Immunological characteristics of experimental murine infection with Leishmania (Leishmania) amazonensis

The murine models of Leishmania infection are well-studied and suitable models for studying this disease, which, despite its incidence of nearly 2 million new cases worldwide per year and its prevalence of 12 million cases, has been a somewhat neglected disease. Data obtained using such models are important for a better understanding of the disease in humans due to similarities in physiology and the advantage provided by the uniform infection profile within each mouse strain. In this review, we focus on studies of experimental murine infection with Leishmania (Leishmania) amazonensis, a species that has been associated with infections exhibiting various clinical features in humans. Mainly, we point out and discuss reports on: the effects of variations of the inoculum (such as strain, site, and size) in the establishment and development of the infection; characteristics of the infection in distinct mouse strains; and, the effects and subversions of the infection on components of the host innate and adaptive immune responses. The results obtained in these studies show that L. (L.) amazonensis infection in mice presents some unique features and immunoregulatory mechanisms, making it an interesting model for obtaining further knowledge of potential drugs targets and immunotherapy in Leishmania infection.     

Lack of postexposure analgesic efficacy of low concentrations of eugenol in zebrafish

Objective

To test the postexposure analgesic efficacy of low doses of eugenol in zebrafish.

Comparison of nutrient uptake and antioxidative response among four Labiatae herb species under salt stress condition

Herbs of the Labiatae have relatively low salt tolerance. They are widely grown in drylands, but salt stress there is a typical problem and may reduce yields. To examine their salt tolerance mechanisms, we grew basil, sage, thyme, and oregano in nutrient solution containing 50 mM NaCl and determined the biomass; contents of Na, K, and Mg in leaf blades, stems, and roots; contents of total chlorophyll, malondialdehyde (MDA), hydrogen peroxide in leaf blades; and activities of antioxidative enzymes in leaf blades. The salt tolerance decreased in the order of basil ≈ sage > thyme > oregano. The good salt tolerance of basil was explained by a significant increase in the activity of catalase, in addition to the low Na/K ratio of leaf blades due to the retention of Na in stems and roots and of K in leaf blades. The good salt tolerance of sage was explained by the low Na/K ratio in leaf blades and the prevention of lipid peroxidation by high antioxidative enzyme activities, despite its poorer management of nutrient uptake. In thyme, although catalase activity increased significantly to alleviate salt-induced oxidative stress caused by Na influx into all parts, low K and Mg allowed shoot weight in particular to decrease. In oregano, antioxidative responses appeared as significant increases in ascorbate peroxidase and glutathione reductase activity, and K was accumulated in leaf blades, but serious salt-induced oxidative stress caused by high Na influx into all parts reduced the growth of all parts. These results show that despite similar responses among species, salt tolerance is not necessarily the same. In this experiment, we revealed the salt tolerance mechanism of each of four Labiatae herbs by revealing their strengths and weaknesses in nutrient uptake and antioxidative responses.     

Damping-off of soybean in southern Brazil can be associated with different species of Globisporangium spp. and Pythium spp.

Damping-off of soybean is a common problem in some fields in southern Brazil. Frequently, Phytophthora sojae has been pointed out as the main causal agent. However, questions were raised as to whether other oomycetes could also be present. Hence, the objectives of this study were to isolate, identify, and determine the pathogenicity and aggressiveness of oomycetes isolated from soybean seedlings. Thirty-three isolates were obtained from 13 fields. Isolates were identified based on morphology and molecular methods. Nine species were recovered: Pythium conidiophorum (n = 14, 42.4%), Globisporangium rostratifingens (n = 7, 21.2%), G. irregulare (n = 2, 6.1%), G. spinosum (n = 1, 3.0%), G. ultimum var. sporangiiferum (n = 2, 6.1%), Pythium acanthicum (n = 2, 6.1%), P. deliense (n = 1, 3.0%), P. inflatum (n = 1, 3.0%), and P. torulosum (n = 3, 9.1%). Pathogenicity and aggressiveness were determined with two soybean cultivars (DM57i52 and NEO530) in a seed rot assay and a root rot assay. All isolates were pathogenic to soybean with variable levels of aggressiveness in both assays. The range of seed disease severity index varied from 7.5 to 100 for DM57i52 and from 23.1 to 100 for NEO530. The mean root disease severity index of the soybean cultivars ranged from 29.4 to 92.8. This is the first official report of G. irregulare, G. rostratifingens, G. spinosum, G. ultimum var. sporangiiferum, P. acanthicum, P. conidiophorum, P. deliense, P. inflatum, and P. torulosum causing damping-off of soybean in the state of Rio Grande do Sul in southern Brazil.     

Castor beans hulls as a replacement for Tifton 85 hay in lamb diets

The objective of this study was to evaluate the effect of replacing Tifton 85 hay (0, 33, 66, or 100 %) with by castor bean hulls on the intake and nutrient digestibility and performance of 28 noncastrated lambs, averaging 7 months in age, 19.5?±?2.0 kg average live weight, fed on diets based on forage cactus. The animals were housed in individual pens for 100 days. The digestibility trial occurred 40 days after the start of the performance using LIPE® as an external indicator for estimating fecal dry matter production. The replacement of hay with Castor beans hulls linearly decreased the intakes of dry matter, neutral detergent fiber, organic matter, and crude protein and reduced the digestibility coefficients of the dry matter, neutral detergent fiber, organic matter content, crude protein, and total digestible nutrient content. The hot carcass and true carcass yields were not influenced by the treatments and the feed conversion increased linearly. The empty body weight, live weight at slaughter, hot and cold carcass weights, total gain, and average daily gain decreased linearly with the replacement. The replacement of Tifton 85 hay with castor bean hulls reduced the intake of dry matter, crude protein and energy, consequently reducing the performance of the animals. However, as the regression coefficients were of low magnitude, data were subjected to analysis of variance and means were compared using the Dunnett test with diet without castor bean hulls set as the control treatment. The Tifton 85 hay can be replaced up to 66 % by castor bean hulls in finishing diets for lambs.     

Antiviral Sulfoquinovosyldiacylglycerols (SQDGs) from the Brazilian Brown Seaweed Sargassum vulgare

Total lipids from the Brazilian brown seaweed Sargassum vulgare were extracted with chloroform/methanol 2:1 and 1:2 (v/v) at room temperature. After performing Folch partition of the crude lipid extract, the lipids recovered from the Folch lower layer were fractionated on a silica gel column eluted with chloroform, acetone and methanol. The fraction eluted with methanol, presented a strong orcinol-positive band characteristic of the presence of sulfatides when examined by TLC. This fraction was then purified by two successive silica gel column chromatography giving rise to fractions F4I86 and F4II90 that exhibited strong activity against herpes simplex virus type 1 and 2. The chemical structures present in both fractions were elucidated by ESI-MS and ¹H/¹³C NMR analysis HSQC fingerprints based on their tandem–MS behavior as sulfoquinovosildiacylglycerols (SQDGs). The main SQDG present in both fractions and responsible for the anti-herpes activity observed was identified as 1,2-di-O-palmitoyl-3-O-(6-sulfo-α-d-quinovopyranosyl)-glycerol.     

Evaluation of the discriminance capacity of RAPD, isoenzymes and morphologic markers in apple (Malus x domestica Borkh.) and the congruence among classifications

Twenty-one apple accessions were characterised and classified taxonomically with randomly amplified polymorphic DNA (RAPD). The results were compared with morphological and isoenzymatic classifications of the same material by calculating the congruence between similarity matrices. The results confirmed the greater discrimination capacity of RAPD compared with isoenzymes, although the taxonomic clusters obtained with both types of markers were congruent. The genetic relationships calculated using RAPD and isoenzymes showed little congruence with the morphological relationships among accessions. Isoenzymatic markers are useful in germplasm banks to evaluate the diversity of newly introduced accessions. The use of RAPD markers are especially beneficial to discriminate between material that is genetically similar, to evaluate genetic variability within a collection and to choose the components of the core collection.     

The assessment of genetic variation in Spanish primitive cultivars of barley, Hordeum vulgare L., by a combination of isozymes and hordeins

Five isozyme and endosperm reserve protein systems were analysed using electrophoretic techniques in order to investigate the genetic diversity of 222 accessions of Spanish, local varieties of barley, Hordeum vulgare L., maintained at the Centro de Conservación de Recursos Fitogeneéticos of the I.N.I.A. (Alcalá de Henares, Spain). The esterase (EST) isozymes provided more information than did the other systems analysed, showing a total of 14 variable markers. The cathodic peroxidases (CPX) and acid phosphatases (ACPH) were also polymorphic. Malate dehydrogenase (MDH) and glutamate oxaloacetate transaminase (GOT) were monomorphic. The hordeins showed patterns of up to 15 bands, the majority of which were very useful in distinguishing genotypes. 17.2% of accessions showed a uniform genotype, 29.8% showed practically identical genotypes and 53% showed mixtures of different genotypes. It is noteworthy that the use of only two systems (EST and hordeins) and the analysis of only six loci (Est-1, Est-2, Est-4, Hor-1, Hor-2 and Hor-3) is sufficient to reveal the genetic diversity of the collection.     

Detection of Fusarium oxysporum f. sp. dianthi in Carnation Tissue by PCR Amplification of Transposon Insertions

ABSTRACT Strains of the carnation wilt pathogen, Fusarium oxysporum f. sp. dianthi, can be distinguished by DNA fingerprint patterns, using the fungal transposable elements Fot1 and impala as probes for Southern hybridization. The DNA fingerprints correspond to three groups of F. oxysporum f. sp. dianthi strains: the first group includes isolates of races 1 and 8; the second group includes isolates of races 2, 5 and 6; and the third group includes isolates of race 4. Genomic DNAs flanking race-associated insertion sites of Fot1 (from races 1, 2, and 8) or impala (from race 4) were amplified by the inverse polymerase chain reaction (PCR) technique. These regions were cloned and sequenced, and three sets of primers overlapping the 3' or 5' end of the transposon and its genomic insertion were designed. Using fungal genomic DNA as template in PCR experiments, primer pairs generated amplification products of 295, 564 and 1,315 bp, corresponding to races 1 and 8; races 2, 5, and 6; and race 4, respectively. When multiplex PCR was performed with genomic DNA belonging to races 1 and 8, 2, or 4, single amplimers were generated, allowing clear race determination of the isolate tested. PCR was successfully performed on DNA extracted from susceptible carnation cv. Indios infected with isolates representative of races 1, 2, 4, and 8.     

Characterisation of Mycoplasma capricolum P60 surface lipoprotein and its evaluation in a recombinant ELISA

This paper reports the identification and characterisation of a 60kDa surface protein antigen (P60) of Mycoplasma capricolum subspecies capricolum (Mcc), and describes its diagnostic application. Genomic localization and presence in P60 of conserved functional domains suggested a structural and functional relationship with the immunodominant antigen P48 of Mycoplasma agalactiae, a basic membrane protein. A rP60-ELISA was developed, and it resulted in a high specificity for Mcc infections after evaluation with 125 goat sera. The comparison with an existent ELISA based on whole Mcc cell lysates showed that the two assays have comparable sensitivities, but the rP60-ELISA has the significant advantage of a greater specificity. Results indicate that P60 is a potential marker of Mcc infection, especially useful in areas where the presence of M. capricolum subspecies capripenumoniae is also reported.