Predicting and testing resource partitioning in a tropical fish assemblage of zooplanktivorous 'barbs': an ecomorphological approach

Morphometrics on 25 critical feeding structures predicted conspicuous specializations in Barbus tanapelagius (pursuit hunting for zooplankton), Labeobarbus brevicephalus (surface dwelling pump-filter-feeder on zooplankton) and Barbus pleurogramma (particulate feeding on tough, benthic food), whereas for Barbus humilis intermediate values predicted few constraints and specializations in feeding. These potential niches, set by fish size and structural constraints, were tested by comparing gut contents collected during a 24 months sampling programme on Lake Tana, Ethiopia. Zooplankton dominated the diet of B. tanapelagius (75% of gut volume) and L. brevicephalus (39%). The guts of B. pleurogramma and juveniles of the large labeobarbs showed an array of benthic food types, whereas B. humilis had the widest food niche, both zooplankton (40% of gut volume) and benthic invertebrates. Although the pelagic species showed the largest spatial overlap, their size, feeding modes and utilization of zooplankters differed: L. brevicephalus preyed predominantly on the larger zooplankton ( Daphnia sp.) and B. tanapelagius also on smaller species ( e.g. Bosmina sp. and cyclopoid copepods). The spatial segregation between B. tanapelagius (pelagic) and the juvenile labeobarbs (littoral) indicated the possibility for a small pelagic barb fishery without negative effects on the labeobarb stocks. The ecomorphological approach using the 'Food-Fish Model' appeared to predict competitive positions and resource partitioning appropriately, and is of major importance to evaluate food web interactions.     

Toothless predators: feeding performance and techniques among the piscivores within Lake Tana's endemic barbus species flock (Pisces:Cyprinidae)

With more than 2000 fish species the Cyprinidae is the largest family of vertebrates. Lake Tana, a large lake (3050 km²) situated in the NW‐ highlands of Ethiopia, harbours, as far as we know the only remaining intact species flock of large (max. 100 cm FL) cyprinid fishes (15 Barbus spp.). One of the most intriguing aspects of this endemic Barbus species flock is the large number of piscivores (8). Cyprinid fishes seem not well designed for piscivory, they lack teeth in the oral jaw, have a small slit‐shaped pharyngeal cavity and all lack a stomach with low pH for digesting large prey. Many barbs are benthivorous species, like the ancestral barb in Lake Tana's isolated system. Why then is piscivory, which is rare among cyprinids, so common in Lake Tana Barbus? The aim of present study was to compare the performance and techniques of these piscivorous Barbus with known piscivores from other fish families. We studied prey handling times over prey size, prey capture using high‐speed movies, and assessed the effect of prey size on performance and prey selection in the field. Performances were explained by functional morphology of their feeding system. Overall, Lake Tana's piscivorous Barbus perform relatively 'poor', compared to piscivores from other fish families. For example, Lake Tana's piscivores are only able to handle prey fish smaller than 16% of their own body length. However, Lake Tana lacks potential piscivorous competitors, rendering the piscivorous Barbus by far the 'best' and apparently highly successful. They have adapted to all available macro‐habitats (littoral, offshore pelagic and offshore benthic), using different techniques (ambush, pursuit and cruising), a unique scenario for barbs.     

Growth response of pikeperch larvae in relation to body size and zooplankton abundance

The most critical period at onset of feeding in pikeperch Stizostedion lucioperca is short (<5 days at 20° C). The larvae are sensitive to prey density during the first week of exogenous feeding. First-feeding larvae of 6.5 mm total length ( L_T ) needed prey densities of >585 prey l⁻¹ to maintain mass (C_maint), whereas 5 days older larvae of 7 mm L _T C_maint= 55 prey l⁻¹ and for 11 mm L _T larvae C_maint<10 prey 1⁻¹. Changes in specific growth rates for larvae of 7 and 11 mm were similar to a type-II functional response curve reaching a specific growth rate of 26 and 30% day⁻¹, respectively, at 1000 prey l⁻¹, whereas the 6·5 mm larvae showed a linear growth response reaching a specific growth rate of only 9% day ⁻¹ at 1000 prey l⁻¹. The results suggest that prey density is a limiting factor, which might contribute to the high variation between year-class Strengths.     

Impact of Bythotrephes longimanus on zooplankton assemblages of Harp Lake, Canada: an assessment based on predator consumption and prey production

1. Following the 1993 invasion of Harp Lake by Bythotrephes longimanus a number of small-bodied zooplankton declined dramatically compared to pre-invasion densities, and some larger species increased. 2. To test whether these changes were caused by Bythotrephes, we measured its consumption of zooplankton in 1995 and compared this to production by the prey species with which it overlapped spatially and temporally. 3. On a seasonal basis Bythotrephes consumption was 199 mg dry mass (DM) m⁻², or 25% of zooplankton production which was 783 mg DW m⁻². However, for some species, such as the small copepod Tropocyclops extensus , consumption greatly exceeded production—this was one of the species noted to decline in Harp Lake following the appearance of Bythotrephes . By contrast, for the larger cladocerans Daphnia galeata mendotae and Holopedium gibberum , consumption was much less than production—these species were observed to increase following the invasion. 4. Our results thus support the hypothesis that zooplankton changes in Harp Lake were caused by Bythotrephes . We speculate that lower quality prey remaining in Harp Lake may lead to reduced densities of Bythotrephes , or impose strong selective forces that lead to new adaptations by this predator.     

Functional response of juvenile pink and chum salmon: effects of consumer size and two types of zooplankton prey

Feeding rate experiments were conducted for pink salmon Oncorhynchus gorbuscha fry [mean fork length ( L _F) 39 mm], juveniles (103–104 mm L _F) and juvenile chum salmon Oncorhynchus keta (106–107 mm L _F). Fishes were presented with small copepod ( Tisbi sp.) or larger mysid shrimp ( Mysidopsis bahia ) prey at varying densities ranging from 1 to 235 prey l⁻¹ in feeding rate experiments conducted at water temperatures ranging from 10·5 to 12·0° C under high light levels and low turbidity conditions. Juvenile pink and chum salmon demonstrated a type II functional response to mysid and copepod prey. Mysid prey was readily selected by both species whereas the smaller bodied copepod prey was not. When offered copepods, pink salmon fry fed at a higher maximum consumption rate (2·5 copepods min⁻¹) than larger juvenile pink salmon (0·4 copepods min⁻¹), whereas larger juvenile chum salmon exhibited the highest feeding rate (3·8 copepods min⁻¹). When feeding on mysids, the maximum feeding rate for larger juvenile pink (12·3 mysids min⁻¹) and chum (11·5 mysids min⁻¹) salmon were similar in magnitude, and higher than feeding rates on copepods. Functional response models parameterized for specific sizes of juvenile salmon and zooplankton prey provide an important tool for linking feeding rates to ambient foraging conditions in marine environments, and can enable mechanistic predictions for how feeding and growth should respond to spatial-temporal variability in biological and physical conditions during early marine life stages.     

Food limitation of planktonic rotifers: field experiments in two mountain ponds

1. Resource competition is thought to be important in controlling zooplankton population dynamics and structuring zooplankton communities. Resource competition requires that resources are limiting. Ten field experiments were conducted to determine the presence and intensity of resource limitation of herbivorous planktonic rotifers in two mountain ponds. The intensity of food limitation was measured as Δ r , the difference between population growth rates in food supplemented versus control enclosures.
2. Rotifers were significantly food limited (Δ r  > 0) in most experiments. The intensity of food limitation varied between species. In Grady Lake, mean Δ r was 0.26 day⁻¹ for Polyarthra vulgaris and 0.43 day⁻¹ for an unidentified Synchaeta sp. In L1 Pond, mean Δ r was 0.07 day⁻¹ for Keratella cochlearis and 0.28 day⁻¹ for S. oblonga . The frequent and intense food limitation suggests that edible phytoplankton in both ponds were often present at low densities, were of low nutritional quality, or both. The intensity of food limitation often changed rapidly over time, indicating rapid temporal variation in resource availability.
3. For P. vulgaris , Δ r was not correlated with its population growth rate in Grady Lake. Thus, although food limitation was common, population dynamics may have been more strongly affected by other factors. For K. cochlearis , Δ r increased as population density increased, suggesting the existence of intraspecific resource competition that may regulate population dynamics.     

Scaling up the functional response for spatially heterogeneous systems

Scale transition theory is a framework for predicting regional population dynamics from local process functions and estimates of spatial heterogeneity. Using this framework, we estimated regional scale functional responses for a benthic predator–prey system in the Baltic Sea. Functional responses were based on laboratory experiments or field observations of stomach contents, and prey densities measured at a local scale (0.1 m²) or a regional scale (300 km²). Laboratory data overestimated consumption at high prey densities, whereas predictions based on local scale data tallied closely with consumption observed at the regional scale. The predicted regional functional response was different for increasing and decreasing prey densities, reflecting that predator and prey densities, as well as the covariance between them, exhibit oscillatory dynamics. We conclude that it is important to validate laboratory data with small-scale field observations and that scale transition is a powerful tool for scaling-up process functions in heterogeneous systems.     

Response surface methodology to optimize the nutritional parameters for enhanced production of jasmonic acid by Lasiodiplodia theobromae

Aims:  To find out the cumulative effect of the nutritional parameters and to enhance the production of jasmonic acid (JA) in static fermentation by Lasiodiplodia theobromae using response surface methodology (RSM).
Method and Results:  Malt extract, sucrose, NaNO₃ and MgSO₄.7H₂O were analysed by a 30-trial central composite design using RSM for optimizing their concentrations in the medium and the effect of their mutual interaction on JA production. Sucrose and NaNO₃ were found highly significant in influencing the JA production. Malt extract and MgSO₄.7H₂O showed an effect on the JA production in interaction with other variables. When the optimum values of the parameters obtained through RSM (19·95 g l⁻¹ malt extract, 50 g l⁻¹ sucrose, 7·5 g l⁻¹ NaNO₃ and 3·51 g l⁻¹ MgSO₄.7H₂O) were applied, 32% increase in JA production (299 mg l⁻¹) was observed in comparison with 225 mg l⁻¹ of JA produced with same media components not analysed by RSM and subsequently validated the statistical model.
Conclusions:  Increase in JA production was achieved by optimizing the nutritional parameters.
Significance and Impact of the Study:  This is the first report of using RSM for optimizing a medium for JA production. It resulted in an increase in JA production without augmentation of costly additives.     

In vitro regeneration and genetic transformation of the berberine-producing plant,Thalictrum flavum ssp. glaucum

Protocols have been developed for the in vitro regeneration and Agrobacterium -mediated genetic transformation of meadow rue, Thalictrum flavum ssp. glaucum . Ten-day-old seedlings were bisected along the embryonic axis and the cotyledons were co-cultured with various Agrobacterium tumefaciens strains for 3 days. The cotyledons were cultured on a shoot induction medium (B5 salts and vitamins, 30 g l⁻¹ sucrose, 2 mg l⁻¹ kinetin, and 3 g l⁻¹ Gelrite) containing 25 mg l⁻¹ hygromycin B as the selection agent and 250 mg l⁻¹ timentin to facilitate the elimination of Agrobacterium . Only the oncogenic A. tumefaciens strains A281 and C58 produced transgenic T. flavum callus tissues. A281 was the most effective strain producing hygromycin-resistant callus on 85% of the explants. Transgenic callus was subcultured on the shoot induction medium every 2 weeks. After 12 weeks, hygromycin-resistant shoots that formed on explants exposed to strain A281 were transferred to a root induction medium (B5 salts and vitamins, 25 mg l⁻¹ hygromycin B, 250 mg l⁻¹ timentin, and 3 g l⁻¹ Gelrite). Detection of the β -glucuronidase ( GUS ) gene using a polymerase chain reaction assay, the high levels of GUS mRNA and enzyme activity, and the cytohistochemical localization of GUS activity confirmed the genetic transformation of callus cultures and regenerated plants. The transformation process did not alter the normal content of berberine in transgenic roots or cell cultures; thus, the reported protocol is valuable to study the molecular and metabolic regulation of protoberberine alkaloid biosynthesis.     

Effect of starvation time on the prey capture behaviour, functional response and population growth of Asplanchna sieboldi (Rotifera)

1. We examined the effect of different periods of prior starvation(from 30 min to 16 h) on the prey capture behaviour, and functional and numerical responses of the predatory rotifer Asplanchna sieboldi using the rotifer Brachionus calyciflorus as prey.
2. Feeding activity (i.e. encounter, attack, capture and ingestion) by Asplanchna increased significantly with increasing prey densities from 2 to 16 mL⁻⁻¹ and with increasing prior starvation periods from 0.5 to 8 h.
3.  Asplanchna sieboldi showed a type II functional response at all the prior starvation periods tested. The asymptotic prey density was highest after 8 h of starvation.
4. The instantaneous population growth rate of A. sieboldi ranged from 0.089 ± 0.044 (when starved for 8 h in every 24 h and at a prey density of 2 individuals mL⁻⁻¹ for the other 16 h) to 1.015 ± 0.142 in the control (no starvation and at a prey density of 16 individuals mL⁻⁻¹). The effect of starvation time on the numerical response was evident only at the higher prey density.     

Shape and sources of variations of the functional response of wildfowl: an experiment with mallards, Anas platyrhynchos

Understanding the variations of the functional response of an organism, i.e. the predation rate in relation to prey density, is necessary to understand the interactions between the animal and its food supply. This has received little attention in dabbling ducks so we investigated experimentally the shape of the functional response of mallard feeding on poultry pellets, and assessed the influence of several factors such as the size of food items, sex or individual performance on this functional response. Individual differences in intake rate are of crucial importance in group or gregarious foraging species. We used two approaches of the functional response: 1) the relation between feeding rate (pellets/s) and pellet densities (pellets/m²), and 2) the relationship between instantaneous intake rate (g/s) and biomass density (g/m²). For both approaches, we found that the Type II functional response gave better estimates than a Type I linear functional response but explained only a third of the variance. Our results show that pellet size has a large effect on instantaneous intake rate. The comparison of the functional response parameters suggest that handling time per prey may not reflect the real constraints on intake rate, but that handling time per gram ingested may be more appropriate to integrate the effect of item size in the functional response. We then discuss the possible mechanisms involved. We also found individual variations in the functional response for each of the experiments, with some consistency in the hierarchy regarding feeding efficiency. We did not find any differences between males and females. Our results provide an evaluation of individual variations in intake rate in interference-free conditions, which has rarely been done, and call for more controlled experiments to allow a finer understanding of the mechanisms of food acquisition in dabbling ducks.     

Attraction of zebrafish, Brachydanio rerio, to alanine and its suppression by copper

Preference responses of zebrafish to 10⁻³, 10⁻⁴ and 10⁻⁵M alanine (Ala) were concentration- dependent. Behavioural responses to copper (Cu) and Cu + Ala mixtures were also assessed. Zebrafish avoided 100 and 10 μg Cu l⁻¹, but not 1 μg l⁻¹. Mixtures of 10⁻³ m Ala+ 100 μg Cu l⁻¹ and 10 ⁴ M Ala + 10 μg Cu 1⁻¹ were avoided as intensely as was Cu alone. Responses to 10⁻³ M Ala + 10 or 1 μg Cu l⁻¹ and 10 ⁴ M Ala +1 μg Cu l⁻¹ did not differ statistically from controls (no detectable preference or avoidance). These results demonstrate, firstly, that a concentration of a pollutant avoided by itself (10 μg Cu l⁻¹) may not be avoided when encountered with an attractant chemical stimulus (Ala) and may suppress the preference for an attractant stimulus, and secondly, that a concentration of a pollutant not avoided by itself and not considered deleterious (1 μg Cu l⁻¹) suppresses attraction to Ala (an important constituent of prey odours for many fishes).     

Inhibition of the tempe mould, Rhizopus oligosporus, by ammonia

The hyphal extension rate of Rhizopus oligosporus NRRL 2710 was slowed in the presence of 0·42 and 0·84 mmol NH3 l⁻¹ and inhibited by 1·3 mmol l⁻¹. Sporulation was prevented at NH3 concentrations of 0·42 mmol l⁻¹ andabove. There was no evidence of toxicity due to NH⁺₄ at concentrations up to 300 mmol l⁻¹.Independent of the concentrations of NH3 or NH⁺₄, the lower the pH value, in therange 6·0–9·0, the higher was the rate of hyphal extension. It is suggested that accumulationof toxic levels of NH3 could be responsible for the cessation of mould growth in tempe.     

Optimization and biochemical characterization of a bacteriocin from a newly isolated Bacillus subtilis strain 14B for biocontrol of Agrobacterium spp. strains

Aims:  The identification of a new compound active against Agrobacterium tumefaciens .
Methods and Results:  The culture conditions of a newly isolated Bacillus subtilis strain, designed 14B, were optimized, as a first step, to produce its bacteriocin (termed Bac 14B) for the biocontrol of Agrobacterium spp., the causal agents of the crown gall disease. Bac 14B was then partially purified and biochemically characterized. Bacillus subtilis 14B was observed to produce an antibacterial compound having a protinaceous nature. As estimated by sodium dodecyl sulfate-polyacrilamide gel electrophoresis (SDS-PAGE), the semi-purified bacteriocin substance was found to be a monomeric protein with a molecular weight of 21 kDa. While the latter's antimicrobial activity was completely stable during exposure to a temperature range of up to 100°C for 2 h, its initial activity was totally lost at 121°C for 20 min. The maximum bacteriocin production (4096 AU ml⁻¹) was recorded after 96 h-incubation in an optimized Luria Bertani medium supplemented with 10 g l⁻¹ glucose, 15 g l⁻¹ K₂HPO₄ and 5 g l⁻¹ MgSO₄ 7H₂O at 30°C in a shaking flask culture. Interestingly, the B. subtilis 14B culture supernatant that contained the bacteriocin under study was proved efficient in reducing both the percentage of galled plants and the number of galls in tomato.
Conclusion:  The findings revealed that B. subtilis 14B and its bacteriocin are efficient in reducing the percentage of infections in plants caused by Ag. tumefaciens .
Significance and Impact of the Study:  The results could be useful for the nurserymen who are particularly interested in the biocontrol of the crown gall disease.     

Functional response of Picromerus bidens: effects of host plant

Abstract:  The influence of three solanaceous plants (tomato, sweet pepper and eggplant) on the functional response of the predatory bug Picromerus bidens to densities of fourth-instar larvae of the beet armyworm Spodoptera exigua was assessed. Logistic regression indicated a type II functional response on all host plants. Over all prey densities, P. bidens killed significantly fewer fourth instars of S. exigua on tomato than on sweet pepper or eggplant (1.96 ± 0.17 vs. 4.37 ± 0.19 and 3.90 ± 0.15 larvae per predator per 24 h respectively). A higher theoretical maximum predation rate was estimated on sweet pepper (11.1 prey larvae per day) and eggplant (7.4) than on tomato (5.4). The mean number of prey killed per day by P. bidens females ranged from 0.78 at a density of one prey on tomato to 8.45 at a density of 24 prey on sweet pepper. The data indicated that the estimates of handling time ( T _h) and attack rate ( a ) were highly affected by host plant. Based on asymptotic 95% confidence intervals, a lower attack coefficient was found on tomato (0.02 h⁻¹) than on sweet pepper or eggplant (0.07 and 0.11 h⁻¹ respectively). On the other hand, handling times were significantly longer on tomato (4.42 h) and eggplant (3.23 h) than on sweet pepper (2.15 h). This laboratory study suggests that plant characteristics influence the ability of P. bidens to respond to changes in prey density.     

Isolation and characterization of an intracellular esterase from Lactobacillus casei subsp. casei IFPL731

An intracellular esterase from Lactobacillus casei subsp. casei IFPL731 was purified 1000-fold by ion exchange chromatography and gel filtration chromatography. The relative molecular mass of the native enzyme was 105 kDa, while the subunit molecular mass was estimated to be 38 kDa. The esterase hydrolysed tributyrin and had a preference for esters of short-chain fatty acids (butyrate, caproate and caprylate), while it did not hydrolyse palmitate and sterate esters. The apparent Michaelis-Menten constant of the enzyme on p -nitrophenyl butyrate was 0·3 mmol l⁻¹ while on p -nitrophenyl caprylate, it was 0·04 mmol l⁻¹. The esterase was active over a broad range of pH and temperature values, and retained about 50% of maximal activity at pH 5·0 and 12 °C. Activity was strongly inhibited by 5 mmol l⁻¹ phenylmethylsulphonyl fluoride, β-mercaptoethanol and N -ethylmaleimide, and was stimulated by Zn²⁺ at 1 mmol l⁻¹.     

Effects of feeding and induction strategy on the production of BmR1 antigen in recombinant E. coli

Aim:  To investigate the effects of feeding and induction strategies on the production of Bm R1 recombinant antigen.
Methods and Results:  Fed-batch fermentation was studied with respect to the specific growth rate and mode of induction to assess the growth potential of the bacteria in a bioreactor and to produce high yield of Bm R1 recombinant antigen. Cells were grown at a controlled specific growth rate (μ_set) during pre-induction, followed by constant feeding postinduction. The highest biomass (24·3 g l⁻¹) was obtained during fed-batch process operated at μ_set of 0·15 h⁻¹, whereby lower μ_set (0·075 h⁻¹) gave the highest protein production (9·82 mg l⁻¹). The yield of Bm R1 was increased by 1·2-fold upon induction with 1 mmol l⁻¹ IPTG (isopropyl-β- d -thiogalactoside) compared to using 5 mmol l⁻¹ and showed a further 3·5-fold increase when the culture was induced twice at the late log phase.
Conclusions:  Combination of feeding at a lower μ_set and twice induction with 1 mmol l⁻¹ IPTG yielded the best result of all variables tested, promising an improved method for Bm R1 production .
Significance and Impact of the Study:  This method can be used to increase the production scale of the Bm R1 recombinant antigen to meet the increasing demand for Brugia Rapid^™, a commercial diagnostic test for detection of brugian filariasis.     

Fed-batch culture of Candida guilliermondii FTI 20037 for xylitol production from sugar cane bagasse hydrolysate

A fed-batch culture system was used to study xylitol production by Candida guilliermondii FTI 20037 in a synthetic and a sugar cane bagasse hydrolysate medium. The values achieved for xylitol yield and volumetric productivity were, respectively, 0 · 84 g g⁻¹ and 0 · 64 g l⁻¹ h⁻¹ using the synthetic medium and 0 · 78 g g⁻¹ and 0 · 62 g l⁻¹ h⁻¹ using the hydrolysate medium.     

Interactions between humic matter and bacteria when disinfecting water with UV light

Aims:  To investigate the impact of aquatic humic matter on the inactivation of Escherichia coli and Bacillus subtilis by ultraviolet (UV) light.
Methods and Results:  A bench-scale study investigated the potential for Aldrich^® humic acid (AHA) and Suwannee River natural organic matter (SR-NOM) to coat the surface of E. coli and B. subtilis and offer protection from low-pressure UV light. UV doses of 5 and 14 mJ cm⁻² were applied using a collimated beam at four concentrations of humic matter (0, 10, 50 and 120 mg l⁻¹) in reagent grade water. Both AHA and SR-NOM were found to offer statistically significant protection of both E. coli and B. subtilis at concentrations of 50 and 120 mg l⁻¹ for a UV dose of 14 mJ cm⁻².
Conclusions:  Both E. coli and B. subtilis are susceptible to coating by humic matter which can reduce the sensitivity of the cells to UV light.
Significance and impact of the study:  Micro-organisms in the environment may acquire characteristics through interaction with humic matter that render them more resistant to UV disinfection than would be predicted based on laboratory inactivation studies using clean cells.     

The linamarase of Mucor circinelloides LU M40 and its detoxifying activity on cassava

Mucor circinelloides LU M40 produced 12·2 mU ml⁻¹ of linamarase activity when grown in a 3 l fermenter in the following optimized medium (g l⁻¹ deionized water): pectin, 10·0; (NH₄)₂SO₄,
1·0; KH₂PO₄, 2·0; Na₂HPO₄, 0·7; MgSO₄.7H₂O, 0·5; yeast extract, 1·0; Tween-80,
1·0, added after 48 h of fermentation. The purified linamarase was a dimeric protein with a molecular mass of 210 kDa; the enzyme showed optimum catalytic activity at pH 5·5 and 40 °C and had a wide range (3·0–7·0) of pH stability. The enzyme substrate specificity on plant cyanogenic glycosides was wide; the K_m value for linamarin was 2·93 mmol l⁻¹. The addition, before processing, of the fungal crude enzyme to cassava roots facilitated and shortened detoxification; after 24 h of fermentation, all cyanogenic glycosides were hydrolysed.