Apoptosis-mediated anticancer activity in prostate cancer cells of a chestnut honey (Castanea sativa L.) quinoline-pyrrolidine gamma-lactam alkaloid.

Prostate cancer (PCa) is the most common malignancy in men and represents the second leading cause of cancer deaths in Western countries. PCa is initially androgen-dependent, however, this tumor inevitably progresses as castration-resistant prostate cancer (CRPC), which represents the most aggressive phase of the pathology. In this work, in two CRPC cell lines (DU145 and PC3), we studied the in vitro inhibitory properties of the tryptophan-derived endogenous metabolite kynurenic acid (KYNA) and of the lactam form of 3-2'-pyrrilonidinyl-kynurenic acid (3-PKA-L), alkaloids usually present in combination in chestnut honey. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell colony formation assay, and Western blot analysis of the major mediator proteins involved in apoptotic processes. In all experiments, KYNA was scarcely or not active while 3-PKA-L showed anticancer activity in the high concentration range (0.01 mM - 1 mM) from 24 to 72 h. The results obtained showed that cell death was induced by extrinsic apoptotic pathway, by cell morphological changes and reduction of cell colonies number. These novel results represent the first promising step to the accurate description of 3-PKA-L cytotoxic effect, not observed with KYNA, paving the way to the search of new anticancer agents, as well as to the better understanding of the physiopathological role of this interesting natural product.

Supplementation of diets with tannins from Chestnut wood or an extract from Stevia rebaudiana Bertoni and effects on in vitro rumen fermentation, protozoa count and methane production.

The aim of the trial was to evaluate the effect of dietary additions of Stevia rebaudiana Bertoni extract (SB) and Chestnut wood tannin (CWT) on the in vitro rumen fermentability, protozoal population and methane yield. Both plant products were tested at 3 different levels of inclusion (0.75, 1.50 and 3.00% of incubated dry matter, DM) in a total mixed ration (TMR) for ruminants by using rumen batch culture systems and a rumen inoculum collected from sheep. Total volatile fatty acid concentration, their proportions and gas production were not modified by the plant extracts inclusion, except a significant linear increment of gas production at 24 hr for SB (p = .049). Ammonia concentration decreased (p < .05) of about 17% when 1.50 or 3.00% of CWT were included into TMR. Rumen protozoa population was depressed by the SB inclusion (p = .002) with a maximum reduction of 40% at the highest SB dosage, whereas CWT negatively affected total protozoa counts (-19%) only at the dose of 3.00%. In vitro DM and NDF degradability were not affected by the supplementation of SB and CWT, as well as the methane yield. Thus, the addition of SB and CWT decreases the in vitro protozoa population of the rumen with different intensity and without effects on fermentation parameters, apart from a reduction of nitrogen degradability caused by CWT. Despite the effect on protozoa, no decreasing effect on methane production was detected.

Mechanisms underlying detection of seed dormancy by a scatter-hoarding rodent.

The mechanism underlying detection of seed dormancy by scatter-hoarding rodents is unclear, although previous work suggests that the pericarp plays an important role in signaling dormancy status. Eastern gray squirrels (Sciurus carolinensis) consume early germinating seeds as they are more likely to perish immediately, whereas dormant seeds tend to be cached. To examine the mechanisms underlying dormancy detection, we characterized physical and chemical differences between germinating and dormant pericarps of northern red oak (Quercus rubra), American chestnut (Castanea dentata) and the BC3 hybrid of Chinese chestnut and American chestnut (Castanea mollissima × C. dentata) using scanning electron microscopy and gas chromatography- mass spectrometry. We found that, as seeds break dormancy, the wax layer on the pericarp degrades and is accompanied by the escape of lower molecular weight kernel compounds or lipid metabolism byproducts. Our field experiments showed that squirrels were 4-8 times more likely to consume seeds that were altered to remove pericarp wax coating or that were sprayed with seed chemicals. We argue that dormancy detection by scatter-hoarding rodents is a complex process involving physical cues such as loss of pericarp wax and chemical cues such as emission of olfactory cues.

A new leaf-mining moth, Caloptilia aesculi, sp. nov. (Lepidoptera: Gracillariidae: Gracillariinae) feeding on Aesculus chinensis Bunge (Hippocastanaceae) from China.

A new species of the genus Caloptilia associating with the Chinese horse chestnut, Aesculus chinensis Bunge (Hippocastanaceae) from China is described. The photographs of the adults, male and female genitalia, larva and pupa, the leaf mines and leaf shelters (rolls and stacks) are given. This is the first report of host association with Hippocastanaceae in the subfamily Gracillariinae. The sequence of mitochondrial COI barcoding region of this species is provided and its phylogenetic position is analyzed with other Caloptilia species.

Development of a natural preservative obtained from male chestnut flowers: optimization of a heat-assisted extraction technique.

The aim of this work was to optimize the conditions for the extraction of phenolic compounds (PC) from male chestnut flowers using heat-assisted extraction in developing extracts rich in PC for potential industrial application as a natural ingredient. The study conditions of time (t), temperature (T), solvent (S, water-ethanol mixtures) and solid-to-liquid ratio (S/L) were optimized. The responses used were obtained from the quantification of the fourteen major individual PC identified by HPLC-DAD-ESI/MS (seven hydrolysable tannins and seven flavonoids). The recovery of hydrolysable tannins was higher than that of flavonoids, with trigalloyl-HHDP-glucoside being the major one. The conditions that maximized the PC content were t = 20.0 ± 37.7 min, T = 25.0 ± 5.7 °C, S = 0.0 ± 8.7% ethanol and S/L = 82.8 g L-1, producing an extract with 86.5 mg PC g-1 of extract. The results highlight the potential of valorising chestnut flower agro-residues as a productive source of PC for the development of bio-based ingredients for food/pharmaceutical/cosmeceutical industrial applications able to compete with synthetic compounds.

Comparison of Physicochemical Properties of Starches from Nine Chinese Chestnut Varieties.

Chestnut is a popular food in many countries and is also an important starch source. In previous studies, physicochemical properties of starches have been compared among different Chinese chestnut varieties growing under different conditions. In this study, nine Chinese chestnut varieties from the same farm were investigated for starch physicochemical properties to exclude the effects of growing conditions. The dry kernels had starch contents from 42.7 to 49.3%. Starches from different varieties had similar morphologies and exhibited round, oval, ellipsoidal, and polygonal shapes with a central hilum and smooth surface. Starch had bimodal size distribution and the volume-weighted mean diameter ranged from 7.2 to 8.2 µm among nine varieties. The starches had apparent amylose contents from 23.8 to 27.3% but exhibited the same C-type crystalline structure and similar relative crystallinity, ordered degree, and lamellar structure. The gelatinization onset, peak, and conclusion temperatures ranged from 60.4 to 63.9 °C, from 64.8 to 68.3 °C, and from 70.5 to 74.5 °C, respectively, among nine starches; and the peak, hot, breakdown, final, and setback viscosities ranged from 5524 to 6505 mPa s, from 3042 to 3616 mPa s, from 2205 to 2954 mPa s, from 4378 to 4942 mPa s, and from 1326 to 1788 mPa s, respectively. The rapidly digestible starch, slowly digestible starch, and resistant starch ranged from 2.6 to 3.7%, from 5.7 to 12.7%, and from 84.4 to 90.7%, respectively, for native starch, and from 79.6 to 89.5%, from 1.3 to 3.8%, and from 7.1 to 17.4%, respectively, for gelatinized starch.

Isolation of quercetin and mandelic acid from Aesculus indica fruit and their biological activities.

BACKGROUND: In this study Aesculus indica fruit was subjected to isolation of phytochemicals. Two antioxidants quercetin and Mandelic acid were isolated in pure state. The free radical scavenging and acetyl choline esterase inhibitory potential of the crude extract and sub fractions were also determined. RESULTS: The antioxidant capacity of crude extract, fractions and isolated compounds were determined by DPPH and ABTS methods. Folin-Ciocalteu reagent method was used to estimate the total phenolic contents and were found to be 78.34 ± 0.96, 44.16 ± 1.05, 65.45 ± 1.29, 37.85 ± 1.44 and 50.23 ± 2.431 (mg/g of gallic acid) in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fractions respectively. The flavonoid concentration in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fraction were; 85.30 ± 1.20, 53.80 ± 1.07, 77.50 ± 1.12, 26.30 ± 1.35 and 37.78 ± 1.25 (mg/g of quercetin) respectively. The chloroform fraction was more potent against enzymes, acetyl choline esterase and butyryl choline esterase (IC50 = 85 and 160 µg/ml respectively). The phenolic compounds in the crude extract and fractions were determined using HPLC standard method. Chlorogenic acid, quercetin, phloroglucinol, rutin, mandelic acid and hydroxy benzoic acid were detected at retention times 6.005, 10.062, 22.623, 30.597, 35.490 and 36.211 in crude extract and different fractions. The ethyl acetate fraction was rich in the targeted compounds and was therefore subjected to column isolation. The HPLC chromatogram of isolated compounds showed single peak at specified retention times which confirms their isolation in pure state. The isolated compounds were then characterized by FTIR and NMR spectrophotometric techniques. CONCLUSION: The Aesculus indica fruit extracts showed antioxidant and anticholine esterase inhibitory potentials. Two bioactive compounds were isolated in the pure form ethyl acetate fraction. From results it was concluded that the fruit of this plant could be used to minimize oxidative stress caused by reactive oxygen species.

Overexpression of DEMETER, a DNA demethylase, promotes early apical bud maturation in poplar.

The transition from active growth to dormancy is critical for the survival of perennial plants. We identified a DEMETER-like (CsDML) cDNA from a winter-enriched cDNA subtractive library in chestnut (Castanea sativa Mill.), an economically and ecologically important species. Next, we characterized this DNA demethylase and its putative ortholog in the more experimentally tractable hybrid poplar (Populus tremula × alba), under the signals that trigger bud dormancy in trees. We performed phylogenetic and protein sequence analysis, gene expression profiling, and 5-methyl-cytosine methylation immunodetection studies to evaluate the role of CsDML and its homolog in poplar, PtaDML6. Transgenic hybrid poplars overexpressing CsDML were produced and analysed. Short days and cold temperatures induced CsDML and PtaDML6. Overexpression of CsDML accelerated short-day-induced bud formation, specifically from Stages 1 to 0. Buds acquired a red-brown coloration earlier than wild-type plants, alongside with the up-regulation of flavonoid biosynthesis enzymes and accumulation of flavonoids in the shoot apical meristem and bud scales. Our data show that the CsDML gene induces bud formation needed for the survival of the apical meristem under the harsh conditions of winter.

Triterpene saponins from Billia rosea.

Five previously undescribed triterpene saponins, billiosides A-E, and a known analogue, were isolated from the seeds of Billia rosea (Planch. & Linden) C. Ulloa & P. Jørg. Their structures were elucidated on the basis of extensive 1D and 2D NMR experiments (1H, 13C, DEPT, COSY, TOCSY, NOESY, ROESY, HSQC, and HMBC) and mass spectrometry as (3ß,21ß,22α)-3-[(2-O-ß-D-glucopyranosyl-O-[α-L-arabinopyranosyl-(1 â†’ 4)]-ß-D-glucopyranosyl)oxy]-21-[((2E,6S)-2,6-dimethyl-6-hydroxyocta-2,7-dienoyl)oxy]-22-(acetyloxy)-24-hydroxyolean-12-en-28-oic acid, (3ß,21ß,22α)-3-[(2-O-ß-D-galactopyranosyl-ß-D-glucopyranosyl)oxy]-21,22-dihydroxyolean-12-en-28-yl O-α-L-arabinopyranosyl-(1 â†’ 4)-ß-D-glucopyranoside, (3ß,21ß,22α)-3-[(2-O-ß-D-galactopyranosyl-O-[α-L-arabinopyranosyl-(1 â†’ 4)]-ß-D-xylopyranosyl)oxy]-21,22-dihydroxyolean-12-en-28-yl O-ß-D-glucopyranoside, (3ß,21ß,22α)-3-[(2-O-ß-D-galactopyranosyl-O-[α-L-arabinopyranosyl-(1 â†’ 4)]-ß-D-glucopyranosyl)oxy]-21,22-dihydroxyolean-12-en-28-yl O-ß-D-glucopyranoside, (3ß,21ß,22α)-3-[(2-O-ß-D-galactopyranosyl-O-[α-L-arabinopyranosyl-(1 â†’ 4)]-ß-D-glucopyranosyl)oxy]-21,22-dihydroxyolean-12-en-28-yl O-ß-D-glucopyranosyl-(1 â†’ 6)-ß-D-glucopyranoside, and dipteroside A. Billiosides B and C exhibited moderate effects when tested as hepatic glucose-6-phosphatase inhibitors and as glucose intestinal absorption inhibitors, using in situ rat intestinal segments.

Testing the monophyly of Aesculus L. and Billia Peyr., woody genera of tribe Hippocastaneae of the Sapindaceae.

Hippocastaneae is a well-supported clade in Sapindaceae that comprises 15+ species; 12+ in Aesculus, two in Billia, and one in Handeliodendron Rehder. The monophyly of Aesculus and Billia were widely assumed, but a recent molecular phylogenetic study of Sapindanceae used seven species of Aesculus and one each of Billia and Handeliodendron and showed that Billia and Handeliodendron were nested within Aesculus. Here, we tested the hypothesis that Aesculus and Billia are mutually monophyletic using phylogenetic analyses of seven molecular markers and 31 accessions of Hippocastaneae representing 14 species. We performed phylogenetic analyses using a dataset of concatenated genes as well as with coalescent method for constructing a species tree from individual gene trees. The analysis of seven concatenated markers and the species tree strongly supported the mutual monophyly of Aesculus and Billia. We also recovered support for the traditional arrangement of genera within Hippocastaneae: Aesculus and Billia comprising a clade that is sister to Handeliodendron. However, the relationships among the genera remain incompletely resolved.

Procyanidins, from Castanea mollissima Bl. shell, induces autophagy following apoptosis associated with PI3K/AKT/mTOR inhibition in HepG2 cells.

Procyanidins from Castanea mollissima Bl. shell (CSPCs) induced autophagy and apoptosis in HepG2 cells and its mechanism remains to be examined. In this paper, autophagy was measured by the lipid modification of light chain-3 (LC3) and the formation of autophagosomes. Hoechst 33258 staining and flow cytometer analysis were used to measure apoptosis. The western blot analysis was used to examine the effects of CSPCs on the expression of LC3, PI3K, phosphorylation of AKT, mTOR, Bcl-2, Bad, Bax, BID and cleaved caspase 3 in HepG2 cells. The results showed that 3-methyladenine (3-MA) and apoptosis inhibitor (Z-VAD) could inhibited the death of HepG2 induced by CSPCs for 48h (150µg/mL). CSPCs induced the accumulation of autophagosomes and microtubule-associated proteins light chain 3-II (LC3-II, a marker of autophagy). P-AKT, PI3K and mTOR were significantly decreased on CSPCs exposure. However, these phenomena were not observed in the group pretreated with the autophagy inhibitor 3-MA and Z-VAD. CSPCs also induced the expression of Bad, Bax and Beclin-1 proteins and decreased the expression of Bcl-2, which was inhibited by 3-MA and Z-VAD. Moreover the apoptotic cell death could be inhibited by 3-MA. In addition, inhibition of LC3-II by siRNA-dependent knockdown attenuated the cleavage of caspase 3. These results suggested CSPCs could trigger autophagy via inhibition of the PI3K/AKT/mTOR signaling pathway, enhanced apoptosis in HepG2 cells which may be associated with the mitochondria-dependent signaling way.

Kinetics of co-pyrolysis of sawdust, coal and tar.

Two coals, sawdust and coal tar were selected to prepare briquettes. Thermogravimetric analyses at three heating rates (i.e. 10, 20 and 30°C/min) and up to 1000°C were carried out with the briquette components. Four blends were prepared and the experimental decomposition profiles were compared with the calculated data taking into account the amount of each component in the blend. No interaction was found when comparing the experimental and calculated decomposition profiles of the blends. Isoconversional models OFW (Ozawa-Flynn-Wall) and KAS (Kissinger-Akahira-Sunose) were used to obtain the activation energies of the blend components. The activation energies obtained were introduced in the Coats-Redfern (CR) model to derive the pre-exponential factors. The thermal decomposition profiles calculated using the kinetic parameters were in good agreement with the experimental results in the case of the briquette components, but worse results were obtained in the case of the blends due to their greater complexity.

Comparative analysis of the antioxidant and anticancer activities of chestnut inner shell extracts prepared with various solvents.

BACKGROUND: The chestnut inner shell (CIS) has long been used as a medicinal herb for strengthening the antioxidant in the cosmetic industry. However, little is known about the antioxidant and anticancer effects of the CIS. RESULTS: The antioxidant and anticancer effects of CIS extracts (CISEs) were investigated by the use of various methods and cancer cell lines, respectively. The total polyphenol content of CISEs using ethanol, methanol, butanol, ethyl acetate and water were 53.30, 43.98, 32.16, 26.79 and 11.53 mg gallic acid equivalents g(-1), respectively. The CISEs using ethanol and methanol exhibited high antioxidant activities in the 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, ß-carotene bleaching and ferric reducing ability of plasma assays; the effects were equivalent to those of butylated hydroxytoluene. All CISEs at 2.5 mg mL(-1) were shown to have a cytotoxic effect over 50%, and the CISE using ethyl acetate at 0.6 mg mL(-1) was proved to have 90% cytotoxic effect against the tested cancer cells. CONCLUSION: The ethanol and methanol CISEs had potent antioxidant effects, and the ethyl acetate CISE had the highest cytotoxicity. These results suggest that CISEs could be used as functional ingredients for antioxidant and anticancer effects in foods as extraction solvents.

Plantation forestry under global warming: hybrid poplars with improved thermotolerance provide new insights on the in vivo function of small heat shock protein chaperones.

Climate-driven heat stress is a key factor affecting forest plantation yields. While its effects are expected to worsen during this century, breeding more tolerant genotypes has proven elusive. We report here a substantial and durable increase in the thermotolerance of hybrid poplar (Populus tremula×Populus alba) through overexpression of a major small heat shock protein (sHSP) with convenient features. Experimental evidence was obtained linking protective effects in the transgenic events with the unique chaperone activity of sHSPs. In addition, significant positive correlations were observed between phenotype strength and heterologous sHSP accumulation. The remarkable baseline levels of transgene product (up to 1.8% of total leaf protein) have not been reported in analogous studies with herbaceous species. As judged by protein analyses, such an accumulation is not matched either by endogenous sHSPs in both heat-stressed poplar plants and field-grown adult trees. Quantitative real time-polymerase chain reaction analyses supported these observations and allowed us to identify the poplar members most responsive to heat stress. Interestingly, sHSP overaccumulation was not associated with pleiotropic effects that might decrease yields. The poplar lines developed here also outperformed controls under in vitro and ex vitro culture conditions (callus biomass, shoot production, and ex vitro survival), even in the absence of thermal stress. These results reinforce the feasibility of improving valuable genotypes for plantation forestry, a field where in vitro recalcitrance, long breeding cycles, and other practical factors constrain conventional genetic approaches. They also provide new insights into the biological functions of the least understood family of heat shock protein chaperones.

CsRAV1 induces sylleptic branching in hybrid poplar.

• Sylleptic branching in trees may increase significantly branch number, leaf area and the general growth of the tree, particularly in its early years. Although this is a very important trait, so far little is known about the genes that control this process. • This article characterizes the Castanea sativa RAV1 gene, homologous to Arabidopsis TEM genes, by analyzing its circadian behavior and examining its winter expression in chestnut stems and buds. Transgenic hybrid poplars over-expressing CsRAV1 or showing RNA interference down-regulated PtaRAV1 and PtaRAV2 expression were produced and analyzed. • Over-expression of the CsRAV1 gene induces the early formation of sylleptic branches in hybrid poplar plantlets during the same growing season in which the lateral buds form. Only minor growth differences and no changes in wood anatomy are produced. • The possibility of generating trees with a greater biomass by manipulating the CsRAV1 gene makes CsRAV1 transgenic plants promising candidates for bioenergy production.

CYP1, a hypovirus-regulated cyclophilin, is required for virulence in the chestnut blight fungus.

Cyclophilins are peptidyl-prolyl cis-trans isomerases that are highly conserved throughout eukaryotes and are the cellular target of the immunosuppressive drug cyclosporin A (CsA). We cloned cyp1, a cyclophilin A-encoding gene in the phytopathogenic fungus Cryphonectria parasitica, and showed that this gene was downregulated following infection by a virulence-attenuating hypovirus. The function of cyp1 was further investigated by construction of a cyp1 deletion mutant. Although the wild-type C. parasitica strain EP155 was sensitive to CsA, the Δcyp1 strain was highly tolerant to CsA, indicating that CYP1 was the target of CsA. Deletion of cyp1 resulted in reduced virulence when inoculated to chestnut stems. Transcriptional analysis revealed that deletion of cyp1 also reduced transcript levels for genes encoding key components of the heterotrimeric guanosine triphosphate-binding protein signalling pathway that are essential for sensing environmental cues and are involved in C. parasitica development and virulence.

Diversity and infection prevalence of endosymbionts in natural populations of the chestnut weevil: relevance of local climate and host plants.

Many insects are ubiquitously associated with multiple endosymbionts, whose infection patterns often exhibit spatial and temporal variations. How such endosymbiont variations are relevant to local adaptation of the host organisms is of ecological interest. Here, we report a comprehensive survey of endosymbionts in natural populations of the chestnut weevil Curculio sikkimensis, whose larvae are notorious pests of cultivated chestnuts and also infest acorns of various wild oaks. From 968 insects representing 55 localities across the Japanese Archipelago and originating from 10 host plant species, we identified six distinct endosymbiont lineages, namely Curculioniphilus, Sodalis, Serratia, Wolbachia, Rickettsia and Spiroplasma, at different infection frequencies (96.7%, 12.8%, 82.3%, 82.5%, 28.2% and 6.8%, respectively) and with different geographical distribution patterns. Multiple endosymbiont infections were very common; 3.18±0.61 (ranging from 1.74 to 5.50) endosymbionts per insect on average in each of the local populations. Five pairs of endosymbionts (Curculioniphilus-Serratia, Curculioniphilus-Wolbachia, Sodalis-Rickettsia, Wolbachia-Rickettsia and Rickettsia-Spiroplasma) co-infected the same host individuals more frequently than expected, while infections with Serratia and Wolbachia were negatively correlated to each other. Infection frequencies of the endosymbionts were significantly correlated with climatic and ecological factors: for example, higher Sodalis, Wolbachia and Rickettsia infections at localities of higher temperature; lower Wolbachia and Rickettsia infections at localities of greater snowfall; and higher Curculioniphilus, Sodalis, Serratia, Wolbachia and Rickettsia infections on acorns than on chestnuts. These patterns are discussed in relation to potential host-endosymbiont co-evolution via local adaptation across geographical populations.

Seed predation by mammals in forest fragments in Monteverde, Costa Rica.

Few studies have evaluated seed predation in fragmented landscapes, in which lower species diversity is expected to modifying ecological interactions. The rates of seed removal by mammals were investigated in a continuous forest and two fragmented patches of Premontane Tropical Moist Forest, in Monteverde, Costa Rica. The composition of mammalian seed-predators in each site was recorded during 16 months. The removal of four native tree species of experimental seeds: Ocotea valeriana and Ocotea whitei (Lauraceae), Panopsis costaricensis (Proteaceae) and Billia colombiana (Hippocastanaceae) in forest understories was followed during two annual fruiting seasons for each species. Results indicated similar species composition of seed-predators between continuous forest, the largest fragment (350 ha). However the smaller fragment (20 ha), had fewer seed predators. In this fragment, the specialized seed predator Heteromys desmarestianus (Rodentia) was more abundant. Unexpectedly, seed-predation in the two forest fragments and the continuous forest did not differ statistically for any of the seed species. Apparently, the higher abundance of small seed-predators in the fragments was compensated by the absence of medium and large seed-predators, like Agouti paca, Dasyprocta punctata (both Rodentia) and Pecari tajacu (Artiodactyla) recorded in continuous forest. Removal of experimentally-placed seeds was higher when the number of naturally occurring seeds in the sites was lower. This result could best be attributed to differential satiation of seed predators rather than differences in richness or abundance of seed predators.

Seed predation by mammals in forest fragments in Monteverde, Costa Rica

Few studies have evaluated seed predation in fragmented landscapes, in which lower species diversity is expected to modifying ecological interactions. The rates of seed removal by mammals were investigated in a continuous forest and two fragmented patches of Premontane Tropical Moist Forest, in Monteverde, Costa Rica. The composition of mammalian seed-predators in each site was recorded during 16 months. The removal of four native tree species of experimental seeds: Ocotea valeriana and Ocotea whitei (Lauraceae), Panopsis costaricensis (Proteaceae) and Billia colombiana (Hippocastanaceae) in forest understories was followed during two annual fruiting seasons for each species. Results indicated similar species composition of seed-predators between continuous forest, the largest fragment (350 ha). However the smaller fragment (20 ha), had fewer seed predators. In this fragment, the specialized seed predator Heteromys desmarestianus (Rodentia) was more abundant. Unexpectedly, seed-predation in the two forest fragments and the continuous forest did not differ statistically for any of the seed species. Apparently, the higher abundance of small seed-predators in the fragments was compensated by the absence of medium and large seed-predators, like Agouti paca, Dasyprocta punctata (both Rodentia) and Pecari tajacu (Artiodactyla) recorded in continuous forest. Removal of experimentally-placed seeds was higher when the number of naturally occurring seeds in the sites was lower. This result could best be attributed to differential satiation of seed predators rather than differences in richness or abundance of seed predators. Rev. Biol. Trop. 57 (3): 865-877. Epub 2009 September 30.

Pocos estudios han evaluado la depredación de semillas en ambientes fragmentados, en éstos la menor diversidad de especies debe estar modificando las interacciones ecológicas. Se investigó la remoción de semillas por mamíferos en un bosque continuo y dos fragmentos en Monteverde, Costa Rica. Se registró la composición de mamíferos en cada sitio durante 16 meses y se observó la remoción de semillas experimentales del suelo del bosque. Se utilizó semillas de cuatro especies de árboles: Ocotea valeriana, Ocotea whitei, Panopsis costaricensis y Billia colombiana, durante dos periodos anuales de fructificación cada una. Los resultados indican una composición de depredadores de semillas similar entre el bosque continuo y el fragmento de 350 ha, más diversos que el fragmento de 20 ha, en el cual el especialista en semillas Heteromys desmarestianus fue más abundante. La depredación de semillas entre el bosque continuo y los fragmentos no fue estadísticamente diferente al considerar todas las especies de semillas, debido a la mayor abundancia de depredadores pequeños, que pudo compensar la ausencia de depredadores de talla mediana y grande que sí están presentes en el bosque continuo. La depredación sobre semillas fue mayor cuando la oferta de semillas no experimentales fue menor, indicando que la saciedad de los depredadores puede estar determinando la remoción de semillas en fragmentos, más que la riqueza o abundancia de especies depredadoras.