Authors:Ángela Lacombe-Antoneli, Segundo Píriz, Alberto Quesada and Santiago Vadillo
Footrot is widely considered the most severe and most common foot pathology in small ruminants. This study tested the ability of a molecular typing system based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay of the 16S rRNA gene to discriminate between the strict anaerobe genera most commonly isolated from footrot (
Bacteroides, Dichelobacter, Fusobacterium, Porphyromonas
) in goats in Extremadura (Spain), with a view to facilitating identification for diagnostic purposes and thus providing a useful tool for future epidemiological studies. Although the electrophoretic patterns obtained with the enzyme
1I were more readily interpreted, and may thus be the best initial option, results may be confirmed by a second enzyme (
I). The PCR-RFLP assay of the 16S rRNA gene may therefore prove a useful addition to conventional biochemical identification techniques, providing taxonomic information at genus level.
Furlanello , T. (2008) : Babesia canis canis and Babesia canis vogeli clinicopathological findings and DNA detection bymeans of PCR-RFLP in blood from Italian dogs suspected of tick-borne disease . Vet. Parasitol . 157 , 211 – 221
Authors:Judit Bíró, Noémi Erdei, Ibolya Székely and L. Stipkovits
Increasing use of Mycoplasma gallisepticum (MG) live vaccines has led to a need for the differentiation of MG strains. The MG strains MK-7, MS-16, S6, FS-9 and R strains and the MG live vaccine strain F were compared by random amplification of polymorphic DNA (RAPD) in this study. Using RAPD, different patterns were found among the MG strains. In addition to this, we examined the differentiating potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) primers targeted at the crmA, crmB, crmC, gapA, mgc2 and pvpA genes encoding cytadherence-related surface proteins. These proteins may take part in the pathogenesis of MG-induced disease. Differentiation of strain F is based on the identification of restriction enzyme sites in the PCR amplicons. Using HphI enzyme, crmC PCR amplicons produced different RFLP patterns. Digestion of amplicons of gapA-specific PCR with MboI enzyme also produced distinct patterns. Differences were observed among strains R and F by digestion of mgc2 PCR amplicons with HaeIII and VspI enzymes and digestion of pvpA PCR amplicons with AccI, PvuII and ScrFI endonucleases. This method can be used for the rapid differentiation of vaccine strain from wild strains. Differentiation of MG strains is a great advantage for diagnosticians or practitioners and it is useful for epidemiological studies.
Authors:I. Anton, A. Zsolnai, I. Komlósi, A. Király and L. Fésüs
The effect of the porcine myogenin (Myog) 3' polymorphism on birth weight, growth rate, carcass weight, lean weight, lean meat percentage and backfat thickness has been investigated in Hungarian Large White pigs. MYOG genotypes were determined by PCR-RFLP assay. The obtained MYOGA frequency value was 0.6275. Due to the small number of BB piglets the effect of the MYOG genotypes on birth weight was not significant; however, an increasing tendency was observed from genotype AA to BB. The growth rate difference between MYOG genotypes was significant: BB animals showed the highest growth rate values during the fattening period. Since few results are available on the possible use of MYOG gene polymorphism in selection to improve carcass and growth traits, by this study the authors hope to provide additional data on this particular subject.
Authors:J. Kundu, J. Jarošová, S. Gadiou and G. Cervená
One-step RT-PCR followed by a RFLP assay was developed for the typing of
Barley yellow dwarf virus
(BYDV) member of
genera. RFLP analysis performed on 23 samples including one lab isolate showed three types of
II restriction profile. A partial coat protein (CP) gene sequencing was carried out and confirmed the RFLP analysis. Both sequencing and RFLP analysis identified the presence of 3 BYDV-PAV (including the lab isolate Blatno85), 10 BYDV-MAV and 10 BYDV-PAS isolates among the field samples. One-step RT-PCR together with RFLP presents an easy and reliable assay for routine BYDV typing. These methods revealed that PAS and MAV are more dominant than PAV in the Czech Republic.
Authors:S.ibel Başkurt, E. Doğaç, V. Taşkın and Belg.in Taşkın
In the present study, the frequencies of three organophosphate (OP) resistance-associated mutations in acetylcholinesterase gene of Bactrocera oleae (BoAce) populations collected from 8 different important olivegrowing areas in the west part of Turkey were determined. Populations were sampled from the areas that have been treated with only the pyrethroid α-cypermethrin; pyrethroids plus OPs; deltamethrin with pheromone eco-traps, and no insecticide treatment applied areas for many years. For Ile214Val and Gly488Ser point mutations PCR-RFLP and for Δ3Q deletion mutation PCR diagnostic tests were carried out. Seventy-two percent of the total individuals analyzed in the study were exhibited heterozygous genotype (RS) for both Ile214Val and Gly488Ser point and homozygous susceptible genotype (SS) for Δ3Q deletion mutations. This RS/RS/SS combination together with RS/RR/SS with the frequency of 13% were the most common two combinations observed in all of the populations under different insecticide regimes, even in the populations under no insecticide pressure for many years. Independent evaluation of the three mutations resulted in 0.450, 0.534 and 0.037 frequency values for the resistant alleles of 214Val, 488Ser and Δ3Q mutations, respectively. Among the studied populations, the frequencies of resistant alleles for the positions of 214 and 488 were not differed from each other. However, in 3 of the populations the frequency of the R allele of Δ3Q was zero and it changed between 0.025 and 0.100 in the remaining five populations. Results of this study contributed to the distribution pattern of the two point mutations in Europe and a pattern for Δ3Q mutation was determined for the first time in the field collected olive fly samples.
Leung, E. T. Y., Kam, K. M., Chiu, A., Leung, P. H., Seto, W. H., Yuen, K. Y., Yam, W. C.: Detection of KatGSer315Thr substitution in respiratory specimens from patients with isoniazid-resistant
using PCR-RFLP. J. Med
Authors:István Anton, Katalin Kovács, László Fésüs, József Várhegyi, László Lehel, Zoltán Hajda, J. Polgár, Ferenc Szabó and Attila Zsolnai
The objective of this study was to estimate the effect of the thyroglobulin (TG) locus on beef quality traits in some beef cattle breeds and to investigate the effect of the DGAT1 locus on milk production traits in the Hungarian Holstein Friesian population. TG and DGAT1 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. At the TG locus TT bulls showed the highest fat percentage values in the longissimus dorsi muscle (m. longissimus dorsi); the difference between CC and TT genotypes was significant. DGAT1 GC/GC cows had the highest milk, fat and protein yield values. Due to the relatively small number of GC/GC cows the difference proved to be significant only between AA/AA and AA/GC genotypes.