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

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.     

Use of culture-derived Fusarium oxysporum f. sp. cubense, race 1 filtrates for rapid and non-destructive in vitro differentiation between resistant and susceptible clones of field-grown banana

Banana and plantain are among the most important food crops in developing countries but production is threatened by increasing virulent forms of Fusarium oxysporum f. sp. cubense. Chemical control is not economically effective and,therefore, breeding programs are necessary. Traditional field studies of new genotype resistance to this disease are time-consuming and destructive. Therefore,we developed a rapid and non-destructive procedure to differentiate field-grown banana resistant from susceptible clones. This procedure implicates application of culture filtrates of Fusarium oxysporum f. sp. cubense race 1 onto banana leaves. The relationship between duration of the fungal in vitro incubation, and the fungal culture fresh mass, the culture filtrate absorbency, and the Gross Michel (susceptible cultivar)leaf lesion area (after application of the culture filtrate) were similar and at 24day-incubation the highest values of the recorded indicators were observed. A comparison between Gross Michel and FHIA-01(resistant) was also performed. The most relevant differences between cultivars were observed at 48 hours after application of the culture filtrate, and in the middle-aged leaves. The position of the culture filtrate application in the leaf limb (distal, middle, proximal) was not determinant. A wider comparison among banana cultivars confirmed previous results informed by other researchers using different systems to study this plant-fungus interaction. Such a confirmation validates the effectiveness of the procedure described here to select rapid and non-destructively banana resistance to this disease at field level. This revised version was published online in August 2006 with corrections to the Cover Date.     

Detection of Flexibacter maritimus in fish tissue using nested PCR amplification

A nested polymerase chain reaction (PCR) system was developed for the detection of Flexibacter maritimus from fish tissue. The total procedure for the diagnosis of marine flexibacteriosis, from the point of DNA extraction to the electrophoretic analysis, can be performed in < 4 h. This was achieved by the combination of a short thermal cycling programme with a rapid DNA extraction procedure. The assay was extremely sensitive, capable of detecting as few as 75 cfu mg(-1) fish tissue. The accuracy of the nested PCR was confirmed under field conditions using tissue samples recovered during 1993-2002 from fish suffering marine flexibacteriosis. The nested PCR method proved to be efficient for the rapid and sensitive detection of F. maritimus from fish tissues and can be used for routine diagnosis of the disease caused by this pathogen.     

Shedding light on the migratory patterns of the Amazonian goliath catfish,Brachyplatystoma platynemum,using otolith 87Sr/86Sr analyses

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Oral cavity evaluation and dental chart registration of coati (Nasua nasua) in captivity

The aims of this study were to develop a dental evaluate any oral cavity disease, develop gypsum models of the dental arches, and to register the occlusions found in coatis (Nasua nasua) in captivity Formulation of the dental chart was assisted by intraoral radiographs from the head of an adult coati cadaver of the same species with the following dental formula.: I 3/3, C 1/1, P 4/3, M 2/2. Seven live coatis of the Nasua nasua species were evaluated. Five of the seven coatis presented with various dental abnormalities as follows: dental plaque (71.4 0%), gingivitis (71.4 %), periodontitis (57.1 %), dental stain (42.9 %), dental abrasion (57.1 %), dental fracture (57.1 %), pulp exposure (42.9 %), malocclusion (57.1 %) and supernumerary teeth (14.2 %).     

Resistance to common bacterial blight in Phaseolus vulgaris L. recombinant inbred lines under natural infection of Xanthomonas axonopodis pv. phaseoli

Among the main causes of poor yield in common beans are fungal, viral and bacterial diseases. Common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli (Xap), is one of the major bacterial diseases leading to significant losses in Brazil. Chemical control is ineffective, therefore, the use of resistant varieties becomes an interesting alternative. The objective of the present work was to evaluate disease resistance under natural infection of the pathogen in 109 recombinant inbred lines (F₇) of P. vulgaris originated from the cross HAB-52 (susceptible — snapbean) × BAC-6 (resistant — common bean) in two different environments, as well as to calculate genetic parameters to assist in the selection of promising materials to be used in the CBB resistance breeding program. The data of the genetic parameters were compared to those calculated for the F₃ generation originated from the same cross. The heritability results for DI (disease index) and VI (variation index) in F₃ were 26.85% and 0.26, respectively, whereas in F₇ they were 91.77% and 1.36, respectively. These results demonstrate a potential to be explored for this advanced population, that in the future, along with other pathogen variability studies and tests in other environments, may provide more information regarding a more precise evaluation of promising genotypes to be used in common bean breeding programs aiming to obtain CBB resistant varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Irrigation of paddy soil with industrial landfill leachate: impacts in rice productivity,plant nutrition,and chemical characteristics of soil

Treated industrial effluents have high levels of nutrients and dissolved organic matter. The irrigation of rice by flooding can increase nutrient uptake and grain yield. Therefore, this study evaluated the nutrient contents in the shoots and grain of the rice crop and also the chemical of the soil after irrigation of the crop with leachate of the treated industrial effluent. A greenhouse experiment was conducted using pots filled with 20 kg of soil in a randomized block design with three replications. The treatments consisted of control (irrigation with distilled water) and four concentrations of the leachate (25, 50, 75, and 100 %) for irrigation. At the end of the experiment, the nutrient contents in tissues of rice plants, sterility of spikelets, and grain mass were evaluated. Results showed that irrigation with the leachate at 25 % content increased the macro- and micronutrients’ concentrations in the shoot biomass and grain, except for potassium and iron. Irrigation with the industrial leachate decreased tillering and grain yield; however, it increased chlorophyll content, sterility of spikelets, and sodium intake at this leachate concentration. The potassium and sodium levels and the electrical conductivity values of soils irrigated with treated industrial leachate were increased. The use of the treated leachate from industrial effluents is an alternative that reuses the nutritional load, but the volume of leachate should be limited and monitored to prevent the sodicity in the soil and problemsdue to eutrophication.     

Manipulation of Ovarian Function for the Reproductive Management of Dairy Cows

Veterinary Research Communications -     

Evaluation of Magnetic Coagulant (α-Fe<Subscript>2</Subscript>O<Subscript>3</Subscript>-MO) and its Reuse in Textile Wastewater Treatment

The textile industries are characterized as one of the biggest consumers of potable water and chemical products throughout its process, being responsible for the elevated wastewater generation with intense coloration and wide polluting potential. In this context, the present study proposes the development and application of a new coagulant material for textile wastewater treatment. The proposed coagulant (α-Fe₂O₃-MO) was composed by hematite nanoparticles (α-Fe₂O₃) obtained by a simple non-pollutant methodology, associated with Moringa oleifera (MO) seeds saline extract compounds. Coagulation/flocculation (CF) efficiency was evaluated by removal of physicochemical parameters such as apparent color, turbidity, and compounds with absorption at UV_254nm (UV_254nm) through CF tests carried out on Jar test equipment and sedimentation carried out in the presence and absence of external magnetic field (600 k Am^?1). Kinetics sedimentation was from 0 to 90 min. The use of this new coagulant allowed the removal of 92.37% for apparent color, 91.43% for turbidity, and 46.09% for UV_254nm, indicating that the proposed coagulant association was efficient in the treatment of this type of wastewater under external magnetic field with only 10 min of sedimentation. In addition, the resulting sludge from CF process was tested as base material for a new coagulant synthesis, demonstrating great reuse potential. Therefore, the new proposed coagulant, composed of α-Fe₂O₃ and the compounds present in the seed extract of MO, has applicability for textile wastewater treatment demonstrating high removal rate for all evaluated parameters with cost reduction in the proposed treatment for this wastewater.