AF the average content was 2.6 µg/kg with peanuts and maize as principal substrates. The incidence of OTA was low, with a mean level of 6.4 µg/kg recorded. The majority (79%) of samples contained more than one mycotoxin and the most frequent co-occurrence found was FB1 + ZEA + DON, detected in 21% of samples (mainly maize) analysed. Co-contamination with FB1 + ZEA + DON + AF was found in 11% of the samples. Although a large proportion of samples had fairly low levels of individual mycotoxins, this should be of concern as the co-occurrence of mycotoxins may generate additive or synergistic effect in humans, especially if the respective commodities are consumed almost on a daily basis.

Studies on carcinogenic and toxic effects of ochratoxin A in chicks.

Carcinogenic/toxic effects of ochratoxin A (OTA) in various internal organs of Plymouth Rock chicks were determined. The number of OTA-induced neoplasms was similar in chicks given 25 ppm L-ß-phenylalanine (PHE) in addition to 5 ppm OTA compared to chicks given only 5 ppm OTA, which showed that PHE cannot be used as a real protector against the carcinogenic or toxic effects of OTA in chicks. OTA was found to provoke strong degenerative changes in liver and kidneys, degenerative changes and depletion of cells in lymphoid organs, oedematous and degenerative changes in the brain, muscular haemorrhages and fatty changes in the bone marrow. The target organs for carcinogenic effect of OTA in chicks were found to be kidneys and liver.

Complex etiology, prophylaxis and hygiene control in mycotoxic nephropathies in farm animals and humans.

Various etiological factors contributing to the development of mycotoxic nephropathy in farm animals and humans are reviewed. The possible synergistic effect between ochratoxin A (OTA) and other mycotoxins, as penicillic acid (PA) and fumonisin B(1) (FB(1)), contributing to this nephropathy is also considered and discussed. The most convenient ways of prophylaxis and various preventive measures against OTA contamination of feeds or foods are reviewed. A reference is made concerning the most successful methods of veterinary hygiene control in the slaughterhouses in order to prevent the entering of OTA in commercial channels with a view to human health. The economic efficacy of these prophylactic procedures is also considered. An evaluation of human exposure to OTA is made.

Experimental cadmium poisoning in sheep.

Pathomorphological investigations of internal organs were made in sheep given sublethal doses of cadmium. Early histological damages in kidneys were established by renal biopsy in some experimental animals. The main histological changes were characterized by granular degeneration in proximal tubules and glomerular endothelial proliferation in kidneys, granular degeneration in hepatocytes, pericapillary oedema and activation of capillary endothelium and Kupffer cells in liver, oedematous and degenerative changes in cerebrum and in the region of Purkinje cells of cerebellum, hyperplasia and proliferation of alveolar epithelium and perivascular or peribronchial mononuclear cell infiltration in lung, and degenerative changes in the medulla and zona glomerulosa of adrenal glands.

Studies on some feed additives giving partial protection against ochratoxin A toxicity in chicks.

Significant protective effects of the feed additives: water extract of artichoke, sesame seed, Roxazyme-G and L-beta phenylalanine against the growth inhibitory effect of ochratoxin A (OTA) and associated pathomorphological changes were seen. Similarly, there was less OTA-induced decrease in serum total protein and increase of serum creatinine and urea in the chicks. Whereas OTA induced strong degenerative changes and an increase in weight of kidneys and liver as well as a decrease of the weight of lymphoid organs the additives variously gave protection against these changes. The protection of Roxazyme-G and sesame seed was better expressed in kidneys and liver, whereas the phenylalanine better protected the weight changes in gizzard, heart and the changes in differential WBC count. Notably, sesame seed gave strong protection against 5 ppm OTA-induced suppression of humoral immune response, for which artichoke also had some beneficial effect, whereas phenylalanine had hardly any effect.

Spontaneous mycotoxic nephropathy in Bulgarian chickens with unclarified mycotoxin aetiology.

Histopathological, biochemical and toxicological investigations of tissues and blood of normally slaughtered chickens exhibiting different frequencies (1-2%, 40-50% and above 80%) of nephropathy changes (congested or pale and enlarged kidneys) at the slaughtering meat inspection were carried out to elucidate the aetiology of nephropathies of chickens encountered in Bulgaria. A close relationship was observed between the frequency of this nephropathy and the rate of nephrotoxic mycotoxin ochratoxin A in muscles, kidneys and livers of chickens, but the levels of ochratoxin A in corresponding feed samples (0.1-0.3 ppm) were significantly lower than the levels (2-4 ppm) required to reproduce such nephropathy. Clinicomorphological changes such as nervous symptoms, vascular and oedematous changes in various internal organs and the brain, and subcutaneous or liver and kidney haemorrhages in addition to known degenerative changes in the kidneys, liver and lymphoid organs differed from the classical description of the nephropathy made in Scandinavia. The conclusion is that the Bulgarian chicken nephropathy may have a multitoxic aetiology because it cannot be explained by the concentration of ochratoxin A alone.