Characterization of digestive protease activity in the bluefin tuna Thunnus thynnus (Linnaeus, 1758)

The digestive proteases of bluefin tuna Thunnus thynnus (Linnaeus, 1758) were studied. Protease activity in stomach extracts showed two optima, at pH 2.0 and 3.5. Optimum activity for intestinal proteases was found at pH 10.0 and 12.0. Protease activity in stomach extracts was found to be stable at several pH, except for pH 12.0. Alkaline protease activity of intestine extracts was highly sensitive to acidic pH. Optimal temperatures found were 50 ºC for acid proteases and 60 ºC for alkaline proteases. The use of specific protease inhibitors indicated that the main presence of acid proteases was in stomach extracts, and serine proteases in intestine extracts. Active fractions in extracts were identified with SDS-PAGE.Se estudió la actividad proteasa digestiva de Thunnus thynnus (Linnaeus, 1758). Se encontraron dos óptimos de pH, tanto para la actividad proteasa ácida de los extractos estomacales (2,0 y 3,5) como para la actividad proteasa alcalina de los intestinales (10,0 y 12,0). La primera se mantuvo muy estable en un rango amplio de pH, a excepción de pH muy alcalinos (12,0). Por su parte, la actividad proteasa alcalina se vio afectada por pH muy ácidos. Los óptimos de temperatura se determinaron en 50 y 60 ºC para la actividad proteasa ácida y alcalina, respectivamente. Mediante el uso de inhibidores de proteasa se comprobó la presencia de proteasas aspárticas en los extractos estomacales y de serin-proteasas en los intestinales. La realización de zimogramas permitió identificar las distintas fracciones con actividad proteolítica presentes en cada extracto.Instituto Español de Oceanografí

Effect of phytase supplements on in vitro hydrolysis of proteins by rainbow trout Oncorhynchus mykiss (Walbaum, 1792)

The present paper evaluates the impact on plant meal and on an experimental feed product of supplementing them with four levels of phytase (0, 2 000, 4 000 and 6 000 FTU/kg), by assessing in vitro hydrolysis of protein in rainbow trout Oncorhynchus mykiss (Walbaum, 1792) using a pH-stat system. The effect on fish meal's digestibility of adding phytic acid was also evaluated. The highest degree of hydrolysis (DH) was obtained when phytase was added at 4 000 FTU/kg. The experimental diet's DH diminished when phytase was added at 6 000 FTU/kg. However, protein hydrolysis dropped as the percentage of phytic acid in the in vitro reaction mixture increased.En este trabajo se evaluó el efecto de la suplementación con 4 niveles de fitasa (0, 2 000, 4 000 y 6 000 FTU/kg) en un pienso experimental y sobre una harina vegetal, evaluando la hidrólisis in vitro de la proteína por pH-stat con extractos enzimáticos de trucha arcoiris Oncorhynchus mykiss (Walbaum, 1792). Además, se evaluó el efecto que ejerce la adición de ácido fítico sobre la digestibilidad de una harina de pescado. Se observó que, para el pienso experimental, el mayor grado de hidrólisis se obtiene cuando la inclusión de fitasa es de 4 000 FTU/kg, disminuyendo ligeramente cuando la inclusión es de 6 000 FTU/kg. En cambio, se observó una disminución de la hidrólisis proteica a medida que se incrementaba el porcentaje de ácido fítico en la mezcla de reacción in vitro.Instituto Español de Oceanografí

Effect of increased rearing temperature on digestive function in cobia early juvenile

The present study is focused to elucidate the main characteristics of the digestive function of this carnivorous fast-growing fish living at high temperatures. With this aim, we have examined the effects of an increased temperature from 30 to 34 °C on the daily pattern of gastrointestinal pH, enzymatic proteolytic digestive activity and the feed transit time in early juveniles of cobia (Rachycentron canadum), a species living in tropical and subtropical waters with an increasing aquaculture production. Fish were fed two meals a day. Gastric luminal pH was permanently acidic (mean pH values: 2.76-4.74) while the intestinal pH increased from neutral/slightly acidic to slightly alkaline when the digesta was present, with an increasing alkalinity from proximal to distal intestine (mean pH values: 6.05 to 7.69). The temperature did not affect the gastric pH but a slightly higher acidity was induced in the intestine at 34 °C. Pepsin activity showed a daily rhythm at 30 °C with maximum in the middle of the light period, while at 34 °C some hourly changes coinciding with feed adding without a clear daily trend during the 24-h period were observed. The trypsin activity exhibited a daily rhythm at both temperatures with an increase after morning feeding to reach a maximum several hours later. Average pepsin activity during the daily cycle was slightly higher at 34 °C (6.1 and 7.3 U mg-1 BW at 30 and 34 °C respectively), but values were significantly different only at 8 and 24 h after the morning meal. Similarly, the trypsin activity was significantly affected by the temperature only at 8 and 16 h after the morning meal, but daily activity averages were similar (1.20 and 1.29 U g-1 BW at 30 and 34 °C respectively). The partial transit rates of the first meal in the stomach for each period inter-samplings were higher during the first 4-h period and decreased progressively along the rest of the 24-h cycle at both temperatures, but no significant differences were detected at 30 °C. In addition, the transit was notably faster at 34 °C particularly during the first 8 h after feeding, with rates between 100 and 65% of total volume displaced (intake or released) during each 4-h period. In the intestine the transit rate was relatively constant and similar at both temperatures during 12 h after feeding. Then the rates remained very low during the following 12 h. Residence time of the first meal was longer at 30 than at 34 °C, particularly in the stomach (12 h:02 min vs 4 h:54 min respectively). In the intestine the difference was not so large (8 h:18 min vs 6 h:24 min respectively). In a parallel study under same conditions, cobia reared at 30 °C grew faster and showed a more favorable feed conversion ratio than those at elevated temperature (34 °C). The present results indicate that at 34 °C, a subtle increase of proteolytic activity cannot compensate for the faster gut transit rate. Therefore, 30 °C is more appropriate temperature for the early on-growing of cobia because at higher temperatures the digestion efficiency decrease being one of the causes for a lower growth.Agência financiadora European Union (EU) 691150 MINECO, Spain + FEDER/ERDF contribution EFISHDIGESTAGL2014-52888 European Social Fund, the Operational Programme Human Potential IF/00482/2014/CP1217/CT0005 Portuguese Foundation for Science and Technology Portuguese Foundation for Science and Technology UDI/Multi/04326/2013 Norwegian Agency for Development Cooperation NORHED QZA-0485 SRV-13/0010info:eu-repo/semantics/publishedVersio

Burden and risk factors for Pseudomonas aeruginosa community-acquired pneumonia:a Multinational Point Prevalence Study of Hospitalised Patients

Pseudornonas aeruginosa is a challenging bacterium to treat due to its intrinsic resistance to the antibiotics used most frequently in patients with community-acquired pneumonia (CAP). Data about the global burden and risk factors associated with P. aeruginosa-CAP are limited. We assessed the multinational burden and specific risk factors associated with P. aeruginosa-CAP. We enrolled 3193 patients in 54 countries with confirmed diagnosis of CAP who underwent microbiological testing at admission. Prevalence was calculated according to the identification of P. aeruginosa. Logistic regression analysis was used to identify risk factors for antibiotic-susceptible and antibiotic-resistant P. aeruginosa-CAP. The prevalence of P. aeruginosa and antibiotic-resistant P. aeruginosa-CAP was 4.2% and 2.0%, respectively. The rate of P. aeruginosa CAP in patients with prior infection/colonisation due to P. aeruginosa and at least one of the three independently associated chronic lung diseases (i.e. tracheostomy, bronchiectasis and/or very severe chronic obstructive pulmonary disease) was 67%. In contrast, the rate of P. aeruginosa-CAP was 2% in patients without prior P. aeruginosa infection/colonisation and none of the selected chronic lung diseases. The multinational prevalence of P. aeruginosa-CAP is low. The risk factors identified in this study may guide healthcare professionals in deciding empirical antibiotic coverage for CAP patients

Three short-period Jupiters from TESS: HIP 65Ab, TOI-157b, and TOI-169b

We report the confirmation and mass determination of three hot Jupiters discovered by the Transiting Exoplanet Survey Satellite (TESS) mission: HIP 65Ab (TOI-129, TIC-201248411) is an ultra-short-period Jupiter orbiting a bright (V = 11.1 mag) K4-dwarf every 0.98 days. It is a massive 3.213 ± 0.078 MJ planet in a grazing transit configuration with an impact parameter of b = 1.17-0.08+0.10. As a result the radius is poorly constrained, 2.03-0.49+0.61RJ. The planet's distance to its host star is less than twice the separation at which it would be destroyed by Roche lobe overflow. It is expected to spiral into HIP 65A on a timescale ranging from 80 Myr to a few gigayears, assuming a reduced tidal dissipation quality factor of Qs′ = 107 - 109. We performed a full phase-curve analysis of the TESS data and detected both illumination- and ellipsoidal variations as well as Doppler boosting. HIP 65A is part of a binary stellar system, with HIP 65B separated by 269 AU (3.95 arcsec on sky). TOI-157b (TIC 140691463) is a typical hot Jupiter with a mass of 1.18 ± 0.13 MJ and a radius of 1.29 ± 0.02 RJ. It has a period of 2.08 days, which corresponds to a separation of just 0.03 AU. This makes TOI-157 an interesting system, as the host star is an evolved G9 sub-giant star (V = 12.7). TOI-169b (TIC 183120439) is a bloated Jupiter orbiting a V = 12.4 G-type star. It has a mass of 0.79 ±0.06 MJ and a radius of 1.09-0.05+0.08RJ. Despite having the longest orbital period (P = 2.26 days) of the three planets, TOI-169b receives the most irradiation and is situated on the edge of the Neptune desert. All three host stars are metal rich with [Fe / H] ranging from 0.18 to0.24

Evaluation of the ontogeny of digestive enzymes in larvae of theft, Pagrus auriga (Pisces: Sparidae)

La hurta, Pagrus auriga, es un pez de elevado valor comercial en el Sur de España, cuyo cultivo está comenzando a ser desarrollado. En este trabajo, se presentan los primeros resultados obtenidos acerca de la evolución de las principales enzimas digestivas desde la eclosión de las larvas hasta que alcanzan una edad de 40 días. La actividad de la tripsina aumentó gradualmente a lo largo del desarrollo larvario, hasta alcanzar un valor estacionario en torno a 40 UF/individuo,en larvas a partir de 25 días. La quimotripsina presentó una actividad similar en las larvas de más edad. Sin embargo, hasta alcanzar los 10 días de edad, la actividad de la quimotripsina creciómucho más rápidamente. La actividad de proteasa ácida se detectó a partir de los 30 días de edad. La actividad amilasa también se incrementó a lo largo del desarrollo larvario, pero lo hizo con una velocidad muy inferior a la encontrada para las proteasas. Las esterasas se incrementaron en dos órdenes de magnitud desde el inicio de la alimentación hasta los 40 días de edad, alcanzando máximos cercanos a 100 U/individuo