Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

Significant improvement of shrimp growth performance by growth hormone‐releasing peptide‐6 immersion treatments

Growth hormone‐releasing peptide‐6 (GHRP‐6) is one of the earliest developed synthetic peptidyl growth hormone secretagogue receptor agonists. These compounds mimic the effect of the endogenous ligand ghrelin. In vertebrates, ghrelin is a potent circulating orexigenic hormone with functional roles in controlling food intake, energy expenditure, adiposity, growth hormone secretion and immunity. Ghrelin has been studied mainly in vertebrates; thus, little is known about its role in invertebrates, including crustaceans. We first evaluated the effect of GHRP‐6 injection over feed intake in shrimp and its effects on shrimp growth when the peptide was administrated by successive immersion baths. GHRP‐6 increased feed intake, body weight and size, the number of rostral spines and gill branches, protein concentration and haemocyte number in treated shrimps. We also evaluated the peptide uptake and clearance in a pharmacokinetics, using [H³]GHRP‐6 administered to postlarvae. Given a limited exposure and efficient clearance of the peptide‐associated radioactivity from larvae, our findings suggested that GHRP‐6‐treated Litopenaeus vannamei can be consumed safely by humans after aquaculture applications. These results propose that GHRP‐6 could be an additional tool to study growth physiology in crustaceans and also a promising candidate for development into a new biotechnology product for improving shrimp growth and quality.     

Genetics of resistance to bean golden mosaic virus in Phaseolus vulgaris L.

Summary The genetics of resistance to bean golden mosaic virus (BGMV) was studied in an 8×8 complete diallel cross of common bean, Phaseolus vulgaris L. The 28F₁ hybrids, their reciprocals, and eight parents were artificially inoculated with BGMV under glasshouse conditions. Data were recorded and analyzed for foliar yellowing, plant dwarfing, flower abortion, and pod formation, using a randomized complete block design with two replications.General combining ability (GCA) mean squares were highly significant (P<0.01) and larger than values for specific combining ability for all traits. Reciprocal and maternal effects were nonsignificant. Cultivars Royal Red and Alubia Cerrillos possessed significant negative and positive GCA for all traits, respectively. Porrillo Sintetico also had negative GCA for all traits except plant dwarfing. None of the GCA effects were significant for Great Northern 31 and PVA 1111. Positive associations existed among all traits studied.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Ileal digestibility of amino acids,phosphorus, phytate and energy in pigs fed sorghum‐based diets supplemented with phytase and Pancreatin®

The effects of phytase supplementation on the apparent ileal digestibility (AID) of amino acids (AA) have been inconsistent. Two experiments evaluated the effect of providing a mixture of pancreatic enzymes (Pancreatin^®) to growing pigs fed sorghum–soybean meal diets supplemented with phytase on the AID of AA, energy, and phosphorus (P), as well as the ileal digestibility (ID) of phytate; there were four periods per experiment. In Experiment 1, eight pigs (BW 22.1 ± 1.3 kg) were fitted with a T‐cannula at the distal ileum. Each period consisted of 9 days; 7 days for diet adaptation, and 2 days for digesta collection. Treatments (T) were: (i) basal sorghum–soybean meal diet, (ii) basal diet plus Pancreatin^®, (iii) basal diet plus phytase and (iv) basal diet plus phytase and Pancreatin^®. Phytase increased the digestibilities of phytate and P (p < 0.001), but did not affect the AID of AA and energy (p > 0.10). Except for methionine (p = 0.07), Pancreatin^® did not affect the AID of AA. Phytase and Pancreatin^® did not interact (p > 0.10). Experiment 2 was similar to Experiment 1, but Pancreatin^® was infused into duodenum. Pancreatin^® infusion did not affect the AID of AA (p > 0.10); and tended to reduce (p = 0.09) the AID of lysine. Phytase × Pancreatin^® interactions were not observed (p > 0.10). In conclusion, phytase and Pancreatin^® did not improve the AID of AA in growing pigs fed sorghum–soybean meal diets indicating that phytates did not affect AA digestibility.     

Radial variation in sap flow in five laurel forest tree species in Tenerife,Canary Islands

Variations in radial patterns of xylem water content and sap flow rate were measured in five laurel forest tree species (Laurus azorica (Seub.) Franco, Persea indica (L.) Spreng., Myrica faya Ait., Erica arborea L. and Ilex perado Ait. ssp. platyphylla (Webb & Berth.) Tutin) growing in an experimental plot at Agua García, Tenerife, Canary Islands. Measurements were performed around midday during warm and sunny days by the heat field deformation method. In all species, water content was almost constant (around 35% by volume) over the whole xylem cross-sectional area. There were no differences in wood color over the whole cross-sectional area of the stem in most species with the exception of E. arborea, whose wood became darker in the inner layers. Radial patterns of sap flow were highly variable and did not show clear relationships with tree diameter or species. Sap flow occurred over the whole xylem cross-sectional area in some species, whereas it was limited to the outer xylem layers in others. Sap flow rate was either similar along the xylem radius or exhibited a peak in the outer part of the xylem area. Low sap flow rates with little variation in radial pattern were typical for shaded suppressed trees, whereas dominant trees exhibited high sap flow rates with a peak in the radial pattern. Stem damage resulted in a significant decrease in sap flow rate in the outer xylem layers. The outer xylem is more important for whole tree water supply than the inner xylem because of its larger size. We conclude that measurement of radial flow pattern provides a reliable method of integrating sap flow from individual measuring points to the whole tree.     

Dynamics of violaxanthin and lutein epoxide xanthophyll cycles in Lauraceae tree species under field conditions

Two xanthophyll cycles have been described in higher plants: the violaxanthin xanthophyll (V or VAZ) cycle, which is present in all species, and the taxonomically restricted lutein epoxide xanthophyll (Lx) cycle, which involves the light-induced de-epoxidation of Lx to lutein (L) and its epoxidation back to Lx in low light. Laboratory experiments indicate that the first reaction occurs quickly, but the second reaction is much slower. We investigated the Lx cycle under field conditions in several tree species of the Lauraceae family to determine its relationship with the ubiquitous V cycle. The field study was conducted in two natural laurel forests: one in the Canary Islands, where Laurus azorica (Seub.) Franco, Ocotea foetens (Aiton.) Benth, Apollonias barbujana (Cav.) Bornm. and Persea indica (L.) Spreng were studied; and one in the Basque Atlantic coast where Laurus nobilis L. was studied. The results were complemented by a taxonomic study. The presence of Lx was widespread among Lauraceae species, but its concentration varied even among closely related species. The V pool size correlated positively with growth irradiance, whereas the relationship between Lx pool size and growth irradiance varied with species. A functional Lx cycle was confirmed under field conditions only in O. foetens and L. nobilis. Furthermore, in O. foetens, a correlation between Lx de-epoxidation and photoinhibition suggested a protective role for this cycle. We conclude that, unlike the V cycle, which is normally correlated with irradiance, the operation and light dependence of the Lx cycle is species-dependent.