Haemolymph inorganic ion composition, physiology and behaviour dependent on temperature and ambient magnesium concentration in early life history stages of Paralomis granulosa (Decapoda, Anomura, Lithodidae)

Marine brachyuran and anomuran crustaceans are completely absent from the extremely cold (-1.8 °C) Antarctic continental shelf, but caridean shrimps are abundant. This has at least partly been attributed to low capacities for magnesium excretion in brachyuran and anomuran lithodid crabs ([Mg2+]HL = 20-50 mmol/L) compared to caridean shrimp species ([Mg2+]HL = 5-12 mmol/L). Magnesium has an anaesthetizing effect and reduces cold tolerance and activity of adult brachyuran crabs. We investigated whether the capacity for magnesium regulation is a factor that influences temperature-dependent activity of early ontogenetic stages of the Sub-Antarctic lithodid crab Paralomis granulosa. Ion composition (Na+, Mg2+, Ca2+, Cl-, [SO4]2-) was measured in haemolymph withdrawn from larval stages, the first and second juvenile instars (crabs I and II) and adult males and females. Magnesium excretion improved during ontogeny, but haemolymph sulphate concentration was lowest in the zoeal stages. Neither haemolymph magnesium concentrations nor Ca2+:Mg2+ ratios paralleled activity levels of the life stages. Long-term (3 week) cold exposure of crab I to 1 °C caused a significant rise of haemolymph sulphate concentration and a decrease in magnesium and calcium concentrations compared to control temperature (9 °C). Spontaneous swimming activity of the zoeal stages was determined at 1, 4 and 9 °C in natural sea water (NSW, [Mg2+] = 51 mmol/L) and in sea water enriched with magnesium (NSW + Mg2+, [Mg2+] = 97 mmol/L). It declined significantly with temperature but only insignificantly with increased magnesium concentration. Spontaneous velocities were low, reflecting the demersal life style of the zoeae. Heart rate, scaphognathite beat rate and forced swimming activity (maxilliped beat rate, zoea I) or antennule beat rate (crab I) were investigated in response to acute temperature change (9, 6, 3, 1, -1 °C) in NSW or NSW + Mg2+. High magnesium concentration reduced heart rates in both stages. The temperature-frequency curve of the maxilliped beat (maximum: 9.6 beats/s at 6.6 °C in NSW) of zoea I was depressed and shifted towards warmer temperatures by 2 °C in NSW + Mg2+, but antennule beat rate of crab I was not affected. Magnesium may therefore influence cold tolerance of highly active larvae, but it remains questionable whether the slow-moving lithodid crabs with demersal larvae would benefit from an enhanced magnesium excretion in nature.

Haemolymph inorganic ion composition, physiology and behaviour dependent on temperature and magnesium in adult Paralomis granulosa (Decapoda, Anomura, Lithodidae)

A low capacity for regulation of extracellular Mg2+ has been proposed to exclude reptant marine decapod crustaceans from temperatures below 0°C and thus to exclude them from the high Antarctic. To test this hypothesis and to elaborate the underlying mechanisms in the most cold-tolerant reptant decapod family of the sub-Antarctic, the Lithodidae, thermal tolerance was determined in the crab Paralomis granulosa (Decapoda, Anomura, Lithodidae) using an acute stepwise temperature protocol (-1°, 1°, 4°, 7°, 10°, and 13°C). Arterial and venous oxygen partial pressures (Po2) in hemolymph, heartbeat and ventilation beat frequencies, and hemolymph cation composition were measured at rest and after a forced activity (righting) trial. Scopes for heartbeat and ventilation beat frequencies and intermittent heartbeat and scaphognathite beat rates at rest were evaluated. Hemolymph [Mg2+] was experimentally reduced from 30 mmol/L to a level naturally observed in Antarctic caridean shrimps (12 mmol/L) to investigate whether the animals remain more active and tolerant to cold (-1°, 1°, and 4°C). In natural seawater, righting speed was significantly slower at -1° and 13°C, compared with acclimation temperature (4°C). Arterial and venous hemolymph Po2 increased in response to cooling even though heartbeat and ventilation beat frequencies as well as scopes decreased. At rest, ionic composition of the hemolymph was not affected by temperature. Activity induced a significant increase in hemolymph [K+] at -1° and 1°C. Reduction of hemolymph [Mg2+] did not result in an increase in activity, an increase in heartbeat and ventilation beat frequencies, or a shift in thermal tolerance to lower temperatures. In conclusion, oxygen delivery in this cold-water crustacean was not acutely limiting cold tolerance, and animals may have been constrained more by their functional capacity and motility. In contrast to earlier findings in temperate and subpolar brachyuran crabs, these constraints remained insensitive to changing Mg2+ levels.

Seawater carbonate chemistry, physiology, morphology of larval sea urchins, Strongylocentrotus purpuratus in a laboratory experiment

Ocean warming and ocean acidification, both consequences of anthropogenic production of CO2, will combine to influence the physiological performance of many species in the marine environment. In this study, we used an integrative approach to forecast the impact of future ocean conditions on larval purple sea urchins (Strongylocentrotus purpuratus) from the northeast Pacific Ocean. In laboratory experiments that simulated ocean warming and ocean acidification, we examined larval development, skeletal growth, metabolism and patterns of gene expression using an orthogonal comparison of two temperature (13°C and 18°C) and pCO2 (400 and 1100 µatm) conditions. Simultaneous exposure to increased temperature and pCO2 significantly reduced larval metabolism and triggered a widespread downregulation of histone encoding genes. pCO2 but not temperature impaired skeletal growth and reduced the expression of a major spicule matrix protein, suggesting that skeletal growth will not be further inhibited by ocean warming. Importantly, shifts in skeletal growth were not associated with developmental delay. Collectively, our results indicate that global change variables will have additive effects that exceed thresholds for optimized physiological performance in this keystone marine species.

Response of the calcifying coccolithophore Emiliania huxleyi to low pH/high pCO2: from physiology to molecular level, 2011

The emergence of ocean acidification as a significant threat to calcifying organisms in marine ecosystems creates a pressing need to understand the physiological and molecular mechanisms by which calcification is affected by environmental parameters. We report here, for the first time, changes in gene expression induced by variations in pH/pCO2 in the widespread and abundant coccolithophore Emiliania huxleyi. Batch cultures were subjected to increased partial pressure of CO2 (pCO2; i.e. decreased pH), and the changes in expression of four functional gene classes directly or indirectly related to calcification were investigated. Increased pCO2 did not affect the calcification rate and only carbonic anhydrase transcripts exhibited a significant down-regulation. Our observation that elevated pCO2 induces only limited changes in the transcription of several transporters of calcium and bicarbonate gives new significant elements to understand cellular mechanisms underlying the early response of E. huxleyi to CO2-driven ocean acidification.

Coralline algal physiology is more adversely affected by elevated temperature than reduced pH

In this study we analyzed the physiological responses of coralline algae to ocean acidification (OA) and global warming, by exposing algal thalli of three species with contrasting photobiology and growth-form to reduced pH and elevated temperature. The analysis aimed to discern between direct and combined effects, while elucidating the role of light and photosynthesis inhibition in this response. We demonstrate the high sensitivity of coralline algae to photodamage under elevated temperature and its severe consequences on thallus photosynthesis and calcification rates. Moderate levels of light-stress, however, were maintained under reduced pH, resulting in no impact on algal photosynthesis, although moderate adverse effects on calcification rates were still observed. Accordingly, our results support the conclusion that global warming is a stronger threat to algal performance than OA, in particular in highly illuminated habitats such as coral reefs. We provide in this study a quantitative physiological model for the estimation of the impact of thermal-stress on coralline carbonate production, useful to foresee the impact of global warming on coralline contribution to reef carbon budgets, reef cementation, coral recruitment and the maintenance of reef biodiversity. This model, however, cannot yet account for the moderate physiological impact of low pH on coralline calcification.

Acid-base physiology response to ocean acidification of two ecologically and economically important holothuroids from contrasting habitats, Holothuria scabra and Holothuria parva

Sea cucumbers are dominant invertebrates in several ecosystems such as coral reefs, seagrass meadows and mangroves. As bioturbators, they have an important ecological role in making available calcium carbonate and nutrients to the rest of the community. However, due to their commercial value, they face overexploitation in the natural environment. On top of that, occurring ocean acidification could impact these organisms, considered sensitive as echinoderms are osmoconformers, high-magnesium calcite producers and have a low metabolism. As a first investigation of the impact of ocean acidification on sea cucumbers, we tested the impact of short-term (6 to 12 days) exposure to ocean acidification (seawater pH 7.7 and 7.4) on two sea cucumbers collected in SW Madagascar, Holothuria scabra, a high commercial value species living in the seagrass meadows, and H. parva, inhabiting the mangroves. The former lives in a habitat with moderate fluctuations of seawater chemistry (driven by day-night differences) while the second lives in a highly variable intertidal environment. In both species, pH of the coelomic fluid was significantly negatively affected by reduced seawater pH, with a pronounced extracellular acidosis in individuals maintained at pH 7.7 and 7.4. This acidosis was due to an increased dissolved inorganic carbon content and pCO2 of the coelomic fluid, indicating a limited diffusion of the CO2 towards the external medium. However, respiration and ammonium excretion rates were not affected. No evidence of accumulation of bicarbonate was observed to buffer the coelomic fluid pH. If this acidosis stays uncompensated for when facing long-term exposure, other processes could be affected in both species, eventually leading to impacts on their ecological role.

Effects of increasing levels of pea hulls in the diet on productive performance, development of the gastrointestinal tract, and nutrient retention of broilers from one to eighteeen days of age

The effects of inclusion of pea hulls (PH) in the diet on growth performance, development of the gastrointestinal tract and nutrient retention were studied in broilers from 1 to 18d of age. There were a control diet based on low fibre ingredients (69.3 total dietary fibre (16.1g crude fibre/kg)) and three additional diets that resulted from the dilution of the basal diet with 25, 50 and 75g PH/kg (81.2, 93.2, and 105.1g total dietary fibre/kg diet, respectively). Each treatment was replicated six times and the experimental unit was a cage with 12 chicks. Growth performance, development of the gastrointestinal tract and the coefficients of total tract apparent retention (CTTAR) of nutrients were recorded at 6, 12 and 18d of age. In addition, jejunal morphology was measured at 12 and 18d and the coefficients of apparent ileal digestibility (CAID) of nutrients at 18d of age. Pea hulls inclusion affected all the parameters studied. The inclusion of 25 and 50g PH/kg diet improved growth performance as compared to the control diet. The relative weight (g/kg body weight) of proventriculus (P≤0.01), gizzard (P≤0.001) and ceca (P≤0.05) increased linearly as the level of PH in the diet increased. The inclusion of PH affected quadratically (P≤0.01) villus height:crypt depth ratio with the highest value shown at 25g PH/kg. In general, the CTTAR and CAID of nutrients increased linearly and quadratically (P≤0.05) with increasing levels of PH, showing maximum values with PH level between 25 and 50g/kg diet. We conclude that the size of the digestive organs increases with increasing levels of PH in the diet. In general, the best performance and nutrient digestibility values were observed with levels of PH within the range of 25 and 50g/kg. Therefore, young broilers have a requirement for a minimum amount of dietary fibre. When pea hulls are used as a source of fibre, the level of total dietary fibre required for optimal performance is within the range of 81.2–93.2g/kg diet (25.6–35.0g crude fibre/kg diet). An excess of total dietary fibre (above 93.2g/kg diet) might reduce nutrient digestibility and growth performance to values similar to those observed with the control diet.

The relevance of seed size in modulating leaf physiology and early plant performance in two tree species

he size of seeds and the microsite of seed dispersal may affect the early establishment of seedlings through different physiological processes. Here, we examined the effects of seed size and light availability on seedling growth and survival, and whether such effects were mediated by water use efficiency. Acorns of Quercus petraea and the more drought-tolerant Quercus pyrenaica were sowed within and around a tree canopy gap in a sub-Mediterranean forest stand. We monitored seedling emergence and measured predawn leaf water potential (Ψpd), leaf nitrogen per unit area (Na), leaf mass per area, leaf carbon isotope composition (δ13C) and plant growth at the end of the first summer. Survival was measured on the next year. Path analysis revealed a consistent pattern in both species of higher δ13C as Ψpd decreased and higher δ13C as seedlings emerged later in the season, indicating an increase in 13C as the growing season is shorter and drier. There was a direct positive effect of seed size on δ13C in Q. petraea that was absent in Q. pyrenaica. Leaf δ13C had no effect on growth but the probability of surviving until the second year was higher for those seedlings of Q. pyrenaica that had lower δ13C on the first year. In conclusion, leaf δ13C is affected by seed size, seedling emergence time and the availability of light and water, however, leaf δ13C is irrelevant for first year growth, which is directly dependent on the amount of seed reserves.

Factores nutricionales que influyen sobre las variables productivas y el desarrollo del tracto gastrointestinal en pollitas comerciales rubias = Nutritional factors affecting productive performance and gastrointestinal tract traits in commercial brown-egg laying pullets

El objetivo general de esta Tesis doctoral fue estudiar la influencia de diversos factores nutricionales sobre los parámetros productivos y el desarrollo del tracto digestivo de pollitas rubias destinadas a la producción de huevos comerciales. Para alcanzar este objetivo se realizaron tres experimentos donde se estudió el cereal principal, el tamaño de partícula del cereal y el nivel de energía y la presentación de los piensos. En el experimento 1 se estudió la influencia del cereal (piensos con enzimas) y la presentación del pienso sobre los parámetros productivos y las características del tracto digestivo en 576 pollitas rubias de 1 a 120 d de edad. De 1 a 45 d de la edad, se utilizaron 4 piensos experimentales organizados de forma factorial con 2 cereales al 50% de inclusión (maíz vs. trigo) y 2 presentaciones del pienso (harina vs. gránulo de 2- mm de diámetro). Cada tratamiento se replicó 6 veces (24 pollitas por réplica). De 46 a 120 d de edad todas las dietas (maíz o trigo) se ofrecieron en harina y por tanto, la única diferencia entre tratamientos fue el cereal utilizado. De 1 a 120 d de edad, las pollitas que recibieron los piensos basados en maíz tuvieron una ganancia de peso vivo (PV) superior (P < 0,05) que las que recibieron los piensos basados en trigo, pero el índice de conversión (IC) fue similar para ambos grupos. De 1 a 45 d de edad, las pollitas alimentadas con gránulo consumieron más pienso (P < 0,001) y tuvieron una ganancia de peso superior (P < 0,001) que las pollitas alimentadas con harina. Gran parte de los efectos beneficiosos de la granulación sobre los parámetros productivos se mantuvieron al final de la prueba (120 d de edad). A los 45 d de edad, el peso relativo de la molleja (PR; g/kg PV) fue superior (P < 0,01) en pollitas alimentadas con maíz que en pollitas alimentadas con trigo. La alimentación en gránulo redujo el PR del tracto gastro intestinal (TGI) y de la molleja (P < 0,001), así como la longitud relativa (LR; cm/kg PV) del intestino delgado (P< 0.01) a ambas edades (45 y 120 d de edad). El tipo de cereal utilizado no afectó al pH del contenido de la molleja a 120 d de edad pero fué inferior (P < 0,01) en las pollitas que recibieron el pienso en harina de 1 a 45 d de la edad que en las que recibieron el pienso en gránulo. Se concluye que el trigo puede substituir al maíz en piensos para pollitas si se acepta una ligera reducción en la ganancia de peso. Asímismo, la alimentación en gránulo de 1 a 45 d de edad aumentó la ganancia de peso a esta edad y al final de la prueba, así como el pH de la molleja a 120 d de edad. La presentación del pienso en gránulo redujo el PR de la molleja y la LR del TGI a 120 d de edad. En el experimento 2 se utilizaron un total de 864 pollitas rubias Hy-Line de 1 d de edad para estudiar la influencia del cereal de la dieta (500 g de maíz o trigo/kg) y el tamaño de partícula del mismo (molienda con molino de martillos con un diámetro de criba de 6, 8, o 10-mm) sobre los parámetros productivos y las características del TGI de 1 a 120 d de edad. Cada uno de los 6 tratamientos se replicó 6 veces (24 pollitas por réplica). De 1 a 45 d de edad, la ganancia de PV aumentó (P< 0,001) y el IC se mejoró (P < 0,05) al reducir el tamaño de partícula del cereal, pero no se observaron diferencias en el periodo crecimiento de 45 a 120 d de edad. A los 45 d de vida, las pollitas alimentadas con maíz tendieron (P < 0,10) a tener un mayor PR del TGI y del proventrículo y una mayor LR del intestino delgado que las pollitas alimentadas con trigo. Asímismo, el PR del TGI a esta edad, aumentó (P < 0,05) a medida que aumentaba el tamaño de partícula del cereal utilizado. A los 120 d de edad, el tratamiento no afectó el PR de ninguno de los órganos del TGI ni al pH de la molleja. Sin embargo, la LR del intestino delgado fue superior (P < 0,05) para las pollitas alimentadas con trigo que para las pollitas alimentadas con maíz. La LR del TGI se redujó (P < 0,05) al aumentar el tamaño de partícula del cereal. Se concluye que el trigo puede incluirse 500 g/kg en piensos de pollitas de 1 a 120 días de edad y que el tamaño de partícula de los cereales afecta el crecimiento de las pollitas durante los primeros 45 d de vida, pero no después. Por lo tanto, se recomienda moler el cereal utilizado al inicio del período de recría (1 a 45 d de edad) con una criba de diámetro igual o inferior a 8 mm. En el experimento 3 se utilizaron un total de 1.152 pollitas rubias Hy-Line de 1 d de edad para estudiar la influencia del nivel de energía de la dieta y la presentación del pienso sobre la productividad y las características del TGI. De 1 a 45 d de edad se utilizaron 6 piensos organizados de forma factorial con 3 concentraciones energéticas (baja: 11,44 MJ; media: 12,05 MJ y alta: 12,66 MJ/kg) y 2 presentaciones del pienso (harina vs. gránulo). De 45 a 120 d todos los piensos experimentales se suministraron en forma de harina y por tanto, la única diferencia entre tratamientos fue el nivel de EMAn utilizado. Cada uno de los 6 tratamientos se replicó 8 veces y la unidad experimental fue la jaula con 24 pollitas. De 1 a 120 d de edad, la ganancia de PV y el IC mejoraron a medida que aumentó la EMAn del pienso (P < 0,001). Las pollitas alimentadas con gránulo de 1 a 45 d de edad comieron mas y tuvieron una ganancia de peso superior (P < 0,001) que las alimentadas con harina. En el global de la prueba, la ganancia de PV fue mayor (P < 0,01) para las pollitas alimentadas con piensos en gránulo. A los 45 d de edad, el PR de todos los segmentos del TGI estudiados fue inferior para las pollitas alimentadas con piensos de alta energía que para las pollitas alimentadas con piensos de media o baja energía. A 120 d de edad, el PR de la molleja fue superior (P < 0,01) para las pollitas alimentadas con piensos de baja energía que con los otros piensos. Sin embargo, la LR del TGI no se vió afectada por el nivel de energía de los piensos. A los 45 d de edad, la alimentación con gránulo redujo el PR del proventrículo (P < 0,05), de la molleja (P < 0,001) y del TGI (P < 0.001), así como la LR del intestino delgado (P < 0,05) y de los ciegos (P < 0,001). A pesar de que las pollitas solo recibieron los piensos en gránulo durante los primeros 45 d de vida, la alimentación con gránulos redujo el PR de la molleja y del proventrículo a 120 d de edad. Se concluye que la alimentación con gránulos durante los primeros 45 d de vida mejora el consumo de pienso y el PV de las pollitas a 120 d de edad. Un aumento del nivel de energía de la dieta de 12,0 a 12,7 MJ/kg mejora los parámetros productivos de 1 a 120 d de edad pero reduce el tamaño del proventrículo y de la molleja. En base de estos resultados concluimos que maíz y trigo con enzimas pueden utilizarse indistintamente en piensos para pollitas de 1 a 120 d de edad con sólo una ligera disminución del PV final con trigo. La granulación y la reducción del tamaño de partícula del cereal del pienso de primera edad (1 a 45 d de vida) y el uso de piensos de alta densidad energética, mejoran los PV a 120 d de edad. Por lo tanto, es recomendable moler los cereales con cribas de no más de 8-mm de diámetro. También, la granulación del pienso y el uso de piensos de alta energía (pobres en fibra bruta) pueden reducir el desarrollo del TGI especialmente de la molleja, lo que puede perjudicar el consumo posterior de pienso durante el inicio del ciclo de puesta. ABSTRACT The general objective of this Thesis was to study the effect of different nutritional factors on productive performance and the development of the gastrointestinal tract (GIT) of commercial brown egg-laying pullets from 1 to 120 d of age. In this respect, the influence of type and particle size of the cereal, and feed form, and energy content of the die,t were studied in 3 experiments. In experiment 1, the influence of the main cereal and feed form of the diet on performance and GIT traits was studied in 576 brown-egg laying pullets from 1 to 120 d of age. From 1 to 45 d of age, 4 diets arranged factorially with 2 cereals (maize vs. wheat) and 2 feed forms (mash vs. pellets) were used. Each treatment was replicated 6 times (24 pullets per replicate). From 46 to 120 d of age, all diets were offered in mash form and therefore, the only difference among diets was the cereal used. Cumulatively, pullets fed the maize diets had higher body weight (BW) gain (P< 0.05) but similar feed conversion ratio (FCR) than pullets fed the wheat diets. From 1 to 45 d of age, pullets fed pellets consumed more feed (P < 0.001) and had higher BW gain (P < 0.001) than pullets fed mash. Most of the beneficial effects of pelleting on productive performance of the birds were still evident at 120 d of age. At 45 d of age, gizzard relative weight (RW; g/kg BW) was higher (P < 0.01) in pullets fed maize than in pullets fed wheat. Feeding pellets reduced the RW of the GIT and the gizzard (P < 0.001) as well as the relative length (RL; cm/kg BW) of the small intestine (SI, P < 0.01) at both ages. The pH of the gizzard contents at 120 d of age was not affected by the main cereal of the diet, but was lower in pullets fed mash from 1 to 45 d of age (P < 0.01) than in pullets fed pellets. We conclude that wheat supplemented with enzymes can be used in substitution of maize in pullet diets with only a slight reduction in BW gain at 120 d of age. Also, feeding pellets from 1 to 45 d of age increased BW gain and pH of the gizzard, and reduced the RW of the gizzard and the RL of the GIT at 120 d of age. In experiment 2, a total of 864 brown-egg laying pullets was used to study the effects of the main cereal of the diet (500 g maize or wheat/kg) and particle size of the cereal (hammer milled to pass through a 6, 8, and 10-mm screen) on growth performance and GIT traits from 1 to 120 d of age. Each of the 6 treatments was replicated 6 times (24 pullets per replicate). Type of cereal did not affect pullet performance at any age. From 1 to 45 d of age, BW gain was increased (P < 0.001) and FCR was improved (P < 0.05) as the particle size of the cereal was reduced, but no effects were observed after this age. At 45 d of age, pullets fed maize tended (P < 0.10) to have a heavier RW of the GIT and proventriculus and a higher relative length (RL, cm/kg BW) of the SI than pullets fed wheat. Also at this age, the RW of the GIT increased (P < 0.05) with increases in particle size of the cereal. At 120 d of age, dietary treatment did not affect the RW of any of the organs studied or gizzard pH, but the RL of the SI was higher (P < 0.05) for pullets fed wheat than for pullets fed maize. Also, the RL of the SI was reduced (P < 0.05) as the particle size of the cereal increased. We conclude that 500 g wheat/kg can be included in pullet feeds from 1 to 120 d of age, and that particle size of the cereal affects pullet performance during the first 45 d of life but not thereafter. Therefore, it is recommended to grind the cereal used in this period with a screen size of no more than 8-mm. In experiment 3, a total of 1,152 one-day-old Hy-Line Brown egg laying pullets were used to study the influence of the energy content of the diet and feed form on productive performance and on several GIT traits. From 1 to 45 d of age, there were 6 diets arranged factorially with 3 concentrations of AMEn (low: 11.66 MJ/kg, medium: 12.05 MJ/kg and high: 12.66 MJ/kg) of the diet and 2 feed forms (mash vs. pellets). From 45 to 120 d all diets were fed in mash form and therefore, the only difference among treatments in this period was the energy content of the diets. Each of the 6 treatments was replicated 8 times and the experimental unit was formed by 24 pullets. Cumulatively, BW gain and FCR improved as the AMEn of the diet increased (P < 0.001). Also, pullets fed pellets from 1 to 45 d of age had higher feed intake and BW gain (P < 0.001) in this period and higher cumulative BW gain (P < 0.01) than pullets fed mash. At 45 d of age, the RWof all the segments of the GIT was lower for pullets fed the high- than for pullets fed the medium- or low- energy diets. At 120 d of age, the RW of the gizzard was higher (P < 0.01) for pullets fed the low energy diets than for pullets fed the other diets. However, the RL of the GIT was not affected by the energy content of the diet. Feeding pellets reduced the RW of the proventriculus (P < 0.05), gizzard (P < 0.001), and GIT (P < 0.001), as well as the RL of the small intestine (P < 0.05) and the ceaca (P < 0.001) at 45 d of age. The effects of feeding pellets on RW of gizzard and proventriculus were still evident at 120 d of age. We concluded that feeding pellets from 1 to 45 d of age improved feed intake and BW of pullets at 120 d of age and that an increase in the energy content of the diet increased pullet performance at all ages but reduced the RW of the proventriculus and gizzard. We conclude that maize and wheat can be used indistinctly in diets for egg laying pullets from 1 to 120 d of age, with only a slight reduction in final BW when wheat is used. Also, particle size of the cereal affects pullet performance during the first 45 d of life but not thereafter. Pelleting of the feeds, and grinding the cereal with a screen size of no more than 8-mm from 1 to 45 d of age, and the use of high density energy diets are recommended in order to achieve adequate target BW at 120 d of age. However, pelleting of the feed, very fine grinding, and the use of high AMEn diets might hinder the development of the GIT, especially that of the gizzard, which might affect feed intake of laying hens especially at the beginning of the production cycle.

Transport of Pru p 3 across gastrointestinal epithelium - an essential step towards the induction of food allergy?

Background Since intestinal absorption of food protein can trigger an allergic reaction, the effect of plant food allergen on intestinal epithelial cell permeability and its ability to cross the epithelial monolayer was evaluated. Objective To study the interaction of Pru p 3 with intestinal epithelium, its natural entrance, analyzing transport kinetics and cellular responses that trigger. Methods This was achieved using Pru p 3, the peach LTP, as a model. Enterocytic monolayers were established by culturing Caco 2 cells, as a model of enterocytes, on permeable supports that separate the apical and basal compartments. Pru p 3 was added to the apical compartment, the transepithelial resistance (TEER) was measured, and the transport was quantified. Results The peach allergen that crossed the cell monolayer was detected in the cell fraction and in the basal medium by immunodetection with specific antibodies and the quantity was measured by ELISA assay. Pru p 3 was able to cross the monolayer without disturbing the integrity of the tight junctions. This transport was significantly higher than that of a non-allergenic peach LTP, LTP1, and occurred via lipid raft pathway. The incubation of Caco 2 cells with Pru p 3 and LTP1 produced the expression of epithelial-specific cytokines TSLP, IL33 and IL25. Conclusion These results suggest that Pru p 3 was able to cross the cell monolayer by the transcellular route and then induce the production of Th2 cytokines. The results of the present study represent a step towards clarifying the importance of Pru p 3 as a sensitizer. Clinical relevance The capacity of food allergens to cross the intestinal monolayer could explain their high allergenic capacity and its fast diffusion through the body associating to severe symptoms.

Molecular physiology of nickel and cobalt homeostasis in Rhizobium leguminosarum.

Transition metals such as Fe, Cu, Mn, Ni, or Co are essential nutrients, as they are constitutive elements of a significant fraction of cell proteins. Such metals are present in the active site of many enzymes, and also participate as structural elements in different proteins. From a chemical point of view, metals have a defined order of affinity for binding, designated as the Irving-Williams series (Irving and Williams, 1948) Mg2+ menor que Mn2+ menor que Fe2+ menor que Co2+ menor que Ni2+ menor que Cu2+mayor queZn2+ Since cells contain a high number of different proteins harbouring different metal ions, a simplistic model in which proteins are synthesized and metals imported into a ?cytoplasmic soup? cannot explain the final product that we find in the cell. Instead we need to envisage a complex model in which specific ligands are present in definite amounts to leave the right amounts of available metals and protein binding sites, so specific pairs can bind appropriately. A critical control on the amount of ligands and metal present is exerted through specific metal-responsive regulators able to induce the synthesis of the right amount of ligands (essentially metal binding proteins), import and efflux proteins. These systems are adapted to establish the metal-protein equilibria compatible with the formation of the right metalloprotein complexes. Understanding this complex network of interactions is central to the understanding of metal metabolism for the synthesis of metalloenzymes, a key topic in the Rhizobium-legume symbiosis. In the case of the Rhizobium leguminosarum bv viciae (Rlv) UPM791 -Pisum sativum symbiotic system, the concentration of nickel in the plant nutrient solution is a limiting factor for hydrogenase expression, and provision of high amounts of this element to the plant nutrient solution is required to ensure optimal levels of enzyme synthesis (Brito et al., 1994).

Fish physiology put into practice: A robotic fish model

Underwater creatures are capable of high performance movements in water. Thus, underwaterrobot design based on the mechanism of fish locomotion appears to be a promising approach.Over the past few years, researches have been developing underwater robots based on underwatercreatures swimming mechanism.

Lectures on plant physiology. Authorized English translation by R.J. Harvey Gibson.

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