The protective effects of the herbal feed supplements Silybum marianum, Silymarin, Withania somnifera, and Centella asiatica against ochratoxin A (OTA) toxicity were studied in 48 New Zealand White rabbits (37-day-old) during an 80-day experiment. OTA was given at 2 ppm, whereas Silybum marianum, Silymarin, Withania somnifera, and Centella asiatica were given at feed levels of 5000 ppm, 25,000 ppm, 4000 ppm, and 4600 ppm, respectively. All rabbits were immunized against Rabbit Hemorrhagic Disease Virus (RHDV). OTA was found to induce an immunosuppressive effect on the humoral immune response. Reliable protection against OTA-provoked immunosuppression by Silimarin and Withania somnifera was found. The OTA-induced immunosuppression was responsible for secondary bacterial infection (pasteurellosis) and the death of two rabbits from the OTA-exposed group and one rabbit each from the groups protected with Silybum marianum and Centella asiatica. A decreased body weight was found in rabbits exposed to OTA, but the decrease was slighter in the rabbits protected with herbal supplements. The target organs damaged by OTA exposure were the liver, kidneys, and spleen, while weaker lesions were found in other internal organs, except in the cases of secondary pasteurellosis, in which the strongest damage was found in the lung. All investigated herbal supplements appeared to have stronger protective effects against OTA-induced damage to the kidneys and liver, with slightly protective effects observed in the lungs, myocardium, spleen, brain, intestine, testicles, and ovaries.

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.

This review paper highlights the most prevalent mycotoxins in agricultural commodities and possible hazard of mycotoxin occurrence in foods/feeds in regards to foodborne diseases. The underestimated hazard of multiple mycotoxin exposure of humans or animals is also briefly evaluated. The risk assessment and management of the risk of mycotoxin contamination of foods/feeds by some appropriate preventive measures against mycotoxin contamination of raw materials and food commodities in the pre-harvesting and post-harvesting time is elucidated. The most common chemical or physical methods of mycotoxins decontamination of agricultural commodities and raw materials are evaluated. The practical use of feed/food additives as a possible method for preventing the adverse effects of some target mycotoxins or mycotoxins combination is shortly reviewed. The adequate processing of some raw materials and agricultural commodities with regards to possible mycotoxin decontamination is evaluated. A survey is also made in regard to the veterinary preventive measures and the possible hygiene control and risk assessment in some practical cases of foodborne mycotoxicoses in order to prevent mycotoxins contamination of target food commodities from animal origin and to avoid a possible entering of some mycotoxins in commercial channels with a view to ensure adequate public health.

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.

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.

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.

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.

Protective effects of herbal feed additive Silymarin against the deleterious toxic effects of ochratoxin A (OTA) on internal organs and blood biochemistry was seen. The observed histopathological and biochemical changes were well expressed in OTA-exposed chicks without Silymarin supplementation, followed by chicks treated supplementary with Silymarin in addition to OTA treatment, whereas no pathological changes were seen in the control chicks or the chicks treated with Silymarin only. The observed increase in the serum levels of uric acid, glucose and the enzyme activity of AST and ALT in OTA treated chicks and the found decrease of the same biochemical indices in the chicks protected by Silymarin in addition to OTA treatment also supported the protective effects of this herbal additive on the kidneys and liver. The same herbal substance Silymarin could be used as a practical approach for safely utilizing of OTA-contaminated feed. Key words: ochratoxin A, protection, herbal additive, Silymarin, blood biochemistry, pathomorphology

A protective effect of two herbs,Glycyrrhiza glabraandTinospora cordifolia, given as feed additives was observed against the growth inhibitory effect of ochratoxin A (OTA) and associated immunosuppression and biochemical or pathomorphological changes. The feed levels of 3 mg/kg OTA and fine powder of one of both herbs were given during a period of 32 days to female broiler chicks divided into 3 experimental and 1 control groups (14 chicks per group). The observed pathological and biochemical changes, the changes in relative organs’ weight and body weight, and the decrease of antibody titer against Newcastle disease were more pronounced in the OTA-treated chicks without herbal supplementation, and less pronounced in the chicks treated additionally withG. glabraorT. cordifoliaas was shown by the better feed performance and the higher body weight in the chicks treated with the herbs. The higher relative weight of lymphoid organs of the chicks supplemented with both herbs revealed their beneficial effects on the immune system. The hepatoprotective effect of both herbs was evident, being stronger in the chicks additionally supplemented withG. glabrashown by the pathomorphological findings and by the lower levels of aspartate transaminase (131.1 U/l) compared to chicks given only OTA (156.0 U/l). A protective effect ofT. cordifoliaon the bone marrow and kidneys was found as was shown by the lower levels of uric acid (382.9 μmol/l) compared to chicks given only OTA (466.9 μmol/l).

Abstract The aim of the study was to provide an in-depth characterization of the methanol-aqueous extract from the aerial parts of Gypsophila glomerata Pall. Ex Adams (Caryophyllaceae) (EGG) and to assess its protective potential on carbon tetrachloride (CCl4)-induced liver and kidney damage in male Wistar rats. Twenty-two flavonoid C-, O- and C,O--glycosides in EGG were annotated by mass spectrometry--based molecular networking; nine of them are reported in this species for the first time. Fourteen-day oral administration of EGG at a dose 200 mg kg−1 bm prevented significantly CCl4-induced liver injury, discerned by an amelioration of the markers of oxidative stress (GSH and MDA) and transaminase activity. EGG decreased the serum level of urea and creatinine as well. The observed improvement of biochemical parameters was supported by histopathological observations. The protective hepatorenal effects of EGG, rich in 2“-О-pentosyl-6-С-hexosyl-apigenin/luteolin/ methylluteolin and their acetyl- and methoxycinnamoyl-derivatives, were comparable with the effects of the positive control silymarin.

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.

This review is designed to initiate a discussion we believe is necessary for the biomedical community, because of some recent evidences for existing of a new body anatomical system, or the primo vascular system (PVS), which could be the missing link in the scientific explanation of the unknown mechanism of action of acupuncture. Some important questions for the medical society, (eg, "What is the main source of the mistrust of Western medicine toward traditional Oriental medicine and could it be overcome?" or "Is the PVS a real one and what is its distribution, formation, and function?" or "Are there scientific proofs for intimate relationships of the PVS with meridian system and whether the PVS would be the physical basis of meridians?") are deeply studied and appropriately answered. Various pieces of knowledge are now combined to achieve a better understanding and to provide an acceptable explanation about the functions of such new system and to explain the functional path used by traditional Eastern medicine to cure diseases. Some possibilities to use this PVS for development of some innovative therapies to treat some diseases are also discussed (eg, pharmacopuncture as a new innovative drug delivery method that combines acupuncture therapy with medication by injecting pharmacological substances into target acupoints).

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.

The protective effects of herbal feed additives Silybum marianum, Withania somnifera and Centella asiatica against the toxic effects of ochratoxin A (OTA) were studied in 70 broiler chicks aged from 11 to 42 days. OTA was given with the feed at levels 5 mg/kg, whereas S. marianum , W. somnifera and C. asiatica were given at levels of 1,100, 4,000 and 4,600 mg/kg, respectively. All chicks were immunised at the age of 14 days against Newcastle disease. A protective effect of all studied herbal additives against the immunosuppressive effect of OTA and associated biochemical or pathomorphological changes was seen. The intensity of macroscopical and histopathological changes, the deviations in relative organs’ weight or body weight, the biochemical changes and the decrease of antibody titer were strongest in the OTA-exposed chicks without herbal supplementation; followed by chicks treated additionally with C. asiatica , whereas the same changes were significantly slighter or not seen in chicks additionally treated with the herbal additives W. somnifera or S. marianum . The slight increase in the serum levels of uric acid and the enzyme activity of aspartate transaminase and alanine transaminase also supported the protective effects of both herbs on the kidneys and/or liver. The strong immunosuppressive effect of OTA on humoral immune response against Newcastle disease was completely prevented in the chicks taking the herbal additives W. somnifera or S. marianum , which was supported by the higher relative weight of immunocompetent (lymphoid) organs in the same chicks. A hepatoprotective effect was found in OTA-exposed chicks treated additionally with W. somnifera and S. marianum, whereas a nephroprotective effect was only found in the chicks additionally treated with S. marianum as observed from the biochemical and pathomorphological findings. The same herbs could be used as a practical approach for safely utilising of OTA-contaminated feed.

The importance of Tinospora cordifolia extract was investigated for its possible protective effect in spleen and blood in male ICR albino mice against ochratoxin A-induced toxicity (OTA). OTA instigates oxidative changes and results in the overproduction of free radicals, changes in body/organ weight, and the food consumption and is considered as a risk factor for animals and humans. Reactive oxygen species (ROS), reactive nitrogen species (RNS), and consequent lipid-peroxidative damages caused by OTA are considered to be the main mechanisms leading to oxidative stress disorders in organs and blood. TC oral administrations prevent the physiological status of animals and improve the biochemical parameters of the urine and plasma. Moreover, EPR analysis show that TC administration decreased Asc• and NO• radicals and ROS productions in the spleen and serum, even in OTA-treated group. The significant decrease in MDA formation in the spleen and serum and the increased expression of SOD activity in TC and TC + OTA groups confirm the positive modulatory effect of TC extract on the cellular antioxidant system. TC and TC + OTA treatment caused significant reduction of genotoxic potential (8-OHdG) of OTA, followed by a decreased oxidative activity and gradual recovery of ROS-induced DNA damage. The results suggested that TC extract protect against OTA-induced oxidative disorders and other abnormalities. As typical antioxidant TC could protect splenic macrophages and regain intracellular antioxidant capacity.

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.

The possible interaction between Pasteurella multocida and the mycotoxin fumonisin B1 (FB1), recognised as one of the most often food/feed contaminant, was studied with the aim to evaluate whether and how FB1 can influence and/or complicate the development and severity of various pathological damages provoked by Pasteurella multocida in some internal organs of pigs. Heavier lung pathology was seen in pigs experimentally infected with Pasteurella multocida, when the same were exposed to 20ppm dietary levels of fumonisin B1 (FB1) as was assessed by gross pathology, pathomorphological examinations, clinical biochemistry and some immunological investigations. The most typical damages in FB1 treated pigs were the strong oedema in the lung and the slight oedema in the other internal organs and mild degenerative changes in the kidneys, whereas the typical pathomorphological findings in pigs infected with Pasteurella multocida was broncho-interstitial pneumonia. FB1 was found to aggravate pneumonic changes provoked by P. multocida in the cranial lobes of the lung and to complicate pneumonic damages with interstitial oedema in the lung. No macroscopic damages were observed in the pigs infected only with Pasteurella multocida. It can be concluded that the feed intake of FB1 in pigs may complicate or exacerbate the course of P. multocida serotype A infection.

Authors’ addresses: 1 Radiation Biotechnology Group, Institute of Nuclear Medicine and Allied Sciences, Brig. S.K. Mazumdar Marg, Defence Research and Development Organization, Delhi-110054, India. 2 Department Chemistry and Biochemistry, Medical Faculty, Trakia University, 6000 Stara Zagora, Bulgaria. 3 Department Physiology, Pathophysiology and Pharmacology, Medical Faculty, Trakia University, 6000 Stara Zagora, Bulgaria. 4 Department of General and Clinical Pathology, Faculty of Veterinary Medicine, Trakia University, 6000 Stara Zagora, Bulgaria.

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.