Evaluation of ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid for tolerance to Xanthomonas axonopodis pv. citri and Xylella fastidiosa

Asiatic citrus canker (ACC), caused by Xanthomonas axonopodis Starr & Garces pv. citri (Hasse) Vauterin et al., and citrus variegated chlorosis (CVC), caused by Xylella fastidiosa Wells et al., are considered the main diseases affecting sweet orange scion varieties in Brazil. Among commercial varieties, mandarins and tangerines are recognized as tolerant to these pathogens. We report herein the production of ‘Hamlin’ sweet orange (Citrus sinensis L. Osbeck) + ‘Montenegrina’ mandarin (Citrus deliciosa Ten.) allotetraploid somatic hybrid plants by protoplast fusion with improved disease tolerance that could be used as a donor of resistance genes in interploid hybridisation. Somatic hybridisation was confirmed by leaf morphology, flow cytometry and RAPD analyses. The somatic hybrid was propagated by grafting and cultivated in a screenhouse for tolerance assays. For X. axonopodis pv. citri assays, buds were collected from both ‘Hamlin’ sweet orange and the somatic hybrid and grafted onto ‘Cleopatra’ mandarin (Citrus reshni hort. ex Tanaka). As a negative control, buds from ‘Mexerica Tardia’ mandarin (C. deliciosa) were collected and grafted onto ‘Cleopatra’ mandarin. Two-month old plants with at least one young vegetative flush were individually spray-inoculated with a 10⁶ CFU mL⁻¹X. axonopodis pv. citri suspension and incubated in a growth chamber, at 27 °C, under 16-h photoperiod. The somatic hybrid showed a statistically significant reduction in susceptibility to ACC 30 days after inoculation. Compared to ‘Hamlin’ sweet orange, disease severity was reduced by 70%, with similar tolerance to that of the mandarin negative control. For X. fastidiosa assays, buds were collected from the somatic hybrid and its parental plants and grafted onto ‘Rangpur’ lime (Citrus limonia Osbeck). The developed plants were needle-inoculated with a X. fastidiosa suspension (8.7 × 10¹⁰ CFU mL⁻¹) into the new growth flush stem. Bacterial population was quantified both at 4 (at the inoculation point) and 8 months (50 cm above the inoculation point) after inoculation. The first evaluation detected X. fastidiosa in 63% of ‘Hamlin’ sweet orange and ‘Hamlin’ + ‘Montenegrina’ mandarin samples. In the second evaluation, X. fastidiosa was detected in 47.4% of ‘Hamlin’ sweet orange and 10.5% of ‘Hamlin’ + ‘Montenegrina’ somatic hybrid samples, suggesting that bacterial movement was restricted in the somatic hybrid. X. fastidiosa was not detected in both evaluations in samples collected from leaves of ‘Montenegrina’ mandarin. These results indicate that the ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid has potential for improved disease tolerance that should enhance its value regarding future use in citrus breeding programs.     

Distribution of mycorrhizal fungal spores in soils under agroforestry and monocultural coffee systems in Brazil

Deep-rooting trees in agroforestry systems may promote distribution of spores of arbuscular mycorrhizal fungi (AMF) at deeper soil levels. We investigated the vertical distribution of AMF spores in Oxisols under agroforestry and monocultural (unshaded) coffee systems in on-farm experiments (Brazil). The number of AMF spores was considered as an indicator of mycorrhiza incidence in soil. Spores were extracted from 0–1, 2–3, 5–7.5, 10–15, 20–30, 40–60 cm soil-depths in agroforestry and monocultural coffee systems, of three different age groups (young, medium-aged and old), using centrifugation methods, and counted. Fine roots were collected and dry-weighed from 0–30 cm in young and old systems and from several depths in medium-aged systems. Soils were characterised with respect to texture, pH, organic matter, calcium, magnesium, phosphorus and potassium. Agroforestry had a higher percentage of spores (12–21% of the total number of spores) and roots (on average 1.5 g L^–1 soil) in deeper layers (20–60 cm), and a lower percentage (79–88%) closer to the surface (0–15 cm) than the monocultural fields (respectively 3–12%, 0.6 g L^–1 soil and 88–97%). Greater numbers of spores in the deeper soil layers may be explained by greater amounts of roots and may be an indicator of greater incidence of mycorrhiza in agroforestry than in monocultural coffee systems. Greater mycorrhizal incidence at deeper soil layers in the agroforestry system may change the dynamics of phosphorus cycling in soil, making this nutrient more available to plants.This revised version was published online in November 2005 with corrections to the Cover Date.     

Phosphorus pools in Oxisols under shaded and unshaded coffee systems on farmers' fields in Brazil

Phosphorus (P) is a primary limiting nutrient for crop production in weathered tropical soils. The deficiency is mainly caused by sorption of phosphate onto Al- and Fe- (hydr)oxides. We hypothesise that the distribution of soil P among various pools is influenced by land use. Our objective was to characterise the soil inorganic (Pi) and organic P (Po) pools and to compare the various pools at different depths in agroforestry (shaded) and monocultural (unshaded) coffee cultivation systems. The study was carried out in the Atlantic Coastal Rainforest domain, Brazil, with Oxisols as the dominant soil type. Soils were collected from four farmers' coffee (Coffea arabica L.) fields, two agroforestry and two monocultural systems. Three profiles were sampled per field, at depths of 2–3, 10–15 and 40–60 cm. A simplified sequential P fractionation was carried out, using resin, 0.5 M NaHCO₃, 0.1 M NaOH, 1 M HCl and concentrated HCl as extractants. Sum-P (resin, NaHCO₃ NaOH, 1 M HCl and concentrated HCl) ranged from 370 to 830 mg kg^–1. Concentrated HCl extracted the largest portion (74%), followed by NaOH (22.5%). Labile (sum of resin, NaHCO₃ and NaOH) P ranged from 13 to 40% of Sum-P. The major part (62%) of the labile fraction was Po. In the agroforestry fields, the amount of Po decreased less with depth and the percentage of Po in labile pools was higher than in monocultural fields. This suggests that agroforestry maintains larger fractions of P available to agricultural crops by influencing the dynamics of P through the conversion of part of the Pi into Po, thereby reducing P losses to the unavailable pools.This revised version was published online in November 2005 with corrections to the Cover Date.     

Probiotic dietary supplementation in Nile tilapia as prophylaxis against streptococcosis

This study evaluated the effects of the probiotic Lactobacillus plantarum as dietary supplement on growth performance, haemato‐immunological responses, microbiology, histology and transmission electron microscopy of the intestinal epithelium of Nile tilapia challenged with Streptococcus agalactiae. Fish were distributed into two groups: control (unsupplemented) group and the group fed L. plantarum supplemented diet for a period of 58 days. We observed an increase in the concentration of lactic acid bacteria and a reduction in the number of Vibrionaceae in supplemented fish. A significant increase in the final weight, specific growth rate and feed efficiency was also observed in supplemented fish. After challenge, the number of thrombocytes and neutrophils also increased in supplemented animals. Transmission electron microscopy showed damage to the intestinal mucosa and the presence of bacteria similar to S. agalactiae in both infected groups. L. plantarum colonized the intestines of fish, enhanced the growth performance and modulated some haematological parameters.     

Physiological and pharmacokinetic responses in neotropical Piaractus mesopotamicus to the essential oil from Lippia sidoides (Verbenaceae) as an anesthetic

International Aquatic Research - The goal of the study was to evaluate Lippia sidoides essential oil as an anesthetic for the tropical fish pacu Piaractus mesopotamicus. The physiological and...     

Conditions for the Production of Carotenoids by Thraustochytrium sp. ATCC 26185 and Aurantiochytrium sp. ATCC PRA-276

There is an intensive search for alternative sources of bioactive molecules, including carotenoids, due to their potent antioxidant activities. Some aquatic microorganisms, such as thraustochytrids, have the ability to synthesize large amounts of carotenoids. The objective of this investigation was to study the effect of growth conditions (carbon: total nitrogen ratio and cultivation system) on the production of carotenoids by Thraustochytrium sp. and Aurantiochytrium sp. For Thraustochytrium sp. culture, carotenoid production was lowered by increasing the initial glucose concentration or continuous supply of glucose and nitrogen. For Aurantiochytrium sp. culture, restraining nitrogen supply or especially using a continuous supply of glucose and total nitrogen decreased total carotenoid productivity. The highest total carotenoid production (2.22 mg/L, w/v) was obtained after 192 h of cultivation of Thraustochytrium sp. using initial concentrations of 30 g/L glucose and 2.4 g/L total nitrogen (batch system). The highest production of astaxanthin was achieved in the fed-batch system with a C/N ratio of 6.2 for Thraustochytrium sp. and 40 for Aurantiochytrium sp. Therefore, it was found that the carotenoid profile can be changed by modifying the growth conditions, which can be useful for the food industry and biotechnological applications.     

Land use change drives the spatio-temporal variation of ecosystem services and their interactions along an altitudinal gradient in Brazil

Landscape Ecology - Land use and land cover (LULC) changes may affect the provision of ecosystem services. However, little is known how LULC changes influence the spatio-temporal variation in...     

Testicular alterations in cryptorchid/orchiopexic rats chronically exposed to acrylamide or di-butyl-phthalate

Exposure of Sprague-Dawley (SD) rats to acrylamide (AA) or di-butyl-phthalate (DBP) from the 12th gestational day to the 16th postnatal week (PNW) has been shown to reduce the effectiveness of orchiopexy in recovering the testicular alterations associated with experimental cryptorchidism established at weaning. Herein, we provide information about the long-term effects of AA or DBP on the testes of cryptorchid/orchiopexic rats. Male offspring exposed in utero to 10 mg/kg/day AA or 500 mg/kg/day DBP underwent bilateral surgical cryptorchidism at the 3rd PNW and orchiopexy at the 6th week, with continuous exposure to the chemicals through diet until the 58th week. Regardless of the test chemical, there were severe qualitative/quantitative alterations in the seminiferous tubules and increased numbers of Leydig cells. There was an increase and decrease in the number of tubules with c-Kit- and placental alkaline phosphatase-labeled germ cells, respectively, as compared to those in the control group, suggesting an imbalance between apoptosis and cell proliferation processes. The histological scores of the testicular lesions at the end of this one-year study were higher than those in the previous 16-week study, indicating that exposure of rats to the toxicants AA or DBP enhanced the testicular alterations induced by the chemicals beginning at the intra-uterine life, and impaired the effectiveness of orchiopexy in restoring the testes to normal morphology. Although the present experimental protocol does not completely replicate the natural human undescended testes, our findings may contribute to understanding the alterations occurring in cryptorchid/orchiopexic testes potentially exposed to exogenous chemicals for extended periods.     

Molecular subtyping of Staphylococcus aureus isolates from cases of bovine mastitis in Brazil.

Sixty-six isolates of Staphylococcus aureus obtained from milk samples of dairy cows suffering from subclinical mastitis in southern Brazil were analysed by five different molecular typing methods. These included the analysis of plasmid profiles, the analysis of coagulase (coa) gene polymorphisms by PCR amplification of the 3' terminal region of the coa gene, the PCR-based detection of polymorphisms in the X region of the protein A gene (spa), the PCR-directed analysis of variations in the spacer region between 16S and 23S rRNA, and the comparison of pulsed-field gel electrophoretically separated genomic SmaI fragment patterns. The molecular typing methods were supplemented with the biochemical characterization of the isolates and the determination of their in-vitro susceptibility to 14 different antibiotics. All genotypic and phenotypic typing methods were analyzed for their ability to discriminate between the isolates. Macrorestriction analysis proved to be the most discriminatory single method (D = 0.96) followed by rRNA spacer typing (D = 0.85), coa PCR (D = 0.82), and spa PCR (D = 0.80).