Invertebrate food supply and breeding success of Mallards Anas platyrhynchos at flooded gravel quarries in southern Britain

Reproductive parameters of Mallard Anas platyrhynchos were investigated in relation to vegetative development, physical characteristics and invertebrate food abundance of a complex of flooded gravel quarries in southern Britain. Breeding pair density over the study area was particularly high with 2.2 pairs km(-1) of shoreline. Across all lakes breeding success was limited to an average of 0.9 ducklings fledged pair(-1), yet an associated study showed that very little of the mortality could be attributed to predation. The results of stepwise multiple regressions indicated that the availability of emerging insects was important in determining the breeding density and breeding success of Mallards rather than habitat structure. We suggest that the overall great availability of suitable nest-sites combined with the variable but unmeasured stocking densities of fish may explain the lack of the latter correlations.

Fluorimetric determination of intra- and extracellular free amino acids in the microalgae Tetraselmis gracilis (Prasinophyceae) using monolithic column in reversed phase mode

This paper describes the development and application of an RP HPLC method using a C(18) monolithic stationary phase for the separation and quantification of extra- and intracellular amino acids in a batch cultivation of the marine alga Tetraselmis gracilis. Fluorimetric detection was made after separation of the o-phthaldialdehyde 2-mercaptoethanol (OPA-2MCE) derivatives using a binary gradient elution. Separation of 19 amino acids was achieved with resolution >1.5 in about 39 min at a flow rate of 1.5 mL/min. RSD of analyses in seawater medium ranged from 0.36% for Orn (0.50 mu mol/L) to 12% for Ile (0.10 mu mol/L). The main constituents of the intracellular dissolved free amino acids (DFAAs) in the exponential growth phase were arginine (Arg), asparagine (Asn), alanine (Ala), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), glycine (Gly), glutamine (Gln), and leucine (Leu). The major amino acids excreted to the media were valine (Val), Ala, Ser, and Gly. The monolithic phase facilitates the analysis by shortening the separation time and saving solvents and instrumentation costs (indeed conventional HPLC instrumentation can be used, running at lower pressures than those ones used with packed particle columns).

Implementing stepwise solvent elution in sequential injection chromatography for fluorimetric determination of intracellular free amino acids in the microalgae Tetraselmis gracilis

The concept of sequential injection chromatography (SIC) was exploited to automate the fluorimetric determination of amino acids after pre-column derivatization with ophthaldialdehyde (OPA) in presence of 2-mercaptoethanol (2MCE) using a reverse phase monolithic C(18) stationary phase. The method is low-priced and based on five steps of isocratic elutions. The first step employs the mixture methanol: tetrahydrofuran: 10 mmol L(-1) phosphate buffer (pH 7.2) at the volumetric ratio of 8:1:91; the other steps use methanol: 10 mmol L-1 phosphate buffer (pH 7.2) at volumetric ratios of 20:80, 35:65, SO:SO and 65:35. At a flow rate of 10 mu L s(-1) a 25 mm long-column was able to separate aspartic acid (Asp), glutamic acid (Glu), asparagine (Asn), serine (Ser), glutamine (Gln), glycine (Gly), threonine (Thr), citruline (Ctr), arginine (Arg), alanine (Ala), tyrosine (Tyr), phenylalanine (Phe), ornithine (Orn) and lysine (Lys) with resolution >1.2 as well as methionine (Met) and valine (Val) with resolution of 0.6. Under these conditions isoleucine (Ile) and leucine (Leu) co-eluted. The entire cycle of amino acids derivatization, chromatographic separation and column conditioning at the end of separation lasted 25 min. At a flow rate of 40 mu L s(-1) such time was reduced to 10 min at the cost of resolution worsening for the pairs Ctr/Arg and Orn/Lys. The detection limits varied from 0.092 mu mol L(-1) for Tyr to 0.51 mu mol L(-1) for Orn. The method was successfully applied to the determination of intracellular free amino acids in the green alga Tetraselmis gracilis during a period of seven days of cultivation. Samples spiked with known amounts of amino acids resulted in recoveries between 94 and 112%. (C) 2008 Elsevier B.V. All rights reserved.

Development of twenty-four novel microsatellite markers for the freshwater crayfish, Geocharax gracilis, using next generation sequencing

The crayfish Geocharax gracilis is an important inhabitant of natural and agricultural drainage systems in south-eastern Australia. To investigate population structure, genetic diversity and patterns of connectivity in natural and human-altered ecosystems, we isolated and characterised 24 microsatellite loci using next generation sequencing. Loci were initially tested for levels of variation based on 12 individuals from across the species’ geographical range. A further 33 individuals from a single population were used to test for departures from Hardy–Weinberg equilibrium and linkage disequilibrium. We detected high to moderate levels of genetic variation across most loci with a mean allelic richness of 8.42 and observed heterozygosity of 0.629 (all samples combined). We found no evidence for linkage disequilibrium between any loci and only three loci (Geo01, Geo24 and Geo47) showed significant deviations from Hardy–Weinberg expectations. These same three loci, plus two additional loci (Geo06 and Geo28), also showed the presence of null alleles. These 24 variable markers will provide an important tool for future population genetic assessments in natural and human altered environments.

Transfer of maternal antibodies against avian influenza virus in mallards (Anas platyrhynchos)

Maternal antibodies protect chicks from infection with pathogens early in life and may impact pathogen dynamics due to the alteration of the proportion of susceptible individuals in a population. We investigated the transfer of maternal antibodies against avian influenza virus (AIV) in a key AIV host species, the mallard (Anas platyrhynchos). Combining observations in both the field and in mallards kept in captivity, we connected maternal AIV antibody concentrations in eggs to (i) female body condition, (ii) female AIV antibody concentration, (iii) egg laying order, (iv) egg size and (v) embryo sex. We applied maternity analysis to the eggs collected in the field to account for intraspecific nest parasitism, which is reportedly high in Anseriformes, detecting parasitic eggs in one out of eight clutches. AIV antibody prevalence in free-living and captive females was respectively 48% and 56%, with 43% and 24% of the eggs receiving these antibodies maternally. In both field and captive study, maternal AIV antibody concentrations in egg yolk correlated positively with circulating AIV antibody concentrations in females. In the captive study, yolk AIV antibody concentrations correlated positively with egg laying order. Female body mass and egg size from the field and captive study, and embryos sex from the field study were not associated with maternal AIV antibody concentrations in eggs. Our study indicates that maternal AIV antibody transfer may potentially play an important role in shaping AIV infection dynamics in mallards.

Fuelling rates of garganey (Anas querquedula) staging in the Camargue, Southern France, during spring migration

Most species of long-distance migratory birds put on energy stores to fuel their travels. However, recent studies have highlighted the potential costs associated with carrying too much fuel, either through increased predation risk or decreased flight efficiency. Consequently, it is now widely accepted that migratory birds should carry optimal rather than maximum fuel loads. Information from 372 garganey (Anas querquedula) ringed and recaptured at least once during the same spring in the Camargue, southern France, was used to document fuelling rates of individual ducks in relation to environmental variation and individual variation in condition. On average, garganey added very little fuel stores in the Camargue (mean gain per day = 0.33 g, less than 0.5% of mean body-mass in total over an average stay of 5 days). Fuelling rates were negatively correlated with body mass at capture, but it cannot be excluded that this pattern was a statistical artefact. Given their body-mass at ringing, garganey could potentially still fly long distances when they stop in the Camargue. It is therefore likely that the aim of their stay in southern France is more for resting than refuelling, a finding that may have implications for the proper management of stop-over sites.

Wingbeat frequency and the body drag anomaly: wind-tunnel observations on a thrush nightingale (Luscinia Luscinia) and a teal (Anas crecca)

A teal (Anas crecca) and a thrush nightingale (Luscinia luscinia) were trained to fly in the Lund wind tunnel for periods of up to 3 and 16 h respectively. Both birds flew in steady flapping flight, with such regularity that their wingbeat frequencies could be determined by viewing them through a shutter stroboscope. When flying at a constant air speed, the teal's wingbeat frequency varied with the 0.364 power of the body mass and the thrush nightingale's varied with the 0.430 power. Both exponents differed from zero, but neither differed from the predicted value (0.5) at the 1 % level of significance. The teal continued to flap steadily as the tunnel tilt angle was varied from -1° (climb) to +6° (descent), while the wingbeat frequency declined progressively by about 11%. In both birds, the plot of wingbeat frequency against air speed in level flight was U-shaped, with small but statistically significant curvature. We identified the minima of these curves with the minimum power speed (Vmp) and found that the values predicted for Vmp, using previously published default values for the required variables, were only about two-thirds of the observed minimum-frequency speeds. The discrepancy could be resolved if the body drag coefficients (CDb) of both birds were near 0.08, rather than near 0.40 as previously assumed. The previously published high values for body drag coefficients were derived from wind-tunnel measurements on frozen bird bodies, from which the wings had been removed, and had long been regarded as anomalous, as values below 0.01 are given in the engineering literature for streamlined bodies. We suggest that birds of any size that have well-streamlined bodies can achieve minimum body drag coefficients of around 0.05 if the feet can be fully retracted under the flank feathers. In such birds, field observations of flight speeds may need to be reinterpreted in the light of higher estimates of Vmp. Estimates of the effective lift:drag ratio and range can also be revised upwards. Birds that have large feet or trailing legs may have higher body drag coefficients. The original estimates of around CDb=0.4 could be correct for species, such as pelicans and large herons, that also have prominent heads. We see no evidence for any progressive reduction of body drag coefficient in the Reynolds number range covered by our experiments, that is 21600-215 000 on the basis of body cross-sectional diameter.

Large scale laboratory cultures of Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) and Diaphanosoma biergei Korinek, 1981 (Cladocera)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cultivo de Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) em laboratório utilizando meio CHU12 e de macrófita com NPK

Two different culture media, namely CHU12 and inorganic fertilizer NPK (20-5-20) associated with macrophyte (Eichhornia crassipes) extract, were used to evaluate the development of A. gracilis. Growth rate, development, nutritional value and medium water quality were analyzed. A. gracilis in macrophyte+NPK medium had mean cell density (333x10(5) cells/mL) higher than medium CHU12 (302x10(5) cells/mL). Protein rate (12.52% PS), dry weight (397x10(7) pg/cell) and ashes (4.08x10(7) pg/cell) in A. gracilis was higher (p<0.05) than medium macrophyte+NPK. on the other hand, lipids, carbohydrates and fibers had the same rate (p>0.05) in both media. When hydrological variables in the culture medium of A. gracilis are taken into account, dissolved oxygen and free CO2 alone failed to have any significant difference (p>0.05) between the media employed. A. gracilis in the macrophyte+NPK medium had the same nutritional rate as the commercial medium CHU12, with excellent results for alga growth. Macrophyte medium may be used for large scale alga culture in the feed of fish larvae.

Structural features of the epididymal region of the domestic duck (Anas plathyrynchos)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Emprego da microscopia eletrônica na avaliação pós-vacinal de epitélio traqueal de patos (Anas platyrhynchos) imunizados contra a doença de Newcastle

Avaliou-se o emprego da microscopia eletrônica de varredura no estudo da reação respiratória pós-vacinal em epitélio traqueal de patos (Anas platyrhynchos) imunizados contra a doença de Newcastle. Foram utilizadas 48 aves, distribuídas em quatro grupos: T1 - grupo de aves-controle (não vacinadas), T2 - grupo de aves vacinadas com a estirpe Ulster 2C, T3 - grupo de aves vacinadas com a estirpe B1 e T4 - grupo de aves vacinadas com a estirpe LaSota. Independente do grupo experimental, as aves não apresentaram sinais clínicos detectáveis de reação respiratória pós-vacinal. Ao microscópio eletrônico de varredura, observou-se que os animais vacinados com as estirpes B1 e LaSota desenvolveram descamação epitelial da traqueia, enquanto os vacinados com a estirpe Ulster 2C não, mostrando um epitélio traqueal íntegro, semelhante ao do grupo-controle. Os patos vacinados com a estirpe B1 mostraram evidências de regeneração epitelial da traqueia decorridos 21 dias pós-vacinação, o que não ocorreu com os vacinados com a amostra LaSota.

Cyclic-loading of the human gracilis and semitendinosus muscle tendons: Study of young adult cadavers

During knee ligament reconstruction, the tendon graft is tensioned to prevent the occurrence of excessive graft elongation during the postoperative period. Tensioning may be achieved by applying a cyclic or static load to the graft during fixation. Although this procedure is part of the surgery, there is no consensus in international literature regarding ideal tension levels to be used in this procedure. This study was conducted on 10 tendons of the human gracilis muscle and 10 tendons of semitendinosus muscle removed from five male cadavers whose mean age was 20.8 years. These tendons underwent 10 in vitro strain cycles at three levels of deformation (2.5, 3, and 4%) and the value of the deforming load used for each cycle was recorded. The statistical analysis demonstrated that in order to attain the same level of deformation during the 10 cycles there was a reduction in the value of strain applied to the graft, observed at the three levels of deformation. It was concluded that the semitendinosus tendon presents a more uniform mechanical behavior and that there is a need for new graft tensioning protocols that consider the force associated with deformation.