Ontogenetic changes in chemical alarm cue recognition and fast‐start performance in guppies (Poecilia reticulata)

Risk recognition and fast‐start performance are critical to fish survival when faced with predators. Many fish species have been shown to recognize risks associated with chemical cues released by injured conspecifics. However, little is known about the ontogeny of “risk” recognition via damage‐released chemical alarm cues and fast‐start performance in prey fishes. The objectives of this study were to determine whether risk recognition and fast‐start performance in guppies (Poecilia reticulata) exhibit ontogenetic variation, and whether there is a trade‐off between risk recognition and fast‐start performance. To achieve these objectives, individual guppies from replicate groups were assayed on one of the 1st, 7th, 14th, 21st, or 28th day after their birth. We found that both the risk recognition and fast‐start performance in guppies exhibited ontogenetic variation, as on days 1 and 7, fish did not exhibit risk recognition when exposed to alarm cues from conspecifics, but by day 14, such recognition was evident. Noticeable increases in maximum linear velocity (V_max), maximum linear acceleration (A_max), and escape distance (S_120 ms) were concurrent with progressive ontogenetic stage, and no significant correlations were found between risk recognition and fast‐start performance at any ontogenetic stage. Our findings reveal ontogenetic variation in damage‐released chemical cue recognition and fast‐start performance in guppies.     

Effects of two parasites, Schistocephalus solidus (Cestoda) and Bunodera spp. (Trematoda), on the escape fast-start performance of three-spined sticklebacks

The fast‐start performance of three‐spined sticklebacks Gasterosteus aculeatus infected with Schistocephalus solidus and Bunodera spp. was determined and two distinct fast‐start responses (A and B) were observed. ‘A’ starts were of higher flexibility than B and three‐way ANOVA showed significant effects of A and B starts (P < 0·05), time (P < 0·05) and per cent standard body length, L_S (P < 0·05) on the orientation angle (angle of an individual segment of the fish with respect to the direction of travel). Schistocephalus solidus infection reduced maximum velocity (P < 0·05) and maximum acceleration (P < 0·05) of infected fish. Uninfected fish and fish infected with S. solidus up to a parasite index (parasite mass divided by the sum of fish and parasite mass) of 0·1 executed both types. Infected fish exclusively executed B starts for parasite index between 0·1 and 0·2. This was not due to a reduction in body flexibility associated with mechanical obstruction caused by S. solidus as no significant difference in the ratios of body width (P > 0·05) or depth (P > 0·05) to L_S were found between uninfected and infected fish. At a parasite index >0·2, infected fish were unable to perform escape fast‐starts increasing the likelihood of predation by their definitive hosts such as loons or belted kingfishers. Three‐spined sticklebacks infected with S. solidus with a parasite index of c. 0·2–0·3, however, were compromised by a suite of behavioural (e.g. increased foraging activity and amount of food consumed, increased risk associated with feeding and increased response latency to predatory stimuli), physiological (e.g. increased rate of oxygen consumption, slower growth, delayed sexual maturation and breeding success) and biomechanical (e.g. decreased fast‐start performance) factors. Bunodera spp. did not affect the escape fast‐start performance of three‐spined sticklebacks and no significant difference for maximum velocity (P > 0·05) and maximum acceleration (P > 0·05) was found.     

Evaluation of a chromogenic agar medium for the detection of extended-spectrum ß-lactamase-producing Enterobacteriaceae

Aim: To compare the performance of a new chromogenic agar medium CHROMagar ESBL (KC‐ESBL) to chromID ESBL (SB‐ESBL) for the detection and presumptive identification of extended‐spectrum β‐lactamase (ESBL)‐producing Enterobacteriaceae directly from clinical specimens. Methods and Results: A total of 256 specimens were screened for ESBL producers. Also, the genotypes of the ESBLs and plasmid‐mediated AmpC β‐lactamases (pAmpCBLs) were characterized by PCR and sequencing. Among the 256 specimens, 17 (6·6%) ESBL producers were isolated on both media. The sensitivity, specificity, positive predictive value and negative predictive value were higher for KC‐ESBL (100, 93·3, 51·5 and 100%, respectively) than for SB‐ESBL (88·2, 92·9, 46·9 and 99·1%, respectively) (P = 0·72). Enterobacteriaceae harbouring pAmpCBL genes as well as chromosomal cephalosporinase‐ and penicillinase‐hyperproducing Enterobacteriaceae and Pseudomonas aeruginosa accounted for the false‐positive results. Conclusion: KC‐ESBL can detect ESBL producers from clinical specimens with good selectivity and rapid presumptive identification by means of colony colour at 24 h. Significance and Impact of the Study: This is the first study that has evaluated the performance of KC‐ESBL that enables the detection and presumptive identification of ESBL producers from clinical specimens.     

CULTIVATION OF ARTHROSPIRA (SPIRULINA) PLATENSIS (CYANOPHYCEAE) BY FED‐BATCH ADDITION OF AMMONIUM CHLORIDE AT EXPONENTIALLY INCREASING FEEDING RATES1

Arthrospira (Spirulina) platensis (Nordstedt) Gomont was cultivated under light‐limited conditions in 5‐L open tanks by daily supplying NH₄Cl as nitrogen source. Exponentially increasing feeding rates were adopted to prevent ammonia toxicity. The total feeding time (T) was varied between 12 and 20 days, and the starting (m₀) and total (m_T) quantities of the nitrogen source per unit reactor volume were varied in the ranges 0.19–1.7 mM and 2.3–23.1 mM, respectively. This intermittent addition of the nitrogen source prevented ammonia from reaching inhibitory levels and ensured final cell concentrations (X_m) and cell productivities (P_x) comparable with those of batch runs with KNO₃. Moreover, the lower nitrogen addition due to the use of NH₄Cl rather than KNO₃ allowed for higher nitrogen‐to‐cell conversions (Y_x/n). These results were evaluated using three‐factor, five‐level, central composite experimental planning, combined with the response surface methodology, selecting T, m₀, and m_T as the independent variables and X_m, P_x, and Y_x/n as the response variables. This approach allowed us to identify, through the simultaneous optimization of the variables, T=16 days, m₀=1.7 mM, and m_T=21.5 mM as the best conditions for A. platensis cultivation at 72 μmol photons·m^?2·s^?1. Under these conditions, a maximum cell concentration of 1239 mg ·L^?1 was obtained, which is a value comparable with that obtained using KNO₃ as nitrogen source and nearly coincident with the theoretical one estimated by the response surface methodology.     

Diverse modes of 5′‐[4‐(aminoiminomethyl)phenyl]‐[2,2′‐bifuran]‐5‐carboximidamide (DB832) interaction with multi‐stranded DNA structures

The modes of binding of 5′‐[4‐(aminoiminomethyl)phenyl]‐[2,2′‐Bifuran]‐5‐carboximidamide (DB832) to multi‐stranded DNAs: human telomere quadruplex, monomolecular R‐triplex, pyr/pur/pyr triplex consisting of 12 T*(T·A) triplets, and DNA double helical hairpin were studied. The optical adsorption of the ligand was used for monitoring the binding and for determination of the association constants and the numbers of binding sites. CD spectra of DB832 complexes with the oligonucleotides and the data on the energy transfer from DNA bases to the bound DB832 assisted in elucidating the binding modes. The affinity of DB832 to the studied multi‐stranded DNAs was found to be greater (K_ass ≈ 10⁷M^?1) than to the duplex DNA (K_ass ≈ 2 × 10⁵M^?1). A considerable stabilizing effect of DB832 binding on R‐triplex conformation was detected. The nature of the ligand tight binding differed for the studied multi‐stranded DNA depending on their specific conformational features: recombination‐type R‐triplex demonstrated the highest affinity for DB832 groove binding, while pyr/pur/pyr TTA triplex favored DB832 intercalation at the end stacking contacts and the human telomere quadruplex d[AG₃(T₂AG₃)₃] accommodated the ligand in a capping mode. Additionally, the pyr/pur/pyr TTA triplex and d[AG₃(T₂AG₃)₃] quadruplex bound DB832 into their grooves, though with a markedly lesser affinity. DB832 may be useful for discrimination of the multi‐sranded DNA conformations and for R‐triplex stabilization. © 2009 Wiley Periodicals, Inc. Biopolymers 93: 8–20, 2010. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com     

Effects of the timing of initial feeding on growth and survival of spotted mandarin fish Siniperca scherzeri larvae

The effects of delayed first feeding on growth and survival of spotted mandarin fish Siniperca scherzeri larvae were examined under controlled conditions. Morphometric characters [yolk‐sac volume, oil globule volume, head depth (H_D), body depth (B_D), eye diameter (E_D), musculature height (M_H), mouth diameter (M_D) and total length (L_T)], body mass (M), specific growth rate (S_GR) and survival were evaluated under different first‐feeding time (2, 3, 4 and 5 days after hatching). Larvae began to feed exogenously at 2 days after hatching (DAH) and the point of no return (P_NR) occurred between 5 and 6 DAH at 23° C, range ±1·0° C. The yolk volume of larvae first‐fed at 2 days had a significant difference compared with that of larvae first‐fed at 3, 4 and 5 days on 3 and 4 DAH. The larvae first‐fed at 2 days achieved comparatively better growth performance than that of 3, 4 and 5 days. On 5 DAH, all morphometric characters had significant differences between 2 and 5 days and 2 and 4 days initial feeding, respectively. Total mortality was recorded on 9 DAH for the larvae first‐fed at 5 days. On 12 DAH, significant differences were observed between 2 and 4 days and 3 and 4 days initial feeding for all morphometric characters. From 16 DAH to the end of experiment, all growth variables of the larvae first‐fed at 2 days were significantly higher than those in other treatments. The S_GR (2–9 DAH) first‐fed at 2 and 3 days were significantly higher than 4 and 5 day treatments, and the S_GR (9–16 DAH) first‐fed at 2 days was significantly higher than 3 and 4 day treatments. There was no significant difference, however, of S_GR (16–28 DAH) among treatments. Survival rate was significantly higher at 2 days initial feeding (27·42%) when compared with 3 (15·96%) and 4 days (7·92%) initial feeding at the end of experiment. The present study suggests that the first feeding of S. scherzeri larvae should be initiated at 2 days after hatching for achieving good growth and survival.     

Diurnal rhythms of plasma growth hormone,somatostatin, thyroid hormones,cortisol and glucose concentrations in rainbow trout,Oncorhynchus mykiss,during progressive food deprivation

Abstract

The diurnal patterns of changes in plasma cortisol, growth hormone (GH), somatostatin‐14 (SRIF), thyroid hormones (L‐thyroxine, T₄ and triiodo‐L‐thyronine, T₃) and glucose were investigated in rainbow trout, Oncorhynchus mykiss, both during single meal‐feeding and during a progressive fast of 13 weeks. All measured variables exhibited a diurnal pattern in fed rainbow trout, most of which appeared to be correlated with the time of feeding (30–60 min after the onset of light), while additional changes, associated with the scotophase were also found for cortisol. Although fasting had no affect on mean daily plasma cortisol or SRIF concentrations, there was a progressive increase in mean daily plasma GH concentrations and a progressive decrease in mean daily plasma thyroid hormone and glucose concentrations associated with fasting. However, for GH, significant changes were not evident until week 10 of the fast. In addition, fasting appeared to phase‐shift the diurnal patterns of plasma GH, cortisol and glucose concentrations; the consequence of such a shift is that conclusions as to the effects of fasting, if based on a single time of sampling, may be flawed.     

Swimming in four goldfish Carassius auratus morphotypes: understanding functional design and performance employing artificially selected forms

Four goldfish Carassius auratus morphotypes of similar length (50 mm): common (streamlined, full complement of paired and median fins, bifurcated caudal fin), comet (‘common like’ but with a long, deeply forked caudal fin), fantail (short, deep body with twinned caudal and anal fins) and eggfish (similar to the fantail but lacking a dorsal fin) were compared. Drag, steady swimming kinematics, energetics, fast‐start performance, stability in yaw and roll and propulsive muscle ultrastructural characteristics were measured. A performance ‘pairing’ (common and comet; fantail and eggfish) was a recurrent theme for most performance variables. Fantail and eggfish drag were higher (requiring more thrust at any given velocity) than those for the more streamlined common and comet. This was reflected in kinematics; tailbeat frequency and stride length at any given velocity for the common and comet were lower and higher, respectively, than that of the fantail and eggfish. Common and comet fatigue times were not significantly different from those of their ancestor, crucian carp Carassius carassius, and higher than the fantail and eggfish. The cost of transport of the common and comet (c. 0· 6 mg O₂ kg^?1 m^?1) was accurately predicted by the mass scaling relationship for fish, but values for the fantail and eggfish (c. 1· 3 mg O₂ kg^?1 m^?1) were not. Rolling and yawing motions in eggfish (dorsal fin absent) during steady swimming were associated with significant energy losses. Eggfish maximum fast‐start acceleration (c. 5 m s^?2) was poor due to the absence of inertial and lifting contributions to thrust from the dorsal fin and energy wasting rolling motions. Common and comet fast‐start performance (average velocity c. 0· 45 m s^?1, maximum velocity c. 1· 2 m s^?1, average acceleration c. 7· 5 m s^?2, maximum acceleration c. 35 m s^?2) was similar to that of other locomotor generalists (e.g. rainbow trout Oncorhynchus mykiss). Artificially selected fishes can contribute to the understanding of form and movement in fishes and complement studies of the role of locomotor adaptations in natural systems.     

Myotube production in fast myotomal muscle is switched-off at shorter body lengths in male than female Atlantic halibut Hippoglossus hippoglossus (L.) resulting in a lower final fibre number

A sampling method is described to determine accurately the number of fast myotomal muscle fibres (N_F) in a large flatfish species, the Atlantic halibut Hippoglossus hippoglossus. An unusual feature of the fast myotomal muscle is the presence of internalized strips of slow muscle fibres. In fish of 1·5–3·5 kg (n = 24), the total cross‐sectional area (A_TC) of fast muscle was 18% greater in the dorsal than ventral myotomal compartments (P < 0·05), whereas there was no significant difference between left‐ and right‐hand sides of the body. Due the bilateral asymmetry, muscle blocks (5 × 5 × 5 mm) were prepared to systematically sample each myotomal quadrant (dorsal, ventral, left‐ and right‐side) and the diameters of 150 fast fibres measured per block. Smooth non‐parametric probability functions were fitted to a minimum of 800 measurements of fibre diameter per quadrant (n = 5). There were no significant differences in the distribution of muscle fibre diameters between myotomal compartments and therefore N_F could be estimated from a single quadrant. The number of blocks required to estimate N_F with a repeatability of ±2·5% increased from six at 300 g body mass to 17 at 96·5 kg, caused by variation within and between blocks. Gompertz curves were fitted to measurements of fibre number and fork length (L_F). The estimated final fibre number was 8·96 × 10⁵ (7·99–9·94 × 10⁵, 95% CI) for males and 1·73 × 10⁶ (1·56–1·90 × 10⁶, 95% CI) for female fish. The estimated L_F for cessation of fibre recruitment in the fast muscle of female fish (1775 mm) was almost twice that in males (810 mm), reflecting their greater ultimate body size.     

Interaction between migration behaviour and estuarine mortality in cultivated Atlantic salmon Salmo salar smolts

Migration behaviour and estuarine mortality of cultivated Atlantic salmon Salmo salar smolts in a 16 km long estuary were studied using two methods: (1) acoustic telemetry and (2) group tagging in combination with trap nets. Progression rates of surviving individuals through the estuary were relatively slow using both methods [0·38 L_T (total length) s^?1 v. 0·25 L_T s^?1]. In 2012, the progression rate was slow from the river to the estuary (0·55 L_T s^?1) and the first part of the estuary (0·31 L_T s^?1), but increased thereafter (1·45–2·21 L_T s^?1). In 2013, the progression rate was fast from the river to the estuary (4·31 L_T s^?1) but was slower thereafter (0·18–0·91 L_T s^?1). Survival to the fjord was higher in 2012 (47%) compared to 2013 (6%). Fast moving individuals were more likely to migrate successfully through the estuary compared to slower moving individuals. Adult recapture of coded‐wire‐tagged S. salar was generally low (0·00–0·04%). Mortality hot spots were related to topographically distinct areas such as the river outlet (in 2012) or the sill separating the estuary and the fjord (in 2013). At the sill, an aggregation of cod Gadus morhua predating on cultivated smolts was identified. The results indicate that slow progression rates through the estuary decreases the likelihood of smolts being detected outside the estuary. The highly stochastic and site‐specific mortality patterns observed in this study highlight the complexity in extrapolating mortality patterns of single release groups to the entire smolt run of wild S. salar.     

Cholecystokinin and tryptic activity in the gut and body of developing Atlantic halibut larvae: evidence for participation in the regulation of protein digestion

At 7 days after first feeding (DAFF), the peptide hormone cholecystokinin (CCK) content (fmol individual^?1) and the tryptic activity [μmol arginine‐methyl‐coumarinyl‐7‐amide (MCA) min^?1 individual^?1] per individual gut of Atlantic halibut Hippoglossus hippoglossus larvae were low: 0·2 ± 0·1 and 0·14 ± 0·10, respectively. Thereafter, both parameters increased with the increase in gut mass and reached 19·67 ± 5·58 and 2·71 ± 0·64 at 26 DAFF, respectively. Due to the small sample size, the dry mass (M_G, mg) of the individual gut could not be determined accurately at 7 DAFF. At 13 DAFF M_G represented 5·5% of whole body dry mass (M_w, mg) while at 26 DAFF it had increased to 23%. The mass specific tryptic activity [μmol MCA min^?1 per mg dry mass (M)] in the gut increased from 2·74 ± 1 ± 98 at 13 DAFF to 5·00 ± 0·78 at 26 DAFF. There was more individual variation in the mass specific CCK content (fmol M^?1) but no significant differences were found, although the data indicated an increase (from 23·38 ± 11·26 at 13 DAFF to 36·27 ± 8·96 fmol M^?1 at 26 DAFF). At 7 DAFF the CCK content of the gut represented c. 2% of the whole body CCK content while it increased to c. 62% of the whole body CCK content at 26 DAFF. This demonstrates that it is necessary to separate neural and gastrointestinal sources of CCK in order to determine its alimentary role in fish larvae. Trypsin activity was only found in the gut compartment. In larvae aged 45 DAFF dietary proteins delivery into the gut by tube‐feeding appeared to stimulate post‐prandial secretion of CCK from the gut as well as stimulate pancreatic trypsin secretion, suggesting that both factors contribute to protein digestion.     

Characterization of Schizothorax prenanti cgnrhII gene: fasting affects cgnrhII expression

In this study, the role of chicken gonadotropin‐releasing hormone II (cgnrhII) in feeding regulation was investigated in Schizothorax prenanti. First, the full‐length S. prenanti cgnrhII cDNA consisted of 693 bp with an open reading frame of 261 bp encoding a protein of 86 amino acids. Next, cgnrhII was widely expressed in the central and peripheral tissues. Last, there were significant changes in cgnrhII mRNA expression in the fasted group compared to the fed group in the S. prenanti hypothalamus during 24 h fasting (P < 0·05). Furthermore, the cgnrhII gene expression presented a significant decrease in the fasted group compared with the fed group (P < 0·05) on days 3, 5 and 7, after re‐feeding, there was no significant changes in cgnrhII mRNA expression level between refed and fed group on day 9 (P > 0·05). Thus, the results suggest that cGnRH II expression is influenced by fasting and the gene may be involved in feeding regulation in S. prenanti.     

Unravelling growth trajectories from complicated otoliths – the case of Brazilian codling Urophycis brasiliensis

Uncertainty regarding the age determination of the Brazilian codling Urophycis brasiliensis has hampered its stock assessment. Transverse sections of otoliths displayed up to seven (in males) and 12 (in females) alternate opaque and translucent bands that could not be conclusively validated as annuli, resulting in unrealistically high ages of first maturity (A₅₀) (A_50male = 4·5 years and A_50female = 6 years). Therefore, growth was described by the von Bertalanffy (VB) model using an alternative approach that combined microstructure data (daily growth increments) and a fixed asymptotic total length (L_∞). This approach was supported by applying it to two other co‐occurring species, the whitemouth croaker Micropogonias furnieri and the king weakfish Macrodon atricauda, for which daily and annual ring formation has previously been validated. The sensitivity to realistic errors associated with the choice of the L_∞ and the daily increment readings was shown to be low. The results show that U. brasiliensis has a fast growth rate (K_male = 1·19 year^?1, K_female = 0·71 year^?1) and early maturation (A_50male = 1·1–1·5 years, A_50female = 1·6–1·8 years); typical life‐history traits for a sub‐tropical coastal gadiform. This novel study offers an alternative approach for age and growth reconstruction for species with complex patterns of opaque and translucent bands provided that daily growth increments in the yearlings can be counted and L_∞ reliably estimated.     

Antiviral activity of Distictella elongata (Vahl) Urb. (Bignoniaceae), a potentially useful source of anti-dengue drugs from the state of Minas Gerais, Brazil

Aims: To investigate the in vitro antiviral activity of Distictella elongata (Vahl) Urb. ethanol extracts from leaves (LEE), fruits (FEE), stems and their main components. Methods and Results: The antiviral activity was evaluated against human herpesvirus type 1 (HSV‐1), murine encephalomyocarditis virus (EMCV), vaccinia virus Western Reserve (VACV‐WR) and dengue virus 2 (DENV‐2) by the 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) colorimetric assay. LEE presented anti‐HSV‐1 [EC₅₀ 142·8 ± 5·3 μg ml^?1; selectivity index (SI) 2·0] and anti‐DENV‐2 activity (EC₅₀ 9·8 ± 1·3 μg ml^?1; SI 1·5). The pectolinarin ( 1 ) isolated from LEE was less active against HSV‐1 and DENV‐2. A mixture of the triterpenoids ursolic, pomolic and oleanolic acids was also obtained. Ursolic and oleanolic acids have shown antiviral activity against HSV‐1. A mixture of pectolinarin ( 1 ) and acacetin‐7‐O‐rutinoside ( 2 ) was isolated from FEE and has presented anti‐DENV‐2 activity (EC₅₀ 11·1 ± 1·6 μg ml^?1; SI > 45). Besides the antiviral activity, D. elongata has disclosed antioxidant effect. Conclusions: These data shows that D. elongata has antiviral activity mainly against HSV‐1 and DENV‐2, besides antioxidant activity. These effects might be principally attributed to flavonoids isolated. Significance and Impact of the Study: Distictella elongata might be considered a promising source of anti‐dengue fever phytochemicals.     

A test of the preference–performance hypothesis with stream insects: selective oviposition affects the hatching success of caddisfly eggs

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Reproductive biology of female striped marlin Kajikia audax in the western Pacific Ocean

Length and mass data for 1260 (536 females, 683 males, 41 sex unknown) striped marlin Kajikia audax were collected at the fish markets of Tungkang, Singkang and Nanfangao from July 2004 to September 2010. Of these samples, 534 gonads (236 females and 298 males) ranging from 95 to 206 cm in eye‐to‐fork length (L _EF ) and 8 to 88 kg in round mass (M _R), were collected. Chi‐square tests indicated sex ratios were homogeneous among months in 2004 and 2006–2008, but not in 2005, 2009 and 2010; and there were significant differences in sex ratio by size. The overall sex ratio (R _S ) differed significantly from the expected 0·5. Kajikia audax are sexually dimorphic and the proportions of females increased with size between 140 and 210 cm L _EF . Reproductive activity was assessed using a gonado‐somatic index (I _G ), external appearance of the gonads and histological examination and results indicated that the spawning season occurred from April to August with a peak in June to July. Based on histological observations and the distribution of oocyte diameters, K. audax are multiple spawners and their oocytes develop asynchronously. The estimated length‐at‐50% maturity (L _EF50 ) was c . 181 cm (c . 4·8 years of age) for females. The proportion of reproductively active females in the spawning season with ovaries containing postovulatory follicles (0·27) indicated that they spawned every 3·7 days on average. The hydrated oocyte method estimated mean ± S.D. batch fecundity (F _B ) to be 4·4 ± 2·02 million eggs; average relative fecundity was 53·6 ± 13·9 oocytes g^?1 M _R; and the average annual fecundity was 181·3 ± 48·3 million eggs. The parameters estimated in this study are key information for stock assessments of K. audax in the north‐western and central Pacific and will contribute to the conservation, management and sustainable yield of this species.     

36° step size of proton‐driven c‐ring rotation in FoF1‐ATP synthase

Synthesis of adenosine triphosphate ATP, the ‘biological energy currency’, is accomplished by F_oF₁‐ATP synthase. In the plasma membrane of Escherichia coli, proton‐driven rotation of a ring of 10 c subunits in the F_o motor powers catalysis in the F₁ motor. Although F₁ uses 120° stepping during ATP synthesis, models of F_o predict either an incremental rotation of c subunits in 36° steps or larger step sizes comprising several fast substeps. Using single‐molecule fluorescence resonance energy transfer, we provide the first experimental determination of a 36° sequential stepping mode of the c‐ring during ATP synthesis.     

Growth, sex differentiation and gonad and plasma levels of sex steroids in male‐ and female‐dominant populations of Dicentrarchus labrax obtained through repeated size grading

Starting from 66 days post hatching (dph), European sea bass Dicentrarchus labrax were graded successively to create a fast growing (L‐extreme) and a slow growing (S‐extreme) population. The L‐extreme population grew significantly larger (ANOVA, n = 89–101, P < 0·01) attaining twice the wet mass of the S‐extreme population at 300 dph (130·9 ± 1·8 v. 66·7 ± 0·9 g, mean ± s .e .). When the two populations were sexed, the L‐extreme consisted of 96·5% and the S‐extreme of 30·2% females, while the ungraded control had 59·2% females. Sex differentiation began first in females at a total length (L_T) of 97 ± 4 mm and wet mass of 9·4 ± 1·2 g (150 dph), and was completed when fish reached 166 ± 6 mm and 53·4 ± 6·4 g (250 dph) in both sexes. Precocious maturation in males was positively correlated to growth. Gonad oestradiol (E₂) was significantly higher in the female‐dominant population at the onset of ovarian differentiation (ANOVA, n = 10, P < 0·05) and in the plasma after the appearance of the first primary oocytes (P < 0·01). Gonad testosterone (T) increased in both populations after sex differentiation (ANOVA, n = 10, P < 0·05), while plasma levels were significantly higher in the male‐dominant population (P < 0·001). Both gonad and plasma 11‐keto testosterone (11‐KT) were significantly higher in the male‐dominant population (ANOVA, n = 10, P < 0·01) reaching maximal values at spermiation. The results suggest that E₂ is closely related with ovarian differentiation and the onset of oogenesis, while T and 11‐KT is more related to spermatogenesis and precocious maturation.     

Catch composition,reproductive biology and diet of the bramble shark Echinorhinus brucus (Squaliformes: Echinorhinidae) from the south‐eastern Arabian Sea

Fishery and biological data are presented for the poorly known bramble shark Echinorhinus brucus (Squaliformes: Echinorhinidae), from the deep waters of the south‐eastern Arabian Sea. A total of 5318 individuals from by‐catch landings of deep‐water bottom set longlines, gillnets and shrimp trawl fisheries operating at depths of 200–1200 m were recorded between January 2008 and December 2011 at the Kochi Fisheries Harbour (Kerala). A total of 431 individuals, from 46 to 318 cm total length (L_T) and 0·8 to 132 kg total mass (M_T), were examined to determine biological data for this species. The L_T at which 50% were mature (L_T50) for females and males was estimated at 189 and 187 cm L_T. Litter size ranged from 10 to 36 and size at birth was between 42 and 46 cm L_T. Dietary analysis of stomach contents revealed E. brucus feeds on a variety of prey including crustaceans (69% index of relative importance, I_RI), teleosts (25·8% I_RI), cephalopods (1·7% I_RI) and elasmobranchs (0·7% I_RI). This study provides the first detailed biological data for this species and also highlights the extent of the by‐catch fishery for this species in Indian waters.