Assessing genetic variability in germplasm of Phaseolus vulgaris L. collected in Northern Portugal     

Rita C. Coelho  Miguel A. Faria  Joana Rocha  Aida Reis  Maria Beatriz P.P. Oliveira  Eugénia Nunes 《Scientia Horticulturae》2009

The common bean (Phaseolus vulgaris L.) is the most consumed legume in the world. In Portugal, the geographic isolation of the region surrounded by the mountainous barrier of Peneda-Gerês, Barroso and Marão is thought to have the safeguard of a large number of adapted bean populations. In order to assess the value of this germplasm to breeding programs, a study of 20 Portuguese landraces collected in this Northern region were evaluated for agronomical (days to flowering, plant height, days to harvest, weight of seeds per plant and 100 seeds weight), chemical (crude protein) and genetic diversity (microsatellite DNA).  相似文献   

Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil     

Macieira Dde B  Messick JB  Cerqueira Ade M  Freire IM  Linhares GF  Almeida NK  Almosny NR 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2005,34(1):44-48

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.  相似文献   

Quantification,risk factors,and health impact of natural congenital infection with bovine viral diarrhea virus in dairy calves     

Muñoz-Zanzi CA  Hietala SK  Thurmond MC  Johnson WO 《American journal of veterinary research》2003,64(3):358-365

OBJECTIVES: To estimate risk and identify risk factors for congenital infection with bovine viral diarrhea virus (BVDV) not resulting in persistent infection and examine effect of congenital infection on health of dairy calves. ANIMALS: 466 calves. PROCEDURES: Calves from 2 intensively managed drylot dairies with different vaccination programs and endemic BVDV infection were sampled before ingesting colostrum and tested with their dams for BVDV and BVDV serum-neutralizing antibodies. Records of treatments and death up to 10 months of age were obtained from calf ranch or dairy personnel. Risk factors for congenital infection, including dam parity and BVDV titer, were examined by use of logistic regression analysis. Effect of congenital infection on morbidity and mortality rates was examined by use of survival analysis methods. RESULTS: Fetal infection was identified in 10.1% of calves, of which 0.5% had persistent infection and 9.6% had congenital infection. Although dependent on herd, congenital infection was associated with high BVDV type 2 titers in dams at calving and with multiparous dams. Calves with congenital infection had 2-fold higher risk of a severe illness, compared with calves without congenital infection. CONCLUSIONS AND CLINICAL RELEVANCE: The unexpectedly high proportion of apparently healthy calves found to be congenitally infected provided an estimate of the amount of fetal infection via exposure of dams and thus virus transmission in the herds. Findings indicate that congenital infection with BVDV may have a negative impact on calf health, with subsequent impact on herd health.  相似文献   

Effect of multiple sampling on diagnostic sensitivity.     

Mark C Thurmond  Wesley O Johnson 《Journal of veterinary diagnostic investigation》2004,16(3):233-236

Overall diagnostic sensitivity is the probability that a diagnostic procedure will detect an agent if the tested animal is indeed infected. The overall or effective sensitivity is a function of both the probability that the assay will detect the agent if it is present in the sample tested and the probability that the agent will be present in the sample tested if the animal is infected with the agent. Thus, even with a highly sensitive assay, the probability of detecting an infected animal may be low or nil if the sampling procedure failed to capture the agent in samples tested by the assay. In this article, it is demonstrated how increased frequency of testing, such as testing multiple subsamples, can have a profound effect on increasing the overall sensitivity of a diagnostic procedure.  相似文献   

Evaluation of bluetongue virus diagnostic tests in free-ranging bighorn sheep     

Randall S. Singer  Walter M. Boyce  Ian A. Gardner  Wesley O. Johnson  Amy S. Fisher 《Preventive veterinary medicine》1998,35(4):6857-282

Five bluetongue virus (BTV) diagnostic tests were evaluated for use in free-ranging bighorn sheep. We sampled one bighorn sheep population four times between 1989 and 1995. The tests evaluated included virus isolation (VI), polymerase-chain reaction (PCR), serum neutralization (SN), agar-gel immunodiffusion (AGID), and competitive enzyme-linked immunosorbent assay (c-ELISA). The c-ELISA, AGID and SN tests had high levels of agreement in determining serogroup exposure in bighorn sheep. We used maximum-likelihood algorithms to estimate the parameters of each diagnostic test used. Although the c-ELISA and AGID had high sensitivity and specificity, the SN had perfect specificity but lower apparent sensitivity. Due to the potential of cross-reactions among multiple serotypes, results of the SN must be interpreted with caution when assessing serotype exposure in an area where multiple serotypes are endemic. The PCR assay delineated convalescent antibody titers from more-recent infections, and consequently, was pivotal in distinguishing a different exposure pattern between the bighorn sheep and cattle in an adjacent herd. Based on an increasing seroprevalence (50% to 100%), BTV circulated through this bighorn sheep population between 1989 and 1993. This increase in seroprevalence coincided with a bighorn die-off due to BTV infection in June, 1991. An adjacent cattle herd was sampled in 1995 for comparison. The bighorn sheep and adjacent cattle had different patterns of exposure to BTV between 1994 and 1995. There was no evidence that BTV circulated through the bighorn sheep population from 1994 to 1995. In 1995, seroprevalence to BTV decreased to 72%, none of yearling bighorn was seropositive, and all of the 39 bighorn sheep were PCR-negative. In contrast, all adult cattle were seropositive to BTV by c-ELISA and SN, and 4 of the calves were seropositive; 11 of the 24 cattle were PCR-positive, including all five calves. Overall, the pattern of temporal herd immunity in the bighorn sheep appeared to follow a classic epidemic curve, with the appearance and subsequent disappearance of herd immunity coinciding with the 1991 die-off in this population. As low levels of herd immunity and high proportions of susceptible animals are key factors in the development of epidemics, this population of bighorn sheep may be at increased risk for a BTV epidemic in the future.  相似文献   

Relationships Between Angular Leaf Spot, Healthy Leaf Area, Effective Leaf Area and Yield of Phaseolus vulgaris     

W.C. Jesus Junior  F.X.R. Vale  R.R. Coelho  P.A. Paul  B. Hau  A. Bergamin Filho  L. Zambolim  R.D. Berger 《European journal of plant pathology / European Foundation for Plant Pathology》2003,109(6):625-632

Three field experiments were carried out with the bean cultivar Carioca Comum to investigate the relationships among visual and virtual severity of angular leaf spot (caused by Phaeoisariopsis griseola), area under visual and virtual disease progress curves (AUDPC), healthy leaf area index on any given day (HLAI), healthy leaf area duration (HAD), healthy leaf area absorption (HAA), effective leaf area duration (ELAD), effective leaf area absorption (ELAA) and yield of Phaseolus vulgaris. To obtain a wide range of disease severities, the plots were sprayed with fungicide at different stages of plant growth (before, during and after flowering). Visual and virtual severity and AUDPC showed no significant correlation with yield. However, HAD, HAA, ELAD and ELAA were significantly correlated with yield. Variables that considered the effective leaf area (ELAD and ELAA) provided similar or better coefficients of determination (R²) than those that considered the remaining green leaf area only (HAD and HAA). Single-point models with HLAI, effective leaf area index (ELAI), intercepted radiation by healthy leaf area (HRI) and intercepted radiation by effective leaf area (EHRI) to estimate yield at various times during the crop season were developed. The slope of the relationship between yield and HLAI, ELAI, HRI and EHRI proved to be stable, regardless of planting date and bean growth stage (from R6 to R8).  相似文献   

Following the dynamics of strobilurin resistance in Blumeria graminis f.sp. tritici using quantitative allele-specific real-time PCR measurements with the fluorescent dye SYBR Green I   总被引：1，自引：1，他引：1  

B. A. Fraaije †  J. A. Butters  J. M. Coelho  D. R. Jones  D. W. Hollomon 《Plant pathology》2002,51(1):45-54

Strobilurin-resistant isolates of Blumeria ( Erysiphe ) graminis f.sp. tritici , the cause of wheat powdery mildew, were more than 10-fold less sensitive to azoxystrobin than sensitive isolates. In all resistant isolates, a mutation resulting in the replacement of a glycine by an alanine residue at codon 143 (G143A) in the mitochondrial cytochrome b gene was found. Allele-specific primers were designed to detect this point mutation in infected wheat leaves. Using quantitative fluorescent allele-specific real-time polymerase chain reaction (PCR) measurements, strobilurin-resistant A143 alleles could be detected amongst strobilurin-sensitive G143 alleles at a frequency of at least 1 in 10 000, depending on the amount of target and nontarget DNA. Most isolates tested were dominant homoplasmic for either the A143 or G143 allele, although mixed populations of alleles could be detected in some isolates. In some of these isolates, strobilurin resistance was not always stable when they were maintained for many generations in the absence of selection. The allele-specific real-time PCR assay was also used to follow the dynamics of A143 alleles in field populations of B . graminis f.sp. tritici before and after application of fungicides. As expected, the A143 allele frequency only increased under selection pressure from a strobilurin fungicide. After three sprays of azoxystrobin, a pronounced selection for the strobilurin-resistant allele, with an increase in average frequency from 2·2 to 58%, was measured. The use of quantitative real-time PCR diagnostics for early detection of fungicide resistance genes at low frequency, coupled with risk evaluation, will be invaluable for further resistance risk assessment and validation of antiresistance strategies.  相似文献   

Transfer of Bovine Blastocysts Derived from Short-term In Vitro Culture of Low Quality Morulae Produced In Vivo     

RH Alvarez  M Meneghel  AC Martinez  RML Pires  EA Schammass 《Reproduction in domestic animals》2008,43(3):257-260

The aim of this study was to evaluate if blastocysts arising from in vitro culture of Grade 3 bovine morulae produced in vivo can promote acceptable pregnancy rates when transferred into recipients. Embryos of different stages and qualities were recovered from superovulated Bos taurus and B. indicus donors. Grade 3 morulae were cultured in either Holding Plus® or TCM‐199 (supplemented with 10% bovine fetal serum) media for 24 h at 38.5°C. After this culture period, the resulting blastocysts were morphologically classified (Grades 1, 2 and 3) and transferred into recipients previously synchronized with the donors. Non‐cultured Grades 1 and 3 morulae were used as control. Pregnancy diagnosis was carried out 60 days after embryo transfer and the data were analysed by logistic regression, considering variables, such as embryo quality (Grade), donor breed, culture medium, donor‐recipient synchrony and seasonality. Embryo quality was the only variable, showing significant effect on the pregnancy rate. Pregnancy rates for non‐cultured Grade 1 and 3 morulae, and blastocysts arising from cultured Grade 3 morulae were 58.1% (n = 31), 17.1% (n = 35) and 51.1% (n = 47), respectively (p < 0.05). There were no statistical differences between non‐cultured Grade 1 morulae and cultured blastocysts. Pregnancy rates for Grades 1 (65.0%) and 2 (60.0%) were higher than Grade 3 (29.4%) cultured blastocysts (p < 0.05). It was concluded that short‐term in vitro culture is a very convenient method of identifying morphologically low quality morulae with higher chances of continuing development after the transfer into recipients.  相似文献   

Assessment of primers designed for the subspecies-specific discrimination among Babesia canis canis, Babesia canis vogeli and Babesia canis rossi by PCR assay     

Duarte SC  Linhares GF  Romanowsky TN  da Silveira Neto OJ  Borges LM 《Veterinary parasitology》2008,152(1-2):16-20

Canine babesiosis is an infectious disease caused by either Babesia gibsoni or Babesia canis protozoans. The latter is also classified under three different phylogenetic groups, referred to as subspecies B. canis canis, B. canis vogeli and B. canis rossi. The objective of the present study was to validate and standardize a PCR assay to discriminate the organisms at the subspecies level. First, the reference sequences of the 18S rRNA, 5.8S rRNA and 28S rRNA genes, including the internal transcribed spacer 1 (ITS1) and 2 (ITS2) of the most common species and subspecies of the genus Babesia were retrieved from the GenBank database. Subspecies-specific primers (BAB3, BAB4 and BAB5) and one genus-specific primer were designed from the alignment of the sequences. The PCR assays were evaluated in three different combinations of primer pairs in order to assure complete specificity for each reaction. The results of the tests had demonstrated effectiveness of the novel primer pairs BAB1/BAB3, BAB1/BAB4 and BAB1/BAB5 for the amplification of the subspecies-specific target fragments of 746 bp (B. c. canis), 546 bp (B. c. vogeli) and 342 bp (B. c. rossi) by PCR. The original enzymatic amplification assays with novel primers reported in this paper were confirmed to be a reliable tool for the specific discrimination among B. canis subspecies by single-step PCR assays.  相似文献   

Differentiation between canine cutaneous and visceral leishmaniasis by the detection of immunoglobulin G specific for Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi antigens using flow cytometry     

Santiago Mde A  Ribeiro FC  Mouta-Confort E  Nascimento LD  Schubach Ade O  Madeira Mde F  Bertho AL 《Veterinary parasitology》2008,154(3-4):341-346

Flow cytometry employing Leishmania (L.) chagasi (Lc) and L. (Viannia) braziliensis (Lb) antigen was used to establish the differential diagnosis between visceral (VL) and cutaneous leishmaniasis (CL) in dogs. Flow cytometry permitted the detection of Leishmania-specific immunoglobulin G in sera from 19 dogs: nine with CL and 10 with VL. A significant difference in the percentage of positive staining was observed in sera from dogs with CL between the homologous antigen (69% for Lb) and the heterologous antigen (42% for Lc). However, this difference was not significant in sera from dogs with VL (61% for Lb and 73% for Lc). No significant staining was observed in control sera (0.6% for Lb and 0.4% for Lc) consisting of samples from healthy dogs, or in the group with sporotrichosis (1.8% for Lb and 1.5% for Lc), a differential diagnosis of CL. The results suggest that flow cytometry might be useful for the differentiation between CL and VL in dogs, with practical applications in areas where the two infections overlap.  相似文献