Species-specific DNA markers for identification of two root-knot nematodes of coffee: Meloidogyne arabicida and M. izalcoensis     

Valdir Ribeiro Correa  Marcilene Fernandes Almeida dos Santos  Maria Ritta Alves Almeida  José Ricardo Peixoto  Philippe Castagnone-Sereno  Regina Maria Dechechi Gomes Carneiro 《European journal of plant pathology / European Foundation for Plant Pathology》2013,137(2):305-313

Seven root-knot nematodes (RKN), including Meloidogyne exigua, M. incognita, M. paranaensis, M. enterolobii, M. arabicida, M. izalcoensis and M. arenaria are major pathogens of coffee crop in the Americas. Species-specific primers for their identification have been developed for five of them and constitute a fast and reliable method of identification. Here we report a PCR-based assay for specific detection of M. arabicida and M. izalcoensis. Random Amplified Polymorphic DNA fragments specific for these two species were converted into sequence characterized amplified region (SCAR) markers. PCR amplification using the SCAR primers produced a specific fragment of 300 bp and 670 bp for M. arabicida and M. izalcoensis, respectively, which were absent in other coffee-associated Meloidogyne spp. tested. SCAR primers also allowed successful amplification of DNA from single second-stage juveniles (J2), males and females. In addition, these primers were able to unambiguously detect the target species in nematode suspensions extracted from soil and roots samples, in different isolates of the same species or when used in multiplex PCR reactions containing mixtures of species. These results demonstrated the effectiveness of these SCAR markers and their multiplex use with those previously developed for M. exigua, M. incognita, M. paranaensis, M. enterolobii and M. arenaria constitute an essential detection tool. This diagnostic kit will contribute for specific J2 identification of the major RKN infecting coffee from field samples in the Americas.  相似文献   

Guava decline: effect of root exudates from Meloidogyne enterolobii-parasitized plants on Fusarium solani in vitro and on growth and development of guava seedlings under controlled conditions     

Vicente Martins Gomes  Ricardo Moreira Souza  Silvaldo Felipe da Silveira  Alexandre Macedo Almeida 《European journal of plant pathology / European Foundation for Plant Pathology》2013,137(2):393-401

In guava decline, Fusarium solani-immune guava trees become susceptible to extensive root rot caused by this fungus after parasitism by Meloidogyne enterolobii. To understand the mechanisms involved in this disease, root exudates were collected from nematode-inoculated (NI) or uninoculated (UN) guava plants cultivated in sand. After filtration through a Millipore® membrane, NI and UN exudates were used: i) to prepare media to assess their effect on mycelial growth and production of propagules of F. solani isolate UENF/CF 163, and ii) to incubate macro- and microconidia to assess their effect on germination. NI exudates promoted (P?<?0.05) more mycelial growth and production of propagules than UN exudates or water. NI and UN exudates were used to water guava seedlings laid over seed germination paper inside plastic boxes. Half of the seedlings had an agar plug colonized by the fungus positioned in the collar region. Upon watering with NI exudates the fungus caused (P?<?0.05) extensive rotting of the seedlings’ rootlets. NI and UN exudates, either unlyophilized or lyophilized and re-suspended to the original concentration, were used to water guava seedlings grown in sterile sand before being inoculated (or left uninoculated) as described before. Solely upon watering with NI exudates, in its unlyophilized form or after lyophilization, the fungus caused a reduction (P?<?0.05) of shoot and root biomass associated with rotting of roots. These results suggest that M. enterolobii induces chemical changes in the root exudates of guava trees, which are necessary for root invasion causing root rotting by F. solani.  相似文献   

Decomposition of Medicago sativa Debris Incubated at Different Depths under Mediterranean Climate     

Pere Rovira  V. Ramon Vallejo 《Arid Land Research and Management》2013,27(3):265-280

Reports on the dynamics of carbon and nitrogen at different depths in soil profiles usually describe a higher mineralization of both at upper horizons. Nevertheless, under Mediterranean climate, upper soil horizons are strongly affected by drought, especially in summer, and may offer pedoclimatic conditions less favorable to microbial activity than deep soil layers. Therefore, decomposition could be slower in the upper soil layers. To test this hypothesis, mixtures of Medicago sativa ground plants and soil material were incubated in the field, at 5, 20, and 40 cm depth, in nylon mesh bags. Mineralization of carbon and nitrogen was studied for two years. At 5 cm, mineralization of both elements was lower, and no differences were found between 20 and 40 cm. After two years of field incubation, the remaining carbon (as a percentage of the initial content) was 27.95 % +/- 0.88 at 5 cm depth, 19.87% +/- 0.77 at 20 cm, and 18.78 % +/- 1.19 at 40 cm. Mineralization of nitrogen exceeded that of carbon. After two years of field incubation, the remaining nitrogen (as a percentage of the initial content) was 17.62% +/- 3.06 at 5 cm depth, 12.17% +/- 0.94 at 20 cm, and 11.26% +/- 0.99 at 40 cm. The biodegradation rate in upper layers was lower for all biochemical fractions (water-soluble compounds, lipids, polysaccharides, lignin). This contrasts with the usual findings on this topic, but is consistent with our previous results with forest litter samples. Mineralization clearly followed double-exponential kinetics, with a labile and a recalcitrant pool of both carbon and nitrogen. The labile pool of carbon accounted for about 50% of the total initial carbon, whereas that of nitrogen accounted for about 60%. No clear effect of depth on the proportion between the labile and the recalcitrant pool was observed, neither for carbon nor for nitrogen. It was not possible to identify both pools with the biochemical fractions, suggesting that these pools should be interpreted in physical terms (unprotected vs. protected ). No direct effect of depth on the global retention of N was detected.  相似文献   

The pyrethroid resistance status and mechanisms in Aedes aegypti from the Guerrero state,Mexico     

H. Angélica Aponte  R. Patricia Penilla  Felipe Dzul-Manzanilla  Azael Che-Mendoza  Alma D. López  Francisco Solis  Pablo Manrique-Saide  Hilary Ranson  Audrey Lenhart  Philip J. McCall  Américo D. Rodríguez 《Pesticide biochemistry and physiology》2013

Dengue is one of the most important vector-borne diseases worldwide and is a public health problem in Mexico. Most programs in dengue endemic countries rely on insecticides for Aedes control. In Mexico, pyrethroid insecticides (mainly permethrin and deltamethrin) have been extensively used over a decade as adulticides and represented a strong selection for insecticide resistance for dengue vectors in several parts of the country. We studied the type, frequency and distribution of insecticide resistance mechanisms in Aedes aegypti from six municipalities in the state of Guerrero selected on the basis of historically intense chemical control and a high risk for dengue transmission. Ae. aegypti eggs were collected from October 2009 to January 2010 using ovitraps. F₁ adults, emerged from these collections, were exposed to permethrin, deltamethrin and DDT in WHO diagnostic tests and showed high resistance levels to both pyrethroids and DDT. This was consistent with the presence of increased metabolic enzyme activities and target site insensitivity due to kdr mutations. Biochemical assays showed elevated esterase and glutathione S-transferase activities in the six municipalities. The V1016I kdr mutation on the IIS6 domain of the sodium channel gene was present in an overall frequency of 0.80. A second mutation, F1534C on the IIIS6 domain of the same gene was also detected, being the first report of this mutation in Guerrero. The multiple resistance mechanisms present in Ae. aegypti from Guerrero state represent a warning for the efficacy of the pyrethroid usage and consequently for the success of the dengue control program.  相似文献   

Characterization of sulfated quercetin and epicatechin metabolites     

Dueñas M  González-Manzano S  Surco-Laos F  González-Paramas A  Santos-Buelga C 《Journal of agricultural and food chemistry》2012,60(14):3592-3598

Different monosulfates of quercetin and epicatechin with metabolic interest were obtained by hemisynthesis and characterized regarding their chromatographic behavior and absorption and mass spectra. Three of these compounds were further isolated, and their structures were elucidated by mass spectrometry and (1)H and (13)C nuclear magnetic resonance using one- and two-dimensional techniques (heteronuclear single-quantum coherence and heteronuclear multiple-bond correlation). The calculation of the proton and carbon shifts caused by sulfation allowed for the assignment of the position of the sulfate group in the flavonoids, so that the compounds were identified as quercetin-3'-O-sulfate, quercetin 4'-O-sulfate, and epicatechin 4'-O-sulfate. It was found that sulfation at position 3' induced a large upfield shift in the carbon bearing the sulfate group and downfield displacements of the adjacent carbons, whereas no significant upfield or downfield shifts were observed with respect to the parent flavonoid when sulfation was produced at position 4'.  相似文献   

Control of Paratuberculosis in Sheep and Goats     

Ramon A. Juste  Valentin Perez 《Veterinary Clinics of North America: Food Animal Practice》2011,27(1):127-138

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Quality and aromatic sensory descriptors (mainly fresh and dry fruit character) of Spanish red wines can be predicted from their aroma-active chemical composition     

San-Juan F  Ferreira V  Cacho J  Escudero A 《Journal of agricultural and food chemistry》2011,59(14):7916-7924

A satisfactory model explaining quality could be built in a set of 25 high quality Spanish red wines, by aroma-active chemical composition. The quality of the wines was positively correlated with the wine content in fruity esters, acids, enolones, and wood derived compounds, and negatively with phenylacetaldehyde, acetic acid, methional, and 4-ethylphenol. Wine fruitiness was demonstrated to be positively related not only to the wine content on fruity esters and enolones, but to wine volatile fatty acids. Fruitiness is strongly suppressed by 4-ethylphenol, acetic acid, phenylacetaldehyde, and methional, this involved in the perception of dry-fruit notes. Sensory effects were more intense in the presence of β-damascenone and β-ionone. A satisfactory model explaining animal notes could be built. Finally, the vegetal character of this set of wines could be related to the combined effect of dimethylsulfide (DMS), 1-hexanol, and methanethiol.  相似文献   

Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil     

Ribeiro MG  Takai S  Guazzelli A  Lara GH  da Silva AV  Fernandes MC  Condas LA  Siqueira AK  Salerno T 《Research in veterinary science》2011,91(3):478-481

The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15–17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.  相似文献   

Prevalence of antibodies against Toxoplasma gondii among Brazilian white-eared opossums (Didelphis albiventris)     

Fornazari F  Teixeira CR  da Silva RC  Leiva M  de Almeida SC  Langoni H 《Veterinary parasitology》2011,179(1-3):238-241

Considering that Toxoplasma gondii is a parasite of global importance which affects several animal species including humans, the current study aimed to investigate the prevalence of antibodies against T. gondii among 72 white-eared opossums (Didelphis albiventris) from Botucatu Municipality (22°53'S 48°26'W), S?o Paulo State, Brazil. The investigation was carried out from January 2008 to December 2009, when the animals had their blood samples collected and subjected to the modified agglutination test (MAT); 12 specimens had brain samples bioassayed in mice. Seroprevalence was 5.5% (n=4) and bioassays were negative. Older animals had higher prevalence of antibodies against T. gondii. Opossums in closer contact with the urban environment are likely more exposed to T. gondii than animals from the sylvatic environment.  相似文献   

Phylogenetic identification of Cystoisospora spp. from dogs, cats, and raccoon dogs in Japan     

Matsubayashi M  Carreno RA  Tani H  Yoshiuchi R  Kanai T  Kimata I  Uni S  Furuya M  Sasai K 《Veterinary parasitology》2011,176(2-3):270-274

Cystoisospora spp. from feces in dogs, cats, and raccoon dogs were isolated, sequenced at the small subunit ribosomal RNA gene locus and compared to other Cystoisospora spp. Cystoisospora oocysts from dogs and raccoon dogs were morphologically similar with those of C. ohioensis, and cat isolates were similar with those of C. felis. The sequences from dogs and raccoon dogs, and cats have a homology with C. ohioensis and C. felis, respectively. Phylogenetic analysis of the DNA sequences showed that the dog and raccoon dog isolates were nested in a clade with other Cystoisospora spp. including C. ohioensis, C. belli, and C. orlovi. The cat isolate formed a sister group with C. felis that was a separate clade from the dog and raccoon dog group. We report sequence variation in these Cystoisospora sequences and have identified raccoon dogs as another carnivore host for Cystoisospora spp. infecting dogs.  相似文献