Tree density and site quality influence on Pinus halepensis Mill. reproductive characteristics after large fires

In Spain, many Pinus halepensis Mill. forests have been seriously affected by significant forest fires in the past decade, in 1994 alone, more than 100 000 ha were burned in Eastern Spain. In order to study the reproductive characteristics of P. halepensis after forest fires, six locations were selected in four areas affected by serious fires in the summer of 1994, and the following different precipitation zones were studied: dry-subhumid, dry and semi-arid. Ten years after the fires, data relevant to the production of pine cones: serotinous (grey), mature (brown), immature (green) and opened cones, was collected from areas with natural post-fire regeneration. Various cone and seed characteristics, such as pine cone seed number and weight, and germination percentage, were measured. The results showed greater production of cones and strobili in high-density sites. The biggest cone sizes (volume) and seed number per cone were related to site quality (dry-subhumid precipitation zone). Also, viability and germination percentages were higher with better site quality, with significant differences in values for serotinous and non-serotinous cones. Despite these differences, the canopy seed bank was large enough to ensure regeneration in this area for this age stand.     

Nutrient deficiency in the production of artemisinin, dihydroartemisinic acid, and artemisinic acid in Artemisia annua L

Artemisia annua became a valuable agricultural crop after the World Health Organization recommended artemisinin as a component of ACT (artemisinin-combination based therapies) for malaria in 2001. A cloned, greenhouse-grown, A. annua (Artemis) subjected to an acidic soil and macronutrient deficit was evaluated for artemisinin production. Lack of lime (L) and macronutrients (N, P, and K) reduced leaf biomass accumulation. When L was provided (pH 5.1), the highest average leaf biomass was achieved with the "complete" (+N, +P, +K, and +L) treatment (70.3 g/plant), and the least biomass was achieved with the untreated (-N, -P, -K, and -L) treatment (6.18 g/plant). The nutrient least required for biomass accumulation per plant (g) was K (49.0 g), followed by P (36.5 g) and N (14.3 g). The artemisinin concentration (g/100 g) was significantly higher (75.5%) in -K plants when compared to plants under the complete treatment (1.62 vs 0.93%). Although the artemisinin total yield (g/plant) was 21% higher in -K plants, it was not significantly different from plants under the complete treatment (0.80 vs 0.66 g/plant). There were no marked treatment effects for concentration (g/100 g) or yield (g/plant) of both dihydroartemisinic acid and artemisinic acid, although higher levels were achieved in plants under the complete or -K treatments. There was a positive and significant correlation between artemisinin and both artemisinic acid and dihydroartemisin acid, in g/100 g and g/plant. This is the first report where potassium deficiency significantly increases the concentration (g/100 g) of artemisinin. Thus, under a mild potassium deficiency, A. annua farmers could achieve similar gains in artemisinin/ha, while saving on potassium fertilization, increasing the profitability of artemisinin production.     

Antioxidant capacity and phenolic content of grapes, sun-dried raisins, and golden raisins and their effect on ex vivo serum antioxidant capacity

Grapes and raisins provide phenolic antioxidants, which contribute to their potential health benefits. The objectives of this study were to compare the antioxidant capacity and phenolic content of green Thompson seedless grapes (the most common variety of grapes consumed in the United States), sun-dried raisins, and golden raisins (both produced from Thompson seedless grapes) and to observe the effects of their consumption over 4 weeks in 15 healthy human males with a cross-over design. The oxygen radical absorbance capacity (ORAC) (positive statistical significance for grapes after 2 weeks and golden raisins after 3 weeks), serum oxidation (positive statistical significance for golden raisin lag time after 4 weeks), total phenolics (no significant effects), and C-reactive protein (no significant effects) were monitored. Immediately postconsumption, there were some significant nonpositive changes. It is hypothesized that these negative results may be explained by postprandial oxidation, a known effect after carbohydrate consumption. Golden raisins had the highest antioxidant capacity and phenolic content. The consumption of a serving of grapes or raisins each day, in addition to a typical diet, may not be sufficient to overcome postprandial oxidation when consumed with other high carbohydrate foods but may have beneficial antioxidant effects over time.     

Effect of a Bowman-Birk proteinase inhibitor from Phaseolus coccineus on Hypothenemus hampei gut proteinases in vitro

The coffee berry borer, Hypothenemus hampei (Ferrari), is an important devastating coffee pest worldwide. Both trypsin and chymotrypsin enzyme activities from H. hampei larval midgut can be inactivated by proteinaceous enzyme-inhibitors. A serine proteinase inhibitor belonging to the Bowman-Birk class was purified from a wild accession of Phaseolus coccineus L. seeds. The inhibitor (PcBBI1) is a cysteine-rich protein that is heat-stable at alkaline pH. MALDI-TOF/MS analysis showed that PcBBI1 occurs in seeds as a monomer (8689 Da) or dimer (17,378 Da). Using in vitro inhibition assays, it was found that PcBBI1 has a high inhibitory activity against H. hampei trypsin-like enzymes, bovine pancreatic chymotrypsin, and trypsin. According to this, PcBBI1 could be a promising tool to make genetically modified coffee with resistance to coffee berry borer.     

Effects of Artemisia annua and Foeniculum vulgare on chickens highly infected with Eimeria tenella (Phylum Apicomplexa)

Background

Intensive poultry production systems depend on chemoprophylaxis with anticoccidial drugs to combat infection. A floor-pen study was conducted to evaluate the anticoccidial effect of Artemisia annua and Foeniculum vulgare on Eimeria tenella infection. Five experimental groups were established: negative control (untreated, unchallenged); positive control (untreated, challenged); a group medicated with 125 ppm lasalocid and challenged; a group medicated with A. annua leaf powder at 1.5% in feed and challenged; and a group treated with the mixed oils of A. annua and Foeniculum vulgare in equal parts, 7.5% in water and challenged. The effects of A. annua and oil extract of A. annua + F. vulgare on E. tenella infection were assessed by clinical signs, mortality, fecal oocyst output, faeces, lesion score, weight gain, and feed conversion.

Results

Clinical signs were noticed only in three chickens from the lasalocid group, six from the A. annua group, and nine from the A. annua + F. vulgare group, but were present in 19 infected chickens from the positive control group. Bloody diarrhea was registered in only two chickens from A. annua group, but in 17 chickens from the positive control group. Mortality also occurred in the positive control group (7/20). Chickens treated with A. annua had a significant reduction in faecal oocysts (95.6%; P = 0.027) and in lesion score (56.3%; P = 0.005) when compared to the positive control. At the end of experiment, chickens treated with A. annua leaf powder had the highest body weight gain (68.2 g/day), after the negative control group, and the best feed conversion (1.85) among all experimental groups.

Conclusions

Our results suggest that A. annua leaf powder (Aa-p), at 1.5% of the daily diet post-infection, can be a valuable alternative for synthetic coccidiostats, such as lasalocid.     

Usefulness of the murine model to study the immune response against Histoplasma capsulatum infection

The present paper is an overview of the primary events that are associated with the histoplasmosis immune response in the murine model. Valuable data that have been recorded in the scientific literature have contributed to an improved understanding of the clinical course of this systemic mycosis, which is caused by the dimorphic fungus Histoplasma capsulatum. Data must be analyzed carefully, given that misinterpretation could be generated because most of the available information is based on experimental host–parasite interactions that used inappropriate proceedings, i.e., the non-natural route of infection with the parasitic and virulent fungal yeast-phase, which is not the usual infective phase of the etiological agent of this mycosis.     

Distribution patterns of acorns after primary dispersion in a fragmented oak forest and their consequences on predators and dispersers

Forest fragmentation affects seed production, but little is known about how seeds are distributed in fragmented forests after primary dispersion and how this affects their interactions with other organisms. This study addressed these issues in a fragmented oak forest of Quercus laeta where four habitat types were recognized: interior and edge of patches, deforested matrix around patches, and an abandoned field. Areas and distances to the abandoned field were measured for 40 patches, and acorns were sampled in all habitats. Only samples from the interior and edge of patches had acorns, which were classified as aborted, viable, and parasitized by insects or fungi. Acorns in all these classes were regressed against patch area and distance to the abandoned field. Field experiments were also conducted to determine whether acorn removal rates varied across habitats. Aborted acorns within patches were not related to area or distance, but they decreased with increasing values of both variables at patch edges. Within patches, viable acorns were positively related to increases in area and distance, while the converse occurred for insect-damaged acorns; fungal-infested acorns were not related to patch variables in this habitat. Complex interactions between area and distance explained how acorns in these classes were distributed across patch edges. Acorn removal was higher within patches and their edges when compared to human-disturbed habitats; both mice and birds were identified as potential acorn dispersers. These results suggest that forest fragmentation affects acorn-mediated processes and that this may influence the recovery of patch connectivity in the landscape.     

The genetic structure of Mytilus chilensis (Hupé 1854) populations along the Chilean coast based on RAPDs analysis

The Chilean blue mussel Mytilus chilensis is an important commercial species. However, little has been published on the population genetics of this species, despite the need to implement management and conservation policies. Randomly amplified polymorphic DNA‐polymerase chain reaction analysis was used to estimate genetic variation within and between eight natural populations along the whole range of its Chilean natural distribution (ca. 1900 km from Arauco (VIII Region) to Punta Arenas (XII Region)). The values of Nei's unbiased genetic distance, D (0.030–0.107), among populations were small, despite the large geographic separation. A mantel test using 50 000 randomizations showed evidence for a significant correlation (r=0.74, P<0.05) between genetic and geographic (coastal) distance. Punta Arenas population was the most genetically differentiated from the others, although the scale of differentiation was not large (D=0.076–0.107). The levels of gene flow (Nm=1.55) found in this study prevent differentiation among populations by genetic drift. This is the result of the long‐lived planktotrophic larvae of M. chilensis, which provides this species with considerable dispersal ability throughout its range, which is favoured by the ocean currents along the Chilean coast. A restricted larval dispersal towards the north due to the Cape Horn Current derived from the West Wind Drift could be the cause of the higher genetic differentiation of Punta Arenas population from the northern populations. For management purposes of the M. chilensis fishery, the results provide no evidence for discrete stocks, with the possible exception of the Punta Arenas population. The present study provides the baseline data in order to continue further characterization of these mussel populations, considering the great increase in aquaculture of this species.     

Development and Deterioration of Spermatophores in Pond-Reared Penaeus vannamei

Spermatophore deterioration in pond-reared Penaeus vannamei was studied, and spermatophore quality was evaluated and improved. Sperm production of males collected from grow-out ponds was increased by two techniques: eyestalk ablation using a fresh frozen maturation diet (13% body weight); and laboratory culture using a diet of the formulated feed Nicovita Plus (3% body weight/ d) and frozen squid (2%). Findings are complementary to previous reports that eyestalk ablation improves quality of spermatophores in young (25.7 g) males. The timing of eyestalk ablation for activation of the endocrine mechanism, leading to improved spermatophore quality was also explored. After eyestalk ablation, performed between postmolt and intermolt stages, 26 g males required a minimum of three spermatophore regenerations or 42 d to significantly increase spermatophore size and sperm count. On the other hand, the laboratory culture (2.5 mo) technique improved the quality of spermatophores in successive regenerations for non-ablated males. In the present study, subadult P. vannamei produced spermatophores which, if not transferred or manually ejaculated, gradually deteriorated (successive stages are described), while a new compound spermatophore was being synthesized.     

Sperm maturation and capacitation in the open thelycum shrimp Litopenaeus (Crustacea: Decapoda: Penaeoidea)

Transmission electron microscopy was applied to sperm removed from males and females belonging to Litopenaeus vannamei, L. stylirostris and L. occidentalis. It was discovered that a region named filamentous meshwork (FM), located between the nucleus and the hemispherical cap, develops differently in these three closely related species. In L. vannamei, the FM is synthesized in the male reproductive system, but seems to complete its formation after mating. In L. stylirostris, the FM region was not present in spermatophores collected from males or in sperm from the thelycum. In L. occidentalis, the FM region is fully developed in male sperm. It is suggested that completion of the FM is required for acrosome maturation, and the process continues after mating in some species of Litopenaeus. In vitro induction of the acrosome reaction in sperm from males and females of L. occidentalis demonstrated for the first time that reactivity is significantly superior in sperm cells that have been attached to the open thelycum for some hours, as compared to sperm in males (prior to transfer). This finding suggests that matured sperm cells of L. occidentalis become capacitated to react against egg water after mating.