Growth, stress response and free amino acid levels in Senegalese sole (Solea senegalensis Kaup 1858) chronically exposed to exogenous ammonia

Stressful husbandry conditions are likely to affect growth and amino acid metabolism in fish. In this study, chronic ammonia exposure was used to test the effects of a stressor on growth and amino acid metabolism of Senegalese sole juveniles. The fish were exposed for 52 days to 11.6 mg L⁻¹ [low‐TAN (L‐TAN)] or 23.2 mg L⁻¹ [high‐TAN (H‐TAN)] of total ammonia nitrogen (TAN), or to 0 mg L⁻¹ (Control). Growth in L‐TAN groups was slightly but significantly different from the Control groups [relative growth rate (RGR=0.35±0.13 and 0.52±0.23% day⁻¹ respectively)]. In H‐TAN groups, growth was severely affected (RGR=0.01±0.13% day⁻¹). Stress parameters (plasma cortisol and glucose) showed slight or no significant differences between treatments. Plasma free amino acid (FAA) concentrations were affected after H‐TAN treatment. Increases in glutamine and aspartate concentrations in H‐TAN fish suggest alterations in amino acid metabolism related to nitrogen excretion processes. Some of the changes in FAA concentrations also suggest mobilization to energy supply and synthesis of metabolites related to stress response. Therefore, Senegalese sole seem to adapt to the L‐TAN concentration tested, but the H‐TAN concentration reduced growth and affected amino acid metabolism.     

Feeding responses of juvenile shrimp Litopenaeus vannamei (Boone) fed at different frequencies under laboratory conditions

In shrimp farms, food partitioning during the course of the day is contradictory, ethology studies may help in determining the proper frequency. Litopenaeus vannamei juveniles were placed in 30 L aquariums (41 m⁻²), exposed to a 12:12 h light–dark cycle. Feeding was provided at 10% of biomass three, four and seven times per day, with observations made in 15 min h⁻¹ windows, initiated after the ration was offered. Latency to access the feeding tray and to start eating was recorded using instantaneous focal sampling and digestive tract filling (DTF) by the continuous focal method. Weight gain was recorded at the end of the experiment. We performed seven repetitions, with 28 individuals observed for 33 days for each treatment (490 h of observation). The three-times offering induced less latency for accessing the tray and for ingesting the feed as well as a higher DTF when compared with the other frequencies. Animals fed three and four times had similar weight gains, and were greater than those fed seven times. Our results indicate that a more spaced food offering stimulates the search for and ingestion of feed. As compared with other frequencies, the three-times-per-day option assumes lower labour costs and a more efficient use of the feed.     

Method of determination of nitrosamines in sausages by CO2 supercritical fluid extraction (SFE) and micellar electrokinetic chromatography (MEKC)

In this paper, the use of supercritical fluid extraction (SFE) and micellar electrokinetic capillary chromatography (MEKC) is proposed for the complete analysis of volatile nitrosamines in sausages. The extraction fluid used was CO2 and variables such as density, temperature of thimbles, extraction time, modifier, fluid flow, and kind of traps were investigated. Several experiments were carried out to obtain the most favorable conditions for analysis of volatile nitrosamines in sausages. The recoveries ranged from 21 to 82% for the five nitrosamines studied. The optimal condition of extraction was 0.2 g of sample fortified with 10 mg/kg, using dynamic extraction during 20 min and with adsorbent Florisil in the trap. The solvent selected for the elution of the analytes was methanol.     

Inhibition of trypsin by condensed tannins and wine

Phenolic compounds are abundant vegetable secondary metabolites in the human diet. The ability of procyanidin oligomers and wine polyphenols to inhibit trypsin activity was studied using a versatile and reliable in vitro method. The hydrolysis of the chromogenic substrate N-benzoyl-d,l-arginine-p-nitroanilide (BApNA) by trypsin was followed by spectrophotometry in the presence and absence of condensed tannins and wine. A clear relationship between the degree of polymerization of procyanidins and enzymatic inhibition was observed. Trypsin activity inhibition was also detected in several types of wine. In general, the inhibition increased with the concentration of phenolic compounds in wines. These results may be relevant when considering these compounds as antinutritional factors, thereby contributing to a reduced absorption of nutrients.     

Effect of Leptin on In Vivo Goat Embryo Production

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133 mg follicle‐stimulating hormone (FSH) i.m. in six descending doses at 12‐h intervals. The goats received 4.8 μg/kg human recombinant leptin s.c. (leptin group, n = 5) or phosphate‐buffered saline (PBS) (control group, n = 5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6 days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick‐end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E₂) and progesterone (P₄) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p < 0.005), fewer embryos classified as degenerated (p < 0.001) and fewer TUNEL‐positive cells/blastocyst (p < 0.001). The number of transferable embryos was positively correlated with E₂ concentrations on day of oestrus (r = 0.562; p < 0.01) and P₄ concentrations on the day of embryo collection (r = 0.912; p < 0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.     

Escherichia coli challenge and one type of smectite alter intestinal barrier of pigs

An experiment was conducted to determine how an E. coil challenge and dietary clays affect the intestinal barrier of pigs. Two groups of 32 pigs （initial BW： 6.9 ± 1.0 kg） were distributed in a 2 x 4 factorial arrangement of a randomized complete block design （2 challenge treatments： sham or E. coil, and 4 dietary treatments： control, 0.3% smectite A, 0.3% smectite B and 0.3% zeolite）, with 8 replicates total. Diarrhea score, growth performance, goblet cell size and number, bacterial translocation from intestinal lumen to lymph nodes, intestinal morphology, and relative amounts of sulfo and sialo mucins were measured. The E. coli challenge reduced performance, increased goblet cell size and number in the ileum, increased bacterial translocation from the intestinal lumen to the lymph nodes, and increased ileal crypt depth. One of the clays （smectite A） tended to increase goblet cell size in ileum, which may indicate enhanced protection. In conclusion, E. coli infection degrades intestinal barrier integrity but smectite A may enhance it.     

Intestinal phytase I: Detection and preliminary characterization of activity in the intestinal brush border membrane of hybrid striped bass Morone saxatilis × M. chrysops

While numerous studies have evaluated intestinal phytase activity in terrestrial monogastric species, little work has been conducted investigating the enzyme's activity in finfish. This experiment was conducted to determine if hybrid striped bass Morone saxatilis × M. chrysops possess intestinal phytase activity, and to perform a preliminary characterization of this activity in terms of pH, metal ion dependence and substrate kinetics. Hybrid bass do exhibit low levels of intestinal phytase activity. The activity has an acidic pH optimum between 3.5 and 4.5. Low concentrations of Mn²⁺ slightly enhance this activity. The divalent cations Mg²⁺ and Zn²⁺ were shown to either have no effect on the enzyme activity or to be inhibitory, particularly at high concentrations. The present results indicate that intestinal phytase activity in hybrid bass does follow Michaelis-Menten kinetics, with estimated K_m and V_max values of 2.5 mM phytate and 4.8 units mg⁻¹ protein, respectively. The physiologic significance of the enzyme's presence in hybrid striped bass is questionable due to the low activity as compared to terrestrial monogastrics, the acidic pH optimum and the low affinity the enzyme appears to have for phytate. These observations also raise the question of whether the intestinal phytase activity detected in hybrid bass is merely a manifestation of non-specific acid phosphatase activity. Further investigation is needed to determine the practical significance of phytase activity in hybrid bass in terms of phytate phosphorus utilization. Additional experiments should also be conducted which evaluate intestinal phytase activity in other important aquacultured finfish species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Fish reproductive activity reveals temporal variations predominating spatial heterogeneity in maintaining high functional diversity of a Neotropical reservoir

The reproductive activity of the fish assemblage in a large Neotropical reservoir was evaluated by surveying the occurrence of ichthyoplankton in response to spatial and temporal variation in environmental factors. Fish reproduction was investigated by capturing larvae along representative spatial gradients in the main body (longitudinal) and arms (lateral) of the reservoir. After identification, the captured larvae were classified into different groups according to the functional traits of their parental stock. Sampling occurred monthly at night from October to March over five reproductive periods between 2009 and 2016, using conical-cylindrical plankton nets with flowmeter. Water samples were taken simultaneously allowing the measurement of environmental factors related to reproductive activity. The relationship between the environmental factors and the functional traits was assessed using RLQ and fourth-corner analyses. An evaluation of 51,995 larvae revealed there was greater larval abundance in the lacustrine and transition zones of both longitudinal and lateral gradients. Occurrences often correlated with temporal instead of spatial gradients. Higher rainfall correlated with abundances of detritivorous and migratory species. Higher water levels correlated with higher abundances of non-native species, species without parental care, with external fecundation, and partial spawning. Dry periods, with lower water and higher dissolved oxygen, correlated with higher abundances of insectivores and sedentary species. Although the presence of a dam-free stretch of river upstream of the reservoir had a positive influence on fish larvae in the fluvial zone, the results did not show differentiation along either gradient. Thus, the composition of the functional groups of fish species that reproduce in the Itaipu Reservoir appears to be predominantly influenced by the temporal variation in environmental factors rather than the formation of spatial gradients.     

Tree performance and fruit yield and quality of ‘Okitsu’ Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The ‘Okitsu’ Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars, and it is considered to be tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of ‘Okitsu’ fruits are well suited for the local market in the summer (December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for ‘Okitsu’. Consequently, a field trial was carried out in Bebedouro, São Paulo State, to evaluate the horticultural performance of ‘Okitsu’ Satsuma mandarin budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid Rangpur lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’ and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the mandarins ‘Sun Chu Sha Kat’ (C. reticulata Blanco) and ‘Sunki’ (Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (Citrus sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 6 m × 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of ‘Okitsu’ mandarin. The ‘Flying Dragon’ trifoliate had a unique effect over the ‘Okitsu’ trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The ‘Cravo Limeira’ and ‘Cravo FCAV’ Rangpur limes induced early-ripening of fruits, with low fruit quality. ‘Sun Chu Sha Kat’ and ‘Sunki’ mandarins and the ‘Orlando’ tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock.