Mycotoxic nephropathy was induced in 18 young pigs by diets contaminated with strains of Aspergil... more Mycotoxic nephropathy was induced in 18 young pigs by diets contaminated with strains of Aspergillus ochraceus containing ochratoxin A (OTA) and penicillic acid (PA) at levels corresponding to those naturally encountered in animal feeds in Bulgaria. Haematological and biochemical parameters, as well as the morphological and ultrastructural changes in various internal organs, and especially in the kidneys, were examined at different stages of development of the disease. A mottled surface of the kidneys was only seen in pigs exposed to a mouldy diet containing 180 ppb OTA for 3 months, but microscopic lesions, as well as changes in various haematological and biochemical parameters, were observed in all groups exposed to the same mouldy diet containing only 90 or 180 ppb OTA. Histological examination showed two types of change: degenerative changes affecting the epithelial cells of the proximal tubules, which predominated at the initial stage, and proliferative changes in the interstit...
Spontaneous nephropathy in pigs seen in South Africa was found to have multi-mycotoxic etiology i... more 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picture of this nephropathy was found to differ from the classical description of mycotoxic porcine nephropathy as originally made in Scandinavia by the extensive vascular changes.
A survey was made of the literature concerning the occurrence and incidence of mycotoxic nephropa... more 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.
In a mycological study, a total of 95 human food samples were investigated to evaluate the incide... more In a mycological study, a total of 95 human food samples were investigated to evaluate the incidence of fungal contamination in Cameroon by conventional identification method and partly confirmed by DNA sequencing. The isolated fungal spp. were further studied to determine their toxigenic potentials. The investigation revealed the predominance of Aspergillus and Penicillium with 96% of samples contaminated with at least one species of these fungi, whereas the incidence of co-contamination of samples was 85%. Aspergillus flavus and Aspergillus parasiticus (Flavi section) were the most predominant species contaminating mainly maize and peanuts. In addition, P. crustosum and P. polonicum were the most common contaminants belonging to the genus Penicillium. On the other hand, A. ochraceus (Circumdati section) registered a low incidence rate of 5%, including other members of the Aspergillus group. Other members of the genera Rhizopus and Alternaria spp. were also registered in the study. A majority of fungal strains of A. ochraceus, A. parasiticus, P. crustosum and P. polonicum isolated were toxigenic, producing the mycotoxins tested for, while none was detected in cultures of A. fumigatus. The high incidence rate of fungi contamination coupled with their potentials in producing mycotoxins gives a strong indication that the samples tested may likely be contaminated with various mycotoxins. There is need for further study to assess the incidence of mycotoxins contamination in similar food samples.
Histopathological, biochemical and toxicological investigations of tissues and blood of normally ... more 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.
The combined toxic effect of ochratoxin A (OTA) and penicillic acid (PA) on the body mass, the we... more The combined toxic effect of ochratoxin A (OTA) and penicillic acid (PA) on the body mass, the weight and pathomorphology of some internal organs was studied in 85 broiler chickens fed a mouldy diet containing 130, 300 or 800 ppb OTA and 1000-2000 ppb PA. The main pathomorphological changes were cloudy swelling and granular degeneration in the epithelium and mononuclear cell proliferation and activation of capillary endothelium in the kidney and liver; degenerative changes and depletion of lymphoid cells in lymphoid organs (bursa of Fabricius, thymus and spleen) were also seen. Protective effects of 5% total water extract of artichoke and a new natural phytosubstance Rosallsat against these pathomorphological changes were observed. A significant decrease in body mass and relative weight of lymphoid organs was found after 6 weeks of exposure and a greater decrease after 10 weeks of exposure to OTA and PA, and a protective effect of artichoke extract and a slight effect of Rosallsat against that decrease was observed. A significant increase in relative weight of liver and kidneys was also observed as well as a protective effect of artichoke extract against that increase. The quantity of OTA and the percentage of positive samples were significantly lower in tissues of chickens treated with artichoke extract or Rosallsat in addition to OTA than in those treated with only OTA.
Haematological, biochemical and toxicological investigations of blood and urine of normally slaug... more Haematological, biochemical and toxicological investigations of blood and urine of normally slaughtered pigs exhibiting different frequency (1-2 O/o, 10-20 % and 5 M 0 ' 1 . ) of changes characterized as "enlarged mottled kidneys"; at the slaughtering meat inspection were carried out to elucidate the nature of nephropathies encountered in Bulgaria. A content of ochratoxin A, higher in the spring than the autumn, was found in the serum and urine samples. The mean contamination levels of ochratoxin A in consumed feeds ranged from 114 k 36 ppb for 1994 to 207 k 65 ppb for 1993.
Significant protective effects of the feed additives: water extract of artichoke, sesame seed, Ro... more Significant protective effects of the feed additives: water extract of artichoke, sesame seed, Roxazyme-G and L-(3 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.
The progression of coccidiosis provoked by Eimeria acervulina was followed in chicks fed on OTA-c... more The progression of coccidiosis provoked by Eimeria acervulina was followed in chicks fed on OTA-contaminated as well as on OTAfree diets. More heavy progress of duodenal coccidiosis, including mortality, occurred in OTA-treated chicks as can be seen from the higher value of lesion (3.50) and oocyst (31.65) indices. A stronger decrease of serum total protein was found in OTA-treated chicks (22.80 g/l) than in chicks infected with E. acervulina(24.20 g/l), but that decrease was strongest in chicks treated with OTA and simultaneously infected with E. acervulina (19.71 g/l). The serum concentration of uric acid was significantly increased in all chicks exposed to OTA, most notably in those additionally infected with E. acervulina (1020.6 (l mol/L), whereas the serum enzyme activity of AST was increased only in chicks infected with E. acervulina and highest in those fed OTA contaminated diet (122.2 U/L). OTA induced degenerative changes in kidneys, liver and heart as well as a depletion of lymphoid tissue in the lymphoid organs and a decrease of body weight. Coccidiosis induced only a slight growth depression and duodenal hemorrhages in addition to characteristic duodenal damages. The impairment of kidney function, histopathological changes and general growth depression were stronger when chicks infected with E. acervulina were also given OTA.
In a mycological study, a total of 95 human food samples were investigated to evaluate the incide... more In a mycological study, a total of 95 human food samples were investigated to evaluate the incidence of fungal contamination in Cameroon by conventional identification method and partly confirmed by DNA sequencing. The isolated fungal spp. were further studied to determine their toxigenic potentials. The investigation revealed the predominance of Aspergillus and Penicillium with 96% of samples contaminated with at least one species of these fungi, whereas the incidence of co-contamination of samples was 85%. Aspergillus flavus and Aspergillus parasiticus (Flavi section) were the most predominant species contaminating mainly maize and peanuts. In addition, P. crustosum and P. polonicum were the most common contaminants belonging to the genus Penicillium. On the other hand, A. ochraceus (Circumdati section) registered a low incidence rate of 5%, including other members of the Aspergillus group. Other members of the genera Rhizopus and Alternaria spp. were also registered in the study. A majority of fungal strains of A. ochraceus, A. parasiticus, P. crustosum and P. polonicum isolated were toxigenic, producing the mycotoxins tested for, while none was detected in cultures of A. fumigatus. The high incidence rate of fungi contamination coupled with their potentials in producing mycotoxins gives a strong indication that the samples tested may likely be contaminated with various mycotoxins. There is need for further study to assess the incidence of mycotoxins contamination in similar food samples.
Mild mycotoxic nephropathy was induced in 6 pigs by a diet containing ochratoxin A at 800 ppb, se... more Mild mycotoxic nephropathy was induced in 6 pigs by a diet containing ochratoxin A at 800 ppb, several times higher than that naturally encountered in some feed for pig production in Bulgaria. The nephropathy was expressed only as slightly hypertrophied kidneys with a faintly mottled surface, discernible at the end of the experiment to a skilled observer but probably not recognisable in routine slaughterhouse processing.
Stable radical 1, 1-dipheny-2-picrylhydrazyl (DPPH) is widely used at in vitro models to investig... more Stable radical 1, 1-dipheny-2-picrylhydrazyl (DPPH) is widely used at in vitro models to investigate antioxidant and radical scavenging abilities of natural extracts. This work presents comparative study on DPPH radical scavenging capacity before and after UV irradiation of aqueous extract of Glycyrrhiza Glabra, a plant species belonging to the Indian flora. DPPH scavenging activities of different extract concentrations (at different incubation time intervals) were analyzed and compared by in vitro spectrophotometry and electron paramagnetic resonance (EPR) spectroscopy. 9.93% and 16.79%, DPPH scavenging activities before and after UV irradiation respectively were found by spectrophotometry. By the EPR spectroscopy study statistical significant increase in DPPH radical scavenging for the Glycyrrhiza Glabra extracts was established after UV irradiation (78.39± 0.001%) comparing to the non irradiated samples (14.02± 0.02).
Mycotoxic nephropathy was induced in 18 young pigs by diets contaminated with strains of Aspergil... more Mycotoxic nephropathy was induced in 18 young pigs by diets contaminated with strains of Aspergillus ochraceus containing ochratoxin A (OTA) and penicillic acid (PA) at levels corresponding to those naturally encountered in animal feeds in Bulgaria. Haematological and biochemical parameters, as well as the morphological and ultrastructural changes in various internal organs, and especially in the kidneys, were examined at different stages of development of the disease. A mottled surface of the kidneys was only seen in pigs exposed to a mouldy diet containing 180 ppb OTA for 3 months, but microscopic lesions, as well as changes in various haematological and biochemical parameters, were observed in all groups exposed to the same mouldy diet containing only 90 or 180 ppb OTA. Histological examination showed two types of change: degenerative changes affecting the epithelial cells of the proximal tubules, which predominated at the initial stage, and proliferative changes in the interstit...
Spontaneous nephropathy in pigs seen in South Africa was found to have multi-mycotoxic etiology i... more 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.
A survey was made of the literature concerning the occurrence and incidence of mycotoxic nephropa... more 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.
In a mycological study, a total of 95 human food samples were investigated to evaluate the incide... more In a mycological study, a total of 95 human food samples were investigated to evaluate the incidence of fungal contamination in Cameroon by conventional identification method and partly confirmed by DNA sequencing. The isolated fungal spp. were further studied to determine their toxigenic potentials. The investigation revealed the predominance of Aspergillus and Penicillium with 96% of samples contaminated with at least one species of these fungi, whereas the incidence of co-contamination of samples was 85%. Aspergillus flavus and Aspergillus parasiticus (Flavi section) were the most predominant species contaminating mainly maize and peanuts. In addition, P. crustosum and P. polonicum were the most common contaminants belonging to the genus Penicillium. On the other hand, A. ochraceus (Circumdati section) registered a low incidence rate of 5%, including other members of the Aspergillus group. Other members of the genera Rhizopus and Alternaria spp. were also registered in the study. A majority of fungal strains of A. ochraceus, A. parasiticus, P. crustosum and P. polonicum isolated were toxigenic, producing the mycotoxins tested for, while none was detected in cultures of A. fumigatus. The high incidence rate of fungi contamination coupled with their potentials in producing mycotoxins gives a strong indication that the samples tested may likely be contaminated with various mycotoxins. There is need for further study to assess the incidence of mycotoxins contamination in similar food samples.
Histopathological, biochemical and toxicological investigations of tissues and blood of normally ... more 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.
The combined toxic effect of ochratoxin A (OTA) and penicillic acid (PA) on the body mass, the we... more The combined toxic effect of ochratoxin A (OTA) and penicillic acid (PA) on the body mass, the weight and pathomorphology of some internal organs was studied in 85 broiler chickens fed a mouldy diet containing 130, 300 or 800 ppb OTA and 1000-2000 ppb PA. The main pathomorphological changes were cloudy swelling and granular degeneration in the epithelium and mononuclear cell proliferation and activation of capillary endothelium in the kidney and liver; degenerative changes and depletion of lymphoid cells in lymphoid organs (bursa of Fabricius, thymus and spleen) were also seen. Protective effects of 5% total water extract of artichoke and a new natural phytosubstance Rosallsat against these pathomorphological changes were observed. A significant decrease in body mass and relative weight of lymphoid organs was found after 6 weeks of exposure and a greater decrease after 10 weeks of exposure to OTA and PA, and a protective effect of artichoke extract and a slight effect of Rosallsat against that decrease was observed. A significant increase in relative weight of liver and kidneys was also observed as well as a protective effect of artichoke extract against that increase. The quantity of OTA and the percentage of positive samples were significantly lower in tissues of chickens treated with artichoke extract or Rosallsat in addition to OTA than in those treated with only OTA.
Haematological, biochemical and toxicological investigations of blood and urine of normally slaug... more Haematological, biochemical and toxicological investigations of blood and urine of normally slaughtered pigs exhibiting different frequency (1-2 O/o, 10-20 % and 5 M 0 ' 1 . ) of changes characterized as "enlarged mottled kidneys"; at the slaughtering meat inspection were carried out to elucidate the nature of nephropathies encountered in Bulgaria. A content of ochratoxin A, higher in the spring than the autumn, was found in the serum and urine samples. The mean contamination levels of ochratoxin A in consumed feeds ranged from 114 k 36 ppb for 1994 to 207 k 65 ppb for 1993.
Significant protective effects of the feed additives: water extract of artichoke, sesame seed, Ro... more Significant protective effects of the feed additives: water extract of artichoke, sesame seed, Roxazyme-G and L-(3 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.
The progression of coccidiosis provoked by Eimeria acervulina was followed in chicks fed on OTA-c... more The progression of coccidiosis provoked by Eimeria acervulina was followed in chicks fed on OTA-contaminated as well as on OTAfree diets. More heavy progress of duodenal coccidiosis, including mortality, occurred in OTA-treated chicks as can be seen from the higher value of lesion (3.50) and oocyst (31.65) indices. A stronger decrease of serum total protein was found in OTA-treated chicks (22.80 g/l) than in chicks infected with E. acervulina(24.20 g/l), but that decrease was strongest in chicks treated with OTA and simultaneously infected with E. acervulina (19.71 g/l). The serum concentration of uric acid was significantly increased in all chicks exposed to OTA, most notably in those additionally infected with E. acervulina (1020.6 (l mol/L), whereas the serum enzyme activity of AST was increased only in chicks infected with E. acervulina and highest in those fed OTA contaminated diet (122.2 U/L). OTA induced degenerative changes in kidneys, liver and heart as well as a depletion of lymphoid tissue in the lymphoid organs and a decrease of body weight. Coccidiosis induced only a slight growth depression and duodenal hemorrhages in addition to characteristic duodenal damages. The impairment of kidney function, histopathological changes and general growth depression were stronger when chicks infected with E. acervulina were also given OTA.
In a mycological study, a total of 95 human food samples were investigated to evaluate the incide... more In a mycological study, a total of 95 human food samples were investigated to evaluate the incidence of fungal contamination in Cameroon by conventional identification method and partly confirmed by DNA sequencing. The isolated fungal spp. were further studied to determine their toxigenic potentials. The investigation revealed the predominance of Aspergillus and Penicillium with 96% of samples contaminated with at least one species of these fungi, whereas the incidence of co-contamination of samples was 85%. Aspergillus flavus and Aspergillus parasiticus (Flavi section) were the most predominant species contaminating mainly maize and peanuts. In addition, P. crustosum and P. polonicum were the most common contaminants belonging to the genus Penicillium. On the other hand, A. ochraceus (Circumdati section) registered a low incidence rate of 5%, including other members of the Aspergillus group. Other members of the genera Rhizopus and Alternaria spp. were also registered in the study. A majority of fungal strains of A. ochraceus, A. parasiticus, P. crustosum and P. polonicum isolated were toxigenic, producing the mycotoxins tested for, while none was detected in cultures of A. fumigatus. The high incidence rate of fungi contamination coupled with their potentials in producing mycotoxins gives a strong indication that the samples tested may likely be contaminated with various mycotoxins. There is need for further study to assess the incidence of mycotoxins contamination in similar food samples.
Mild mycotoxic nephropathy was induced in 6 pigs by a diet containing ochratoxin A at 800 ppb, se... more Mild mycotoxic nephropathy was induced in 6 pigs by a diet containing ochratoxin A at 800 ppb, several times higher than that naturally encountered in some feed for pig production in Bulgaria. The nephropathy was expressed only as slightly hypertrophied kidneys with a faintly mottled surface, discernible at the end of the experiment to a skilled observer but probably not recognisable in routine slaughterhouse processing.
Stable radical 1, 1-dipheny-2-picrylhydrazyl (DPPH) is widely used at in vitro models to investig... more Stable radical 1, 1-dipheny-2-picrylhydrazyl (DPPH) is widely used at in vitro models to investigate antioxidant and radical scavenging abilities of natural extracts. This work presents comparative study on DPPH radical scavenging capacity before and after UV irradiation of aqueous extract of Glycyrrhiza Glabra, a plant species belonging to the Indian flora. DPPH scavenging activities of different extract concentrations (at different incubation time intervals) were analyzed and compared by in vitro spectrophotometry and electron paramagnetic resonance (EPR) spectroscopy. 9.93% and 16.79%, DPPH scavenging activities before and after UV irradiation respectively were found by spectrophotometry. By the EPR spectroscopy study statistical significant increase in DPPH radical scavenging for the Glycyrrhiza Glabra extracts was established after UV irradiation (78.39± 0.001%) comparing to the non irradiated samples (14.02± 0.02).
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