EuSANH workshop "Reasons behind the differences in national vaccination schedules for under-five", European Public Health pre-conference workshop, Malta, 8 November 2012.

Vaccination schedules for under-five children in the EU member states differ markedly, mainly as a consequence of differences in programme organization, decision making and history, and to a limited extent by epidemiological differences. There is little willingness towards unification since little evidence exists to prefer one schedule over the others, but the differences might impact on public confidence. Monitoring key determinants influencing individual decision making on immunization ('soft impacts') is thus as important as other existing monitoring systems of the 'hard' impacts of immunization programmes, and both should focus on the impact of these schedule differences. Harmonization of vaccination schedules is not the main issue, but the reasons behind the differences should be explained in an understandable and coherent way to the public. Scientists and advisory bodies should look over the country borders and communicate any crucial information, in order to improve scientific consensus on immunization schedules and programmes. These were the main conclusions of a members' experts panel of the European network of independent science advisory bodies on health (EuSANH), at a workshop in November 2012.

Equal allergenic potency of beta-lactam antibiotics produced by chemical or enzymatic manufacturing--mouse IgE test.

BACKGROUND: Cephalexin and amoxicillin are semisynthetic beta-lactam antibiotics with a broad spectrum of antibacterial activity against gram-positive and gram-negative microorganisms. Both antibiotics are produced by a new 'green' process in which enzyme technology is used to combine the intermediate structure and the side chain in an aqueous medium to yield cephalexin or amoxicillin, thus avoiding the use of several chemical reagents and volatile organic solvents. As a result of the enzyme technology a new residual protein impurity has been identified. To check for the sensitizing capacity of the residual protein, a mouse IgE test was used to detect differences in the production of specific IgE by chemical or enzymatic preparations of the antibiotics. METHODS: Balb/c female mice were immunized intraperitoneally with alum and conjugates of different amoxicillins or cephalexins with ovalbumin (OVA). After 16 days, the amoxicillin mice were injected with one half the original amount of antigen. After 19-23 days, the sera were tested for specific IgE by the passive cutaneous anaphylaxis assay in Sprague-Dawley rats. The greatest dilution of sera which resulted in a positive response was the titer of specific IgE. RESULTS: No significant differences were found between the titers of specific IgE caused by the chemically and enzymatically produced beta-lactam antibiotics, indicating that the antibiotics are equal in allergenicity. CONCLUSIONS: The data show that a residual level of 35 ppm protein did not affect the allergenic potency of these beta-lactam antibiotics as determined by the mouse allergenicity model.

Safety evaluation of a lactase enzyme preparation derived from Kluyveromyces lactis.

Neutralact(R), the DSM brand name of a lactase enzyme preparation, obtained from a homologous rDNA strain of Kluyveromyces lactis, was subjected to a series of toxicological tests to document the safety for use as a processing aid in the dairy industry. The enzyme preparation was examined for subacute oral toxicity and mutagenic potential. As a result of these tests, no evidence of oral toxicity, mutagenicity or clastogenicity was found. Administration of the lactase enzyme preparation at doses of 500, 3000 and 10,000 mg/kg body weight/day for 28 days did not induce noticeable signs of toxicity. The no-observed-adverse-effect level (NOAEL) of the enzyme preparation in the acute toxicity study was 10,000 mg/kg body weight/day (equivalent to 114,000 NL units/kg body weight/day). It can be concluded that no safety concerns were identified in the studies conducted with this lactase enzyme preparation derived from Kluyveromyces lactis under controlled fermentation conditions.

Safety evaluation of amino peptidase enzyme preparation derived from Aspergillus niger.

An amino peptidase enzyme preparation obtained from Aspergillus niger was subjected to a series of toxicological tests to document the safety for use as a processing aid for food. The enzyme preparation was examined for subacute and subchronic oral toxicity, and mutagenic potential. No evidence of oral toxicity or mutagenicity was found. Administration of the amino peptidase enzyme preparation at doses of 500, 1000 and 2000 mg/kg body weight/day for 90 days did not induce noticeable signs of toxicity. The no-observed-adverse-effect level (NOAEL) of the enzyme preparation in the subchronic toxicity study was 2000mg/kg body weight/day (equivalent to 1152 PHEA units/kg body weight/day). It can be concluded that no safety concerns were identified in the studies conducted with this amino peptidase enzyme preparation derived from Aspergillus niger and produced under controlled fermentation conditions.

Safety evaluation of lipase derived from Rhizopus oryzae: summary of toxicological data.

A lipase enzyme, obtained from Rhizopus oryzae produced by a fermentation process was subjected to a series of toxicological tests to document the safety for use as a food additive. The enzyme product was examined for acute, subacute and subchronic oral toxicity, and mutagenic potential. An extensive literature search on the production organism has also been conducted. No evidence of (sub)acute oral toxicity or mutagenic potential was found. Administration of the lipase at dosages of 50, 200 and 1000 mg/kg body weight/day for 90 days did not induce noticeable signs of toxicity. A few minor changes in the chemical composition of the blood in the highest dose group were of no toxicological significance. The no-observed-adverse-effect level of the tox-batch in the subchronic toxicity study was 1000 mg/kg body weight/day. It can be concluded that no safety concerns were identified in the studies conducted with this lipase preparation derived from R. oryzae and produced under controlled fermentation conditions.

Safety evaluation of beta-glucanase derived from Trichoderma reesei: summary of toxicological data.

Barlican, a beta-glucanase enzyme obtained from Trichoderma reesei, was produced by a fermentation process and subjected to a series of toxicological tests to document its safety for use as a feed additive. The enzyme product was examined for general oral toxicity, inhalation toxicity, irritation to eye and skin, skin sensitization and mutagenic potential. An extensive literature search on the production organism was also conducted. Furthermore, safety for target species was assessed in a 28-day oral toxicity study with broilers. A strong skin-sensitizing potential of the beta-glucanase enzyme was detected, but no other evidence of oral or inhalation toxicity, mutagenic potential, eye or skin irritancy was found. Feeding of the beta-glucanase enzyme at dietary levels up to 10,000 ppm in the 90-day subchronic toxicity study in rats did not induce noticeable signs of toxicity. In addition, no adverse effects were observed when broiler chicks were fed dietary concentrations of the beta-glucanase enzyme up to eight times the daily recommended dose. It is therefore concluded that this beta-glucanase preparation is safe for use in feed of the intended target species. However, some occupational health precautions should be taken to avoid skin contact and inhalation, as is the case for almost all enzyme proteins.

Modified suspension Ames test for testing proteinaceous substances: an initial step.

The basic Salmonella/microsome assay (Ames test) is a valuable primary tool by which to discriminate mutagens from non-mutagens. For a variety of chemical test substances this test is easily conducted according to international guidelines for genotoxicity testing. However, the testing of proteinaceous substances in the basic Ames test may generate false positives owing to the presence of growth-promoting constituents in the test sample, such as histidine or its precursors. It was hypothesized that the growth-promoting capacities of biological test samples might be overcome by testing according to the 'suspension variant' of the Ames test, which uses very rich growth conditions thereby overwhelming any growth-enhancing constituents present in a biological test sample. This hypothesis appeared to be correct, although several important modifications had to be made to the suspension assay. The most important aspect of this 'new suspension Ames test' appeared to be the plating of overnight regrown bacteria in the poorest way possible (by omitting histidine and nutrient broth from the overlay agar). This study may comprise an initial step in the development of a modified suspension Ames test for testing proteinaceous substances.

Hematology and serum biochemistry of Japanese quail fed dietary tri-n-butyltin oxide during reproduction.

During a subchronic toxicity and reproduction study with tri-n-butyltin oxide (TBTO) concentrations of 0, 24, 60, and 150 mg/kg diet in Japanese quail, preliminary data on hematology and serum biochemistry were obtained. The absence of serious effects in blood parameters in both adult quail and developing chicks are discussed in view of the adverse effects of TBTO on reproduction.

Effects of bis(tri-n-butyltin)oxide in Japanese quail exposed during egg laying period: an interlaboratory comparison study.

The guideline no. 206 for testing of chemicals of the Organization for Economic Cooperation and Development (OECD) comprising an avian reproduction test using the Japanese quail (Coturnix coturnix japonica; Termminck and Schlegel 1849) as pair-hold test organisms has been applied in a version that reduced the treatment period to 6 weeks without any pretreatment. In the present study bis(tri-n-butyltin)oxide, C.A. No. 56-35-9 (tributyltin oxide, TBTO) was examined by five participants in an interlaboratory comparison test. A comparable regimen of dosing was performed by all participants starting either with 24 or 60 mg/kg TBTO in the feed and ending with 150 or 375 mg/kg. Within this dose range no signs of toxicity in adults were observed. Substance-related effects however were obvious with regard to egg production, fertility, hatching success, and survival of 14 day-old chicks. A clear dose dependency was given regarding effects on egg weight and on hatchability. The no-observed-effect concentrations for these two parameters was 60 mg/kg TBTO, characterizing these parameters as the most sensitive in this investigation. With the presented set of test parameters further aspects of subchronic toxicity in adults and chicks can be assessed as well as the validity of the performed test. Comparing the results for most test parameters consistency is obvious, thus confirming the applicability of the presented test guideline.

Subchronic toxicity and reproduction effects of tri-n-butyltin oxide in Japanese quail.

A subchronic toxicity/reproduction study was performed in Japanese quails that were fed a diet containing 0, 24, 60, and 150 mg tri-n-butyltin oxide (TBTO) per kg basal diet for 6 weeks. Eggs produced during the 6 weeks of treatment were incubated and hatched, and chicks hatched from eggs collected in weeks 5 and 6 of exposure were reared for 2 weeks. In parent quail, neither diminished food consumption nor any overt toxic or histopathologic signs were observed following exposure to TBTO. A statistically significant decrease in hatch-ability and increase in percent of chicks found dead in the shell were observed following TBTO exposure at concentrations of 60 and 150 mg/kg food. However, no significant, adverse effects were recorded on total egg production, eggshell thickness and cracked eggs. Blood chemistry parameters of birds measured at the last day of TBTO treatment revealed a statistically significant decrease in serum aspartate aminotransferase (ASAT) enzyme activity among both sexes in all treatment groups. In addition, a statistically significant dose-related decrease in serum calcium level was observed in females only, while serum follicle stimulating hormone (FSH) levels were statistically significantly reduced in male birds in all treatment groups (approximately 50% of the controls). Moreover, a significant decrease in hepatic microsomal 7-ethoxyresorufin (EROD) activity was recorded in females fed 24 and 60 mg TBTO/kg diet and males fed 60 and 150 mg TBTO/kg diet, whereas pentoxyresorufin-o-deetylase (PROD) activity was only significantly decreased in males fed 150 mg TBTO/kg diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of polyunsaturated fatty acid supplementation and membrane fluidity on ozone and nitrogen dioxide sensitivity of rat alveolar macrophages.

The phospholipid polyunsaturated fatty acid (PUFA) content and the membrane fluidity of rat alveolar macrophages were modified dose-dependently and in different ways. This was done to study the importance of both membrane characteristics for the cellular sensitivity toward ozone and nitrogen dioxide. Cells preincubated with arachidonic acid (20:4) complexed to bovine serum albumin (BSA) demonstrated an increased in vitro sensitivity versus ozone and nitrogen dioxide. The phenomenon was only observed at the highest 20:4 concentrations tested, whereas the membrane fluidity of the 20:4-treated cells already showed a maximum increase at lower preincubation concentrations. Hence it could be concluded that the increased ozone and nitrogen dioxide sensitivity of PUFA-enriched cells is not caused by their increased membrane fluidity, resulting in an increased accessibility of sensitive cellular fatty acid moieties or amino acid residues. This conclusion receives further support from other observations. These results strongly support the involvement of lipid oxidation in the mechanism(s) of toxic action of both ozone and nitrogen dioxide in an intact cell system.