ABSTRACT. This work aimed to improve reproductive maturation in two penaeoid species (Artemesia longinaris and Pleoticus muelleri) by using carotenes added in artificial feeds and comparing antioxidant activity and carotenoids profiles. Immature females were kept for 45 days at salinity 33, temperature at 18°C, pH 7, 12:12 h photoperiod, ammonium <0.2mg L−1 and seawater exchanged at 100% per day. Feeding regimes consisted of 50% fresh frozen ingredients (clam muscle and squid mantle) and 50% dry pellets. For each species, three pelletized feeds were tested, one as a control (C) and two supplemented each one with 300 mg astaxanthin (A) and β-carotene (B) kg−1 of diet. At the end of the trials, ovaries, midgut gland, and integument were sampled. In both species, the addition of carotenoids improved ovarian development: 100% of females fed A or B diets were mature, but of the females fed with diet C only 75% (P. muelleri) and 64% (A. longinaris) matured. Ovaries of mature females exhibited the highest concentration of carotenoids. The abundance of free astaxanthin regarding total carotenes, suggests that excess of free astaxanthin could be biotransformed to esterified astaxanthin. The scavenging properties were evaluated against the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical by electron resonance spectroscopy (EPR). For all feed treatments, the extracts exhibited strong DPPH scavenging activity, without significant differences among them.

Water-soluble extracts from seaweeds Macrocystis pyrifera (M) and Undaria pinnatifida (U) were prepared to use the polysaccharide bioactive fraction as functional food. Chemical analysis showed that the main components of both extracts were mannitol and fucoidans. Also in both cases minor amounts of rhamnose, glucose, xylose and arabinose were determine. Content of fucose+galactose was higher in U. pinnatifida (8.7%) than in M. pyrifera (5.9%). Extracts were used as feed additive to Artemesia longinaris at inclusion levels of 1 and 2% (M1 and M2; U1 and U2), a diet without extract was used as control. Radical scavenging activities were estimated for two extracts and for midgut gland samples of experimental animals. Scavenging activity was detected in both extracts, the signal decayed to 50 and 62% in 18 min for U. pinnatifida and M. pyrifera, respectively. After 30 days of feeding, results did not show variation of survival, but growth was improved in animals fed with U1 and U2. Supplementation with 1 and 2% of U. pinnatifida showed similar quenching capacity of homogenates than those fed with M2 with a rapid 1,1-diphenylpicrylhydrazyl (DPPH) decay at 2 min of reaction. Recommended concentration for culture A. longinaris is 2%, as it is that produce the best antioxidant capacity, although the growth performance was improved also significantly only with U. pinnatifida extract.

The aim of the present study was to determine lethal and sublethal effects of nitrite on the freshwater prawn, Palaemonetes argentinus (Nobili 1901). The 24, 48, 72 and 96 h median lethal concentration (LC50) of nitrite were found to be 103.07; 91.94; 82.39 and 62.53 mg L-1, respectively. The antioxidant activity of superoxide dismutase (SOD) was positively correlated with the concentration of nitrite in the environment, thus increasing the formation of hydrogen peroxide, substrate of catalase (CAT), which also showed an increased activity under the same experimental conditions. Histological samples showed that nitrite exposure caused hyperplasia, necrosis and changes in the lamellar epithelium in gills; while the effects in the hepatopancreas were degenerative desquamation, epithelial deterioration and hyperplasia. In conclusion, nitrite toxicity in P. argentinus increases with time of exposure. Moreover, this study proves that histological analysis of crustacean gills and/or hepatopancreas is a useful tool to determine the presence of toxic concentrations of nitrite in the water. Additionally, the measurement of the activity of antioxidant enzymes (e.g., SOD and CAT) as possible indicators of nitrite pollution should be considered in future studies.

El objetivo del presente estudio fue determinar los efectos letales y subletales del nitrito en el camarón de agua dulce, Palaemonetes argentinus (Nobili 1901). Las concentraciones letales medias (LC50) de nitrito a las 24; 48; 72 y 96 h al nitrito fueron 103,07; 91,94; 82,39 y 62,53 mg L-1, respectivamente. La actividad antioxidante de la superóxido dismutasa (SOD) se correlacionó positivamente con la concentración de nitrito ambiental; este incremento en su actividad eleva la formación de peróxido de hidrógeno, sustrato de la catalasa (CAT), que también mostró un incremento en su actividad bajo las mismas condiciones experimentales. Las muestras histológicas evidenciaron que la exposición al nitrito causa hiperplasia, necrosis y cambios en el epitelio lamelar en las branquias; mientras que en el hepatopancreas los efectos son descamación degenerativa, deterioro epitelial e hiperplasia. En conclusión, la toxicidad del nitrito en P. argentinus se incrementa conforme el tiempo de exposición. Este estudio demuestra que el análisis histológico de las branquias y/o hepatopancreas del camarón P. argentinus es útil para determinar la presencia de concentraciones tóxicas de nitrito en el agua. Además, la medición de las enzimas antioxidantes (por ejemplo, SOD y CAT) debería ser considerado en futuros estudios como posibles indicadores de contaminación por nitrito.

The present study compared the effect of different starch/cellulose ratios (30/0, 20/10, 10/20, 0/30) on the metabolic response and apparent digestibility in two species of penaeoids: Artemesia longinaris and Pleoticus muelleri. Adult animals were used in order to obtain sufficient quantities of haemolymph and faecal material for analysis. No significant differences were found in levels of plasma metabolites in P. muelleri, but in A. longinaris, a significant increase was observed in glucose, total protein, and cholesterol in correlation with increased dietary starch. The apparent digestibility coefficients decreased from 83.7% to 51.2% (A. longinaris) and from 71.9% to 7.6% (P. muelleri) as the dietary starch levels increased. The ratio of amylase activity to protease activity (A/P ratio) declined in A. longinaris when the percentage of dietary starch increased. In contrast, the A/P ratio for P. muelleri increased with higher starch concentrations. These results demonstrated a close relationship between the feeding habits and digestive physiology of the two species studied; they also suggest a more herbivorous behavior for A. longinaris and more omnivorous habits for P. muelleri.

En el presente estudio se comparó el efecto de diferentes concentraciones de almidón/celulosa (30/0; 20/10; 10/20; 0/30) sobre la respuesta metabólica y la digestibilidad aparente en dos especies de peneidos, Artemesia longinaris y Pleoticus muelleri. Se utilizaron animales adultos a fin de obtener cantidades suficientes de hemolinfa y heces para los análisis. No hubo diferencias significativas en los niveles de metabolitos plasmáticos en P. muelleri, en cambio en A. longinaris se observó un incremento significativo de la glucosa, proteínas totales y colesterol en relación con el aumento del almidón en la dieta. Los coeficientes de digestibilidad aparente disminuyeron de 83,7% a 51,2% (A. longinaris) y de 71,9% a 7,6% (P. muelleri) a medida que los porcentajes de almidón en la dieta aumentaron. El cociente entre la actividad de amilasa y proteasa (A/P) se redujo en A. longinaris con los mayores porcentajes de almidón dietario; por el contrario, el cociente A/P en P. muelleri aumentó cuando la concentración fue más alta. Estos resultados demostraron que existe una estrecha relación entre los hábitos alimentarios y la fisiología digestiva de las dos especies estudiadas; sugiriendo un comportamiento más herbívoro para A. longinaris y más omnívoro para P. muelleri.

O objetivo do presente trabalho foi determinar a digestibilidade aparente in vivo da proteína bruta de ingredientes de origem animal (farinhas de peixe, osso e carne e concentrado de proteína de lula) e ingredientes vegetais (farinha de soja) em camarões Artemesia longinaris utilizando rações contendo 0,25% de óxido de cromo. Três grupos de camarões, utilizados como replicatas, foram alimentados e as fezes coletadas. A velocidade de hidrólise da proteína de cada ração foi medida in vitro utilizando extrato enzimático da glândula do intestino médio dos camarões alimentados com a ração correspondente e foi comparado com aqueles obtidos com o extrato enzimático de camarões selvagens. Os coeficientes de digestibilidade aparente in vivo mostraram diferenças significativas entre as rações testadas (P<0,05). A farinha de peixe apresentou a maior digestibilidade (92%), enquanto valores intermediários de digestibilidade (83%) foram encontrados para a farinha de carne e ossos. A ração contendo farinha de soja e concentrado de proteína de lula resultou em menor digestibilidade (63%). Não houve diferença significativa entre os valores de digestibilidade in vitro para as rações testadas. Estes resultados indicam a limitação inerente dos ensaios enzimáticos in vitro, os quais poderiam ser complementados com estudos in vivo.

This study was undertaken to determine the in vivo crude protein apparent digestibility in the prawn Artemesia longinaris, using feeds with 0.25% of chromic oxide and animal (fish meal, meat and bone meal and squid protein concentrate) and plant (soybean meal) ingredients. Three replicate groups of prawn were fed and the feces were collected. The rate of protein hydrolysis was measured in vitro using midgut gland enzyme extract from the prawns fed the respective feeds and was compared with those found with enzyme extract of wild prawn. The in vivo apparent digestibility coefficients showed significant differences among the feeds (P<0.05). Fish meal feed presented the highest digestibility (92%); intermediate digestibility (83%) was found for meat and bone meal feed, and the less digestible feed (63%) was that containing soybean meal and squid proteins concentrate. No significant differences in the in vitro protein digestibility were found among the experimental feeds. The results indicated the limitation of in vitro enzyme assays and that it should be complemented by in vivo studies.

Tobacco is one of the major preventable causes of morbidity and premature mortality in the world. A simple, rapid method for determination of cotinine, major metabolite of nicotine in urine, is described. The assay involves a fast liquid-liquid extraction with chloroform as a solvent in alkaline environment. The extract was dried at 37 °C and the residue was dissolved in 100 µL of standard internal of pyridine in methanol solution, and an aliquot was analyzed by gas-liquid chromatograph equipped with a flame -ionization detector. This technique was performed to test urine of a sample of smokers and non-smokers who were previously asked about their smoking habit. The limit of detection was 3.1 ng/mL, and using two calibration curves, a linearity range from 3 a 5.000 ng/mL was covered. The median and quartiles (Q) of cotinine in smokers were 255.4 ng/mL, 15.9 ng/mL (Q1) -1050.2 ng/mL (Q3) ng/mL and in non-smokers 15.8 ng/mL, 4.1 ng/mL (Q1) - 38.4 ng/mL (Q3). Wilcoxon sumrank: z = 4.865 p < 0.01. There was a relationship between cotinine concentration in urine and number of smoked cigarettes. Spearman's rho = 0.5672 (p < 0.001). This technique could identify smokers, second hand smokers and non-smokers using cut-off of 10-15 ng/mL of cotinine in urine, usually adopted in epidemiological studies.

La exposición a tabaco es una de las causas prevenibles de morbilidad y mortalidad prematura más importante. Se describe un método simple y rápido de detección de cotinina (principal metabolito de nicotina) en orina humana. Se utilizó cromatografía gas-líquido (CGL) con un detector de ionización de llama (FID). Se realizó una extracción líquido-líquido con cloroformo en medio alcalino; el extracto fue secado a 37 °C y redisuelto en 100 µL de solución alcohólica con estándar interno piridina. Se analizó la orina de personas que declararon ser fumadoras y no fumadoras. Mediante la construcción de dos curvas de calibración se cubrió un rango de linealidad de 3 a 5.000 ng/mL. Los fumadores presentaron una mediana y cuartiles (Q) de concentración de cotinina: 255,4 ng/mL, 15,9 ng/mL (Q1) - 1.050,2 ng/mL (Q3), respectivamente, y los no fumadores 15,8 ng/mL, 4,1 ng/mL (Q1) - 38,4 ng/mL (Q3). Wilcoxon sumrank: z = 4,865 p < 0,01. La concentración de cotinina se incrementó con el número de cigarrillos fumados (rho de Spearman = 0,5672 p < 0,001. El límite de detección fue 3,1 ng/mL y permitió la diferenciación entre los no fumadores, fumadores pasivos y activos usando el valor de corte de 10-15 ng/mL de cotinina en orina, frecuentemente utilizado en los estudios epidemiológicos.