Effects of rabbit gut passage on seed retrieval and germination of three shrub species

It has been known for a few decades that European rabbits consume seeds and fleshy fruits of native woody plants, but relevant factors in the endozoochory processes such as seed predation (chewing and digestion), sexual differentiation, or the temporal pattern of seed recovery have been little evaluated until now. In this study, we examined seed dispersal of three Mediterranean shrub species by wild rabbits through monitoring of seed retrieval and germination after gut passage. Twelve adult wild rabbits (Oryctolagus cuniculus; six males and six females) of similar size and age were fed seeds of three shrub species with fleshy fruits (Crataegus monogyna, Myrtus communis and Pistacia lentiscus). After ingestion of fruits, seeds were retrieved from dung every 12 h for a day and a half. The viability and germination of retrieved seeds were tested along with that of uneaten seeds. Between 5% and 76% of ingested seeds were retrieved from dung, with significant differences between species and sex. Most M. communis seeds were retrieved with 12–24 h after ingestion; almost all C. monogyna seeds were recovered with 0–12 h after ingestion; no seeds of P. lentiscus were recovered. Only in the case of M. communis seed was the recovery rate greater in female than in male. Passage through the rabbit gut significantly increased seed germination in M. communis, and decreased it in C. monogyna. In conclusion, wild rabbits acted in this study as predators of C. monogyna and P. lentiscus seeds and potential dispersers of M. communis seeds.     

Do frugivorous birds concentrate their foraging activities on those alien plants with the most abundant and nutritious fruits in the South African Mediterranean-climate region?

Alien plants with abundant and nutritious fruits may compete more effectively for avian dispersal services than native shrubs. This premise was examined by comparing daily foraging activity (visitation frequency, foraging bird number and foraging period) by four frugivorous bird species of different size on fruits of two native and two alien shrub species co-occurring at four different sites. Also, the quantities of seeds consumed daily by each of the four birds species from fruits of the four shrub species were measured and compared with the numbers of fruits and seeds, and the mass of fruits present in the shrub canopies as well as with their fruit monosaccharide concentrations. The quantities of seed consumed daily by four different size bird species (Columba arquatrix, Colius striatus, Pycnonotus capensis, Zosterops pallidus subsp capensis) were positively correlated with the numbers of seeds per m² of canopy area and with fruit mass and fruit monosaccharide content per m² of canopy area, as well as with the monosaccharide concentration of individual fruits, except in the C. arquatrix (African olive pigeon). All four bird species displayed the highest daily visitation frequencies on fruits of the alien Solanum mauritianum which were more abundant and nutritious than fruits of the other alien Lantana camara and fruits the natives Olea europaea subsp africana and Chrysanthemoides monilifera. They also all consumed greater quantities of seed daily from fruits of the alien S. mauritianum than from fruits of the other shrub species. These results corroborate proposals that frugivorous birds concentrate their foraging activities on those alien plants with the most abundant and nutritious fruits.     

Grazing-induced morphological and growth rate changes in Anarthrophyllum rigidum,a Patagonian leguminous shrub

Grazing influences the morphology and growth rate of shrubs, and consequently, their population dynamics. It has been shown that grazing directly affects the growth of shrubs. On the other hand, the reduction of grass biomass by herbivores reduces soil–water competition between grasses and shrubs, and indirectly, could enhance the growth of shrubs. However, the assessment of the long-term effects of grazing on the growth of shrubs in the arid Patagonia has been hampered by the lack of long and homogeneous records of plant population dynamics and primary production. In this study, we combined growth-ring and allometric analyses to assess the long-term effect of grazing on individuals of Anarthrophyllum rigidum, a leguminous shrub widely distributed across the Patagonian steppe. A. rigidum has evergreen leaves rich in proteins that constitute an important complement to the diet of sheep, particularly in winter when the abundance of grasses is reduced. Our observations indicate that individuals of A. rigidum nearby the water source used by livestock were smaller in size (35.5 cm vs. 67.39 cm), presented a larger number of basal branches (23 vs. 12), and showed slower rates of growth (8.2 mm year^?1 vs. 14.3 mm year^?1) than individuals located far from the water source. This first quantification of the long-term effects of grazing on A. rigidum in the dry Patagonian steppe suggests that beneficial effects of grazing through the reduction of grasses that compete with shrubs for soil–water should be more obvious for livestock non-preferred than preferred shrubs     

How do evergreen and deciduous species respond to shade?—Tolerance and plasticity of subtropical tree and shrub species of South-East China

Different plant species make use of resource gradients such as light in different ways. First, plant species specialize in using different parts of the gradient, resulting in niche partitioning, Second, within the section of the gradient used by a species, plants respond to a different resource supply by plasticity. Our study addressed both of these strategies, with the main objective to relate mean responses and plasticity indices of seedlings of woody species to species characteristics such as leaf habit and to variation in branch lengths and local frequency of adult trees in forest communities.A greenhouse experiment with 36 deciduous and 35 evergreen subtropical tree and shrub species was carried out to test the influence of light and nutrient availability on trait expression and plasticity of the species. The greenhouse responses of seedlings were compared with adult individuals in the field, based on a set of 46 species that occurred also in the 27 permanent plots in a secondary subtropical broadleaved forest in Zhejiang Province (SE-China).In the greenhouse experiment, most variables showed significant differences between unshaded (250 μE m⁻² s⁻¹) and shaded (10 μE m⁻² s⁻¹) treatments as well as between high and low nutrient supply. Deciduous species were more plastic than evergreen species in their response to light. No significant correlations were detected in mean responses and plasticity between juvenile plants in the greenhouse and adult trees in the field. However, some trait values such as biomass and node density as well as plasticity of several traits were positively related to the species’ abundance in the field, suggesting that locally abundant species tend to be less “plastic” than locally rare species.     

An appraisal of seed enumeration and videographic techniques for determining seed removal rates by birds

We examined the efficacy of seed enumeration and videographic techniques for determining seed removal by birds from indigenous (Chrysanthemoides monilifera and Olea europaea subsp africana) and alien (Lantana camara and Solanum mauritianum) shrubs at different study sites in the Cape Floristic Region. The seed enumeration technique involved counting the numbers of fruits and associated seeds removed monthly by birds, excluding those naturally abscised, from the shrub canopy. The videographic technique involved visual counts from images of the numbers of fruits and associated seeds consumed by birds over specific time intervals captured by a digital camcorder. Daily seed removal rates by all birds, irrespective of species, measured by both techniques were similar with no significant interactions evident between measuring techniques, site and shrub species. Both techniques displayed higher seed removal from tiny‐seeded S. mauritianum than other shrub species; this was also evident among individual bird species. However, the seed enumeration technique was unable to discriminate between foraging organisms, contamination of traps by wind‐blown fruits abscised from neighbouring branches and fruit theft from the canopy and the traps. In contrast, the videographic technique provided permanent visual and time‐lapse records for individual foraging bird species allowing greater measurement precision and interpretation of fruit removal behaviour by birds. We recommend use of the videographic technique over the seed enumeration technique for studying vertebrates’ seed removal in a detailed manner.     

Fruit nutritional composition and non-nutritive traits of indigenous South African tree species

Frugivorous animals play a major role in dispersing tropical, and to a lesser extent, temperate tree species. In order to attract potential seed dispersers, plants generally offer a reward of fleshy fruit pulp. Criteria for fruit choice by avian frugivores are influenced by a number of non-nutritive (e.g. fruit size and colour) factors; and nutritional composition of the fruit. There is a paucity of nutritional composition and other fruit trait data of indigenous South African fruit. This information is necessary in order to determine which frugivores are likely to ingest which fruits and consequently act as potential seed dispersal agents. This information would provide us with an understanding of the inter-relationships between indigenous fruit and frugivores in South Africa. Consequently nutritional composition was investigated in various indigenous fruit species that avian frugivores feed on. Fruits were collected from 38 indigenous tree species found in KwaZulu-Natal Afromontane and coastal forests. Pulp was freeze-dried to constant mass and then analysed for sugar, lipid and protein content; and for water content determination. Fruit width in this study ranged from 4 mm (Searsia rehmanniana and Trema orientalis) to 40 mm (Annona senegalensis, Ficus sur and Xylotheca kraussiana). Of the fruits examined in this study 29% were black and 43% were red when ripe. Most (84%) fruit species analysed for sugar content were hexose dominant with 50% being fructose and 34% being glucose dominant. Only 16% of the fruit species analysed were sucrose dominant. Fruits in this study were generally observed to be high (mean: 68.1 ± 3.3%; n = 30) in water content; and low in protein and lipid content respectively (mean: 8.2 ± 0.5%; 9.3 ± 2.2%; n = 30) indicating that these fruit species could be considered as nutrient-dilute. Future studies need to determine the nutritional composition of the remaining indigenous South African fruit in order to develop a comprehensive database as well as examining non-nutritive factors.     

Cover as a simple predictor of biomass for two shrubs in Tibet

Knowledge of the quantitative relationship between plant cover and its corresponding biomass for shrubs is not well known, especially for those on the Tibetan Plateau. Based on investigations of 35 sites, 90 plots and 95 standard individuals for two typical shrub species (Rhododendron nivale Hook. f. and Sophora moorcroftiana (Benth.) Baker) across Tibet, we developed allometric models for biomass estimation from measurements of crown diameter and/or height. We found that the parameters of crown projection area (CPA), height and their product (volume) were all significantly (p < 0.01) correlated with dry mass of different organs for both species at individual level. The CPA rather than volume best predicted aboveground dry mass. This is because that the bulk density declined significantly with increasing plant height, leading to the inappropriateness for plant height itself being employed as a parameter in biomass estimation, especially for shrubs in smaller size groups. At community level, cover was tightly correlated with the aboveground, belowground and total biomass (R² = 0.97–0.99). Therefore, biomass for the two shrubs can be simply estimated by measuring plant cover, which enables rapid estimation of shrubland carbon stock at large scales by using satellite data and repeated experiments over time. This non-destructive method using cover to estimate shrub biomass can be applied not only in arid ecosystems but also in alpine or subalpine environment.     

Shrub removal facilitates recovery of wetland species in a rewetted fen

Drainage and shrub expansion are the main threats to the biodiversity of fens and fen meadows, whereas rewetting and the removal of shrubby species are frequently applied restoration measures. We examine whether removal of shrubs enhances recovery of target species in a degraded fen subjected to moderate rewetting. The study was located in the drained fen Ca?owanie (central Poland), where remnants of open fen communities and willow-invaded fens exist in former turf-pits, surrounded by degraded meadows on dried peat. All these three habitat types were included in a monitoring grid, which covered an area of 2.2 ha. Within 55 quadrats of 20 m × 20 m we monitored occurrence of 52 species, i.e. two groups of target species (fen indicators and wet meadow indicators) and indicators of two failure scenarios (degraded fen indicators and eutrophic wetland indicators), during six years following shrub removal, rewetting and re-application of conservational mowing, using a 3-step ordinal abundance scale. NMDS ordination revealed a gradual convergence of shrub removal plots and reference plots. We noticed significant effects of year and habitat type on all indicator groups, but only fen indicators have shown a clear (increasing) trend within shrub removal plots. Degraded fen indicators (ruderal and opportunistic species) initially expanded on shrub removal plots, but this effect disappeared in the following years. We conclude that shrub removal enhances establishment of target species in a moderately drained and then rewetted fen and attribute this effect to lowered competition for light. However, given high costs of this method and long-lasting problems with shrub resprouts, we recommend applying shrub removal only to recently overgrown sites, which still retained high botanical diversity. Heavily degraded fen meadows did not react on the increase of moisture, which indicates that more advanced restoration measures, such as top soil removal are needed there.     

Evaluation of zinc tolerance and accumulation potential of the coastal shrub Limoniastrum monopetalum (L.) Boiss.

The coastal shrub Limoniastrum monopetalum is capable of growth in soil containing extremely high concentrations of heavy metals. A greenhouse experiment was conducted in order to investigate the effects of a range of Zn concentrations (0–130 mmol l⁻¹) on growth and photosynthetic performance, by measuring relative growth rate, total leaf area, plant height, gas exchange, chlorophyll fluorescence parameters and photosynthetic pigment concentrations. We also determined the total zinc, nitrogen, phosphorus, sulphur, calcium, magnesium, sodium, potassium, iron and copper concentrations in the plant tissues. The study species demonstrated hypertolerance to Zn stress, since survival was recorded with leaf concentrations of up to 1700 mg Zn kg⁻¹ dry mass when treated with 130 mmol Zn l⁻¹. L. monopetalum exhibited little overall effects on photosynthetic function at Zn levels of up to 90 mmol l⁻¹. At greater external Zn concentration, plant growth was negatively affected, due in all probability to the recorded decline in net photosynthetic rate, which may be linked to the adverse effect of the metal on photosynthetic electron transport. Growth parameters were virtually unaffected by leaf tissue concentrations as high as 1400 mg Zn kg⁻¹ dry mass thus indicating that this species could play an important role in the phytoremediation of Zn-polluted areas.     

Burial depth and diameter of the rhizome fragments affect the regenerative capacity of a clonal shrub

Clonal plants in highly disturbed habitats are often broken into small fragments of various sizes and buried at various soil depths. As a storage organ, rhizome fragments play an important role in enabling plants to survive in such habitats. But few studies have been concerned about the regenerative capacity of rhizome fragments of clonal shrubs of different rhizome diameter and at different burial depths. Here, we investigated whether deeper burial decreased, and diameter of the rhizome fragment increased, the regenerative capacity of a clonal shrub. Research samples of rhizome fragment (rhizome diameters of 2, 5, 10, 15, and 20 mm) of the clonal shrub Calligonum arborescens were buried at different depths (0, 1, 5, 10, and 20 cm). Increasing the diameter of the rhizome fragments significantly increased the survival rate of fragments, and increased the above-ground, below-ground and total biomass production of fragments. Vegetative reproduction ability also increased with an increase in diameter of the rhizome fragments. With an increase in sand burial depth, above-ground, below-ground, total biomass production and vegetative reproduction ability first decreased and then increased, and no fragments survived at the 0 cm burial depth. These results indicate that sand burial depth and diameter of the rhizome fragments significantly affected the regeneration capacity of C. arborescens. Sand burial is one of the essential prerequisites for C. arborescens rhizome fragments’ survival. Moderate burial depth (5 cm) and larger fragment diameter (20 mm diameter) were more suitable for biomass production and vegetative reproduction. These results indicate that reserves stored in rhizome fragments can contribute greatly to the regeneration capacity of the C. arborescens—responses that are very important for C. arborescens survival and establishment in frequently disturbed habitats.     

Understory cover and biomass indices predictions for forest ecosystems of the Northwestern United States

The understory community is a critical component of many processes of forest ecosystems. Cover and biomass indices of shrubs and herbs of forested ecosystems of Northwestern United States are presented. Various forest data were recorded for 10,895 plots during a Current Vegetation Survey, over the National Forest lands of entire Pacific Northwest. No significant relationships between the percent canopy cover and understory percent cover and biomass indices were found for the 129 ecoclasses analyzed. Disturbance time and type, and the soil characteristics significantly influenced the shrub biomass indices (p-values of <0.001, <0.001, and 0.01, respectively). Only disturbance time and type significantly influenced the shrub percent cover (p-values <0.001). There were no significant interactions between these variables. No significant differences were found for herb biomass indices and cover. Climate variables are reasonable predictors of understory cover and biomass indices. Elevation and slope are also influential: understory cover decreases with altitude, while understory biomass increases with slope. Most models showed weak predictive power (adjusted R-squared ≤ 0.27). However, robust models for the maximum/potential understory biomass indices for the forested areas in the Northwestern United States are reported (adjusted R-squared of 0.76 and 0.51 for shrubs and herbs, respectively). Overall, our study provides conceptual and statistical models for the understory of the National Forest lands of the Pacific Northwest. The results are comparable with other models for the area, suggesting that the predictions regarding understory vegetation are inherently difficult.     

Focal species diversity patterns can provide diagnostic information on plant invasions

In Europe, coastal sandy habitats are considered highly endangered among those included in the EC Directive 92/43/EEC (Habitats Directive). Among the different threats which affect coastal communities, the spread of alien plants has been claimed to induce changes in community diversity and structure. We therefore set out to analyse diversity patterns of native and focal species (diagnostic and characteristic of coastal dune habitats of European conservation interest) in sandy coastal habitats invaded by Carpobrotus aff. acinaciformis, a widespread alien plant. Focal species are a major conservation target for the Habitats Directive and their decline should be considered a serious threat for the whole habitat. The study was performed in the Central Western coast of Italy. We randomly sampled the vegetation of the holocenic dune by 2 m × 2 m plots. First we split the collected data in two sets: invaded and non-invaded. We compared overall native and focal species richness patterns of the two sets by rarefaction curves. Then, in order to describe the singular aspects of species diversity (e.g. richness, Shannon index, Simpson index, Berger–Parker index), we also compared Rènyi's diversity profiles and we tested the significance of the differences between invaded and non invaded sets using a bootstrap procedure. Rarefaction curves of the non-invaded set rise quickly and reach higher accumulation values than the invaded set, but differences between the two curves were not significant. With respect to Rènyi's profiles, the profile for the invaded dataset was always below the non-invaded one, but differences in diversity were significant only when specifically considering the focal species (Shannon, Simpson and Berger–Parker indices). In the analysed case, the invasion is significantly associated with changes in focal species diversity, instead those differences are not evident on the all native species pool. In the case of recent invasions, a consistent decline on focal species diversity may represent an early alarm sign of diversity loss and may help define specific conservation actions to prevent the decrease of overall diversity.     

Changes in traits of shrub canopies across an aridity gradient in northern Patagonia, Argentina

Low and highly variable precipitation pulses exert a strong selective pressure on plant traits and this might provide axes of ecological differentiation among plant species in arid ecosystems. We asked whether aridity contributes to maintain high diversity of species and morphotypes in shrub canopies. We selected eleven study sites evenly distributed across a 400-km transect in northern Patagonia, Argentina. Precipitation is low and highly variable within and between years but almost homogeneous across the transect (125–150 mm). Mean annual temperature varied, however, ranging from 8 °C (west) to 13.5 °C (east) creating a west–east gradient of aridity (aridity index from 3.7 to 7.3, respectively). Sheep grazing commenced in the early 1900s at a similar intensity across the transect. We recorded the richness and cover of shrubs by species and by morphotypes (drought deciduous tall shrubs, evergreen tall shrubs, medium shrubs, and dwarf shrubs), and further calculated the species and morphotype Shannon diversity index at each site. We assessed the presence of spiny leaves, leaf pubescence, thorny stems, and photosynthetic stems in shrub species of all morphotypes and collected green leaves of the dominant shrub species (more than 80% of the total shrub cover) to assess the leaf area, leaf mass per unit area, N-, lignin- and soluble phenolic-concentrations per species at each site. Richness and diversity of shrub species and morphotypes were positively associated with aridity. The richness and diversity of shrub species with pubescent leaves and thorny stems, and nitrogen concentration in green leaves of dominant shrubs increased with increasing aridity. We conclude that our findings on increased diversification in life history traits, species and morhotypes in shrub canopies with increasing aridity support the hypothesis that variability in aridity provides axes of ecological differentiation among shrub species facilitating their coexistence.     

How to efficiently obtain accurate estimates of flower visitation rates by pollinators

Regional declines in insect pollinators have raised concerns about crop pollination. Many pollinator studies use visitation rate (pollinators/time) as a proxy for the quality of crop pollination. Visitation rate estimates are based on observation durations that vary significantly between studies. How observation duration relates to the accuracy of the visitation rate estimate is, however, unknown. We studied this relationship using six day-long observations (06:00 h–19:00 h) in leek-seed production fields (totalling 78 h). We analysed beyond which point in time observing longer did not significantly improve the accuracy of the visitation rate estimate (minimum observation duration). We furthermore explored the relationship between the minimum observation duration and visitation rate, time of day and temperature. We found that the minimum observation duration (mean ± SD: 24 ± 11.9 min) was significantly related to visitation rate, where the observation time required to obtain accurate estimates decreased with increasing visitation rate. Minimum observation duration varied greatly between days and between fields but not within days. Within days, the visitation rates differed significantly only between the hour-intervals 06:00 h–07:00 h (lowest visitation rate) and 09:00 h–11:00 h (highest rate). Minimum observation duration decreased up to around 22 °C beyond which it remained fairly stable. Surprisingly, even after three day-long observations on the same plant we found new pollinator species visiting the flowers, suggesting that species-richness estimates based on plant observations alone probably underestimate true species richness. Because especially between-day variation in visitation rate on single plants can be large, reliable estimates of the pollinator visitation rate during the plant’s flowering time require observations on multiple days. Standardising the number of pollinators rather than the time to observe (standardised pollinator timing approach: time to n-pollinator visits) may provide more consistent accurate assessments of visitation rate, especially for studies that use gradients in visitation rates to examine the contribution of pollinators to crop pollination.     

Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources

This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics – Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole (25 μg), Nalixidic acid (30 μg), Trimethoprim (1.25 μg), Novobiocin (5 μg), Penicilin (10 μg) and Chloramphenicol (10 μg) was tested. The results obtained from this study reveal multi drug resistance pattern among the isolates. All the isolates were completely resistant to Ampicillin, Novobiocin, Sulphamethoxazole and Trimethoprim, respectively but susceptible to Tetracycline (100%), Kanamycin (5.7%), Gentamicin (5.7%) and Oxytetracycline (24.5%). Antibiotics phenotyping of the bacteria revealed 21 different phenotypes among the isolates.     

Factors affecting cactus recruitment in semiarid Chile: A role for nurse effects?

The nurse-protégé hypothesis states that adult plants of one species provide micro-environmental conditions that favor the establishment of seedlings of a second species with no effect for the first species. Several studies suggest this effect should be prevalent in arid and semiarid zones as adult plants often provide shelter from low moisture and high temperature. Echinopsis chiloensis and Eulychnia acida are endemic columnar cacti that inhabit the arid and semiarid zones of Chile. In this study, we examined the pattern of recruitment of both cactus species at Reserva Nacional Las Chinchillas, located ～60 km east from the Pacific coast. We determined number, growth and survivorship of young cacti (<30 cm height) through biannual monitoring between 2009 and 2012 in microhabitats that strongly differ in their abiotic variables (minimum and maximum temperature and mean relative humidity, moisture content, and physical and chemical soil characteristics), under five different shrub species and in open spaces, and examined the association of these cacti with potential nurse plants. Most young cacti occurred under shrubs, the microhabitat having the lowest mean and maximum temperatures and the highest relative humidity. In particular, E. chiloensis and E. acida were found under the shrubs Flourensia thurifera and Bahia ambrosioides, respectively, in a higher frequency than expected by chance, suggesting that these shrub species behave as nurse plants through species-specific effects than are not accounted for by differences in soil nutrients.     

The productive traits,fatty acid profile and nutritional indices of three lupin (Lupinus spp.) species cultivated in a Mediterranean environment for the livestock

The aim of this study was to investigate the productive traits and fatty acid profiles of eight sweet varieties of Lupinus albus, Lupinus angustifolius and Lupinus luteus cultivated in a Mediterranean environment (Sicily, southern Italy). This is being done to identify the most suitable species, from an agronomical and nutritional point of view, for supporting animal husbandry in this area. Seed yield, oil quantity and oil quality were considerably different depending on the genotype. L. albus showed the highest yield (2.0–2.4 t/ha), 100 seed weight (27.0 g, on average), oil content (70.4–75.5 g/kg), and n?3/n?6 polyunsaturated fatty acid ratio (0.39–0.51), L. angustifolius showed the highest content of saturated fatty acids (21.03–26.66 g/100 g) and L. luteus showed the highest levels of n?6 polyunsaturated fatty acids (47.56–50.85 g/100 g), unsaturated/saturated (4.93–5.62) and polyunsaturated/saturated (3.33–3.99) fatty acid ratios and the best atherogenic (0.06) and thrombogenic (0.09–0.11) indices. The data obtained suggests L. albus as the most interesting lupin species for aiding the crop-livestock food chain and L. luteus as a promising crop due to its high nutritive traits for the Mediterranean environment.     

Saxitoxin monitoring in three species of Florida puffer fish

Beginning in April 2002, three species of Florida puffer fish from around the state of Florida, USA were monitored for the presence of saxitoxin (STX). In total, 873 southern (Sphoeroides nephelus), 171 checkered (S. testudineus), and 53 bandtail (S. spengleri) puffer fish were collected between 2002 and 2006 from eight regions: Jacksonville, the Indian River Lagoon, Tequesta, the Florida Keys, Charlotte Harbor, Tampa Bay, Cedar Key, and Apalachicola. Emphasis was placed on collecting specimens from the Indian River Lagoon (IRL), where recreational harvesting of puffer fish led to 28 cases of saxitoxin puffer fish poisoning (SPFP) between January 2002 and May 2004. Southern puffer fish from the northern IRL routinely contained the highest concentrations of STX, with average levels in the skin of 1787 μg STXequiv./100 g tissue. Elevated concentrations were also found in the muscle (1102 μg STXequiv./100 g), gut contents (539 μg STXequiv./100 g), gonads (654 μg STXequiv./100 g), and liver (214 μg STXequiv./100 g). Lower, yet significant (above the action limit of 80 μg STXequiv./100 g tissue), concentrations of STX were also detected in the skin (599 μg STXequiv./100 g), muscle (233 μg STXequiv./100 g), gut contents (197 μg STXequiv./100 g), and gonads (239 μg STXequiv./100 g) of southern puffer fish from Tequesta in the southern IRL, as well as in the gonads (122 μg STXequiv./100 g) of Jacksonville southern puffer fish and the skin (265 μg STXequiv./100 g) of Tampa Bay southern puffer fish. STX concentrations above the action limit were also found in the skin of bandtail puffer fish from the IRL (620 μg STXequiv./100 g), Tequesta (374 μg STXequiv./100 g), and the Florida Keys (230 μg STXequiv./100 g). Checkered puffer fish collected from the IRL, Tequesta, and the Florida Keys on average were nontoxic, containing STX levels below the action limit in all tissues.     

Cellular and nephrotoxicity of selenium species

ProjectBeside its useful functions at very low concentrations, selenium including supplementary Se sources pose a potential toxicological risk. The toxicity of selenium species was tested in HaCaT cell culture and related nephrotoxicity in mice.ProcedureThe apoptotic shrinkage and necrotic expansion of cells were measured by time-lapse image microscopy. Acute nephrotoxicity was estimated upon administration of various selenium species to mice for two weeks. To confirm or to refute the accumulation of Se in the kidney and its potential chronic effect, Se concentration in kidney tissue and histopathlology were tested.ResultsThe comparison of selenium species showed that organic lactomicroSe did not affect cell growth at 5 ppm, but inorganic nanoSe severely hampered it at lower concentration (1 ppm). The in vivo Se treatment (0.5, 5, 50 ppm, corresponding to 4, 40 and 400 μg/kg) was misleading as it did neither affect the outward appearance nor the weight of the kidney. Se accumulation was observed after selenate, selenite, SelPlex, selenite and nanoSe administration, while lactomicroSe caused no traceable accumulation. In vivo, ex vivo and in vitro experiments reflected this order of selenium toxicity: selenate > selenite > SelPlex = nanoSe > lactomicroSe.ConclusionWithin the tested species lactomicroSe was the only non-nephrotoxic selenium source recommended for nutritional Se supplementation.