Use of fungal inocula and pumice for decomposing coffee pulp

A study of decomposition of coffee pulp (from Guatemala City, Guatemala) was carried out with several inocula. Fresh and pressed pulps were studied in field and laboratory experiments. Results showed that the best inoculum among the eight examined in the experiments was the fungus Trichoderma viride, the only one not isolated from the coffee pulp itself. This inoculum, together with pumice, accelerated the decomposition of coffee pulp at ambient temperature during a 45-day test period. © 1985.

Biocompatibility of an adhesive system applied to exposed human dental pulp

Human pulp tissue was directly capped with All Bond 2, or calcium hydroxide and evaluated 7, 30, or 60 days after the procedures. Histological analysis was performed to assess the inflammatory cell response, tissue disorganization, dentin bridging, and the presence of bacteria. At 7 days, with All Bond 2 capping, there was a large area of neutrophilic infiltrate underlying the pulp capping material, and the death of adjacent odontoblasts, was observed. However, with time, the neutrophilic reaction was replaced by fibroblastic proliferation with macrophages and giant cells surrounding globules of resin scattered in the coronal pulp tissue. The persistent inflammatory reaction and hyaline alteration of extracellular matrix inhibited complete pulp repair or dentin bridging. In contrast, at 7 days, the pulp tissue capped with calcium hydroxide exhibited odontoblast-like cells organized underneath coagulation necrosis. Pulp repair evolved into apparent complete dentin bridge formation at 60 days. All Bond 2 did not appear to allow any pulp repair and does not appear to be indicated for direct pulp capping of human teeth. Copyright © 1999 by The American Association of Endodontists.

Water sorption thermodynamic properties applied to persimmon skin and pulp

Moisture equilibrium data of persimmon skin and pulp were determined using the static gravimetric method. Adsorption and desorption isotherms were obtained in the range of 20-70°C, to water activities (a w) from 0.02 to 0.85. The application of the GAB model to the experimental results, using direct nonlinear regression analysis, provided a good agreement between experimental and calculated values. The net isosteric heat of sorption was estimated from equilibrium sorption data, using the Clausius-Clapeyron equation. Isosteric heats of sorption were found to increase with increasing temperature and could be well adjusted by an exponential relationship. The enthalpy-entropy compensation theory was applied to sorption isotherms and plots of ΔH versus ΔS for skin and pulp provided the isokinetic temperatures, indicating an enthalpy controlled sorption process. © 2000 Elsevier Science B.V.

Distribution of Xylella fastidiosa in citrus rootstocks and transmission of citrus variegated chlorosis between sweet orange plants through natural root grafts

To study translocation of Xylella fastidiosa to citrus rootstocks, budsticks from citrus variegated chlorosis (CVC)-affected cv. Pera sweet orange (Citrus sinenesis (L.) Osb.) were top grafted on 15 citrus rootstocks. Disease symptoms were conspicuous 3 months later on all 15 rootstocks tested. The presence of X. fastidiosa was confirmed by light microscopy, double-antibody sandwich enzyme-linked immunosorbent assays, and polymerase chain reaction in rootlets and main roots of CVC-symptomatic Pera sweet orange in 11 of the 15 rootstocks tested. These results suggest that bacterial translocation from the aerial plant parts to the root system occurs but is not essential for X. fastidiosa to induce symptoms in the aerial parts. Bacterial translocation to the roots was not correlated with CVC leaf-symptom severity in the Pera scion. To determine if CVC disease could be transmitted by natural root grafts, two matched seedlings of each of four sweet orange cultivars (Pera, Natal, Valencia, and Caipira) were transplanted into single pots. One seedling rootstock of each pair was inoculated by top grafting with a CVC-contaminated budstick while the other seedling rootstock was cut but not graft inoculated. Transmission of X. fastidiosa from an inoculated plant to a noninoculated plant sharing the same pot was observed in all four sweet orange cultivars tested. Transmission was confirmed by observation of natural roots grafts between the two plants, presence of X. fastidiosa in the root grafts, and disease development in the uninoculated plants. This is the first report of transmission of CVC disease through natural root grafts.

Aspects of IFP for citrus production in Brazil

Brazilian citriculture represents about 25% of the total world citrus production with an area of 851,518 ha and a total production of more than 17 million tons in 1996. Besides its importance to the brazilian economy, represented by more than 1 billion US $ by year from FCOJ exportation, the citriculture has problems related to low productivity, due to several cultural practices and management. The productivity would be improved by an IFP system. The main problems are related to soil, as poor conservation, use of poor drained soils and bad preparation for planting; diseases-canker, CVC, leprosis, Phytophtora gummosis and other fungus diseases; pests - mites, scales, nematodes and others; the use of disease free and improved scion and rootstocks propagative material is usual; cultural practices - as nutrition, irrigation, wind breaks, weed control, pruning, replant and others, as density planting could be improved. Some possible solutions will be discussed for improving the brazilian citrus productivity and quality by IFP, based on research made.

Susceptibility of tangerines to citrus variegated chlorosis

Citrus variegated chlorosis (CVC), a citrus disease first discovered in Brazil in 1987, is caused by the bacterium Xylella fastidiosa and transmitted by sharpshooters and budwood. Since the disease affects almost all sweet orange cultivars, it has become one of the most serious problems for Brazilian citriculture. To evaluate their resistance to CVC disease, fifteen tangerines or mandarins (C. reticulata Blanco) and their hybrids were grafted on Rangpur lime (C. limonia Osb.) and inoculated with CVC-contaminated Pera sweet orange (C. sinensis (L.) Osb.) by twig grafting in a greenhouse. Tangerines and their hybrids Wilking, Fortune, Sunki, Ellendale, Orlando tangelo, Nunes clementine, Nova, Sun Shu Sha Kat, Suenkat, and Batangas showed CVC leaf symptoms and gave positive results on enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) (with specific primers for X. fastidiosa), indicating that they are susceptible to CVC. Although X. fastidiosa bacteria were detected by ELISA and PCR in inoculated plants of tangerines Cravo and Oneco, no CVC leaf symptoms were observed on these two cultivars, suggesting that they are tolerant to the disease. CVC leaf symptoms were not observed and X. fastidiosa was not detected in tangerine Dancy and mandarins Okitsu satsuma and Ponkan after inoculation, showing that they are resistant to the disease.

Evaluation of microsatellite markers in cultivars of sweet orange (Citrus sinensis [L.] Osbeck)

Sweet orange is considered a very important species in the citrus world market and presents wide morphological variability. However, its characterization at the molecular level by random amplified polymorphic DNA (RAPD) and isozyme markers is not appropriate. Microsatellite or simple sequence repeats (SSRs) have been suggested as ideal for studies in cultures of vegetative propagation and as value markers for mapping in several species. However, information on microsatellite polymorphism in citrus species is scarce. In this work, microsatellite markers (AG-repeats) were developed from an enrichment library of genomic DNA of sweet orange cv. Pera (Citrus sinensis [L.] Osbeck), and 31 cultivars of sweet orange were evaluated. Evaluation of 18 microsatellite primers did not permit differentiation of the varieties studied. New microsatellite primers are being evaluated with the aim of detecting polymorphisms among the cultivars and closely related species to be used in genetic mapping programs.

Adubação verde em Citrus: Influência na porosidade e retenção de água de um latossolo vermelho-escuro do noroeste do estado de São Paulo

The objective of this research was to study the porosity, bulk density and retention of water of an Oxisol, located in the Northwestern region of Sn̄o Paulo state, Brazil. The soil was cultivated with Citrus sp., to which green manure was applied between rows for three years. Each of six species of green manure crops (Crotalaria juncea L., Mucuna deeringiana Steph. & Bart., Canavalia ensiformis L. DC., Cajanus cajan L., Lablab purpureum L. and Ricinus communis L.) were seeded for three years (1995, 1996 and 1997) between Citrus rows, plus a treatment with a mix of all six species and a control (natural regrowth af vegetation). The experimental design was a randomized complete block design, with four replications for each of the eight treatments. Water retention, microporosity, macroporosity, total porosity and bulk density were analyzed in the beginning (1995) and end (1997) of the experiment, at three depth ranges (0-0.10; 0.10-0.20 and 0.20-0.40m). We concluded that there were statistically significant differences for bulk density, macroporosity, total porosity and retention of water among the different soil depth ranges; there were no significant differences among treatments though.

Healing process of dog dental pulp after pulpotomy and pulp covering with mineral trioxide aggregate or Portland cement.

Considering several reports about the similarity between the chemical compositions of the mineral trioxide aggregate (MTA) and Portland cement (PC), the subject of this investigation was to analyze the behavior of dog dental pulp after pulpotomy and direct pulp protection with these materials. After pulpotomy, the pulp stumps of 26 roots of dog teeth were protected with MTA or PC. Sixty days after treatment, the animal was sacrificed and the specimens removed and prepared for histomorphological analysis. There was a complete tubular hard tissue bridge in almost all specimens. In conclusion, MTA and PC show similar comparative results when used in direct pulp protection after pulpotomy.

Activity of benomyl for control of postbloom fruit drop of citrus caused by Colletotrichum acutatum

Postbloom fruit drop (PFD) of citrus caused by Colletotrichum acutatum produces orange-brown lesions on petals and induces the abscission of young fruitlets and the retention of the calyces. Despite the fact that C. acutatum is not highly sensitive to benomyl in culture, this fungicide provides good control of the disease under field conditions. This study was undertaken to determine the effect of benomyl on various stages of disease development to understand the basis for its effectiveness in the field. We found that benomyl at 1.0 μg/ml reduced colony area of C. acutatum by about 75% and completely inhibited growth of C. gloeosporioides. Benomyl did not prevent conidial germination even at 100 μg/ml, but reduced germ tube elongation at 10 and 100 μg/ml. When benomyl was applied to flower clusters on screen-house-grown plants before inoculation, disease severity was greatly reduced. Applications at 24 and 48 h, but not at 72 h, after inoculation reduced PFD severity. Application of benomyl to symptomatic petals not bearing conidia did not prevent or reduce production of inoculum. Application to petals bearing conidia reduced viability of these fungal propagules by only about 50%. The viability of appressoria on mature leaves was not affected by benomyl application. Even when appressoria on mature leaves were stimulated to germinate by treatment with flower extracts, subsequent application of benomyl did not reduce propagule numbers below original levels. Benomyl appears to act by preventing infection and early development of the fungus in petals. However, once symptoms have developed, this fungicide has only minimal effects on further disease development and spread.

Effect of Bacillus circulans D1 thermostable xylanase on biobleaching of eucalyptus kraft pulp

The alkalophilic Bacillus circulans D1 was isolated from decayed wood. It produced high levels of extracellular cellulase-free xylanase. The enzyme was thermally stable up to 60°C, with an optimal hydrolysis temperature of 70°C. It was stable over a wide pH range (5.5-10.5), with an optimum pH at 5.5 and 80% of its activity at pH 9.0. This cellulase-free xylanase preparation was used to biobleach kraft pulp. Enzymatic treatment of kraft pulp decreased chlorine dioxide use by 23 and 37% to obtain the same kappa number (κ number) and brightness, respectively. Separation on Sephadex G-50 isolated three fractions with xylanase activity with distinct molecular weights.

Benomyl sensitivity of isolates of Colletotrichum acutatum and C. gloeosporioides from citrus

Postbloom fruit drop (PFD) of citrus, caused by Colletotrichum acutatum, produces orange-brown lesions on petals and results in premature fruit drop and the retention of calyces. C. gloeosporioides is common in groves and causes postharvest anthracnose on fruit. Both diseases are controlled effectively by the fungicide benomyl in research fields and commercial orchards. Highly sensitive and resistant isolates of C. gloeosporioides were found, whereas all isolates of C. acutatum tested were moderately resistant. In preliminary studies conducted in vitro with three isolates of each, mycelial growth of sensitive isolates of C. gloeosporioides was inhibited completely by benomyl (Benlate 50 WP) at 1.0 μg/ml, whereas resistant isolates grew well at 10 μg/ml. Growth of all isolates of C. acutatum was inhibited by about 55% at 0.1 μg/ml and by 80% at 1.0 μg/ml. Spore germination of C. acutatum was inhibited more at 0.1 μg/ml than at 1.0 μg/ml or higher concentrations. In all, 20 isolates of C. acutatum from 17 groves and 20 isolates of C. gloeosporioides from 7 groves were collected from locations with different histories of benomyl usage in São Paulo, Brazil, and Florida, United States. Benomyl at 1.0 μ.g/ml completely inhibited growth of 133 isolates of C. gloeosporioides, with the exception of 7 isolates that were highly resistant to the fungicide, whereas all isolates of C. acutatum were only partially inhibited at 0.1 and 1.0 μg/ml. Analysis of variance indicated that the sensitivity of the isolates of C. acutatum was not affected by benomyl usage or grove of origin, and country of origin had only minor effects. No highly resistant or sensitive isolate of C. acutatum was recovered. Partial sequencing of the β-tubulin gene did not reveal nucleotide substitutions in codons 198 or 200 in C. acutatum that usually are associated with benomyl resistance in other fungi.

In vitro penetration of bleaching agents into the pulp chamber

Aim: To investigate pulp chamber penetration of bleaching agents in teeth following restorative procedures. Methodology: Bovine lateral incisors were sectioned 3 mm apical to the cemento-enamel junction and the coronal pulpal tissue was removed. Teeth were divided into six groups (n = 10): G1, G2 and G3 were not submitted to any restorative procedure, while G4, G5 and G6 were submitted to Class V preparations and restored with composite resin. Acetate buffer was placed in the pulp chamber and treatment agents were applied for 60 min at 37°C as follows: G1 and G4, immersion into distilled water; G2 and G5, 10% carbamide peroxide (CP) exposure; G3 and G6, 35% CP bleaching. The buffer solution was removed and transferred to a glass tube where leuco crystal violet and horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined spectrophotometrically at 596 nm. A standard curve made with known amounts of hydrogen peroxide was used to convert the optical density values of the coloured samples into microgram equivalents of hydrogen peroxide. Data were submitted to ANOVA and Tukey's test (5%). Results: Amounts of hydrogen peroxide found in the pulp chamber of G2 and G5 specimens (0.1833 ± 0.2003 μg) were significantly lower (P = 0.001) when compared to G3 and G6 specimens (0.4604 ± 0.3981 μg). Restored teeth held significantly higher (P = 0.001) hydrogen peroxide concentrations in the pulp chamber than intact teeth. Conclusion: Higher concentrations of the bleaching agent produced higher levels of hydrogen peroxide in the pulp chamber, especially in restored teeth.

Evaluation of systems for timing of fungicide sprays for control of postbloom fruit drop of citrus in Brazil

Postbloom fruit drop (PFD) of citrus, caused by Colletotrichum acutatum, infects petals of citrus flowers and produces orange-brown lesions that induce the abscission of young fruitlets and the retention of calyces. Proper timing of fungicide applications is essential for good disease control. Different systems for timing of fungicide applications for control of PFD in a major citrus-growing region in southern São Paulo state in Brazil were evaluated from 1999 to 2002. The following programs were compared to an unsprayed control using counts of diseased flowers, persistent calyces, or fruit: (i) a phenology-based program currently recommended in Brazil with one application at early and another at peak bloom; (ii) the Florida PFD model; (iii) the postbloom fruit drop-fungicide application decision system (PFD-FAD), a new computer-assisted decision method; and (iv) grower's choice. In 1999, no disease developed, sprays applied with the phenology-based program had no effect, and the Florida PFD model saved two sprays compared with the phenology-based program. In 2000, PFD was moderate and the phenology-based and growers' choice treatments had a significantly lower number of persistent calyces and higher fruit numbers than the control, but no differences were found between those treatments and the PFD model. In 2001, PFD was severe with considerable yield loss. The PFD model, the phenology-based program, and the grower's choice reduced flower blight and the number of persistent calyces, and improved fruit yields with two to three applications, but the PFD-FAD achieved comparable yields with only one spray. In 2002, the disease was mild, with no yield loss, and the Florida PFD model and the PFD-FAD saved one spray compared with the other systems. The PFD model and the PFD-FAD were equally effective for timing fungicide applications to control PFD in Brazil. Scouting of trees is simpler with PFD-FAD; therefore, this system is recommended and should eliminate unnecessary sprays and reduce costs for growers.

Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching

Two extracellular xylanases produced by the thermotolerant fungus Aspergillus caespitosus grown in sugar cane bagasse were purified and characterized. Estimated molecular masses were 26.3 and 27 kDa (xyl I); 7.7 and 17.7 kDa (xyl II) for gel filtration and SDS-PAGE, respectively. Optimal temperature for both xylanases was 50-55°C. Optimal pH was 6.5-7.0 for xyl I, and 5.5-6.5 for xyl II. The thermostability (T half) at 55°C was 27.3 min (xyl I) and >90 min (xyl II). Xylanase activity was inhibited by several ions. β-mercaptoethanol activated 59 and 102% xyl I and xyl II activities, respectively. These enzymes preferentially hydrolyzed birchwood xylan, and the K m and V max values were 2.5 mg/ml and 1679 U/mg protein (xyl I), and 3.9 mg/ml and 113 U/mg protein (xyl II). The action of both xylanases mainly that of xyl II, on kraft pulp reduced kappa number and increased pulp viscosity. © 2004 Elsevier Ltd. All rights reserved.