Assessing mortality changes from size‐frequency curves

The relationship between the instantaneous mortality rate ( Z ) and the instantaneous change in length‐frequency distribution of organisms per unit of animal size (μ _L ) takes the following form: Z  = μ _L k ( L _∞ −  L ) +  k , where k and L _∞ are coefficients of the von Bertalanffy equation, and L is organism size (length). Z and μ_L change coherently when they are measured for a specific size or age class. Therefore, observations of μ _L can provide information on the relative changes in mortality. This is useful when no precise information about animal growth is available and growth curve is assumed to be invariable. This method was tested on a heavily exploited population of St Peter's fish (mango tilapia) Sarotherodon galilaeus in Lake Kinneret, Israel, where large fluctuations in the size structure of the catch have occurred over the past few years. Analysis of the changes in the length‐frequency distributions showed that the changes in μ _L over multiple years estimated for fully exploited fish reflected the respective changes in Z .     

Gastric emptying and the return of appetite in juvenile turbot, Scophthalmus maximus L., fed on artificial diets

Gastric emptying time in Scophthalmus maximus , when fed friable artificial pellets based on fishmeal, is composed of two phases:
(a) a delay time (t_d) during which the meal forms a bolus and which shortens with temperature, and
(b) an emptying phase (duration t_end ) which varies with meal size ( S ), body weight ( W ) and temperature (71 according to:
(where t _end is in h, S is in g, W is in g and T is °C). During the emptying phase, stomach contents decrease curvilinearly according to:
(where S_t , & S_o is in g and t is in h) in which the instantaneous digestion rate, K , varies with fish weight and temperature as:
Food pellets were prepared which remained separate and did not form a bolus in the stomach; K increased if a given meal size was subdivided to increase surface area. If meal size was increased by ingestion of identical pellets, K decreased. After a satiation meal, appetite in young turbot returns in direct relation to the degree of stomach emptiness. When food is regularly available, young turbot feed steadily at a rate which maintains their stomachs at c. 85% maximum fullness. When trained to use demand feeders, the fish interact as a group to feed rhythmically, but feeding rate falls 33% to only two-thirds of the previous rate since stomach fullness, and hence digestion rate (g h⁻¹), is maintained at a lower level. Reduction in dietary energy density below 1 kCal g⁻¹ increases gastric emptying rate and the turbot demonstrate partial compensation by increasing food intake. On energy-rich diets, protein nitrogen and energy assimilation efficiencies remain high (97 5% and 91% respectively) irrespective of feeding rate and frequency.     

Borrelia burgdorferi BBB07 interaction with integrin alpha3beta1 stimulates production of pro-inflammatory mediators in primary human chondrocytes

Borrelia burgdorferi , the causative agent of Lyme disease, activates multiple signalling pathways leading to induction of pro-inflammatory mediators at sites of inflammation. Binding of B. burgdorferi to integrin α₃β₁ on human chondrocytes activates signalling leading to release of several pro-inflammatory mediators, but the B. burgdorferi protein that binds integrin α₃β₁ and elicits this response has remained unknown. A search of the B. burgdorferi genome for a canonical integrin binding motif, the RGD (Arg–Gly–Asp) tripeptide, revealed several candidate ligands for integrins. In this study we show that one of these candidates, BBB07, binds to integrin α₃β₁ and inhibits attachment of intact B. burgdorferi to the same integrin. BBB07 is expressed during murine infection as demonstrated by recognition by infected mouse sera. Recombinant purified BBB07 induces pro-inflammatory mediators in primary human chondrocyte cells by interaction with integrin α₃β₁. This interaction is specific, as P66, another integrin ligand of B. burgdorferi , does not activate signalling through α₃β₁. In summary, we have identified a B. burgdorferi protein, BBB07, that interacts with integrin α₃β₁ and stimulates production of pro-inflammatory mediators in primary human chondrocyte cells.     

The reproduction, age and growth of the spotted rockling

Biology of the Macaronesian endemic rockling Gaidropsarus guttatus was studied in the Azores. The overall sex ratio from the samples was highly in favour of females (1 : 6·33). The growth parameters were L _∞ = 24·23, k  = 1·219 and t ₀ = −0·059. Fish matured at 15 cm L _T and the spawning season was strongly clustered in April.     

A qRT-PCR-based method for the measurement of rrn operon copy number

Aim:  To develop a convenient and accurate method for estimating the rrn operon copy number ( Y _rrn ) in cells of pure prokaryotic cultures based on quantitative real-time polymerase chain reaction (qRT-PCR).
Methods & Results:  Using Escherichia coli, the Y _rrn of which is known to be 7, as a reference, the rrn concentrations of target species and E. coli in sample solutions were measured based on their respective threshold cycle numbers ( C _t ), whereas the cell concentrations of both species were measured by microscopic counting after staining. The Y _rrn of the target species was then calculated from the initial cell concentrations and the rrn concentrations of the target species and E. coli . Using this method, the Y _rrn values of four species, i.e. Xanthomonas campestris , Staphylococcus aureus , Aeromonas hydrophila and Pseudomonas fluorescens , were estimated as 1·80, 4·73, 8·58 and 5·13, respectively, comparable to their respective known values of 2, 5, 10, and 5, resulting in an average deviation of 8%.
Conclusions:  The whole cell qRT-PCR based methods were convenient, accurate and reproducible in quantification of rrn copy number of prokaryotic cells.
Significance and Impact of the Study:  qTR-PCR is a fast and reliable DNA quantification approach. Compared with previous qTR-PCR based methods measuring rrn copy number, the present method avoided the prerequisite for the information on genome size and GC content of target bacteria or a gene with known copy number, thus should be more widely applicable.     

Age, growth and mortality of the Mediterranean amberjack Seriola dumerili (Risso 1810) from the south-eastern Adriatic Sea

The age, growth and mortality of the Mediterranean amberjack Seriola dumerili (Risso 1810) were determined in 298 specimens collected in the south-eastern Adriatic Sea (Donji Molunat Bay) from 17 May to 26 June 1997. The total length ranged from 32.0 to 160.0 cm and weight from 0.5 to 46.5 kg. Ten age classes, ranging from 1° to 10° years were defined by scale readings. Mean total length and weight-at-age data were used to estimate the growth parameters of the von Bertalanffy equation: L _∞=174.6 cm, K =0.190, t ₀=–0.314; W _∞=79.01 kg, K =0.139 and t ₀=–0.746. The length–weight relationship was estimated at: W =0.000123 ×  L ^2.847. The overall sex ratio was 1 : 1.06 in favour of males. Total ( Z ) and natural ( M ) mortality were found to be 0.41 years⁻¹ and 0.30 years^–1, respectively. The exploitation ratio ( E =0.27) indicates that the fishing pressure on the Mediterranean amberjack was low in the investigated region.     

Compensatory growth of juvenile brown flounder Paralichthys olivaceus (Temminck & Schlegel) following thermal manipulation

The compensatory growth of juvenile brown flounder Paralichthys olivaceus (body mass c. 12 g) following different thermal exposure was investigated. Fish were exposed to one of the five temperatures: 8·5 ( T _8·5), 13·0 ( T _13·0), 17·5 ( T _17·5), 22·0 ( T _22·0) and 26·5° C ( T _26·5) for 10 days and fish grew best at 22·0° C. Then the water temperature in all treatments was equably adjusted to 22·0° C over 3 days. At the end of the following 30 days after temperature adjustment, there were no significant differences between body masses of fish in the different treatments (wet body mass at the end of the experiment ranged from 22·13 to 24·56 g). Results indicated that the juvenile P. olivaceus achieved complete compensatory growth. Analysis of the dynamics of the feeding rates and feed conversion efficiencies indicated that compensatory growth of the fish experienced low temperature ( T _8·5, T _13·5 and T _17·5) or high temperature ( T _26·5) exposure was mainly dependent on increasing feed intake (hyperphagia) and possibly by improvement in feed conversion efficiency. The moisture content was not affected by different temperature exposure significantly. The lipid and energy content of juvenile P. olivaceus in T _8·5, however, were significantly lower than other treatment. Results of the current study indicate that a short period of low or high temperature exposure may not affect annual growth, but may affect lipid and energy deposition.     

NIDA522131, a new radioligand for imaging extrathalamic nicotinic acetylcholine receptors: in vitro and in vivo evaluation

A novel radioligand, 6-chloro-3-((2-( S )-azetidinyl)methoxy)-5-(2-fluoropyridin-4-yl)pyridine (NIDA522131), for imaging extrathalamic nicotinic acetylcholine receptors (nAChRs) was characterized in vitro and in vivo using positron emission tomography. The K_d and T_1/2 of dissociation of NIDA522131 binding measured at 37°C in vitro were 4.9 ± 0.4 pmol/L and 81 ± 5 min, respectively. The patterns of radioactivity distribution in monkey brain in vivo was similar to that of 2-[¹⁸F]fluoro-3-(2( S )-azetidinylmethoxy)pyridine (2FA), a radioligand that has been successfully used in humans, and matched the α₄β₂* nAChRs distribution. Comparison between [¹⁸F]NIDA522131 and 2FA demonstrated better in vivo binding properties of the new radioligand and substantially greater radioactivity accumulation in brain. Consistent with [¹⁸F]NIDA522131 elevated affinity for nAChRs and its increased lipophilicity, both, the total and non-displaceable distribution volumes were substantially higher than those of 2FA. Estimated binding potential values in different brain regions, characterizing the specificity of receptor binding, were 3–4 fold higher for [¹⁸F]NIDA522131 than those of 2FA. Pharmacological evaluation in mice demonstrated a toxicity that was comparable to 2FA and is in agreement with a 2300 fold higher affinity at α₄β₂* versus α₃β₄* nAChRs. These results suggest that [¹⁸F]NIDA522131 is a promising positron emission tomography radioligand for studying extrathalamic nAChR in humans.     

Selectivity of diamond, square and hexagonal mesh codends for Atlantic horse mackerel Trachurus trachurus, European hake Merluccius merluccius, and greater forkbeard Phycis blennoides in the eastern Mediterranean

Knowledge on selectivity is considered essential for an adequate management of the trawl fishery. This paper compares the selectivity of conventionally used codends of 44 mm diamond mesh, 40 mm square, and hexagonal mesh for Atlantic horse mackerel, European hake and greater forkbeard in the eastern Mediterranean. Data were collected using the covered codend method and analyzed using the logistic equation with the maximum likelihood method. Mean selection curves were analyzed and compared using the between-haul variations model. The mean L ₅₀ v alues of diamond, square and hexagonal mesh codends are 14.7, 15.9 and 17.1 cm total length (TL) for horse mackerel, 10.4, 14.4 and 11.0 cm TL for hake and 12.2, 15.8 and 12.7 cm TL for greater forkbeard, respectively. For horse mackerel, the differences among the codends are statistically significant (P   <   0.05); however, the L ₅₀ values for all three meshes are higher than the minimum landing size (13 cm TL). For hake and greater forkbeard, the L ₅₀ value for the square mesh codend is significantly different from the values for diamond and hexagonal meshes (P   <   0.01), whereas there is no significant difference between diamond and hexagonal meshes (P   >   0.05). Regardless of the mesh shape, L ₅₀ values were substantially lower than the minimum landing size or size at first maturity for hake and greater forkbeard. To enable more effective solutions, research on species-selective trawls is required for the Mediterranean demersal trawl fishery.     

Anion exchange in the giant erythrocytes of African lungfish

Carbon dioxide transport in African lungfish Protopterus aethiopicus blood conformed to the typical vertebrate scheme, implying a crucial and rate-limiting role of erythrocyte Cl^–/HCO₃^– exchange. The rate coefficient for unidirectional Cl^– efflux via the anion exchanger ( k , s⁻¹) increased with temperature in African lungfish, but values were well below those reported in other species. The erythrocytes of African lungfish were, however, very large (mean cellular volume = 6940 µm³), and the ratio of cell water volume to membrane surface area was high ( V _w A _m⁻¹ = 1·89). Hence, the apparent Cl^– permeability ( P _Cl =  kV _w A _m⁻¹, µm s⁻¹) was close to that in other vertebrates. The plot of ln P _Cl against the inverse absolute temperature was left-shifted in the tropical African lungfish compared to the temperate rainbow trout Oncorhynchus mykiss , which supports the idea that P _Cl is similar among animals when compared at their preferred temperatures. Also, Q ₁₀ for anion exchange calculated from P _Cl values in African lungfish was 2·0, supporting the idea that the temperature sensitivity of erythrocyte anion exchange matches the temperature sensitivity of CO₂ production and transport in ectothermic vertebrates.     

Swimming performance, oxygen consumption and excess post-exercise oxygen consumption in adult transgenic and ocean-ranched coho salmon

Routine oxygen consumption ( M o ₂) was 35% higher in 1 day starved and 21% higher in 4 day starved adult transgenic coho salmon Oncorhynchus kisutch relative to end of migration ocean-ranched coho salmon. Critical swimming speed ( U _crit) and M o ₂ at U _crit ( M o _2max) were significantly lower in 4 day starved transgenic coho salmon (1·25 BL s⁻¹; 8·79 mg O₂ kg⁻¹ min⁻¹) compared to ocean-ranched coho salmon (1·60 BL s⁻¹; 9·87 mg O₂ kg⁻¹ min⁻¹). Transgenic fish swam energetically less efficiently than ocean-ranched fish, as indicated by a poorer swimming economy at U _crit ( M o _2max     ). Although M o _2max was lower in transgenic coho salmon, the excess post-exercise oxygen consumption (EPOC) measured during the first 20 min of recovery was significantly larger in transgenic coho salmon (44·1 mg O₂ kg⁻¹) compared with ocean-ranched coho salmon (34·2 mg O₂ kg⁻¹), which had a faster rate of recovery.     

Purification and biochemical characterization of an endoxylanase from Aspergillus versicolor

An endoxylanase (β-1,4-xylan xylanohydrolase, EC 3.2.1.8) was purified from the culture filtrate of a strain of Aspergillus versicolor grown on oat wheat. The enzyme was purified to homogeneity by chromatography on DEAE-cellulose and Sephadex G-75. The purified enzyme was a monomer of molecular mass estimated to be 19 kDa by SDS-PAGE and gel filtration. The enzyme was glycoprotein with 71% carbohydrate content and exhibited a pI of 5.4. The purified xylanase was specific for xylan hydrolysis. The enzyme had a K _m of 6.5 mg ml⁻¹ and a V _max of 1440 U (mg protein)⁻¹.     

Re-interpretation of the logistic equation for batch microbial growth in relation to Monod kinetics

Aims:  To determine the underlying substrate utilization mechanism in the logistic equation for batch microbial growth by revealing the relationship between the logistic and Monod kinetics. Also, to determine the logistic rate constant in terms of Monod kinetic constants.
Methods and Results:  The logistic equation used to describe batch microbial growth was related to the Monod kinetics and found to be first-order in terms of the substrate and biomass concentrations. The logistic equation constant was also related to the Monod kinetic constants. Similarly, the substrate utilization kinetic equations were derived by using the logistic growth equation and related to the Monod kinetics.
Conclusion:  It is revaled that the logistic growth equation is a special form of the Monod growth kinetics when substrate limitation is first-order with respect to the substrate concentration. The logistic rate constant ( k ) is directly proportional to the maximum specific growth rate constant ( μ _m) and initial substrate concentration ( S ₀) and also inversely related to the saturation constant ( K _s).
Significance and Impact of the Study:  The semi-empirical logistic equation can be used instead of Monod kinetics at low substrate concentrations to describe batch microbial growth using the relationship between the logistic rate constant and the Monod kinetic constants.     

Copper effect on cytochrome b of photosystem II under photoinhibitory conditions

Toxic Cu (II) effect on cytochrome b ₅₅₉ under aerobic photoinhibitory conditions was examined in two different photosystem II (PSII) membrane preparations active in oxygen evolution. The preparations differ in the content of cytochrome b ₅₅₉ redox potential forms. Difference absorption spectra showed that the presence of Cu (II) induced the oxidation of the high-potential form of cytochrome b ₅₅₉ in the dark. Addition of hydroquinone reduced the total oxidized high-potential form of cytochrome b ₅₅₉ present in Cu (II)-treated PSII membranes indicating that no conversion to the low-potential form took place. Spectroscopic determinations of cytochrome b ₅₅₉ during photoinhibitory treatment showed slower kinetics of Cu (II) effect on cytochrome b ₅₅₉ in comparison with the rapid loss of oxygen evolution activity in the same conditions. This result indicates that cytochrome b ₅₅₉ is affected after PSII centres are photoinhibited. The high-potential form was more sensitive to toxic Cu (II) action than the low-potential form under illumination at pH 6.0. The content of the high-potential form of cytochrome b ₅₅₉ was completely lost; however, the low-potential content was unaffected in these conditions. This loss did not involve cytochrome protein degradation. The results are discussed in terms of different binding properties of the heme iron to the protonated or unprotonated histidine ligand in the high-potential and low-potential forms of cytochrome b ₅₅₉, respectively.     

Three parameters comprehensively describe the temperature response of respiratory oxygen reduction

Using an exponential model that relies on Arrhenius kinetics, we explored Type I, Type II and dynamic (e.g. declining Q ₁₀ with increasing temperature) responses of respiration to temperature. Our Arrhenius model provides three parameters: R _REF (the base of the exponential model, nmol g⁻¹ s⁻¹), E ₀ (the overall activation energy of oxygen reduction that dominates its temperature sensitivity, kJ mol⁻¹) and δ (that describes dynamic responses of E ₀ to measurement temperature, 10³ K²). Two parameters, E ₀ and δ , are tightly linked. Increases in overall activation energy at a reference temperature were inversely related to changes in δ . At an E ₀ of ca. 45 kJ mol⁻¹, δ approached zero, and respiratory temperature response was strictly Arrhenius-like. Physiologically, these observations suggest that as contributions of AOX to combined oxygen reduction increase, E ₀( REF ) decreases because of different temperature sensitivities for V _max, and δ increases because of different temperature sensitivities for K _1/2 of AOX and COX. The balance between COX and AOX activity helps regulate plant metabolism by adjusting the demand for ATP to that for reducing power and carbon skeleton intermediates. Our approach enables determination of respiratory capacity in vivo and opens a path to development of process-based models of plant respiration.     

Purification and biochemical characterization of a membrane-bound [NiFe]-hydrogenase from a hydrogen-oxidizing, lithotrophic bacterium, Hydrogenophaga sp. AH-24

Membrane-bound [NiFe]-hydrogenase from Hydrogenophaga sp. AH-24 was purified to homogeneity. The molecular weight was estimated as 100±10 kDa, consisting of two different subunits (62 and 37 kDa). The optimal pH values for H₂ oxidation and evolution were 8.0 and 4.0, respectively, and the activity ratio (H₂ oxidation/H₂ evolution) was 1.61 × 10² at pH 7.0. The optimal temperature was 75 °C. The enzyme was quite stable under air atmosphere (the half-life of activity was c . 48 h at 4 °C), which should be important to function in the aerobic habitat of the strain. The enzyme showed high thermal stability under anaerobic conditions, which retained full activity for over 5 h at 50 °C. The activity increased up to 2.5-fold during incubation at 50 °C under H₂. Using methylene blue as an electron acceptor, the kinetic constants of the purified membrane-bound homogenase (MBH) were V _max=336 U mg⁻¹, k _cat=560 s⁻¹, and k _cat/ K _m=2.24 × 10⁷ M⁻¹ s⁻¹. The MBH exhibited prominent electron paramagnetic resonance signals originating from [3Fe–4S]⁺ and [4Fe–4S]⁺ clusters. On the other hand, signals originating from Ni of the active center were very weak, as observed in other oxygen-stable hydrogenases from aerobic H₂-oxidizing bacteria. This is the first report of catalytic and biochemical characterization of the respiratory MBH from Hydrogenophaga .     

Biology of the salema, Sarpa salpa (L. 1758) (Pisces, Sparidae) from the middle-eastern Adriatic

Salema Sarpa salpa (Linnaeus, 1758), were caught in the middle-eastern Adriatic Sea from August to November 2004. Length range of the samples was between 10.3 and 43.9 cm, with mean values of 25.5 cm in males, 32.6 cm in females, and 11.4 and 20.2cm in immature and hermaphrodite specimens, respectively. Sex ratio (males : females) was 3.1 : 1. Males were observed up to 37 cm length. Hermaphrodites showed lengths between 18 and 21 cm. The species was characterized by protandric hermaphroditism. Size at sexual maturity was 20.6 cm (2 years old) for males. Total length-total weight relationship for the entire population is described by the parameters a  = 0.00893, and b  = 3.1055. Otolith age readings showed that the population consisted of 15 age groups (1–15 years), including a very high proportion of individuals 1–7 years old. The von Bertalanffy growth parameters were L _∞ = 33.11cm, k  = 0.514, and t ₀ = −0.392 years for males and L _∞ = 40.85cm, k  = 0.179, and t ₀ = −2.606 years for females. Survival rate of females ( S  = 0.870) was much greater than for males ( S  = 0.769).     

Age and growth of the striped mullet, Mugil platanus (Actinopterygii, Mugilidae), in a southwestern Atlantic coastal lagoon (37°32'S–57°19'W): a proposal for a life-history model

Species of Mugilidae are an important economic resource supporting several small communities in Argentina and Brazil through fishing. The growth parameters of the von Bertalanffy model, age limit ( t _0.95) and natural mortality ( M ) for the striped mullet ( Mugil platanus ) Günther, 1880, in the Mar Chiquita coastal lagoon (37°32'S–57°19'W) were estimated. These results constitute the first estimated values for the species as: L _∞ (cm) = 563.82; K (years⁻¹) = 0.30; t ₀ (years) = −0.057 (age groups 1–8); t _0.95 = 10.07 years and M  = 0.30. Moreover, a hypothetical model is proposed for the life history of the adult stock of M. platanus from the Mar Chiquita coastal lagoon based on CPUE data, environmental parameters, ovarian maturity stages, gonadosomatic indexes, the allometric growth coefficient b and deposition of hyaline or opaque rings in the otholiths. Mugil platanus is therefore regarded as a species of moderate to rapid growth rate, with a relatively low longevity and a high natural mortality rate, compared to other species of Mugilidae.     

Inheritance of Anthracnose Resistance in the Common Bean Differential Cultivar G 2333 and Identification of a New Molecular Marker Linked to the Co-42 gene

The inheritance of anthracnose resistance of the common bean ( Phaseolus vulgaris L.) differential cultivar G 2333 to Colletotrichum lindemuthianum races 73 and 89 was studied in crosses with the susceptible cultivar Rudá. The segregation ratios of 15 : 1 in the F₂ and 3 : 1 in the backcrosses to Rudá indicate that for each of the races tested there are two independent resistance loci in G 2333. A random amplified polymorphic DNA (RAPD) molecular marker (OPH18_1200C) linked in resistance to race 73 was identified in a BC₃F_2:3 population derived from crosses between Rudá and G 2333. A RAPD molecular marker OPAS13_950C, previously identified as linked to gene Co-4² , was also amplified in this population. Co-segregation analyses showed that these two markers are located at 5.6 (OPH18_1200C) and 11.2 (OPAS13_950C) cM of the Co-4² gene. These markers were not present in BC₁F_2:3 plants resistant to race 89 indicating that this population carries a different resistance gene. DNA amplification of BC₁F_2:3 plants with RAPD molecular marker OPAB_450C, previously identified as linked to gene Co-5 , indicated that this gene is present in this population.