Design and evaluation of nematode 18S rDNA primers for PCR and denaturing gradient gel electrophoresis (DGGE) of soil community DNA     

Ian S. WaiteAnthony G. O'Donnell  Andrew HarrisonJohn T. Davies  Stephanie R. ColvanKlemens Ekschmitt  Hülya DoganVolkmar Wolters  Tom BongersMarina Bongers  Gabor BakonyiPéter Nagy  Efimia M. PapatheodorouGeorge P. Stamou  Sven Boström 《Soil biology & biochemistry》2003,35(9):1165-1173

Consensus nematode 18S ribosomal DNA primers were designed by aligning available 18S sequences and identifying a variable region flanked by highly conserved regions. These primers were then used to amplify nematode 18S rDNA from whole soil community DNA extracted from a range of European grassland types. Cloning of the PCR amplicons (778 bp) followed by restriction digest analysis (RFLP) resulted in the recovery of 34 unique nematode sequences from the four grasslands studied. Comparison of these data with the limited number of 18S rDNA nematode sequences currently held in on-line databases revealed that all of the sequences could be assigned to known nematode taxa albeit tentatively in some cases. Two of the sequences recovered from the site in the Netherlands (wet, hay-grassland) were recovered in a clade that included a sequence of the genus Trichodorus whilst other sequences from this site showed similarity with 18S rDNA sequences of the genus Prismatolaimus (five sequences), Xiphinema (one sequence) and Enoplus (one sequence). Of the remaining sequences, two showed some affinity with Mylonchulus (UK, upland peat), four with Steinernema (UK) and one sequence with Mesorhabditis (Hungary, east European Steppe). Three sequences from the Netherlands and one from Hungary were recovered in a clade that included a sequence of the genus Pratylenchoides whilst three further sequences from the Netherlands and two from Hungary were recovered in a clade encompassing the genus Globodera. Of the remaining nine sequences, two (NL6, NL62) formed a distinct lineage within the Adenophorea with 90% bootstrap recovery in a paraphyletic clade that included sequences of Prismatolaimus and Trichodorus. Seven sequences (three from the Netherlands, three from the UK and one from Greece) were left unassigned though the tree topology suggested some relationship (58% bootstrap recovery) with the genus Cephalobus. To assess whether primers used to amplify 18S rDNA might be used to fingerprint genetic diversity in nematode communities in soil, the environmental sequence data were used to design a second set of primers carrying a GC-clamp. These primers amplified a 469 bp fragment internal to the region flanked by the primer set used to derive the nematode trees and were used to amplify 18S rDNA for subsequent analysis using denaturing gradient gel electrophoresis (DGGE). DGGE analysis of six major European grassland types revealed considerable genetic diversity between sites. However, the relationships seen with the DGGE data were inconsistent with previous studies where the same soils had been characterized with respect to functional and morphological diversity. To confirm that this second set of primers was amplifying nematode sequences, selected bands on the DGGE gels were extracted, PCR amplified and sequenced. The final alignment was 337 bases. These analyses revealed the presence of sequence signatures from the genera Paratrichodorus, Plectus, Steinernema, Globodera, Cephalobus and Pratylenchoides.  相似文献   

A sperm tail defect associated with infertility in a goat--case report.     

A Molnár  P Sarlós  G Fáncsi  J Rátky  S Nagy  A Kovács 《Acta veterinaria Hungarica》2001,49(3):341-348

Semen of an infertile Dutch White (Saanenthal) goat buck was examined. Light and electron microscopic examinations showed aberrations of the sperm tails resembling the so-called Dag or Dag-like defects described in several cattle breeds. Ejaculated semen showed that virtually all of the cells had strongly coiled or broken tails, or fractured midpieces. Ultrastructural investigations by transmission electron microscopy (TEM) showed uneven distribution of the mitochondria in the midpiece. Coiled tails were encapsulated by a common membrane, and dislocated axial fibres and different membranous structures were also present. The ultrastructural characteristics of the defective sperm tails, the missing parts of the axial fibre bundle and the misalignment of the mitochondria indicate that this first case reported in goat is similar to the Dag-like defect in cattle.  相似文献   

Infection experiments with Mycobacterium paratuberculosis strains     

B K?rmendy  S Tuboly  G Nagy 《Acta veterinaria Hungarica》1986,34(1-2):37-47

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Diagnostic value of intravenously administered johnin purified protein derivative (PPD) in bovine paratuberculosis     

B K?rmendy  S Tuboly  G Nagy  J T?r?k  T Abonyi 《Acta veterinaria Hungarica》1990,38(1-2):43-53

The diagnostic value of intravenously administered johnin purified protein derivative (PPD) was studied in 45 cattle of different age, coming from herds infected by, or free from, Mycobacterium paratuberculosis. In addition to observing the clinical symptoms, the animals' sera were assayed for specific antibodies by the complement fixation (CFT) and immunodiffusion (AGID) tests. The blastogenic transformation of peripheral lymphocytes was determined on the basis of 3HTdR incorporation. Changes in the neutrophilic leucocyte/lymphocyte ratio of the blood were also monitored. Detection of the pathogen in the faeces was attempted by microscopic examination and by culturing. Combined evaluation of responses elicited by intravenously administered johnin PPD can be a valuable aid in recognizing infected animals, particularly those among the heifer progeny of infected cows.  相似文献   

New combination for the therapy of canine otitis externa II Efficacy in vitro and in vivo   总被引：1，自引：0，他引：1  

G. Kiss    Sz. Radvanyi    G. Szigeti    B. Lukáts  G. Nagy 《The Journal of small animal practice》1997,38(2):57-60

A new, effective drug combination was developed for the therapy of canlne otitis externa (OE) by taking into consideration the microorganisms most frequently isolated from dogs affected with OE, their combination, their drug sensitivity and the type and incidence of ear canal inflammation. The antlmycotic active ingredient of the combination Is ketoconazoie, Its antlbacteriai component Is gentamlcin suiphate, while its antiphiogistic constltuent is mazipredone hydrochloride. Based upon the results of in vltro pharmacodynamic tests, the antifungal activity of the combination is superior to that of ketoconazoie used alone at the same concentration. A total of 210 dogs affected wlth OE were treated with the combination: 94-2 per cent of them became clinically symptomless and microbioiogicaiiy negative in an average of 8-5 days. No adverse reactions were observed In connection wlth the use of the drug combination. The therapeutic results can be attributed to the hlgh antifungal efficacy of the combhation demonstrated In vltro and to the favourable properties of the solvent mixture.  相似文献   

The supposed involvement of dogs carrying Brucella suis in the spread of swine brucellosis     

B K?rmendy  G Nagy 《Acta veterinaria Academiae Scientiarum Hungaricae》1982,30(1-3):1-7

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Experiences in the production of oil emulsion vaccines     

A Forgács  T Perényi  F Sólyom  B Nagy 《Archiv fuer experimentelle veterinaermedizin》1987,41(5):650-655

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Vaccination of chickens with a recombinant fowlpox virus containing the hemagglutinin-neuraminidase gene of Newcastle disease virus under the control of the fowlpox virus thymidine kinase promoter.          下载免费PDF全文

E Nagy  P J Krell  R A Heckert    J B Derbyshire 《Canadian journal of veterinary research》1994,58(4):306-308

When chickens were vaccinated with a recombinant fowlpox virus (FPV) containing the Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) cDNA under the control of the thymidine kinase (TK) promoter and inserted into the FPV TK gene, the FPV antibody response to the recombinant virus was similar to the response to vaccination with standard FPV, and the recombinant virus protected chickens against challenge with virulent FPV. While the presence of the NDV HN cDNA was demonstrated in the recombinant virus, which was stable on serial passage, expression of HN was not detected by hemagglutination, Western blot analysis or immunoprecipitation of infected cell lysate. Chickens vaccinated with the recombinant virus failed to mount an NDV hemagglutination-inhibition antibody response, and they did not resist challenge with velogenic NDV. It was concluded that the TK promoter was too weak to drive the HN gene, but that the insertion into the FPV TK gene did not reduce the immunogenicity of the virus.  相似文献   

Duration of anti-adhesive and bactericidal activities of milk from vaccinated sows on Escherichia coli O149 in the digestive tract of piglets during the nursing period.     

L K Nagy  B S Bhogal  T Mackenzie 《Research in veterinary science》1979,27(3):289-296

Piglets were exposed orogastrically to Escherichia coli to enable study of the duration of anti-adhesive and bactericidal activities of milk of sows vaccinated with a K88 enriched E coli vaccine. There was a marked increase in the number of the challenge strain in the digestive tract of weaned piglets of all ages (between 888 and 2144 per cent). In contrast, there was a decrease in their number (75 per cent) in the day-old colostrum-fed piglets. When the piglets were two weeks old milk was still capable of reducing the rate of proliferation of the pathogen but at five weeks it proliferated at equal rates in the digestive tract of both suckling and weaned litter-mates. The rate of adhesion of the K88 positive E coli to the small intestine of colostrum deprived piglets was high (5 x 108/g). Rate of adhesion fell gradually in weaned piglets from 5.4 x 107/g at two weeks to 2.0 x 106/g at four to five weeks of age. In contrast, resistance of the small intestine of suckling pigs to adhesion by K88-positive E coli remained relatively stable through the five week period of nursing bacterial counts ranging from 5 x 104/g to 3 x 104/g of tissue.  相似文献   

Brucella allergen for human diagnostic purposes     

G Markos  G Nagy  J Szathmáry 《Acta veterinaria Academiae Scientiarum Hungaricae》1971,21(4):347-353

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