Purification and some properties of β-N-acetyl-D-glucosaminidase from prawn (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

The -N-acetyl-D-glucosaminidase (NAGase, EC 3.2.1.52) from prawn (Penaeus vannamei) was purified by extraction with 30% ethanol solution and ammonium sulfate fractionation, then chromatographed on Sephadex G-100 followed by DEAE-cellulose (DE-32) columns. The purified enzyme determined to be homogeneous by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The specific activity of the purified enzyme was 1,560 U mg^–1. Enzyme molecular weight was determined to be 105,000 Da; it contained two subunits of the same mass (45,000 Da). The pI value was calculated to be 4.8 by isoelectric focusing. The optimum pH and optimum temperature of the enzyme for the hydrolysis of pNP--D-GlcNAc (enzyme substrate) were determined to be pH 5.2 and 45°C, respectively. The behavior of the enzyme during hydrolysis of pNP--D-GlcNAc followed Michaelis–Menten kinetics, with K_m=0.254 mM and V_m=9.438 M min^–1, at pH 5.2 and 37°C. The stability of the enzyme was investigated, and the results showed that the enzyme was stable in a pH range from 4.2 to 10.0 and at temperatures <40°C. The effects of metal ions on the enzyme were also studied. Li⁺, Na⁺ and K⁺ had no influence on enzyme activity. Mg²⁺, Ca²⁺ and Mn²⁺ activated the enzyme, while Ba²⁺, Zn²⁺, Co²⁺, Cd²⁺, Hg²⁺, Pb²⁺ Cu²⁺, Fe³⁺ and Al³⁺ showed various degrees of inhibitory effects on the enzyme.Communicated by O. Kinne, Oldendorf/Luhe     

Reproductive partitioning in communally breeding guira cuckoos,<Emphasis Type="Italic"> Guira guira</Emphasis>

Guira cuckoos, Guira guira, exhibit a rare polygynandrous reproductive system with groups containing several male and female breeders, allowing for important tests of reproductive skew models. Female reproductive strategies involve leaving the group, varying clutch size, egg ejection and infanticide, among others. Here we examined the predictions of reproductive skew models relative to reproductive partitioning among females in groups. We used yolk protein electrophoresis to identify individual females eggs in joint nests. We found that reproductive partitioning favors early-laying females, which lay and incubate more eggs than females that begin laying later. Because the female that lays first tends to switch between repeated nesting bouts, and females do not always contribute eggs to each bout, female reproductive success tends to equalize within groups over time. The pattern of reproductive partitioning differs from that described for anis, another crotophagine joint-nester. We calculated reproductive skew indices for groups in 2 years, for both laying and incubation, as well as an overall population value. These were compared to random skew generated by simulations. Varying degrees of skew were found for different groups, and also across sequential nesting bouts of the same groups. Overall, however, skew did not deviate from random within the population. Nests that reached incubation tended to have lower skew values during the laying phase than nests terminated due to ejection of all eggs followed by desertion. Groups had higher reproductive skew indices in their first nesting bout of the season, and these nests frequently failed. These results illustrate the importance of social organization in determining not only individual, but group success in reproduction, and highlight the flexibility of vertebrate social behavior.Communicated by J. Dickinson     

On the social structure of offspring rearing in the burrower bug,<Emphasis Type="Italic"> Sehirus cinctus</Emphasis> (Hemiptera: Cydnidae)

Maternal-offspring interactions are important in a variety of animals. Understanding the evolution of these interactions requires that we also study the broader social context in which they occur. To date, behavioral studies on burrower bugs, Sehirus cinctus, have focused exclusively on interactions between mothers and offspring. Here we ask whether these interactions occur in a social context that extends beyond the family unit of a mother and her own genetic offspring. Such social structure can arise from behaviors that occur before eggs are laid, or from actions of individuals that occur post-hatching. We present field data showing that lay sites of mothers are spatially aggregated on a scale that would lead to behavioral interactions among families. Microsatellite markers suggest neighboring mothers are unrelated. Laboratory experiments do not support the hypothesis that spatial aggregation results from a direct attraction of females to one another. Other laboratory studies reported here indicate that, after hatching, unrelated clutches sometimes join together to form multifamily groups. Experiments reveal that mothers are not necessary for these joining events to occur. In sum, these data suggest that both mothers and offspring play active, but different, roles in generating the social environment in which offspring rearing occurs.Communicated by N. Wedell     

Baboon loud calls advertise male quality: acoustic features and their relation to rank,age, and exhaustion

Free-ranging adult male baboons give loud two-syllable wahoo calls during dawn choruses, interactions between groups, when chasing females, and in aggressive interactions with other males. Previous research has shown that the rate and duration of these contest wahoos are correlated with a males competitive ability: high-ranking males call more often, call at faster rates, and call for longer bouts than do low-ranking males. Here we report that acoustic features of wahoos also reveal information about male competitive ability. High-ranking males give wahoos with higher fundamental frequencies (F0) and longer hoo syllables. Within-subject analyses revealed that, as males fall in rank, the hoo syllables tend to shorten within a period of months. As males age and continue to fall in rank, F0 declines, hoo syllables shorten, and formant dispersion decreases. Independent of age and rank, within bouts of calling F0 declines and hoo syllables become shorter. Because wahoos are often given while males are running or leaping through trees, variation in these acoustic features may function as an indicator of a males stamina. The acoustic features of contest wahoos thus potentially allow listeners to assess a males competitive ability.Communicated by C. Nunn     

Multimodal signals,imperfect information,and identification of sex in red-backed salamanders (<Emphasis Type="Italic">Plethodon cinereus</Emphasis>)

Theoretical treatments and empirical studies both suggest that signals that occur in multiple sensory modes have superior detectabilities, discriminabilities, and memorabilities. There is also an intuitive link between signal detectability, discriminability, and memorability and the quality of information that is transferred via the signaler. We investigated the role of information quality and sensory modality in the sexual identification of intruding conspecifics by territorial male red-backed salamanders (Plethodon cinereus). Our results imply that these salamanders are able to discriminate between intruding male and female conspecifics (based on the allocation of aggression) in bimodal scenarios (vision and olfaction) even under situations in which the information available about the intruders sexual identities is reduced in quality. In unimodal scenarios (olfaction only), male residents exhibited heightened levels of aggression toward male secretions and reduced levels of aggression toward female secretions. In unimodal scenarios where information pertaining to the sex of the stimulus was reduced in quality, male residents displayed intermediate levels of aggression relative to the responses that male and female stimuli received respectively. Although resident males touched bimodal stimuli significantly more than they touched unimodal stimuli, we were unable to find support for the notion that sensory modality greatly influences how male resident red-backed salamanders allocate aggression toward intruding male versus female stimuli.Communicated by H. Kokko     

The outcome of in-vitro contests between larvae of the endoparasitoid<Emphasis Type="Italic"> Venturia canescens</Emphasis> depends on both their relative and absolute ages

In a laboratory colony of the asexual solitary endoparasitoid Venturia canescens Grav. (Hymenoptera: Ichneumonidae), two genetically and phenotypically distinct lines (RP and RM) appear to coexist sympatrically, with the reproductive success of the RM-line being greater under con-specific superparasitism but lower under self superparasitism. To investigate the basis of the RM-lines advantage under con-specific superparasitism, we examined the outcome of inter-larval physical combat under in-vitro conditions. The results showed that the outcome depended on both the relative and absolute ages of the contestants, and that the competitive abilities of the two lines were not symmetric. In contests involving a larva and an egg, the unhatched wasp lost. In contests involving two larvae, at least one of which was newly hatched, the RP-larva tended to lose. Finally, if both larvae were at least 8–10 h post-hatching when the contest occurred, then the larger larva tended to lose, with the probability increasing with the difference in length between the two larvae. Thus, the higher reproductive success of the RM-line under competing superparasitism with the RP-line is due to a physiological difference between the newly hatched larvae of the two lines, which results in an advantage to the RM-larva independent of the order or time interval between ovipositions.Communicated by P. Heeb     

Triterpenoid and caffeic acid derivatives in the leaves of ragweed, <Emphasis Type="Italic">Ambrosia artemisiifolia</Emphasis> L. (Asterales: Asteraceae), as feeding stimulants of <Emphasis Type="Italic">Ophraella communa</Emphasis> LeSage (Coleoptera: Chrysomelidae)

Summary. The leaf beetle Ophraella communa infests almost exclusively Ambrosia artemisiifolia in the fields of Japan, even though it normally feeds on several Asteraceous plants. A filter paper bioassay showed that the feeding of O. communa is strongly stimulated by methanolic extracts of A. artemisiifolia. The feeding stimulants for O. communa have been isolated from methanolic extracts of A. artemisiifolia. -Amyrin acetate, -amyrin acetate, 5-caffeoylquinic acid (chlorogenic acid) and 3,5-dicaffeoylquinic acid from A. artemisiifolia have been identified as feeding stimulants for O. communa. Triterpenoid derivatives (-amyrin acetate or -amyrin acetate) and caffeic acid derivatives (3, 5-dicaffoylquinic acid or 5-caffeoylquinic acid) showed feeding stimulant activity when mixed together.     

Preferential incubation positions for different sized eggs and their influence on incubation period and hatching asynchrony in Snares crested (<Emphasis Type="Italic">Eudyptes robustus</Emphasis>) and yellow-eyed penguins (<Emphasis Type="Italic">Megadyptes antipodes</Emphasis>)

In crested penguins (Eudyptes spp.), second-laid eggs typically hatch before first eggs. Amongst a variety of factors that have been considered as mechanisms underlying this reversal, has been the idea that crested penguins can adjust the degree of hatching asynchrony by manipulating egg positions (i.e. placing the smaller first egg in the supposedly thermally disadvantaged anterior position) during incubation (termed Preferential Incubation Hypothesis). We tested this in the Snares crested penguin (Eudyptes robustus) and the closely related, but synchronously-hatching, yellow-eyed penguin (Megadyptes antipodes). Snares crested penguins were more likely to place their first eggs, which are smaller than second eggs, in the anterior incubation position than were yellow-eyed penguins, which have a clutch of two similar-sized eggs. But when yellow-eyed penguins, a non-brood reducing species, were provided with an artificial size-dimorphic clutch, they also placed smaller eggs more frequently in the anterior position, suggesting that a general preference exists among penguins to place smaller eggs in the anterior position. Egg temperatures of small first eggs of Snares crested penguins were higher in the anterior than in the posterior position. Large first eggs in lesser size-dimorphic clutches experienced high temperature differences in relation to position, while small first eggs in greater size-dimorphic clutches were incubated at similar temperatures. In yellow-eyed penguins, large eggs within clutches generally had higher egg temperatures than small eggs. Incubation periods of second eggs declined with increasing egg size. Egg-size variation, rather than egg positioning behaviour, influenced hatching patterns in Snares crested penguins. In lesser size-dimorphic clutches, second eggs were more likely to hatch first while in greater size-dimorphic clutches, small first eggs were more likely to hatch at the same time or before the second eggs. This was similar in yellow-eyed penguins, where second eggs hatched earlier in clutches with large first eggs. Our data contradicts the Preferential Incubation Hypothesis and we conclude that this hypothesis is unlikely to explain the reversed hatching asynchrony in crested penguins.Communicated by C. Brown     

Persistence of atrazine impact on aneuploidy in Pacific oysters,<Emphasis Type="Italic"> Crassostrea gigas</Emphasis>

Widespread use of the herbicide atrazine has incited much research on its toxicity in aquatic systems, where it is routinely detected due to runoff from cultivated fields. Moreover, the determination of the genotoxic effect of such pollutants in the marine environment has become a major requirement for ecosystem protection. In the Pacific oyster, Crassostrea gigas, hypodiploid aneuploid cells have regularly been reported. There is a negative correlation between this phenomenon and growth, as well as evidence for a genetic basis. A positive relationship between atrazine and aneuploidy has previously been demonstrated in C. gigas adults and juveniles. To evaluate the persistence of this impact, our study examined the offspring of the same adult population previously treated with different atrazine doses (10 µg l^–1, representing a peak value found in a polluted environment and 100 µg l^–1), and a seawater control. We observed that these offspring exhibited significantly higher aneuploidy levels when their parents had been exposed to atrazine (14.9–16.9% in comparison with the control where the levels ranged from 11.4% to 12.8%). In addition, the present study examined the aneuploidy level of a sample of juveniles, previously exposed for 3.5 months to the same doses of atrazine, then transferred to non-polluted conditions for an additional period of 2.5 months; this aneuploidy level remained significantly different between the treatments applied. These results demonstrate the persistence of an atrazine impact on Pacific oyster aneuploidy in time, within and between generations, indicating that this widely used compound may represent an important factor causing at least medium-term damage to genetic material.Communicated by S.A. Poulet, Roscoff     

Information transfer and gain in flocks: the effects of quality and quantity of social information at different neighbour distances

We assessed experimentally how the quality and quantity of social information affected foraging decisions of starlings (Sturnus vulgaris) at different neighbour distances, and how individuals gained social information as a function of head position. Our experimental set up comprised three bottomless enclosures, each housing one individual placed on a line at different distances. The birds in the extreme enclosures were labelled senders and the one in the centre receiver. We manipulated the foraging opportunities of senders (enhanced, natural, no-foraging), and recorded the behaviour of the receiver. In the first experiment, receivers responded to the condition of senders. Their searching rate and food intake increased when senders foraged in enhanced conditions, and decreased in no-foraging conditions, in relation to natural conditions. Scanning was oriented more in the direction of conspecifics when senders behaviour departed from normal. In the second experiment, responses were dose dependent: receivers increased their searching rate and orientated their gaze more towards conspecifics with the number of senders foraging in enhanced food conditions. In no-foraging conditions, receivers decreased their searching and intake rates with the number of senders, but no variation was found in scanning towards conspecifics. Differences in foraging and scanning behaviour between enhanced and no-foraging conditions were much lower when neighbours were separated farther. Overall, information transfer within starling flocks affects individual foraging and scanning behaviour, with receivers monitoring and copying senders behaviour mainly when neighbours are close. Information transfer may be related to predation information (responding to the vigilance of conspecifics) and foraging information (responding to the feeding success of conspecifics). Both sources of information, balanced by neighbour distance, may simultaneously affect the behaviour of individuals in natural conditions.Communicated by H. Kokko