[Host-pathogen relations in yeasts]. / Wirt-Erreger-Beziehungen bei Hefen.

Allogenic and autogenic factors are decisive for a colonization of warmblooded hosts by yeasts where they can live inside as symbiont, commensal or pathogen. To develop pathogenicity a capacity of yeasts for multiplication inside of humans or homoiothermic animals and resistance to environmental influences is not sufficient. Pathogenicity rather depends on the ability of structural morphological changes, phenomena of adherence and of microbial competition as well as the presence of genes for down-regulation of the host defense and for enzyme production enabling a parasitic life style. Among the yeasts Cryptococcus neoformans and some Candida species meet these requirements.

Adherence of Escherichia coli serogroup O 157 and the Salmonella typhimurium mutant DT 104 to the surface of Saccharomyces boulardii.

The detection of lectin sites for mannose-sensitive adhesion in the outer membrane of Saccharomyces boulardii and the irreversible binding of both enteropathogenic Escherichia coli (EPEC) and salmonellae (serovar Salmonella Typhimurium and Salmonella Enteritidis) provided the motivation to carry out further investigations to find out whether also other enteric bacteria such as entero-haemorrhagic Escherichia coli (EHEC) and the DT 104 mutant of S. Typhimurium have the capacity for binding to the cell wall of this yeast. Reference strains and fresh isolates from clinical cases of EHEC infections as well as salmonellae of the DT 104 mutant were included in this study using the agglutination test. The results first of all showed that EHEC of the serogroup O 157 and the DT 104 mutant of S. Typhimurium were bound to the surface of Saccharomyces boulardii. Because these bacteria do not respond very well to drugs but most of the gastrointestinal infections are caused by them, the use of S. boulardii for treatment and prophylaxis could be an excellent alternative.

[Attitudes toward conception presented by the patients of the Obstetrics and Gynecology Department of the District Hospital in Lublin]. / Postawy wobec prokreacji przyjmowane przez pacjentki oddzialu ginekologiczno-polozniczego Szpitala Wojewódzkiego w Lublinie.

The paper presents the opinions of the women in puerperium about the number of children in a family and the methods applied in family planning. The opinions consider health, psychological and moral aspects. The paper highlights the need for the support to the responsible parenthood provided by the medical units.

[Family planning methods applied by women having their first and next baby]. / Metody planowania rodziny u kobiet rodzacych pierwsze i kolejne dziecko.

The paper focuses on the procreation plans of the women having their first and next babies, their knowledge about the conception regulation methods and their attitudes towards natural family planning methods and contraception.

[Comparative investigations of mycotoxological status of alternatively and conventionally grown crops]. / Vergleichende Untersuchungen zum mykotoxikologischen Status von ökologisch und konventionell angebautem Getreide.

100 samples of rye and 101 samples of wheat coming out of both conventional and alternative or ecological production were investigated for contamination with mycotoxins with interest for our degree of latitude. Deoxynivalenol (DON) was found with thin-layer-chromatography in 131 of 201 samples altogether. A top level of 1250 micrograms DON kg-1 in rye of alternative offspring was detected. The average burden in contaminated rye coming from ecological production was 427 micrograms kg-1 and a mean level of 160 micrograms kg-1 resulting in rye out of conventional growth conditions. In wheat, conventionally grown yield showed slightly lower contamination (mean levels of 420 micrograms DON kg-1 towards 486 micrograms kg-1). The toxins 3-acetyl-deoxynivalenol, nivalenol and fusarenone X were detected in some samples by thin-layer-chromatography. This results could not be confirmed by gas chromatography -mass spectrometry. Zearalenone was found in 40 out of the number of 201 samples of grain by HPLC with fluorescence detection. An average of 6 micrograms and 24 micrograms zearalenone kg-1 in conventionally and alternatively grown wheat and 4 micrograms and 51 micrograms zearalenone kg-1 in conventionally and alternatively produced rye was detected. The highest finding of zearalenone was 199 micrograms kg-1 in alternatively grown rye. Skin toxicity testing did not show any reference of contamination with type-A-trichothecenes. No correlation between contamination of zearalenone or deoxynivalenol and thousand-kernel-weight was detected.

[Comparative studies of the bacterial and mycological status of ecologically and conventionally grown crops]. / Vergleichende Untersuchungen zum bakteriellen und mykologischen Status von ökologisch und konventionell angebautem Getreide.

Wheat and rye derived from conventional and from alternative or ecological production was examined for bacterial and fungal contamination. The overall bacterial and fungal contamination was lower than 10(7.7) germs/g. The amount and type of germs in the grain corresponded to typical contamination of fresh crop. No difference in germ contamination of both conventionally and alternatively grown wheat and rye could be found. No correlation between bacterial or fungal contamination and thousand-kernel-weight was detected.

[Health risks of mycotoxins for humans]. / Gesundheitsgefährdung des Menschen durch Mykotoxine.

Fungi cause diseases in humans in different ways. Due to production of secondary metabolites by molds, dematiaceous hyphomycetes, and primary plant pathogenic fungus species intoxications are included in their pathogenic potency. Mycotoxins mostly enter the body by ingestion of food because they are contaminants of plant products and in case of carry over also of animal products. Recent investigations have shown that mycotoxins detected in feed and food during microbial deterioration can also be produced inside warm-blooded organism, e. g. with aspergillosis in animals (mammals and birds). This means an enhanced risk for humans to suffer from mycotoxicosis, in which the toxic agent has an exogenic or endogenic source. Therefore, attention should be paid to the fact that mycotoxins are not only damaging cells; they very often possess a genotoxic potential.

[Pathologico-anatomic and mycotoxicologic studies of aspergillosis in birds]. / Pathologisch-anatomische und mykotoxikologische Untersuchungen zur Aspergillose bei Vögeln.

4.0% of all birds (n = 11249) examined post mortem at the Institute of Avian Diseases, Oberschleissheim, showed fungal infections of the respiratory tract. Clinical and histopathological findings to date have suggested toxin production in Aspergillus fumigatus infections. To obtain detailed information, preselected tissues from the lungs and air sacs (n = 17) were examined by culture and fluorescence microscopy. In 12 cases gliotoxin (0.1-0.45 mg/kg) was found in the A. fumigatus infected tissues. The importance of gliotoxin, which showed strong cytotoxic effects in cell cultures, is discussed.

[The nutritive efficiency of Bacillus cereus as a probiotic in the raising of piglets. 1. Effect on the growth parameters and gastrointestinal environment]. / Zur nutritiven Wirksamkeit von Bacillus cereus als Probiotikum in der Ferkelaufzucht. 1. Mitteilung--Einfluss auf Wachstumsparameter und gastrointestinales Milieu.

In an experiment with 4 x 12 piglets the probiotic effect of the Bacillus cereus strain FH 1457 S on the performance and changes in the gastrointestinal tract was studied. Beside a negative control the feed was supplemented with 10(7), 10(8) and 10(9) CFU Bacillus cereus/kg. The experiment included two feeding periods, whereby in the first a prestarter and in the second a piglet rearing feed was administrated ad libitum with the animals kept individually in cages. Over the whole feeding trial the piglets with the additions of 10(7), 10(8) und 10(9) CFU Bacillus cereus/kg feed showed higher daily weight gains of 9.5%, 11.1% and 8.6% with an increased feed consumption of 11.1%, 11.0% and 7.9%. The feed conversion rate was not influenced by the additions. In the first period daily weight gains were increased by 8.9%, 14.7% and 7.8%, the feed consumption in this period was 10.2%, 12.1% and 7.2% higher than in the control group. The addition of 10(8) CFU Bacillus cereus had a significantly higher daily weight gain in the first period compared to the control and the feed conversion rate was improved by 2.4%. In the second period the daily weight gains were increased by 9.9%, 8.9% and 9.1%, the feed consumption by 11.4%, 10.5% and 8.8% because of Bacillus cereus addition to the feed. The frequency of diarrhea was not influenced by the treatments. In each Bacillus cereus dosage the concentrations of ammonia, lactate and volatile fatty acids in small intestine contents were mostly diminished. In caecum contents the concentrations of acetic and propionic acid were decreased in the groups supplemented with Bacillus cereus.

[The nutritive effect of Bacillus cereus as a probiotic in the raising of piglets. 2. Effect and microbial count, composition and resistance determination of gastrointestinal and fecal microflora]. / Zur nutritiven Wirksamkeit von Bacillus cereus als Probiotikum in der Ferkelaufzucht. 2. Mitteilung--Einfluss auf Keimzahlen, Zusammensetzung und Resistenzeigenschaften der gastrointestinalen und faecalen Mikroflora.

After a feeding trial of 42 days with 4 x 12 piglets 4 x 6 piglets were slaughtered and the influence of spores of the Bacillus cereus strain FH 1457 S added to the feed on the microorganism counts of Lactobacillus/Bifidobacterium, Eubacteria, Bacteroidaceae, E. coli, Enterococcus and passants (Bacillus cereus) in duodenum, jejunum, ileum, caecum and colon was investigated. Beside a negative control the feed was supplemented with 10(7), 10(8) and 10(9) CFU Bacillus cereus/kg. The feeding trial included two periods each of 21 days. In each period faeces was collected from all animals and the influence on the microorganism counts and also the frequency of resistance of the E. coli and Enterococcus germs against selected antibiotics and chemotherapeutics investigated. The addition of 10(8) CFU Bacillus cereus decreased E. coli counts in duodenum and jejunum, however increased them in ileum, caecum and colon. In the highest dosage the counts of Bacillus cereus in duodenum, caecum, colon and faeces were significantly higher. The addition of Bacillus cereus reduced the counts of E. coli and Enterococcus in faeces in the first period. In the second period the dosage 10(8) CFU showed significant increased counts of E. coli and the sums of microorganisms of the main and satellite flora. The additions had no influence on the frequency of resistance of E. coli and enterococci against the tested antibiotics and synthetic chemotherapeutics. From the addition of Bacillus cereus in this case a selection of factors influencing resistance is not to be expected.

In vitro transformation of the tremorgenic mycotoxin verruculogen.

A quantity of 50 mg of crystalline verruculogen was prepared from batch cultures ofAspergillus fumigatus for the use in thein vitro studies with S-9 liver fractions, feces suspensions, and cultures ofEscherichia coli.Incubation of verruculogen with S-9 liver fractions from swine resulted in the transformation of the parent compound into TR-2 toxin and 3 other more polar.TR-2 toxin was also shown to be the main transformation product when incubat-ing verruculogen with fecal suspensions or pure cultures ofE. coli (0149: K88). Incubation times of more than 2 hours led to a complete degradation of verruculogen as well as TR -2 toxin.

[Regulation of the intestinal flora by food]. / Regulierung der Darmflora über die Nahrung.

The purpose of probiotics is to regulate the bacterial colonisation of the digestive tract. In contrast to antibiotics, with the probiotics it is not a certain metabolic product of bacteria or fungi with a known spectrum of antimicrobial activity which, for example when used as performance enhancers under the conditions permitted according to the EEC guidelines for feed additives, reduce the numbers of bacteria, which appear in the small intestines of domestic animals and compete with the host for nutrients and vitamins, but there it is rather the living micro-organisms themselves which are intended to, to help to improve the performance of domestic animals on the basis of a fundamentally different mechanism of action. Recent findings on the effect of the phenomena of bacterial competition on the processes of an infection, with penetration, adhesion, and multiplication as prerequisites for development of the parasitic mechanism of a pathogen, have shown that by controlling the processes of colonization of the intestines with micro-organisms and also their metabolic activity it is possible to at least reduce the incidence of the diarrhoeal diseases which are responsible for the the greatest economic damage. The numerical displacement of bacterial pathogens is not the only effect of the bacterial competition resulting from the orally administered micro-organisms. The fact that antagonistically acting bacteria or yeasts prevent the pathogenic species from entering the intestinal epithelial cells plays a role here. As a result of which, especially the small intestines are not colonized by pathogens (E. coli and others), the liver is largely relieved of substances accumulating in the course of bacterial protein metabolism, the absorption of fatty acids and other energy-providing compounds is increased, and not readily soluble salts are transformed into soluble compounds. Effects comparable to those obtained with small amounts of antibiotics added to the feed of farm animals as regards increased growth rate and improved feed conversion, can also achieved with the addition of probiotics.

[The protective effect of adsorbents against ochratoxin A in swine]. / Untersuchungen zur protektiven Wirkung von Adsorbentien gegenüber Ochratoxin A beim Schwein.

Adsorption of the mycotoxin ochratoxin A by activated charcoal, various bentonites (acid, alkaline, neutral), and hydrated sodium calcium aluminosilicate was tested in vitro as well as in feeding experiments with pigs. In vitro tests showed that the 1% addition of activated charcoal leads to complete adsorption of ochratoxin A from aqueous solutions. This effect was not influenced by pH-values ranging from 3-8. In contrast, adsorption by bentonite and hydrated sodium calcium aluminosilicate occurred primarily in the acid range (pH 3-4). Dietary addition of hydrated sodium calcium aluminosilicate (1%) and acid bentonite (1%, 10%) to ochratoxin A-contaminated feed (1.0 mg/kg) had no effect on the blood or tissue levels of the toxin in pigs. The addition of 1% activated charcoal caused a slight decrease of ochratoxin A in the blood, whereas a tenfold dosage resulted in a 50% to 80% reduction of ochratoxin A levels in both blood and tissue. Reduction of ochratoxin A absorption via the dietary administration of activated charcoal (5%) was confirmed in a 16 week feeding experiment. However, this experiment also showed the serum level of vitamin E to be lower than in the controls receiving adsorbent-free feed.

Cleavage of zearalenone-glycoside, a "masked" mycotoxin, during digestion in swine.

Comparative analyses of cereal samples pretreated with or without beta-glucosidase indicate the presence of zearalenone-glycoside. To examine the stability of zearalenone-glycoside during digestion, mixed feed was artificially contaminated with synthesized zearalenone-4-beta-D-glucopyranoside (395 micrograms/kg) and fed to a pig over a period of 14 days. The metabolites detected in feces and urine samples were zearalenone and alpha-zearalenol. These results demonstrate that zearalenone-4-beta-D-glucopyranoside is decomposed during digestion and the aglucone, zearalenone, is released. Since zearalenone-glycoside is not detected during routine analysis, but hydrolysed during digestion, it seems likely that such "masked mycotoxins" are involved in cases of mycotoxicoses.

Isolation of a mycotoxin (gliotoxin) from a bovine udder infected with Aspergillus fumigatus.

A bovine udder infected with Aspergillus fumigatus was analysed by physico-chemical methods (thin layer chromatography, high performance liquid chromatography and direct exposure probe-mass spectrometry) for the presence of mycotoxins. Gliotoxin, a fungal metabolite with cytotoxic and immunosuppressive properties was isolated for the first time from naturally infected tissue. The gliotoxin concentration analysed (9.2 mg kg-1 udder) was approximately 100 times higher than the concentration known to produce morphological changes of cells. Gliotoxin may play an important role in the establishment and development of an infection with A fumigatus.

[Determination of ochratoxin A in human milk]. / Bestimmung von Ochratoxin A in Muttermilch.

A method for the determination of ochratoxin A in milk is described. The milk is homogenized in a buffer solution at pH 1.6 to release ochratoxin A from its bond to proteins. Ochratoxin A is extracted with chloroform and the extract cleaned up using a base clean-up step. Analysis is performed by high-pressure liquid chromatography, using a reversed-phase column and fluorescence detection. The detection limit of the method is 0.1 ng/ml and the average recovery rate, tested in the range between 0.5 and 10.0 ng/ml, was found to be 83.1%. Chemical ionization mass spectrometry (direct exposure probe) and an enzyme immunoassay were used as confirmatory tests. Using this method, trace amounts of ochratoxin A were found in 4 of 36 randomly collected human milk samples.

[Changes in the genital tract of female swine after feeding with practice-relevant amounts of zearalenone]. / Veränderungen am Genitaltrakt des weiblichen Schweines nach Verfütterung praxisrelevanter Zearalenonmengen.

Zearalenone is one of the most frequent, naturally occurring mycotoxins in feedstuffs: 13.9% of 710 samples analyzed contained zearalenone in concentrations of 1.0 to 1,725 micrograms/kg. Trial feeding with female pigs demonstrated that a zearalenone concentration of 250 micrograms/kg feed produced distinct redness and swelling of the vulva, slight swelling of the mamma and numerous vesicular follicles and some cystic follicles on the ovaria. The symptoms on the external genital tract were not observed when the zearalenone concentration in the feedstuff, was 50 micrograms/kg but the section also established that vesicular follicles on the ovaria of these animals were more frequent than on the ovaria of the control animal.

On the metabolism of the mycotoxin deoxynivalenol in the isolated perfused rat liver.

An isolated rat liver was perfused with deoxynivalenol (DON) at a dose of 3 mg in a recirculating perfusion system. To identify glucuronide conjugates equal amounts of bile samples, perfusate and liver homogenates were incubated with and without (control) a ß-glucuronidase preparation and analyzed by thin layer chromatography and capillary gas liquid chromatography - chemical ionization mass spectrometry. A total of 40.4% of the administered dose of DON was found to be conjugated with glucuronic acid (perfusate 20.4%, bile 19.2%, liver 0.8%), while only 1.3% of the parent DON (perfusate 1.1%, bile 0.2%) was detected. The cleavage of DON-glucuronide was demonstrated by incubating DON-glucuronide containing bile samples with intestine contents under anaerobic conditions.

Identification of glucuronide metabolites of T-2 toxin and diacetoxyscirpenol in the bile of isolated perfused rat liver.

Isolated rat livers were perfused with either 2 mg T-2 toxin or diacetoxyscirpenol (DAS) in a recirculating perfusion system. To identify glucuronide conjugates, equal amounts of bile samples were incubated with and without (control) a beta-glucuronidase preparation and analyzed by capillary gas liquid chromatography-chemical ionization mass spectrometry. Enzyme treatment of bile obtained from liver perfused with T-2 toxin resulted in the detection of a total of 954 micrograms HT-2 toxin (control 6 micrograms), demonstrating that excretion into the bile was mainly as glucuronide conjugates. Minor metabolites of T-2 toxin in bile were identified as 3'-hydroxy HT-2 toxin (TC-3), 3'-hydroxy-7-hydroxy HT-2 toxin (TC-6), and the glucuronide form of T-2 triol (trace amount). The glucuronide conjugates of monoacetoxyscirpenol (340 micrograms) and scirpenetriol (10 micrograms) were found in bile obtained from liver perfused with DAS, while nonconjugated metabolites were not detected. It is assumed that considerable amounts of T-2 toxin and DAS were metabolized biphasically. In phase I both trichothecenes were deacetylated, in phase II the metabolites were conjugated giving rise to the glucuronic acid adducts.