Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine     

Marcos Mancilla  María-Jesús Grilló  María-Jesús de Miguel  Ignacio López-Go?i  Beatriz San-Román  Ana Zabalza-Baranguá  Ignacio Moriyón 《Veterinary research》2013,44(1):105

Brucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production.  相似文献   

Glacially abraded rock flour from Greenland: Potential for macronutrient supply to plants     

Klara Cecilia Gunnarsen  Lars Stoumann Jensen  Beatriz Gmez-Muoz  Minik Thorleif Rosing  Andreas de Neergaard 《植物养料与土壤学杂志》2019,182(5):846-856

Rock flour (RF) is a fine‐grained material produced naturally by glacial movement and resulting bedrock abrasion. In Greenland fluvial transported RF from the inland ice sheet sediments in riverbeds and marine outflows. This fine‐sized RF (50% < 9.8 µm) has a high reactivity and may therefore potentially be used to rejuvenate nutrient poor soils and provide nutrients to plants. The aim of this study was to evaluate the ability of a RF from Greenland to supply P, K, Mg, and S to plants. A double‐pot system was used, in which ryegrass (Lolium perenne L.) could take up nutrients from both a hydroponic solution and a soil‐compartment with or without RF amendment; a soil mixture or pure sand was used in the soil‐compartment to estimate RF‐soil interaction effects. Omission of single nutrients from the hydroponic solution allowed assessment of which nutrients the RF in the soil‐compartment was able to supply. Ryegrass biomass was harvested four times during 62 days. We found that RF could supply K continuously to plants grown in soil or sand, but insufficient to fully circumvent K deficiency. During 62 days 5.8% and 4.3% of the applied K from RF was accumulated in the aboveground plant tissue in soil and sand, respectively. Mg was supplied from RF to plants in sand, but no significant effects were observed in soil, possibly due to background soil Mg availability. The amounts of P and S supplied to plants were insignificant. These results indicate the potential of Greenland RF to act as a slow release K and Mg fertilizer.  相似文献   

Identification of fructooligosaccharides in different banana cultivars     

Der Agopian RG  Soares CA  Purgatto E  Cordenunsi BR  Lajolo FM 《Journal of agricultural and food chemistry》2008,56(9):3305-3310

Banana has been currently indicated as a good source of fructooligosaccharides (FOS), which are considered to be functional components of foods. However, significant differences in their amounts in bananas have been observed in the literature. This work aims to identify and quantify FOS during ripening in different banana cultivars belonging to the most common genomic groups cultivated in Brazil. Considering that these differences can be due to cultivar, stage of ripening, and the methodologies used for FOS analyses, sugar contents were analyzed by high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) and gas chromatography-mass spectrometry (GC-MS). An initial screening of eight cultivars (Ouro, Nanic?o, Prata, Ma??, Mysore, Pacovan, Terra, and Figo) in a full-ripe stage showed that 1-kestose, the first member of the FOS series (amounts between 297 and 1600 microg/g of DM), was accumulated in all of them. Nystose, the second member, was detected only in Prata cultivar. Five of the cultivars were analyzed during ripening, and a strong correlation could be established with a specific sucrose level ( approximately 200 mg/g of DM), which seems to trigger the synthesis of 1-kestose (the low amounts of FOS, below the functional recommended dose, indicates that banana cannot be considered a good source of FOS).  相似文献   

Organic acid contents in onion cultivars (Allium cepa L.)     

Rodríguez Galdón B  Tascón Rodríguez C  Rodríguez Rodríguez E  Díaz Romero C 《Journal of agricultural and food chemistry》2008,56(15):6512-6519

The following organic acids (glutamic, oxalic, pyruvic, malic, tartaric, citric, and fumaric), pungency, Brix degree, acidity, and pH were determined in onion cultivars (Texas, Guayonje, San Juan de la Rambla, Carrizal Alto, Carrizal Bajo, and Masca) harvested in the same agroclimatic conditions. Glutamic acid was the most abundant organic acid (325 +/- 133 mg/100 g) followed by citric acid (48.5 +/- 24.1 mg/100 g) and malic acid (43.6 +/- 10.4 mg/100 g). There were significant differences between the onion cultivars in the mean concentrations of all of the analyzed parameters. The San Juan de la Rambla and Masca cultivars presented, in general, higher concentrations of the organic acids than the other cultivars. Significant differences in most of the analyzed parameters were observed between the two seed origins for the Masca and San Juan de la Rambla cultivars. The onion samples tended to be classified according to the cultivar and, in the case of San Juan de la Rambla cultivar, according to the precedence of the seeds after applying discriminant analysis.  相似文献   

Spectrophotometric determination of urea in sugar cane distilled spirits     

Labanca RA  Glória MB 《Journal of agricultural and food chemistry》2008,56(13):5211-5215

Urea is an important precursor in the formation of ethyl carbamate, a known carcinogen in alcoholic beverages. Ethyl carbamate has recently been detected at high concentrations in sugar cane distilled spirits, but little is known about the concentration of urea in these beverages. The objectives of this study were to validate methodology for the determination of urea in sugar cane distilled spirits, to determine the levels in 68 samples from different regions within the state of Minas Gerais, Brazil, and to examine the relationship between the concentrations of urea and ethyl carbamate. The method, based on the reaction of urea with 1-phenyl-1,2-propanodione-2-oxime and spectrophotometric quantification at 540 nm, provided linear response from 0.5 to 15.0 mg/L. No purification of the sample was required. The limits of detection and quantification were 0.1 and 0.5 mg/L, respectively. Urea was detected in 69% of the samples at levels varying from 0.50 to 5.10 mg/L. There was no significant difference on the levels of urea in samples from different regions of the state. No significant correlation between the levels of urea and ethyl carbamate was observed for the samples analyzed.  相似文献   

Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples   总被引：1，自引：0，他引：1  

Kim CM  Blanco LB  Alhassan A  Iseki H  Yokoyama N  Xuan X  Igarashi I 《Veterinary parasitology》2008,151(2-4):158-163

We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples were T. equi-positive in the nested PCR assay, while the real-time PCR assay also detected the parasite in all 46 of the nested PCR-positive samples but did not detect T. equi in the remaining 19 negative blood samples. This quantitative real-time PCR assay provides a valuable tool for fast laboratory diagnostic assessment of T. equi infection in horses.  相似文献   

Persistence and molecular evolution of Mycobacterium bovis population from cattle and wildlife in Doñana National Park revealed by genotype variation     

Romero B  Aranaz A  Sandoval A  Alvarez J  de Juan L  Bezos J  Sánchez C  Galka M  Fernández P  Mateos A  Domínguez L 《Veterinary microbiology》2008,132(1-2):87-95

The role of wildlife in tuberculosis epidemiology is being widely studied since it can affect the effectiveness of eradication campaigns in cattle. The health problem is enhanced when it concerns also wildlife welfare and biodiversity conservation. This study was performed to understand the epidemiology of Mycobacterium bovis population affecting livestock and wild animals in the Do?ana National Park using bacteriology and molecular characterisation techniques. Tuberculosis research was performed on 1209 cattle and wild animals (artiodactyla and carnivore) collected over 6 years in the Park. One hundred and sixty-three animals were found to be infected with M. bovis, comprising 7.96% of the cattle and 20.53% of the wild animals tested. Spoligotyping revealed nine patterns, being SB1232 and SB1230 the most prevalent (77.30% and 15.34% of infected animals, respectively). MIRU-VNTR analysis of a selected panel of 92 isolates showed eight different profiles, including several spoligotypes within the same MIRU-VNTR profile. The discriminatory capacity of both techniques in this panel was similar. The results obtained by combination of both techniques corroborate that wildlife species are infected with the M. bovis strains which are more prevalent in cattle and reveal their persistence. Genotype variation between isolates strongly suggests micro-evolutionary events in the M. bovis population in the same area. This study in the Do?ana National Park exposes the risk of introduction of domestic animals into wildlife areas when there is not a warranty of disease freedom, appropriate diagnostic techniques and control measures.  相似文献   

Histopathological findings in the early diagnosis of granulosa cell tumour in bitches     

Ana Carina Heil da Silva Matos  Juliana da Silva Leite  Anglica Consalter  Marcela Freire Vallim de Mello  Maria de Lourdes Gonalves Ferreira  Ana Beatriz Monteiro Fosenca  Ana Maria Reis Ferreira 《Reproduction in domestic animals》2019,54(6):828-834

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call‐Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call‐Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call‐Exner body.  相似文献   

Complications associated with closed castration using the Henderson equine castration instrument in 300 standing equids     

Julien Racine  Beatriz Vidondo  Alessandra Ramseyer  Christoph Koch 《Veterinary surgery : VS》2019,48(1):21-28

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Genetic variability of rhizobacteria from wild populations of four Lupinus species based on PCR-RAPDs§     

José Antonio Lucas García  Augustín Probanza  Beatriz Ramos  Francisco J. Gutiérrez Mañero 《植物养料与土壤学杂志》2001,164(1):1-7

The metabolic capacities of rhizosphere bacteria can depend on intraspecific genetic variability at strain level. We sampled bacteria from the rhizosphere of three populations of four different Lupinus species at two growth stages (flowering: GS1 and fruiting: GS2). Isolates were identified to the genus level by classical biochemical tests. The most abundant genera found were Bacillus, Aureobacterium, Cellulomonas, Pseudomonas, and Arthrobacter. Genetic divergence of rhizobacteria was tested by PCR-RAPDs. The genetic distances were low, with mean values of 37 % for Bacillus, 25 % for Aureobacterium, 46 % for Cellulomonas, 16 % for Pseudomonas, and 23 % for Arthrobacter. Aureobacterium, the most abundant genus, predominated in the rhizosphere of all populations and at both growth stages (GS1 and GS2) of L. angustifolius. The Aureobacterium strains consisted of 11 groups with 90 % similarity indexes. The cluster analysis of these groups shows that strains isolated from different lupin species and sampling times have extraordinary low diversity indexes, or are even identical. This fact, together with the low genetic distance detected in the rhizosphere, reveals a clear specificity in the plant-bacteria interaction. This specificity could be related with several aspects of plant physiology.  相似文献