Identification of 'Ubá' mango tree zygotic and nucellar seedlings using ISSR markers

Polyembryonic seeds are characterized by the development of over one embryo in the same seed, which can be zygotic and nucellar. The objective of this work was to identify the genetic origin, whether zygotic or nucellar, of seedlings of polyembryonic seeds of 'Ubá' mango tree using ISSR markers, and relating them with the vigor of the seedlings. Thus, mangos were harvested in Visconde do Rio Branco (accession 102) and Ubá (accessions 112, 138, 152 and 159), whose seeds were germinated in plastic trays filled with washed sand. Fifty days after sowing, seedlings from five seeds of each one of the accessions 102, 112, 138, 159 and from 10 seeds of the accession 152, were analyzed. These sseedlings were characterized and evaluated for plant height, stem circumference and mass of fresh aerial part and the most vigorous seedling was the one displaying at least two of these traits higher than the other seedlings from seed. Leaves were collected for genomic DNA extraction, which was amplified using seven ISSR primers previously selected based on the amplification profile and considering the number and resolution of fragments. Zygotic seedlings were found in 18 seeds, which were the most vigorous in six seeds. The results evidenced the existence of genetic variability in orchards using seedlings grown from seeds, because the farmer usually uses the most vigorous ones, assuming that this is of nucellar origin. These results also indicate that the most vigorous seedling are not always nucellar, inasmuch as of 20% of the total seeds evaluated, the zygotic seedling was the most vigorous.

Rooting of hardwood cuttings of Roxo de Valinhos fig (Ficus carica L.) with different propagation strategies

The objective of this study was to evaluate the substrate, cuttings collection time, the position and the cutting depth, and the propagation environment on rooting of 'Purple Valinhos' fig tree cuttings in Southwestern Paraná, Brazil. Two experiments were carried out at UTFPR, Câmpus Dois Vizinhos, with hardwoods cuttings from Roxo de Valinhos fig tree. The first experiment used a randomized block design, in 3 x 3 x 2 factorial (substrate x environment x collection time), with four replications of 10 cuttings per plot. The cuttings were collected in the first fifteen days of July and August. The substrates were sand, soil and the mixture of these [1:1 (v / v)]. The environments used were open sky, tunnel with plastic cover and tunnel with half-shade black net cover. The second experiment used a randomized block design, 2 x 2 x 3 factorial (shoot cutting position x soil cover x shoot cutting depth), with four replications of 12 cuttings per plot. In the factor position, the vertically (0 º inclination) and inclined (45 º inclination) shoot cuttings were evaluated. Soil cover was tested with mulching plastic cover or not. The tested depths were 1/3, 1/2 and 2/3 in relation to the total length of the shoot cutting. In both experiments, the following were analyzed: rooting and mortality indices, number of leaves and primary shoots, length of the three largest roots per cutting. It was conclude that, the protected environment with plastic cover on sand as substrate must recommended for the rooting of fig estaca, collecting them in the first half of July. The inclination position and cutting depth of the estaca and the substrate coverage with plastic mulching did not influence the results.

Histological alterations in liver and testis of Astyanax aff. bimaculatus caused by acute exposition to zinc

This study investigated the effect of acute exposition to zinc (Zn) on histology of the liver and testes of yellow tail lambari (Astyanax aff. bimaculatus). The exposure consisted of six concentrations of Zn (0, 3, 5, 10, 15, and 20 mg/L) for 96 hours of exposure. Fragments of liver and testis were routinely processed and embedded in plastic resin based on glycol methacrylate. Fragments of bones, muscles, liver and testis were dehydrated and digested to quantify the absorption levels of Zn in the tissue. Acute exposure to concentrations above 10mg/L has produced structural changes in the liver and gonads. The changes found in the liver were vascular congestion; decrease of cellular volume; displacement of the hepatocyte nucleus; necrosis; disarrangement of cordon structure; leukocyte infiltrate and vacuolization. The changes found in the gonads were ruptured cyst, delayed development of germ cells, pyknotic nucleus, cell cluster, displacement of cyst wall and vacuolization. The histological changes observed were compatible with the increasing concentration of zinc in environment, compromising liver and reproductive functions, because there was an increase in relative frequency of hepatocytes and reduced sperm production

Acclimation of croton and hibiscus seedlings in response to the application of indobultiric acid and humic acid for rooting

The vegetative propagation of ornamental plants can be accelerated by applying plant growth regulators. Amongst them, the use of auxins, plant hormones with physiological effects on cell elongation and rooting have stood out. Alternatively, the application of humic acids, bioactive fraction of soil organic matter, also results in increases in rooting cuttings of ornamental plants. The objective of this work was to study the growth characteristics and the nutritional contents of croton and hibiscus plants during acclimation of seedlings in response to different concentrations of indolebutyric acid (IBA) and humic acid (HA) applied to cuttings for rooting. The experiment was conducted in greenhouse, and the apical stem cuttings were treated with solutions with concentrations of 0, 250, 500, 1000 and 2000 mg L-1of IBA and 0, 10, 20, 30 and 40 mg L-1 of C from HA. At 45 days of rooting in carbonized rice husk, they were individually transferred to plastic bags of 2.0 dm3 containing a mixture of soil: sand: manure (2: 1: 1) as substrate. At 90 days of acclimation, the plants were collected for measurement of growth and nutritional variables. The results showed that the application of the IBA stimulates the absorption of nutrients and growth of croton cuttings and transplanted hibiscus, contributing to formation of vigorous seedlings. A similar response occurred with the application of HA in hibiscus cuttings

Interactions between physical and psychosocial demands of work associated to low back pain

OBJECTIVE: To examine the interaction between physical and psychosocial demands of work associated to low back pain. METHODS: Cross-sectional study carried out in a stratified proportional random sample of 577 plastic industry workers in the metropolitan area of the city of Salvador, Northeast Brazil in 2002. An anonymous standard questionnaire was administered in the workplace by trained interviewers. Physical demands at work were self-rated on a 6-point numeric scale, with anchors at each end of the scale. Factor analysis was carried out on 11 physical demand variables to identify underlying factors. Psychosocial work demands were measured by demand, control and social support questions. Multivariate analysis was performed using the likelihood ratio test. RESULTS: The factor analysis identified two physical work demand factors: material handling (factor 1) and repetitiveness (factor 2). The multiple logistic regression analysis showed that factor 1 was positively associated with low back pain (OR=2.35, 95% CI 1.50;3.66). No interaction was found between physical and psychosocial work demands but both were independently associated to low back pain. CONCLUSIONS: The study found independent effects of physical and psychosocial work demands on low back pain prevalence and emphasizes the importance of physical demands especially of material handling involving trunk bending forward and trunk rotation regardless of age, gender, and body fitness.

Suppression of humoral immunity and lymphocyte responsiveness during experimental trypanosoma cruzi infections

C3H/He and C57B1/6 mice were inoculated with 500 Trypanosoma cruzi trypomastigotes (Strain Y). During the acute phase infected mice presented parasitemia and enlargement of lymph nodes and spleens and intracellular parasites were observed in the heart. Examinations of cells derived from spleen and lymph nodes showed increased numbers of IgM and IgG-bearing cells. During the peak of splenomegaly, about day 17 post-infections, splenic lymphocytes showed a marked decrease in responsiveness to T and B-cell mitogens, parasite antigens and plaque forming cells (PFC) to sheep red blood cells (SRBC). Unfractionated or plastic adherent splenic cells from mice, obtained during the acute phase were able to suppress the response to mitogens by lymphocytes from uninfected mice. During the chronic phase. Disappearance of parasitemia and intracellular parasites in the hearts as well as a decrease in spleen size, was observed. These changes preceded the complete recovery of responsiveness to mitogens and T. cruzi antigens by C57B1/6 splenic lymphocytes. However, this recovery was only partial in the C3H/He mice, known to be more sensitive to T. cruzi infection. Partial recovery of humoral immune response also occurred in both strains of mice during the chronic phase.

A low cost method to produce a gaseous environment for the isolation of Helicobacter pylori

A low cost method (LCM) to produce a gaseous environment for the isolation of Helicobacter pylori, was compared with the standard Gas Park system. The LCM uses a carbonated antacid tablet, a plastic bag with tap water, a candle, and a wide-mouthed glass jar provided with a tight-fitting metalic screw cap and a rubber gasket. Antral gastric biopsies from 153 cases were incubated by duplicate on blood agar plates and treated with the two methods. In 95 cases the agent was isolated from both, and only from the standard method in 10 cases; the opposite condition was found in five cases, and 43 were negative. That difference is not significant (Pearson's X²= 93.25 p > 0,05)

Intestinal parasites in school food handlers in the city of Uberlândia, Minas Gerais, Brazil

In order to verify the presence of intestinal parasites in food handlers, stool samples were collected from 104 cooks and their helpers that were working in food preparation in 20 public elementary schools, in various areas of the city of Uberlândia, Minas Gerais, Brazil. The samples were collected during the months of November and December, 1988, in plastic flasks containing a 10% formaldehyde solution and processed by the Hoffmann, Pons & Janer method. The sediment was examined using triplicate slides. All individuals were females aged between 24 to 69 years. Intestinal parasites were found in 85.0% of the studied schools and 47.1% of the studied food handlers (cooks and helpers) were found to be positive. Among the 49 infected food handlers, 32 (65.3%) carried a single parasite and 17 (34.7%) carried two parasites. The following intestinal parasites were found: Giardia lamblia (21.1%), Entamoeba coli (21.1%), hookworms (9.6%), Ascaris lumbricoides (5.8%), Entamoeba histolytica (2.9%), Hymenolepis nana (1.9%), Strongyloides stercoralis (1.0%). These data emphasize the need for a rigid semi-annual control in all school food handlers, including diagnosis, specific treatment and orientation about the mechanisms of transmission of the intestinal parasites.

A simple and inexprensive method to generate a microaerophilic atmosphere for the isolation of Campylobacter sp

Faeces of 138 chickens were inoculated on Blaser agar plates. One set of plates was incubated in jars with CampyPak envelopes. The others were incubated in "Zip-lock" plastic bags (7 x X in.) and a microaerophilic atmosphere was generated exhaling into the "Zip-lock" plastic bag, after holding the breath for 20 sec. Then, the bag was pressed to evacuate its atmosphere, inflated again, and pressed (4 times), and finally sealed. Campylobacter was isolated from 127 (96.2%) of samples incubated in jars with gas generator envelopes and from 129 (98%) of the specimens incubated into the bags. The proposed methodology offers good savings for cost-conscious laboratories.

Entomophthoramycosis by Conidiobolus coronatus. Report of a case successfully treated with the combination of itraconazole and fluconazole

Rhinoentomophthoramycosis caused by Conidiobolus coronatus in a 61-year old woman was unsuccessfully treated during 8 years with all the antifungals available in the Brazilian market, including potassium iodide for 1 month, sulfamethoxazole plus trimethoprim for 2 months, amphotericin B, total dose of 1130 mg, cetoconazole, 400 mg/day for 6 months, fluconazole, 200 mg/day, for at least 2 months and, itraconazole, 400 mg/day for 2 months, followed by 200 mg/day for 4 more months. Complete clinical and mycological cure was achieved using itraconazol 400 mg/day in association with fluconazol 200 mg/day during 24 months. After cure she was submitted to plastic surgery to repair her facial deformation. Today she remains clinically and mycologically cured after 59/60 months (5 years!) without any specific antifungal. We thus suggest the use of the combination of itraconazole and fluconazole as an additional option for the treatment of this mycosis.

Airborne fungi in the city of Porto Alegre, Rio Grande do Sul, Brazil

Knowledge of anemophilous fungi in a given city or region is important for the ecological diagnosis and specific treatment of allergic manifestations induced by inhaled allergens. In order to diagnose the presence of anemophilous fungi, several qualitative and quantitative techniques are used depending on the study place. This study of fungal air spores was performed with a Rotorod Sampler®, an equipment which samples the air through a plastic rod attached to an electric engine that makes it spin fast enough to collect the particles in the air. The samples were collected once a week during 24 hours using the standard cycle of the manufacturers. A total of 52 samples were obtained from April 2000 through March 2001. The results revealed prevalence of ascosporos (50.49%), Cladosporium (17.86%), Aspergillus/Penicillium (15.03%), basidiosporos (3.84%), rusts (3.82%), and Helminthosporium (2.49%), and a lesser frequency of Botrytis (1.22%), Alternaria (1.19%), smuts (0.90%), Curvularia (0.87%), Nigrospora (0.61%), and Fusarium (0.08%). Also, 1.59% of the spores detected here could not be identified by the systematic key used. More fungal spores were observed during the summer than during the autumn.

Persistence of Vectobac WDG and Metoprag S-2G against Aedes aegypti larvae using a semi-field bioassay in Rio de Janeiro, Brazil

Persistence of Bacillus thuringiensis var. israelensis (Vectobac WDG) and methoprene (Metoprag S-2G) was evaluated against Aedes aegypti late third instar larvae of the Rockefeller strain in a semi-field bioassay. Tests were performed in Rio de Janeiro, using containers made of plastic, iron, concrete and asbestos, placed in a shaded area. The formulations used were 0.2 g of Vectobac-WDG and 1g of Metoprag S-2G per 100 liters of water in house storage containers. Vectobac WDG was tested twice, in March and in April/May, 2002. In March (temperature ranging from 21.5 to 39.3 ºC), 70-100% mortality was observed by the 7th day and declined abruptly thereafter. No significant differences were observed among the container types. In April/May (18.6 to 34.8 ºC) mortality was higher than 70% to 30-36 days in all cases, except in the iron container (40% mortality on the 12th day). Metoprag S-2G was evaluated in April/May, 2002, and induced mortality higher than 70% up to 15 days in the plastic and iron containers and only seven days in the concrete container. In the asbestos container, maximal mortality was achieved on day one post-treatment (66%). Our results point to a low persistence of both formulations in the weather conditions of Rio de Janeiro.

Effect of stalk and leaf extracts from Euphorbiaceae species on Aedes aegypti (Diptera, Culicidae) larvae

The objective of this study was to evaluate the larvicidal activity of essential oil aqueous solutions (hydrolates) obtained by steam distillation of stalks and leaves of Croton argyrophylloides, Croton nepetaefolius, Croton sonderianus and Croton zehntneri against Aedes aegypti larvae. Twenty-five larvae of third instar were placed in plastic beckers, containing the hydrolates (50 mL), in a four repetitions scheme. Water was used as control and the number of dead larvae was counted after 24 hours. The data obtained were submitted to Variance Analysis and Tukey test. Significant differences were observed among the hydrolates from different species and from different parts of each plant (p < 0.001). The hydrolates of stalk and leaf from C. nepetaefolius and C. zehntneri and leaf hydrolate of C. argyrophylloides presented 100% mortality against larvae. The compounds present in C. zenhtneri and C. nepetaefolius are oxygenated phenylpropanoids that are more soluble in water than the monoterpenes and sesquiterpenes detected in the oils of C. argyrophylloides and C. sonderianus. This study showed that all species analyzed presented compounds with larvicidal properties, with differences between each plant parts.

Diterpenos de Copaifera reticulata Ducke com atividade larvicida contra Aedes aegypti (L.) (Diptera, Culicidae)

The objective of this study was to evaluate the larvicidal activity of diterpenoids obtained from the oil-resin of Copaifera reticulata against Aedes aegypti larvae, the principal vector of dengue and urban yellow fever. Four diterpenes were obtained from oil-resin extraction with organic solvents and subsequent chromatographic and spectroscopic procedures allowed to isolation and identification of these compounds as 3-b-acetoxylabdan-8(17)-13-dien-15-oic acid (1), alepterolic acid (2), 3-b-hidroxylabdan-8(17)-en-15-oic acid (3), and ent-agatic acid (4). Each compound was previously dissolved in dimethylsulphoxide, and distilled water was added to obtain the desired concentrations. Twenty larvae of third instars were placed into plastic beckers, containing the solution test (25 mL), in a five repetitions scheme, and their mortality, indicated by torpor and darkening of the cephalic capsule, was recorded after 48h. Probit analyses were used to determine lethal concentrations (LC50 and LC90) and their respective 95% confidence intervals. This study showed that only diterpenoids 1 and 2 exhibited larvicidal properties with LC50 of 0.8 ppm and 87.3 ppm, respectively, revealing the former as the most toxic compound against third instars of Ae. aegypti. Therefore, this compound seems to be an interesting source for new metabolite to be exploited.

TOLL-LIKE RECEPTORS (TLR) 2 AND 4 EXPRESSION OF KERATINOCYTES FROM PATIENTS WITH LOCALIZED AND DISSEMINATED DERMATOPHYTOSIS

There are few studies on the role of innate immune response in dermatophytosis. An investigation was conducted to define the involvement of Toll-Like Receptors (TLRs) 2 and 4 in localized (LD) and disseminated (DD) dermatophytosis due to T. rubrum. Fifteen newly diagnosed patients, eight patients with LD and seven with DD, defined by involvement of at least three body segments were used in this study. Controls comprised twenty skin samples from healthy individuals undergoing plastic surgery. TLR2 and TLR4 were quantified in skin lesions by immunohistochemistry. A reduced expression of TLR4 in the lower and upper epidermis of both LD and DD patients was found compared to controls; TLR2 expression was preserved in the upper and lower epidermis of all three groups. As TLR4 signaling induces the production of inflammatory cytokines and neutrophils recruitment, its reduced expression likely contributed to the lack of resolution of the infection and the consequent chronic nature of the dermatophytosis. As TLR2 expression acts to limit the inflammatory process and preserves the epidermal structure, its preserved expression may also contribute to the persistent infection and limited inflammation that are characteristic of dermatophytic infections.