Feeding habits of the whale shark (<Emphasis Type="Italic">Rhincodon typus</Emphasis>) inferred by fatty acid profiles in the northern Mexican Caribbean

Whale shark (Rhincodon typus, Smith, 1828) is an endangered species with anthropogenic pressures due to increasing demand of encounter tourism activities. Research efforts to identify management and conservation strategies for this species are needed. The Northern Mexican Caribbean is one of the most important feeding aggregation sites of whale sharks worldwide. In this study, Mexican Caribbean whale shark feeding habits are assessed by means of fatty acid (FA) signature analysis, a biochemical non-destructive technique widely applied in trophic ecology studies. Sub-dermal tissue biopsies of 68 whale sharks and samples of their potential prey (zooplankton) were collected during 2010 and 2011 in two areas with high R. typus abundance. Zooplankton samples (n?=?17) were divided in two categories: mixed zooplankton (several groups of zooplankton) and fish eggs (> 95% of sample components were fish eggs). FA profiles of whale shark tissue sampled between years showed significant variability; while there was no intraspecific differences in FA signature related to sex, size and location. FA profiles of whale sharks and their potential prey were dominated by saturated fatty acids (SFA). R. typus FA signature was significantly different from that of mixed zooplankton; on the other hand, whale shark and fish egg FA profiles formed groups with overlapping values and registered high levels of oleic acid. PUFA average ω3/ ω6 ratio on whale shark FA profiles for both years was below 1. Arachidonic acid (ARA) percentage was higher in whale shark biopsies (13.2% in 2010, 6.8% in 2011) compared to values observed in fish eggs (2.0%) and mixed zooplankton (1.4%). Similarity between FA profiles of whale sharks and fish eggs, low levels of bacterial FA found in R. typus biopsies, as well as whale shark feeding behavior observations in the study area, suggest that R. typus is feeding mainly on surface zooplankton in Mexican Caribbean; however, elevated ARA percentages in whale shark samples may indicate that this species has complementary feeding sources, such as demersal zooplankton, which has been reported in other aggregation sites. Results obtained contribute to the knowledge of the whale shark trophic ecology in the area, but are inconclusive. Further studies are recommended to evaluate whale shark FA profiles from different tissues (muscle or blood); also, broader information is needed about zooplankton FA signature in the study area.     

Photosynthesis and antioxidative defense mechanisms in deciphering drought stress tolerance of crop plants

Crop plants are regularly exposed to an array of abiotic and biotic stresses, among them drought stress is a major environmental factor that shows adverse effects on plant growth and productivity. Because of this these factors are considered as hazardous for crop production. Drought stress elicits a plethora of responses in plants resulting in strict amendments in physiological, biochemical, and molecular processes. Photosynthesis is the most fundamental physiological process affected by drought due to a reduction in the CO₂ assimilation rate and disruption of primary photosynthetic reactions and pigments. Drought also expedites the generation of reactive oxygen species (ROS), triggering a cascade of antioxidative defense mechanisms, and affects many other metabolic processes as well as affecting gene expression. Details of the drought stress-induced changes, particularly in crop plants, are discussed in this review, with the major points: 1) leaf water potentials and water use efficiency in plants under drought stress; 2) increased production of ROS under drought leading to oxidative stress in plants and the role of ROS as signaling molecules; 3) molecular responses that lead to the enhanced expression of stress-inducible genes; 4) the decrease in photosynthesis leading to the decreased amount of assimilates, growth, and yield; 5) the antioxidant defense mechanisms comprising of enzymatic and non-enzymatic antioxidants and the other protective mechanisms; 6) progress made in identifying the drought stress tolerance mechanisms; 7) the production of transgenic crop plants with enhanced tolerance to drought stress.     

Reduced impact of ocean acidification on growth and swimming performance of newly hatched tropical sharks (Chiloscyllium plagiosum)

ABSTRACT

Sharks have been facing unprecedented pressure over the last decades, and ocean acidification may represent an additional threat, particularly during their most susceptible life stages. Hence, the present study aimed to investigate the effects of ocean acidification (control pCO₂ ~ 400 μatm; high pCO₂ ~ 900 μatm) on the growth, swimming performance and cholinergic system of juvenile white-spotted bamboo sharks (Chiloscyllium plagiosum). After 45 days of exposure, we observed that high CO₂ did not affect most of the end-points studied. However, somatic growth rate and the percentage of time that sharks spent swimming was significantly reduced under high CO₂ conditions. Moreover, AChE activity decreased in two of the seven brain macroareas analyzed, the telencephalon and optic lobes. As this near-threatened shark species showed small sub-lethal effects to high CO₂ levels, we argue that within a longer time-frame they can potentially reduce individual performance with cascading consequences to shark population dynamics.     

Responses of a coral reef shark acutely exposed to ocean acidification conditions

Anthropogenic ocean acidification (OA) is a threat to coral reef fishes, but few studies have investigated responses of high-trophic-level predators, including sharks. We tested the effects of 72-hr exposure to OA-relevant elevated partial pressures of carbon dioxide (pCO₂) on oxygen uptake rates, acid–base status, and haematology of newborn tropical blacktip reef sharks (Carcharhinus melanopterus). Acute exposure to end-of-century pCO₂ levels resulted in elevated haematocrit (i.e. stress or compensation of oxygen uptake rates) and blood lactate concentrations (i.e. prolonged recovery) in the newborns. Conversely, whole blood and mean corpuscular haemoglobin concentrations, blood pH, estimates of standard and maximum metabolic rates, and aerobic scope remained unaffected. Taken together, newborn blacktip reef sharks appear physiologically robust to end-of-century pCO₂ levels, but less so than other, previously investigated, tropical carpet sharks. Our results suggest peak fluctuating pCO₂ levels in coral reef lagoons could still physiologically affect newborn reef sharks, but studies assessing the effects of long-term exposure and in combination with other anthropogenic stressors are needed.

     

The effect of polyethylene glycol-induced drought stress on photosynthesis,carbohydrates and cell membrane in <Emphasis Type="Italic">Stevia rebaudiana</Emphasis> grown in greenhouse

Drought stress is one of the major environmental stresses that limit crop production in arid regions. A greenhouse culture experiment was conducted to evaluate the response of an agronomically and economically important sweet medical herb (Stevia rebaudiana) to polyethylene glycol (PEG 6000)-induced drought stress (5, 10, and 15% (w/v) PEG, equivalent to leaf water potentials of ??0.49, ??1.40 and ??2.93 MPa, respectively) for 1 month. Plant mass, a major determinant of Stevia yield, showed a reduction after PEG treatments. PEG-reduced photosynthesis traits included the maximal quantum yield of photosystem II (F_v/F_m), efficiency of photosystems I and II (PI_abs), intercellular CO₂, net photosynthesis, chlorophylls, carotenoids and water use efficiency, followed by the reduction of carbohydrates. Under PEG treatment, the reactive oxygen species (ROS) accumulation occurred and plants exhibited an increase in H₂O₂ generation. Consequently, an increase in malondialdehyde and electrolyte leakage was evident in PEG treatment, indicating membrane lipid peroxidation. In PEG-treated plants, the ROS accumulation was accompanied by an increase in activity of some enzymatic and non-enzymatic antioxidants. Leaf extracts of PEG-treated plants showed lower superoxide anion, hydroxyl and nitric oxide radical scavenging activity than control plants. Drought stress also caused the accumulation of the compatible solutes proline and glycine betaine. Collectively, the results demonstrated that PEG-induced oxidative stress, due to insufficient antioxidant mechanisms, provoked damages to cell membrane and photosynthetic apparatus, with consequently reduced carbohydrates and plant growth. These results are of basic importance as vegetative growth is the major determining criterion for Stevia crops and adequate irrigation is crucial for obtaining higher yield.     

Glutathione improves survival of cryopreserved embryogenic calli of <Emphasis Type="Italic">Agapanthus praecox</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis>

The effect of supplementation of reduced glutathione (GSH) to cryoprotectant solution on the generation of reactive oxygen species (ROS) (e.g., H₂O₂, OH^·, and O ₂ ^·? ) and antioxidants (e.g., SOD, POD, CAT, AsA, and GSH), as well as membrane lipid peroxidation (i.e., MDA content) mitigation in cryopreserving of embryogenic calli (EC) of Agapanthus praecox subsp. orientalis was investigated. The vitrification-based cryopreservation method was used in this study. The addition of GSH at a final concentration of 0.08 mM to the cryoprotectant solution has significantly improved cryotolerance of A. praecox EC. The EC post-thaw survival rate increased by 68.34 % using the cryoprotectant solution containing 0.08 mM GSH as compared to the control (GSH-free). EC treated with GSH displayed the reduction in  OH^· generation activity and the contents of H₂O₂ and MDA, as well as enhancement in the inhibition of O ₂ ^·? generation and the antioxidant activity. Treatment with exogenous GSH also increased endogenous AsA and GSH contents after dehydration step. Expression of stress-responsive genes, e.g., peroxidase (POD), peroxiredoxin, ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), and glutathione peroxidase (GPX), was also increased during cryopreservation processes. The expression of DAD1 (Defender against apoptotic cell death) was elevated, while cell death-related protease SBT was suppressed. These results demonstrated that the addition of GSH to cryoprotectant solution affects the ROS level and could effectively improve survival of A. praecox EC through enhancing antioxidant enzyme activities and decreasing cell death.     

Reactivities of Shark 19S and 7S IgM Antibodies to <Emphasis Type="Italic">Salmonella typhimurium</Emphasis>

LOWER vertebrates such as sharks can synthesize humoral antibodies in response to antigenic stimulation with a wide variety of antigens¹. Physicochemical studies have shown that sharks can synthesize both 19S and 7S immunoglobulins and that these two proteins belong to the same immunoglobulin class, which seems to be structurally homologous to IgM as defined for higher animals. Thus the shark immunoglobulins have been designated 19S IgM and 7S IgM^2–4. Because the predominant immunoglobulin (IgG) of most mammals is absent from sharks, the shark monomeric (7S) IgM might be functionally analogous to IgG. One example of the functional differences between IgM and IgG antibodies is the greater reactivity of the former in agglutination and bactericidal reactions^5,6. We have isolated and characterized functionally the relatively high levels of agglutinating antibodies which the nurse shark, Gingly-mostoma cirratum, synthesizes in response to Salmonella typhimurium “O” antigens.     

Post-release fishing mortality of blue (<Emphasis Type="Italic">Prionace glauca</Emphasis>) and silky shark (<Emphasis Type="Italic">Carcharhinus falciformes</Emphasis>) from a Palauan-based commercial longline fishery

Accounting for components of fishing mortality, including post-release mortality (F_r), is necessary for robust assessments of the effects of fishing. Forty-eight blue (Prionace glauca) and 35 silky sharks (Carcharhinus falciformes) were tagged with pop-up satellite archival tags to monitor F_r rates from pelagic longline vessels in the western tropical Pacific Ocean. There is a paucity of F_r studies at low latitudes and identifying factors that significantly explain F_r is critical for understanding fishing mortality. Mean F_r rates were 0.17 [95% CI 0.09–0.30] for blue shark and 0.20 [95% CI 0.10–0.36] for silky shark. When it occurred, F_r was acute with 87% of mortalities within 2 days of release. Several prognostic operational, environmental, biological and handling variables were evaluated to assess their influence on survival outcomes. Using Kaplan–Meier survival curves, logistic regression, accelerated failure time and Cox proportional hazards models to screen variables, the only significant prognostic or risk variable was health condition at haulback. There was close correspondence (~?83% accuracy) between condition at capture and survival outcomes. Reliable methods to classify at-vessel condition represent an inexpensive and simple metric for estimating both F_r and at-vessel (F_c) mortality rates. Examining F_c rates in detail in longline fisheries using capture information on depth, temperature and dissolved oxygen that may act in synergy with condition code and hooking duration is a research priority. Results suggest that a large proportion of shark survive following release and that F_r rates can be increased by improving the haulback condition of captured sharks.     

Odor tracking in sharks is reduced under future ocean acidification conditions

Recent studies show that ocean acidification impairs sensory functions and alters the behavior of teleost fishes. If sharks and other elasmobranchs are similarly affected, this could have significant consequences for marine ecosystems globally. Here, we show that projected future CO₂ levels impair odor tracking behavior of the smooth dogfish (Mustelus canis). Adult M. canis were held for 5 days in a current‐day control (405 ± 26 μatm) and mid (741 ± 22 μatm) or high CO₂ (1064 ± 17 μatm) treatments consistent with the projections for the year 2100 on a ‘business as usual’ scenario. Both control and mid CO₂‐treated individuals maintained normal odor tracking behavior, whereas high CO₂‐treated sharks significantly avoided the odor cues indicative of food. Control sharks spent >60% of their time in the water stream containing the food stimulus, but this value fell below 15% in high CO₂‐treated sharks. In addition, sharks treated under mid and high CO₂ conditions reduced attack behavior compared to the control individuals. Our findings show that shark feeding could be affected by changes in seawater chemistry projected for the end of this century. Understanding the effects of ocean acidification on critical behaviors, such as prey tracking in large predators, can help determine the potential impacts of future ocean acidification on ecosystem function.     

Physiological,biochemical, antioxidant and growth characterizations of gibberellin and paclobutrazol-treated sweet leaf (<Emphasis Type="Italic">Stevia rebaudiana</Emphasis> B.) herb

A greenhouse experiment was designed to study the responses of Stevia rebaudiana herb to paclobutrazol (PBZ) and gibberellin (GA) treatments. GA and PBZ treatments caused no significant impact on photosynthesis pigments while they increased carbohydrates, amino acids and protein metabolites. Stevia showed a potent antioxidant activity through scavenging DPPH, NO^·; O ₂ ^·? and OH^· radicals which was highlighted in GA and PBZ treatments. The enzymatic and non-enzymatic antioxidant system of Stevia plant showed a significant increase in response to PBZ and GA treatments. PBZ treatment decreased plant growth while GA treatment had no significant effect on it. Collectively, both GA and PBZ treatments effectively increased metabolites and antioxidant property of Stevia herb.     

Structural and physiological responses of <Emphasis Type="Italic">Halodule wrightii</Emphasis> to ocean acidification

Coastal areas face high variability of seawater pH. Ocean acidification (OA) and local stressors are enhancing this variability, which poses a threat to marine life. However, these organisms present potential phenotypic plasticity that can offer physiological and structural tools to survive in these extreme conditions. In this study, we evaluated the effects of elevated CO₂ levels and consequent pH reduction on the physiology, anatomy and ultrastructure of the seagrass Halodule wrightii. A mesocosm study was conducted in an open system during a 30-day experiment, where different concentrations of CO₂ were simulated following the natural variability observed in coastal reef systems. This resulted in four experimental conditions simulating the (i) environmental pH (control condition, without CO₂ addition) and (ii) reduced pH by ? 0.3 units, (iii) ? 0.6 units and (iv) ? 0.9 units, in relation to the field condition. The evaluated population only suffered reduced optimum quantum yield (Y(II)), leaf width and cross-section area under the lowest CO₂ addition (? 0.3 pH units) after 30 days of experiment. This fitness commitment should be related to carbon concentration mechanisms present in the evaluated species. For the highest CO₂ level, H. wrightii demonstrated a capacity to compensate any negative effect of the lowest pH. Our results suggest that the physiological behaviour of this primary producer is driven by the interactions among OA and environmental factors, like irradiance and nutrient availability. The observed behaviour highlights that high-frequency pH variability and multifactorial approaches should be applied, and when investigating the impact of OA, factors like irradiance, nutrient availability and temperature must be considered as well.     

Physiological and biochemical changes in different drought-tolerant alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.) varieties under PEG-induced drought stress

Medicago sativa L. cv. Longzhong is a nutritious forage plant in dryland regions of the Loess Plateau with strong drought tolerance and broad adaptability. To understand the adaptation mechanism of alfalfa (M. sativa L. cv. Longzhong) to drought stress, growth, and physiological parameters including levels of chlorophyll content, osmotic adjustment, reactive oxygen species (ROS), and antioxidant enzymes and antioxidants were measured under simulated levels of drought (? 0.40, ? 0.80, ? 1.20, ? 1.60, and ? 2.00 MPa). The changes in M. sativa L. cv. Longzhong were compared with those of plants of M. sativa L. cv. Longdong control (Variety I) suited to moderate rainfall areas and M. sativa L. cv. Gannong No. 3 (Variety II) suited to irrigated areas. The results showed that root–shoot ratio, the chlorophyll (a + b) and osmolytes contents, the degree of lipid peroxidation and ROS production, and the levels of antioxidative enzymes and antioxidants increased significantly with increasing drought stress, whereas plant height, aboveground biomass, chlorophyll a/b ratio, leaf water potential (Ψ₁), and relative water content (RWC) decreased in response to drought. The Longzhong variety responded early to beginning drought stress (between 0 and ? 0.4 MPa) compared with the controls. Under drought stress (between ? 0.4 and ? 2.0 MPa), the Longzhong variety had significantly higher belowground biomass, root–shoot ratio, Ψ₁, RWC, catalase (CAT) activity and reduced glutathione content than those of Varieties I and II, but hydrogen peroxide and hydroxyl free radical (OH·) contents were significantly lower. Step regression analysis showed that OH·, CAT, malondialdehyde, superoxide anion-free radical (O ₂ ^·? ), and superoxide dismutase of Longzhong had the most marked response to drought stress. In conclusion, the stronger drought tolerance of the Longzhong variety might be due to its higher water-holding capacity, root–shoot ratio, and ability to coordinate enzymatic and non-enzymatic antioxidant systems, which coordinate the peroxidation and oxidative systems.     

Juvenile Greenland sharks <Emphasis Type="Italic">Somniosus microcephalus</Emphasis> (Bloch &amp; Schneider, 1801) in the Canadian Arctic

Life-stage-based management of marine fishes requires information on juvenile habitat preferences to ensure sustainable population demographics. This is especially important in the Arctic region given very little is known about the life histories of many native species, yet exploitation by developing commercial and artisanal fisheries is increasing as the ice extent decreases. Through scientific surveys and bycatch data from gillnet fisheries, we document captures of rarely reported juvenile Greenland sharks (Somniosus microcephalus; ≤200 cm total length [TL]) during the ice-free period in the Canadian Arctic. A total of 22 juvenile animals (42 % of total catch; n = 54), including the smallest reliably measured individual of 117 cm TL, were caught on scientific longlines and bottom trawls in Scott Inlet and Sam Ford Trough over three consecutive years. Molecular genetic nuclear markers confirmed species identity for 44 of these sharks sampled; however, two sharks including a juvenile of 150 cm TL were identified as carrying a Pacific sleeper shark (Somniosus pacificus) mitochondrial cytochrome b (cyt b) haplotype. This represents the first record of a Pacific sleeper shark genetic signature in Greenland sharks in Eastern Arctic waters. Juvenile sharks caught as bycatch in gillnet fisheries were only observed offshore in Baffin Bay surrounding a fishery closure area, while larger subadult and mature Greenland sharks (>200 cm TL) were caught in all fishing locations, including areas where juveniles were observed. The repeatable occurrence of juvenile Greenland sharks in a fjord and their presence at two offshore sites indicates that these smaller animals either reside in nurseries or have defined home ranges in both coastal and offshore regions or undertake large-scale inshore–offshore movements.     

Effects of exogenous spermidine on antioxidant system of tomato seedlings exposed to high temperature stress

The effects of foliar spraying with spermidine (Spd) on antioxidant system in tomato (Lycopersicon esculentum Mill.) seedlings were investigated under high temperature stress. The high temperature stress significantly inhibited plant growth and reduced chlorophyll (Chl) content. Application of exogenous 1 mM Spd alleviated the inhibition of growth induced by the high temperature stress. Malondialdehyde (MDA), hydrogen peroxide (H₂O₂) content and superoxide anion (O₂) generation rate were significantly increased by the high temperature stress, but Spd significantly reduced the accumulation of reactive oxygen species (ROS) and MDA content under the stress. The high temperature stress significantly decreased glutathione (GSH) content and activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR), but increased contents of dehydroascorbic acid (DHA), ascorbic acid (AsA), and oxidized glutathione (GSSG) in tomato leaves. However, Spd significantly increased the activities of antioxidant enzymes, levels of antioxidants and endogenous polyamines in tomato leaves under the high temperature stress. In addition, to varying degrees, Spd regulated expression of MnSOD, POD, APX2, APX6, GR, MDHAR, DHAR1, and DHAR2 genes in tomato leaves exposed to the high temperature stress. These results suggest that Spd could change endogenous polyamine levels and alleviate the damage by oxidative stress enhancing the non-enzymatic and enzymatic antioxidant system and the related gene expression.     

The induction of salt stress tolerance by propyl gallate treatment in green microalga <Emphasis Type="Italic">Dunaliella bardawil</Emphasis>, through enhancing ascorbate pool and antioxidant enzymes activity

The effect of propyl gallate (PG), a synthetic antioxidant, on antioxidant responses and salinity tolerance was investigated in the cells of the green microalga, Dunaliella bardawil. Algal suspensions grown at three salinity levels of 1, 2, and 3 M NaCl were incubated with 1 mM of PG. The number of cells was significantly lower in all PG-treated cells compared to untreated controls. Despite PG-induced cell death, the fresh weight of all PG-treated cells was considerably higher than controls. PG-treated cells had enhanced antioxidant capacity because of increased levels of Chlorophyll a, β-carotene, reduced ascorbate, protein, and enzymatic activities, but accumulated lower levels of malonyldialdehyde and hydrogen peroxide compared to untreated cells. The results suggest that PG acts as a signal molecule both directly by reducing of free radical oxidants and indirectly by augmenting ascorbate pool levels, β-carotene production, and antioxidant enzymes activity to boost the capacity of antioxidant systems and radical oxygen species scavenging. Therefore, induction of salt stress tolerance by PG in D. bardawil is associated with metabolic adjustments through activation or synthesis of both enzymatic and non-enzymatic molecules involved in antioxidant systems.     

Selected reactive oxygen species and antioxidant enzymes in common bean after <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">phaseolicola</Emphasis> and <Emphasis Type="Italic">Botrytis cinerea</Emphasis> infection

Phaseolus vulgaris cv. Korona plants were inoculated with the bacteria Pseudomonas syringae pv. phaseolicola (Psp), necrotrophic fungus Botrytis cinerea (Bc) or with both pathogens sequentially. The aim of the experiment was to determine how plants cope with multiple infection with pathogens having different attack strategy. Possible suppression of the non-specific infection with the necrotrophic fungus Bc by earlier Psp inoculation was examined. Concentration of reactive oxygen species (ROS), such as superoxide anion (O₂ ^?) and H₂O₂ and activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) were determined 6, 12, 24 and 48 h after inoculation. The measurements were done for ROS cytosolic fraction and enzymatic cytosolic or apoplastic fraction. Infection with Psp caused significant increase in ROS levels since the beginning of experiment. Activity of the apoplastic enzymes also increased remarkably at the beginning of experiment in contrast to the cytosolic ones. Cytosolic SOD and guaiacol peroxidase (GPOD) activities achieved the maximum values 48 h after treatment. Additional forms of the examined enzymes after specific Psp infection were identified; however, they were not present after single Bc inoculation. Subsequent Bc infection resulted only in changes of H₂O₂ and SOD that occurred to be especially important during plant–pathogen interaction. Cultivar Korona of common bean is considered to be resistant to Psp and mobilises its system upon infection with these bacteria. We put forward a hypothesis that the extent of defence reaction was so great that subsequent infection did not trigger significant additional response.