Morphologic, Molecular, and Pathogenic Characterization of Diaporthe phaseolorum Variability in the Core Soybean-Producing Area of Argentina

ABSTRACT Isolates of the Diaporthe/Phomopsis (D/P) complex were collected in the main soybean producing area of Argentina during the 1996-97, 1997-98, and 1998-99 growing seasons. Twenty-three morphologic characters related to type of colonies, stroma, pycnidia and conidia, presence of perithecia, and asci length were studied by principal component analysis (PCA). Genomic DNA were analyzed by the random amplified polymorphic DNA (RAPD) technique. From both studies, 18 isolates were identified as D/P complex and grouped in four major taxa: (i) Diaporthe phaseolorum var. meridionalis, (ii) D. phaseolorum var. caulivora, (iii) D. phaseolorum var. sojae, and (iv) Phomopsis longicolla. In addition to distinguishing interspecific and intraspecific variability, molecular markers allowed the detection of differences among isolates within the same variety. Pathogenicity was assayed in the greenhouse, by the toothpick method, inoculating the D/P isolates to soybean genotypes carrying different resistance genes (Rdc1, Rdc2, Rdc3, and Rdc4) against soybean stem canker (SSC). Pathogenic analysis distinguished two main groups: (i) the SSC-producing isolates, including D. phaseolorum var. meridionalis and D. phaseolorum var. caulivora, and (ii) the non-SSC-producing isolates, including D. phaseolorum var. sojae and P. longicolla. Cultivar RA-702 (susceptible control) was compatible with both D. phaseolorum var. meridionalis and D. phaseolorum var. caulivora isolates; meanwhile, Tracy-M (Rdc1 and Rdc 2 genes) was incompatible with D. phaseolorum var. meridionalis but compatible with D. phaseolorum var. caulivora isolates. The fact that Rdc1 and Rdc2 together (as in Tracy-M) confer an almost immune reaction to all assayed isolates of D. phaseolorum var. meridionalis but were ineffective against the D. phaseolorum var. caulivora isolates evaluated suggests that the virulence or avirulence genes in D. phaseolorum var. meridionalis and D. phaseolorum var. caulivora are different. Moreover, physiological races of D. phaseolorum var. meridionalis were detected by using differential soybean genotypes carrying distinct single Rdc genes. As far as we know, this is the first report on the existence of physiological races of D. phaseolorum var. meridionalis in South America. Selective pressure due to deployment of resistant host cultivars may have changed the frequency of the virulence or avirulence genes within the population of D. phaseolorum var. meridionalis. On the whole, our results show that pathogenic variability of D. phaseolorum in the core soybean-producing area of Argentina is higher than previously recognized.     

The effects of capsaicin topical therapy in dogs with atopic dermatitis: a randomized,double-blinded,placebo-controlled,cross-over clinical trial

Abstract The efficacy of twice daily topical application of capsaicin (0.025%) for the management of pruritus in dogs with atopic dermatitis (AD) was evaluated in double-blinded, placebo-controlled study. Twelve dogs with AD were randomly assigned to either 0.025% capsaicin or vehicle lotion applied twice daily for 6 weeks. After a 4-week wash-out period, treatments were switched. Significant improvement was reported by owners ( P  = 0.0006), but not by investigators. Owners noted temporary worsening of pruritus after the first week of capsaicin therapy. Overall capsaicin was well tolerated. Substance P (SP) concentrations in the skin did not correlate with the severity of the pruritus and did not change significantly over time and between treatments. Lesional skin had less SP than nonlesional skin ( P  = 0.03). These observations suggest that topical capsaicin should be further evaluated as an adjunctive antipruritic agent in dogs with AD.     

Occurrence and Concentration Levels of Nitro-PAH in the Air of Three Brazilian Cities Experiencing Different Emission Impacts

Nitrated Polycyclic Aromatic Hydrocarbons (NPAH) were determined in the suspended particulates collected in São Paulo (Brazil) and three other cities lying in the São Paulo State, (namely, Araraquara, Piracicaba and Paulínia), during August 2002 and July 2003. São Paulo samples represented a big difference between two groups of days, one of which characterized by heavy injection of pollutants released by industrial and agricultural activities. Most NPAH congeners identified in the atmosphere were among those usually associated to diesel exhaust, with the three important exceptions of 2-nitrofluoranthene, 2- and 4-nitropyrene. Daytime reactions seemed to promote decomposition of primary fluoranthene and pyrene. Trends of NPAH at all sites seemed to be associated to regional transport of polluted air through air masses passing over sugar cane plantations of São Paulo State.     

Breaking the crossability barriers between disomic tetraploid Solanum acaule and tetrasomic tetraploid S. tuberosum

Summary A combination of compatible second pollinations and embryo rescue was applied for systematic production of true tetraploid hybrids from crosses between disomic tetraploid Solanum acaule and tetrasomic tetraploid potato, S. tuberosum. Several genotypes of tetraploid potatoes were pollinated with S. acaule, and the compatible second pollinations were made on the following day, with a genotype of S. phureja, IvP 35 to promote fruit development. Embryo rescue was carried out in 21 families, 14 to 27 days after the first pollination. A total of eight plants were obtained from the embryo rescue and their chromosome numbers were counted in the root tips. Three of the eight plants were identified as tetraploid, and five others as diploid. Morphology, isozyme banding patterns, and pollen stainability, as well as potato spindle tuber viroid (PSTVd) resistance, indicated the hybrid nature of the three plants. This is the first report of successful tetraploid hybrid production between disomic tetraploid S. acaule (4x) and tetrasomic tetraploid potatoes. Seed set from the crosses between one of hybrids and diploid potatoes indicated workable levels of both male and female fertility for introgression of valuable genes from S. acaule into the cultivated potato gene pool. The methodology used may be applied to other disomic tetraploid tuber-bearing Solanum species and with some modifications also to distantly related solanaceous species and genera.     

Accumulation of metals in macrophytes from water reservoirs of a power supply plant, Rio de Janeiro State, Brazil

Aquatic macrophytes are well known accumulators for heavy metals, the reason why they are used as bioindicators for water quality and in phytoremediaton strategies. This study reports on the elemental concentrations in four free-floating aquatic macrophytes (S. auriculata; P. stratiotes; E. crassipes and E. azurea) growing in two water reservoirs (Santana e Vigário, Rio de Janeiro State, Brazil) of an electric power plant that receive input from the polluted Paraíba do Sul River. Filtered water samples and water suspended solids from these environments were also analysed. Inductively coupled plasma mass spectrometry was used as the principal method, allowing the determination of up to 41 elements, including the rare-earth elements (REEs) and other trace metals not assayed before in these macrophytes. The results show that all elements studied are accumulated by the macrophytes with concentration ratios (CR = [plant]: [water]) varying from about 1,000 to 200,000, based on the dry weight of the plant species. With a few exceptions, highest accumulations were observed in E. crassipes in which CRs increase in the sequence: Cu < Mo < Cr < Pb < Tl < Fe < La < Zn < Ce< Mn. Surprisingly high CRs (e.g. Ce: 74,000) and corresponding mass concentrations were observed for the rare-earth elements (e.g. ∑REE: 112 mg kg^?1), also measured in the water suspended particle fraction. The results show that this fraction acts as an effective sink for trace metals in the aquatic system studied and seems to play also an important role in the transfer of metals from water to the plant species.     

Sarcosporidian infection in pigs in Uruguay.

Examination of enzymatic digests of samples of the crux diaphragm obtained at abbatoirs in Montevideo, Uruguay, indicated that 57.2% of 269 pigs weighing 90-140 kg were infected with Sarcocystis sp. The morphology of sarcocystis in H & E stained sections indicated that they were Sarcocystis miescheriana.     

Cellular and cytokine kinetics after epicutaneous allergen challenge (atopy patch testing) with house dust mites in high-IgE beagles

The cellular and cytokine dynamics of reactions triggered by atopy patch testing with house dust mites were studied in six high-IgE beagles. Sites were scored and biopsied at 6, 24, 48, and 96 h, and samples were processed for histopathology, immunohistochemistry, and polymerase chain reaction (PCR). All dogs developed positive reactions at some point in time. Mean clinical scores were significantly higher than baseline at 24, 48, and 96 h. Clinically, one of six dogs had a positive reaction at 6 h; two of six reacted at 24 and 48 h, and five of six at 96 h. Histologically, superficial perivascular mononuclear and granulocytic dermatitis developed (5/6) after 6 h, and progressed in severity at 24 h (6/6). Additionally, at 48 h epidermal spongiosis, hyperplasia and pustules developed (5/6), and were marked at 96 h (6/6). At and beyond 6 h, progressive CD1c-positive epidermal Langerhans cell hyperplasia with cluster formation and dermal dendritic cell infiltration was noted. Cutaneous infiltration of CD3-positive T lymphocytes with epidermal clusters developed over time. mRNA expression for the cytokines gamma-interferon (gamma-IFN), interleukin-6 (IL-6), IL-12p35, IL-13, IL-18, and thymus and activation regulated chemokine (TARC) exhibited significant increases during the challenge compared to baseline, but there was no appreciable alteration in expression for tumour necrosis factor-alpha (TNF-alpha), IL-12p40, IL-10, regulated on activation normal T-cell expressed and secreted (RANTES), IL-5, IL-2, IL-4, and IL-8. No correlation was detected between clinical scores and cytokines. It is concluded that IL-6 plays a role in early reactions followed by an increase of TARC and IL-13, while IL-18 progressively increases in later reactions.     

Pharmacokinetics of pentoxifylline and its 5-hydroxyhexyl metabolite after oral and intravenous administration of pentoxifylline to healthy adult horses

OBJECTIVE: To determine serum pharmacokinetics of pentoxifylline and its 5-hydroxyhexyl metabolite in horses after administration of a single IV dose and after single and multiple oral doses. ANIMALS: 8 healthy adult horses. PROCEDURES: A crossover study design was used with a washout period of 6 days between treatments. Treatments were IV administration of a single dose of pentoxifylline (8.5 mg/kg) and oral administration of generic sustained-release pentoxifylline (10 mg/kg, q 12 h, for 8 days). Blood samples were collected 0, 1, 3, 6, 12, 20, 30, and 45 minutes and 1, 2, 4, 6, 8, and 12 hours after IV administration. For oral administration, blood samples were collected 0, 0.25, 0.5, 0.75, 1, 2, 4, 8, and 12 hours after the first dose and 0, 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, and 24 hours after the last dose. RESULTS: Elimination of pentoxifylline was rapid after IV administration. After oral administration, pentoxifylline was rapidly absorbed and variably eliminated. Higher serum concentrations of pentoxifylline and apparent bioavailability were observed after oral administration of the first dose, compared with values after administration of the last dose on day 8 of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, oral administration of 10 mg of pentoxifylline/kg results in serum concentrations equivalent to those observed for therapeutic doses of pentoxifylline in humans. Twice daily administration appears to be appropriate. However, serum concentrations of pentoxifylline appear to decrease with repeated dosing; thus, practitioners may consider increasing the dosage if clinical response diminishes with repeated administration.