Direct penetration of Rhizopus stolonifer into stone fruits causing rhizopus rot

Rhizopus rot, caused by Rhizopus stolonifer, is a major postharvest disease of stone fruits. The disease is related to the occurrence of mechanical and physical damage; however, observations at a Brazilian wholesale market suggest that direct penetration can occur. Therefore, the penetration mechanisms of R. stolonifer in stone fruits were evaluated. To identify the production of enzymes that help with direct penetration by the pathogen, esterase activity, both in mycelial discs and in spore suspensions of the fungus in water and in modified Van Etten nutrient solution, was measured. Assays were also conducted to evaluate the growth of R. stolonifer on glucose or cutin as a sole carbon source. The pathogen grew on both media, and higher esterase activity was observed in the cutin medium. Wounded and unwounded peaches and nectarines were inoculated with R. stolonifer spore suspensions in water or in modified Van Etten nutrient solution. Wounded fruit inoculated with either of the R. stolonifer spore suspensions developed rhizopus rot, whereas unwounded fruit developed the rot only in the presence of spores in the modified Van Etten nutrient solution. Scanning electron and light microscopic examination showed the fungus can directly penetrate the nectarine cuticle. Diisopropyl fluorophosphate, a serine hydrolase inhibitor, prevented rot development in peaches. The results provide valuable evidence for the ability of R. stolonifer to directly penetrate unwounded stone fruits, probably due to the production of esterase enzymes.     

Fibroblast growth factor-10 maintains the survival and promotes the growth of cultured goat preantral follicles

The aim of the present study was to investigate the effects of fibroblast growth factor-10 (FGF-10) on the survival, activation (transition from primordial to primary follicles), and growth of goat preantral follicles cultured in vitro. Pieces of ovarian cortex were cultured for 1 and 7 d in the absence or presence of FGF-10 (0, 1, 10, 50, 100, and 200 ng/mL). Noncultured and cultured tissues were processed and analyzed by histology, transmission electron microscopy, and viability testing. Results showed that after 7 d, a greater percentage (79.9%) of morphologically normal follicles (containing an oocyte with regular shape and uniform cytoplasm, and organized layers of granulosa cells without a pyknotic nucleus) was observed when cultured with 50 ng/mL of FGF-10 when compared with other concentrations of FGF-10 (0 ng/mL, 67.3%; 1 ng/mL, 68.2%; 10 ng/mL, 63.3%; 100 ng/mL, 64.4%; 200 ng/mL, 52.7%). Ultrastructural analyses and viability testing using fluorescent markers confirmed the follicular integrity of FGF-10 (50 ng/mL)-treated fragments after 7 d of culture. After 7 d, all FGF-10 concentrations reduced the percentage of primordial follicles and increased the percentage of developing follicles. In the presence of 50 ng/mL of FGF-10, follicles increased in diameter after 7 d of culture when compared with other concentrations tested. In conclusion, this study demonstrates that FGF-10 maintains the morphological integrity of goat preantral follicles and stimulates the growth of activated follicles in culture. The culture conditions identified here contribute to the understanding of the factors involved in goat early follicular development.     

Aboveground biomass,transpiration and water use efficiency in eucalypt plantation fertilized with KCl,NaCl and phonolite rock powder

Potassium has important physiological functions in eucalypt plantations, increasing their productivity when applied to soil via mineral fertilizers. There is interest in identifying alternative sources to KCl owing to its high cost and limited reserves. The aim of the study was to test the effect of replacing KCl with NaCl and phonolite rock powder. Two comparisons were made: (1) application of 283 kg ha^?1 of KCl compared with that of 2125 kg ha^?1 of phonolite rock powder (equivalent to 170 kg ha^?1 of K₂O in both treatments); (2) application of 139 kg ha^?1 of NaCl compared with that of 183 kg ha^?1 of KCl (equivalent to 2.33 kmol Na and K, respectively). Radial growth, soil water content, leaf water potential (Ψ), accumulated transpiration, stem volume and biomass increment, as well as water use efficiency (WUE) were evaluated. In the first comparison, both fertilizations presented equal values for all characteristics evaluated. In the second, the accumulated transpiration in trees fertilized with KCl was 17% higher than that in plants fertilized with NaCl. In contrast, the WUE was 20% higher in the trees fertilized with NaCl than in those fertilized with KCl, reflecting the lower water consumption for the same increment in stem volume and biomass. We conclude that phonolite rock powder and NaCl are possible substitutes for conventional K fertilization performed with KCl.

     

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae)

This study investigated the acute toxicity (LC_50-24 hr) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC_50-24 hr) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 μg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC_50-24 hr for Argulus sp. adults was 67.97 μg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 μg/L, while the LC_50-24 hr for Argulus sp. and D. discoidalis was 83.98 μg/L and 82.48 μg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.     

Effects of lipid sources,lysophospholipids and organic acids in maize-based broiler diets on nutrient balance,liver concentration of fat-soluble vitamins,jejunal microbiota and performance

1. Three experiments with a 2 × 2 × 2 factorial arrangement were conducted to evaluate maize-based diets for broilers containing different lipid sources [soybean oil (S) or beef tallow (T)] supplemented with or without lysophospholipids and organic acids on nutrient balance (Experiment I, evaluation period of 10–14 d), on liver concentration of fat-soluble vitamins, on jejunal microbiota (Experiment II, sampling at d 14) and on performance (Experiment III, accumulated periods of 1–14, 1–21 and 1–42 d).

2. A total of 1344 male chicks were used. In each experiment, the birds were allotted in a completely randomised design with 8 replications. The lysophospholipids were mainly composed of lysolecithins and the organic acids blend was constituted by lactic (40%), acetic (7%) and butyric acids (1%).

3. An interaction between lipid sources and lysophospholipids was observed on faecal apparent digestibility of lipid (ADL), which improved with lysophospholipids addition in T diets. Broilers fed on S had higher ADL and faecal apparent digestibility of nitrogen-corrected gross energy (ADGE_N).

4. It was not possible to demonstrate a significant treatment effect on the liver concentration of vitamins A and E, even with the differences in fatty acid profile between S and T.

5. Enterobacteria values were below the detection threshold. Lysophospholipid supplementation reduced gram-positive cocci in T-fed birds. S diets promoted lower total anaerobe counts compared with T diets, independent of additives.

6. S diets increased BW gain and feed:gain ratio in all evaluation periods. Lysophospholipids and organic acids improved feed:gain ratio at 1–21 d in T diets. Furthermore, main effects were observed for lysophospholipids and organic acids at 1–42 d, which increased BW gain and improved feed:gain ratio, respectively.

7. No positive interactions between additives were found.

     

Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus

A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.     

Intensive rearing of juvenile crayfish (Pacifastacus leniusculus,Astacidae) during the first 6 months: effects of size grading

Recent advances in crayfish feeding have enabled the development of size grading studies from the start of first‐feeding. A 180‐day experiment aimed at intensive rearing of Pacifastacus leniusculus was carried out under controlled conditions, evaluating the effects of size grading at two different periods from the onset of exogenous feeding. Stage 2 juveniles were stocked in fibreglass tanks at a density of 100 m², and fed a dry diet for salmonids combined with restricted amounts of Artemia cysts. Five groups were tested: no grading, grading at 60 days (large and small size) and grading at 100 days (large and small size). After 6 months, no significant differences were found in the survival among groups (mean: 73.06%). The highest final growth (pooled results from upper and lower classes: 17.39 mm carapace length, 1.43 g weight) was achieved by the crayfish sorted at 60 days, showing significant differences from the ungraded group. Smaller crayfish graded at 60 days grew significantly faster than smaller crayfish graded at 100 days. The food conversion ratio was lower in the graded groups (mean: 2.64), showing significant differences from the ungraded group (3.23). This study shows that size grading allows a better performance and an improved feeding efficiency.