Comparison of the enzyme linked immunosorbent assay, the haemagglutination inhibition test and the agar gel precipitation test for the detection of antibodies against infectious bronchitis and Newcastle disease in commercial broilers

Serological results as obtained by the agar gel precipitation (AGP) test, haemagglutination inhibition (HI) test (M41, D274, D1466) and an enzyme linked immunosorbent assay (ELISA) from commercial broiler flocks, which had suffered from a clinical infectious bronchitis (IB) infection in the fattening period, were compared with serological findings of control broiler flocks. In addition the Hi-test and ELISA for Newcastle disease (ND) were compared. The AGP results differ significantly between groups. The test had a low sensitivity. The results of the HI-IBV tests were similar for both groups and no difference was seen between flocks with or without clinical IB problems. The IBV ELISA showed a highly significant difference between the two groups. In contrast to the low and not significant correlation coefficients (r) between the IBV HI tests and the ELISA, the value of r between the NDV HI and ELISA was high. The results indicate an ELISA as the preferred test for the diagnosis of IB in broilers.     

K88 fimbriae as carriers of heterologous antigenic determinants.

The K88 fimbriae of enterotoxigenic Escherichia coli are strongly immunogenic antigens that can be used to evoke protective immunity. To find out whether these fimbriae can be used as carriers for foreign epitopes, a highly variable region present in the primary structure of the different K88 variants was replaced with five different heterologous epitopes to investigate to what extent these insertions affected the expression, assembly (biogenesis), stability and immunogenic properties of the resulting hybrid fimbriae. Amino acid residues 163-173, were replaced using site-directed in vitro mutagenesis and the hybrid fimbriae were tested for these aspects using ELISA, immunoelectronmicroscopy and immunoblotting. Replacement of this highly variable region did not affect the biosynthesis of fimbriae, although all mutations tested resulted in a reduced expression depending on the epitope inserted. Testing of the different hybrid fimbriae with a panel of monoclonal antibodies raised against the various K88 serotypes K88ab, K88ac and K88ad indicated that replacement of amino acid sequence 163-173 did not affect conserved or K88ab specific epitopes but the K88ac and K88ad specific conformation was lost. Immunization with hybrid fimbriae raises antibodies specific for the inserted heterologous epitopes.     

Early brain lesions and face-processing development

Studies of functional plasticity after pre- or perinatal brain damage can tell us whether the neural substrate normally involved in the development of a given ability is specific and, if so, when it becomes functionally specified and unique. Development of face processing was investigated in 5- to 17-year-old children who had a unilateral brain injury in the pre-, peri-, or postnatal period. In Studies 1 and 2, patients with a posterior injury involving the temporal regions exhibited a face-processing deficit that was independent of their age at test time. Even though differences were observed between the two hemispheres in face processing during infancy as well as in adults in cases of normal development, no clear differences between right and left injury were observed here in face-processing deficit. Poor postlesional face-processing plasticity seems to contrast with results of several studies on speech development after early unilateral injury. If the difference in the time window for postlesional plasticity between these two areas of competency is confirmed, it would suggest that the two kinds of abilities rely on neural cells which are sensitive to different plasticity factors.     

Comparison of arbitrarily primed PCR with restriction endonuclease and immunoblot analyses for typing Clostridium difficile isolates.

Arbitrarily primed PCR (AP-PCR) was used to genotype 26 clinical isolates of Clostridium difficile previously analyzed by immunoblotting (IB) and 20 isolates typed by restriction endonuclease analysis (REA) with HindIII. Two levels of differentiation were achieved with the AP-PCR approach by use of two different arbitrary primers. With the 19-mer arbitrary primer T-7 (first level of differentiation), a good correlation was found between IB and AP-PCR typing. Twenty isolates grouped into six IB types were separated into seven major AP-PCR types. These seven AP-PCR groups were further discriminated into 12 subtypes after genotyping with the arbitrary primer PG-05 (second level of differentiation). The remaining six isolates, all of different IB types, showed a unique and distinct DNA banding pattern with both of the arbitrary primers, T-7 and PG-05. Twenty isolates representing 20 REA types from 15 REA groups were resolved into 13 AP-PCR DNA profiles with the arbitrary primer T-7. A good correlation was found at this level of differentiation between the major REA groups, Y and M, and AP-PCR typing. While AP-PCR with this primer failed to differentiate isolates in REA groups J, G, R, and B, AP-PCR with PG-05 resolved these four isolates into four distinct AP-PCR types. In addition, one of three M strains and one of four Y strains displayed a slightly different DNA banding pattern by AP-PCR (with PG-05) from that of the other strains in the group. We conclude that AP-PCR is a rapid and sensitive method which not only complements other typing schemes but also may be a substitute and prove to be especially suited for immediate epidemiological tracking of nosocomial infections due to C. difficile.     

Squash Procedure for Protein Immunolocalization in Meiotic Cells

Several techniques have been developed for protein immunolocalization in meiotic cells. However, most of them include treatments that lead to cell disruption and are only suitable for prophase-I cells. We describe a novel squash procedure of cell preparation for protein immunolabelling of different meiotic stages. This procedure is an alternative to both cryosectioning and whole spreading procedures. We present results obtained in mouse spermatocytes with three different antibodies: the MPM-2 mAb against mitotic phosphoepitopes, an anticentromere serum and a polyclonal serum against the SCP3 protein of the axial elements and lateral elements of the synaptonemal complex. The procedure was tested for single and double immunolabelling. With this technique a large number of cells at different meiotic stages can be analysed. Cell stages are easily identified and cell and chromosome structures are preserved. Thus, it allows the study of chromosome behaviour and the relations hips between the different structural elements of the cell throughout meiotic divisions. Our procedure is also suitable for three-dimensional (3D) analyses and proved to be reliable in a wide range of systems including insects and mammals. In addition, the procedure may be interesting to obtain a rapid immunological diagnosis.     

Induction of an adaptive response to quercetin, mitomycin C and hydrogen peroxide by low doses of quercetin in V79 Chinese hamster cells

The adaptive response is a phenomenon by which cells exposedto low, non-cytotoxic doses of a genotoxicant become significantlyresistant to a subsequent higher dose of the same or anothergenotoxic agent. Induction of the adaptive response has beenmainly studied using ionizing radiation and alkylating agentsas genotoxic agents. However, other mutagenic agents may warrantfurther study, since the adaptive response as a whole may bean important general biological mechanism to maintain geneticintegrity and thus could prevent carcinogenic initiation ofcells. The exposure to mutagenic agents present, or formed,in the diet is considered an important factor in the etiologyof human tumors and a considerable number of these agents havenot yet been identified or characterized. Flavonoids are a largegroup of polyphenolic quinoids found in a wide variety of ediblefruits and vegetables and a few, such as quercetin, presentgenotoxic activity in vitro. The mechanisms of mutagenicityof quercetin involve the production of oxygen radicals throughan autoxidation process dependent on pH value and the presenceof oxygen. Although there are few doubts regarding the mutagenicityof quercetin invitro, carcinogenicity of this flavonoid is stillcontroversial. In view of these conflicting results and theradiomimetic nature of the mutagenicity of flavonoids, we addressedthe question of cell exposure to quercetin at the low levelspresent in the diet leading to adaptation to further exposureto mutagens or carcinogens. The work reported here concernsinduction of an adaptive response by low doses of quercetinto challenging doses of quercetin and other compounds, namelyhydrogen peroxide and mitomycin C, using induction of chromosomalaberrations in V79 cells as the end point. ⁴To whom correspondence should be addressed: Tel: +351 1 3610290; Fax: +351 1 3622018; Email: jose.rueff{at}gene.unl.mailpac.pt     

Premières observations sur deux Microsporidies hyperparasites d'Acanthocéphales de Poissons marins et lagunaires

Résumé Microsporidium acanthocephali n. sp. et Microsporidium propinqui n. sp. sont deux espèces de Microsporidies hyperparasites du syncytium tégumentaire d'Acanthocéphales de Poissons marins et lagunaires. Leur étude ultrastructurale met en évidence des caractères communs: mérontes et spores à diplocaryons, filament polaire anisofilaire, mais aussi une vacuole parasitaire autour des mérontes et quelques stades sporogoniques de l'espèce Microsporidium acanthocephali. Les spores des deux parasites sont oviformes; celles de M. acanthocephali, géantes, mesurent 12–14 m de longueur sur 6–7 m de largeur, celles de M. propinqui seulement 3–4 m X 1,25–1,50 m. La méconnaissance de certains aspects de la sporogonie empÊche une assignation générique de ces deux parasites qui peuvent cohabiter chez un mÊme Acanthocéphale.Au cours de recherches sur les Acanthocéphales de Poissons marins et lagunaires (Buron 1986), deux Microsporidies hyperparasites de ces Helminthes ont été observées chez des Poissons Gobiidae et Pleuronectiformes. Les Microsporidies sont déjà connues pour Être des hyperparasites de Grégarines, de Myxosporidies et d'autres Helminthes (Cestodes, Trématodes, Nématodes) (Sprague 1977). Il semble cependant, chez les Acanthocéphales, n'avoir été recensé jusqu'à présent que des «psorospermies» par Moniez (1879) que Balbiani (1884) interprète comme étant des Grégarines (in Dollfus 1946).Le présent travail propose donc de décrire quelques stades du cycle biologique des deux premières Microsporidies indiscutables d'Acanthocéphales.

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Preliminary observations on two microsporidian hyperparasitic in Acanthocephalan of salt water fishes

Microsporidium acanthocephali n. sp. and Microsporidium propinqui n. sp. are two microsporidian species hyperparasitic in the tegumentary syncytium of salt-water fishes. Their ultrastructural study shows that both species have diplocaryotic meronts and spores, anisofilar polar filaments; meronts and some sporogonic stages of M. acanthocephali are surrounded by a parasitic vacuole. Spores of both parasites are oviform; those of M. acanthocephali are gigantic, 12–14 m long and 6–7 m broad, those of M. propinqui are only 3–4 m X 1.25–1.50 m. A poor knowledge of some sporogonic aspects prevents us from all precise generic assigning of these two parasites which may coexist in the same Helminth.

     

High prevalence of tuberculosis in AIDS patients in Spain

A total of 67 cases of tuberculosis was diagnosed in the first 100 cases of AIDS, diagnosed according to the former CDC criteria, at a hospital in Madrid, Spain. This is the highest known prevalence of tuberculosis in AIDS patients both within and outside Spain. The clinical manifestations of tuberculosis were very variable and atypical. The rate of isolation ofMycobacterium tuberculosis from blood was particularly high: of 25 patients in whom blood cultures were performed, 16 were positive. In a third of the patients with proven mycobacteremia, blood was the first or the only positive specimen. In general, therapy resulted in rapid clinical improvement, but in some cases mycobacteria were isolated from clinical or necroscopy specimens months after what was considered adequate therapy.