Causality methods in cosmetovigilance: comparison of Colipa and PLM versus global introspection.

The European Cosmetics Regulation requires a post-marketing system for detection of undesirable effects on human health of cosmetic products. Colipa, the European Cosmetic, toiletry and perfumery association, provided guidelines for causality assessment of these effects. In addition another causality method originally designed for causality rating in Post Launch Monitoring (PLM) of novel foods has been employed to assess causality of cosmetic products. In this study these two causality schemes for consumer cosmetic products were validated against clinical assessment, using the method of global introspection (GI) in 100 reported cases. Causality assessments were performed by three experienced assessors in pharmacovigilance. In the event of discordance between the assessors, an adapted Delphi method was used. The overall Spearman correlation coefficient was 0.74 for comparison of Colipa versus GI, whereas this was 0.50 for PLM versus GI. According to current guidelines, the sensitivity was 0.95 for both the Colipa and PLM method, specificity was 0.84 for Colipa and 0.40 for PLM. From these results it can be concluded the performance of the Colipa causality method yielded better correlation to GI than PLM causality method. The factor identified from comparison of these two schemes as having greatest impact was the course of the reaction.

A general population from Thailand: incidence of common allergens with emphasis on para-phenylenediamine.

BACKGROUND: Most studies on the prevalence of allergy to the permanent hair dye chemical para-phenylenediamine (PPD) are reported from populations of eczema patients attending patch-test clinics, and are assumed to be much higher than in the normal population. No data exist on incidence of senitization to PPD resulting from the use of commercial hair dye preparations over a defined time period. METHOD: A total of 2545 healthy adult volunteers (Bangkok) were screened for PPD allergy through standard patch testing. Volunteers not allergic to PPD were then recruited into two groups: one group applying a commercial hair dye brand as instructed on a monthly basis for 6 months (n=548) and the other group (controls) (n=516) was instructed not to dye their hair for 6 months. Sensitization to PPD resulting from the use of hair dye over this period was then detected by repeat patch testing. RESULTS: The prevalence of PPD allergy in a normal adult population was 2.7% (m=2.4%, f=3.2%). Projected to the adult Thai population, at least 1,000,000 Thai individuals could be allergic to PPD. The incidence of sensitization through the monthly application of standard commercial hair dye preparations over a 6-month period was 1.3%, substantially higher than in controls (0.4%), although numbers were small and not statistically significant. INTERPRETATION: There is a higher prevalence of hair dye allergy among the normal population than previously thought. The incidence of new cases of PPD allergy would indicate that current regulations and practice of hair dye exposure lead to PPD sensitization and allergy, which is a public health problem.

Electronmicroscopy of the surface of Pasteurella haemolytica.

The surfaces of Pasteurella haemolytica, biotype A, serotypes 1, 2, 6, 7 and 9 and of P. haemolytica, biotype T serotypes 3, 4, 10 and 15 were examined by transmission electronmicroscopy with ruthenium red staining and polycationic ferritin labelling, by scanning electronmicroscopy, and by light microscopy. Electronmicroscopy showed that the surface of strains of P. haemolytica biotype A was covered by irregular protrusions which were probably capsular material. The surface and general morphology of P. haemolytica biotype T were distinct from those of biotype A.

Outer membrane proteins of bovine strains of Pasteurella multocida type A and their doubtful role as protective antigens.

Outer membranes were prepared by the Sarkosyl method from 30 strains of Pasteurella multocida and the closely related Taxon 13, which had been isolated from cattle. The patterns of the outer membrane proteins (OMPs) on SDS-PAGE were generally similar to one another, though the four major proteins (a-d) varied somewhat in molecular mass; these patterns allowed the strains to be arranged into 12 groups. Taxon 13 strains and typical P. multocida strains were indistinguishable, both types being found within the same group. Mice were vaccinated with heat-killed bacteria of three strains and challenged with 10 LD50 of homologous and heterologous live bacteria, representing groups based on OMP patterns; the best protection was afforded by strain W674, which protected against nine of the 17 challenge strains; but there was no correlation between protection and PAGE pattern. Pre-vaccination and pre-challenge sera were used in immunoblotting to probe OMPs from protective and non-protective strains. All three vaccines produced antibody to proteins a and d; these proteins appeared to be common to all strains, varying in molecular mass but not in overall antigenic expression. The antibody response to the other two major OMPs appeared to be PAGE-group specific. There was no correlation between protection and the antigen pattern seen by immunoblotting.

Ovine systemic pasteurellosis caused by Pasteurella haemolytica biotype T.

A detailed study was made of lambs aged 5--7 months naturally infected with Pasteurella haemolytica biotype T. In addition to the well known features of such infections, previously unreported necrotic lesions of the tonsil, oesophagus, pharynx and adjacent areas were consistently seen. Large numbers of P. haemolytica were present in the tonsil, oesophageal lesions, lung, liver and spleen, but few or none in other tissues. The evidence indicated that the disease was not a true septicaemia. It is postulated that P. haemolytica biotype T already present in the tonsils multiplies and invades the adjacent tissues of the upper alimentary tract; groups of organisms from this site enter the blood stream as emboli, most of which lodge in the capillary beds of the lung and liver; rapid multiplication of organisms in these tissues leads to death from the effects of endotoxin.

Production of mouse monoclonal antibodies to Pasteurella multocida type A and the immunological properties of a protective anti-lipopolysaccharide antibody.

Eight monoclonal antibodies (MAbs) were produced from mice immunised with whole cells of heat-killed Pasteurella multocida type A which had been cultured under iron-restricted conditions. The MAbs were selected by an enzyme-linked immunosorbent assay (ELISA) in which the antigen consisted of whole bacteria of the immunising strain. Their reactivity was investigated further by immunoblotting, indirect haemagglutination, a complement-mediated bactericidal assay and passive protection of mice. One of the eight MAbs was shown by immunoblotting to react with lipopolysaccharide (LPS), was bactericidal, and completely protected mice against homologous challenge with 10 LD50 of live bacteria. This MAb was selected for further study. Its reaction with LPS of 17 type-A strains and of single strains of types B, D and E was investigated by immunoblotting. Strains that reacted with the anti-LPS MAb in immunoblots were susceptible to its bactericidal activity and gave high ELISA absorbances. Those that did not react were not susceptible to its bactericidal activity and gave low ELISA readings. The relation between bactericidal activity and ELISA absorbance was highly significant (p less than 0.001). Five of the strongly reacting heterologous strains and one non-reacting strain were selected as challenge organisms in a passive protection experiment: only the mice receiving the reacting strains were protected.

Immunity of specific pathogen-free lambs to challenge with an aerosol of Pasteurella haemolytica biotype A serotype 2. Pulmonary antibody and cell responses to primary and secondary infections.

Specific pathogen-free (SPF) lambs previously exposed to an aerosol of P. haemolytica biotype A serotype 2 (A2) were immune to subsequent challenge with an aerosol of P. haemolytica A2. Untreated control lambs were not immune to this challenge. The local immune responses of the lung to these challenges were examined. High IgG and IgA titres to P. haemolytica and high levels of opsonizing antibody against P. haemolytica were present in the lung washings from previously infected immune lambs at autopsy, seven days after the second infection. Lung washings from control lambs, 7 days after challenge with P13 virus and P. haemolytica A2, had no IgG titres, very little opsonizing activity but did have IgA titres which were significantly higher than in unchallenged control lambs. The cellular response of animals challenged with P13 virus and P. haemolytica was significantly greater than that of unchallenged controls or of lambs exposed only to P. haemolytica. However, this finding was complicated by the response to P13 virus. Lymphocytes from lung washings of all lambs failed to respond in a lymphocyte stimulation test to phytohaemagglutinin while blood lymphocytes did respond. There was little specific response to P. haemolytica antigen in the test.

The specificity of the fluorescent antibody test using the sera of rabbits inoculated with strains of myocobacteria.

Sera from experimentally infected rabbits were used to test the specificity of the fluorescent antibody test. It was possible using mono-specific sera to differentiate antigenically Mycobacterium phlei, M fortuitum, M smegmatis, M avium, M intracellulare, M bovis (BCG) and M johnei. The cross-reactivity within the M avium and M intracellulare group was such that one antigen from these groups would detect infection within that group and exclude M johnei infection. The M phlei growth factor independent strain M johnei 316F was shown to be antigenically distinct from a M phlei dependent strain 9N96. There was loss of specificity when M avium infection was superimposed on a previous M johnei infection and when M johnei infection was superimposed on M avium infection.

The specificity and sensitivity of the fluorescent antibody test for Mycobacterium johnei infection in abattoir and culled cattle.

In a series of 100 cattle in which there was no bacteriological or histopathological evidence of Mycobacterium johnei infection there were four positive reactions to the fluorescent antibody (FA) test with M johnei antigen and 20 to the complement fixation (CF) test. In a second series of 118 culled adult cattle M johnei infection was found in 26. The FA test was positive in 16 and the CF in 12 of those infected cattle. In 92 cattle with no evidence of M johnei infection the FA test was positive in seven and the CF test in 22.

The specificity and sensitivity of the fluorescent antibody test in cattle experimentally infected with Mycobacterium avium and Mycobacterium johnei.

In five calves experimentally infected with Mycobacterium avium and in 10 with M johnei it was shown that the serological response by the fluorescent antibody (FA) test was specific. The serological response in the M avium infected calves was transitory and lasted up to five months. The first evidence of a serological response with the FA test in the M johnei dosed calves occurred at four months, and all calves had reacted by nine months after dosing. At the 23 tests carried out between four and 32 months after dosing, when the experiment was terminated, an average of 5-8 animals was positive at any one test. Johne's disease was confirmed bacteriologically and histologically in six of the 10 calves though none had shown clinical signs.

The identification of Moraxella bovis and Neisseria ovis from the eyes of cattle and sheep.

The cultural characteristics of Moraxella bovis and Neisseria ovis from eyes of cattle and sheep were examined to determine which tests precisely identified the isolates. The elongation test to distinguish the bacillary M vovis from the coccal N ovis, the nitrate reduction and the litmus milk tests were found to be the most reliable.

Serotypes of Pasteurella haemolytica in ovine pasteurellosis.

The serotypes of 406 strains of Pasteurella haemolytica were determined. These came from cases of pasteurellosis in sheep of all ages. Serotype A2 was the most frequent and comprised 33 per cent of all strains recovered. Serotypes T3, T4 and T10 comprised 16, 14 and 12 per cent of the total respectively. Serotypes A1 and A6 contributed 5 per cent each, A7, 18, A9, A11 and A12 a total of 8 per cent collectively, serotype A5 was not isolated at all while 6 per cent of strains were serologically untypable. More strains were received during September, October and November than at any other three-month period and in these months the T biotype predominated.

Susceptibility of specific pathogen-free lambs to concentrations of Pasteurella haemolytica serotype A2 in aerosols.

A 100-fold reduction in the numbers of organisms in an aerosol of Pasteurella haemolytica used to infect specific pathogen-free lambs did not alter the number of cases of pneumonia which resulted. In a separate experiment a further 10-fold reduction in the number of organisms in the aerosol did not cause fewer cases of pneumonia.

Studies on strains of Pasteurella haemolytica not typable by the indirect haemagglutination test.

Thirty strains of Pasteurella haemolytica which were untypable by the indirect haemagglutination (IHA) test were examined serologically by rapid plate agglutination (RPA), agar gel diffusion (AGD), crossed immunoelectrophoresis (CIE) and counter current immunoelectrophoresis (CCIE) tests. Nine serogroups were identified by CCIE. Serogroup specificity, dependent on two antigens, was present in heated saline extracts of cells. Single representative strains from two serogroups were not pathogenic for specific pathogen-free lambs.

Characterisation of a new serotype of P haemolytica isolated in Hungary.

Three strains belonging to a new serotype of Pasteurella haemolytica (A16) were isolated from lambs and a wild boar in Hungary. The identity and validity of the new serotype was proved by biochemical tests and by the indirect haemagglutination test using unabsorbed and absorbed hyperimmune sera raised in rabbits.

A serological comparison of Pasteurella haemolytica vaccines containing different adjuvants.

Five adjuvants were compared for their ability to enhance the serological response of sheep to capsule extract of Pasteurella haemolytica biotype A serotype 6. Vaccines of this antigen were inoculated with incomplete Freund's adjuvant, complete Freund's adjuvant, incomplete Freund's adjuvant containing a water soluble extract of Mycobacterium tuberculosis, aluminium hydroxide gel or a combination of aluminium hydroxide gel and incomplete Freund's adjuvant. This latter vaccine induced significantly higher titres of antibody as measured by an indirect haemagglutination test than did any of the other vaccines. The aluminium hydroxide gel alone was shown to be the poorest adjuvant. The local reactions at the sites of inoculations produced by the aluminium hydroxide gel in incomplete Freund's adjuvant vaccine were not severe and were not detectable beyond one month after vaccination in the majority of the sheep.

Experimental infection of lambs with an aerosol of Pasteurella haemolytica.

Specific pathogen free lambs were exposed to an aerosol of Pasteurella haemolytica. Ten lambs vaccinated with an adjuvanted killed P haemolytica vaccine and nine units of P haemolytica administered in an aerosol. Pneumonia histologically indistinguishable from natural pneumonic pasteurellosis occurred in one vaccinated lamb and in four control lambs.

Experimental immunisation of lambs against pneumonic pasteurellosis.

Methods of immunising lambs against pneumonic pasteurellosis, caused by several serotypes of Pasteurella haemolytica, were assessed in specific pathogen free lambs. Lambs were vaccinated intramuscularly with sodium salicylate extract (SSE) of P haemolytica, either alone or in combination with heat-killed organisms (HKO). SSE of P haemolytica type A1 protected vaccinated lambs against pneumonia resulting from challenge with the homologous serotype. SSE of type A2 also provided some protection but this was improved by vaccination with a combination of SSE and HKO.

Pasteurella haemolytica infections in sheep.

Summary Pasteurella haemolytica causes two distinct disease syndromes in sheep. P. haemolytica biotype A causes septicaemia in young lambs and pneumonia in all ages of sheep. Biotype T produces an acute systemic disease affecting principally the upper alimentary tract and lungs in young adult sheep. The bacteriology, epidemiology and clinical and necropsy findings of the two diseases are described and the current situation regarding their experimental reproduction and immunology is reviewed.

Pasteurellosis in sheep.

Pasteurellosis is an important cause of economic loss to the sheep industry. There are two distinct syndromes. The pneumonic form of the disease caused by P haemolytica biotype A occurs as pneumonia in flocks and sporadically in individual sheep. The septicaemic form, caused by P haemolytica biotype T is associated with hyper-acute disease and occurs most commonly in the autumn coinciding with the folding of hoggs on rape, turnips and improved pastures. The factors which predispose sheep to the different forms of the disease are poorly understood but recently it has been possible to reproduce pasteurella pneumonia experimentally.