A scanning electron microscope study of Toxocara genettae Warren, 1972 (Ascaridae), with data on morphometric variation.

We have carried out a morphometric study of the ascaridoid nematode Toxocara genettae, parasite of the small intestine of the genet Genetta genetta, with the aid of scanning electron microscopy (SEM). Host specimens were trapped in Galicia, NW Spain. The most characteristic features of this nematode are the absence of cervical alae, replaced by narrow lateral crests running along the length of the body and the presence of a mucron at the end of the male tail. Other features of interest include the number of labial denticles (90-98 and 96-102 on the dorsal lip, 82-96 and 90-96 on the subventral lip of male and female, respectively), the width of the transverse striations (25-28 microns) and, in many of the specimens examined, the position of the precloacal median papilla at the base of a well-defined cuticular pit.     

Nondestructive assessment of phytopigments in riverbed sediments by the use of instrumental color measurements

Purpose  

The quantification of phytopigments in riverbed sediments deserves further attention because it provides information about eutrophic levels, and therefore about sediment and water quality. Due to the current interest in the study of eutrophication processes, there is a need for the development of a rapid, simple, cost-effective, and nondestructive method of quantifying phytopigment content. We describe one method based on color measurements and without the need for extraction and chemical assay.     

Resveratrol modulates innate and inflammatory responses in fish leucocytes

Resveratrol (RESV; trans-3,5,4'-trihydroxystilbene), a phytoalexin that is produced by some plants, among other effects has well-known antioxidant, anti-inflammatory and immunomodulatory activities in mammals. In the present study, the effects of RESV on several functions of turbot, Psetta maxima (L.), kidney leucocytes (KLs) related to the innate and inflammatory responses were investigated. RESV exerted a dose-dependent inhibitory effect on the migratory response and on the production of reactive oxygen species in KL, after stimulation of the respiratory burst activity with phorbol myristate acetate (PMA). RESV also significantly inhibited the generation of the pro-inflammatory mediator prostaglandin E(2) (PGE(2)) in the supernatant of KL cultures stimulated with acidic sulphated polysaccharides (ASPs) from the seaweed Ulva rigida. The effects of the polyphenol on enzymatic activity and on myeloperoxidase (MPO) gene expression in neutrophils were also tested. It was found that RESV strongly inhibited intracellular and extracellular MPO activity, behaving as a noncompetitive and reversible inhibitor, and also induced a decrease in MPO mRNA levels in turbot neutrophils. These findings indicate that RESV exerts important modulatory effects on inflammatory responses in fish, and considering the importance of innate immunity in these vertebrates and the similarities with mammals, it may be possible to use fish for analysis of the effects of different substances on inflammatory responses.     

Vaccination of turbot, Psetta maxima (L.), against the protozoan parasite Philasterides dicentrarchi: effects on antibody production and protection

The efficacy of a vaccine against the fish pathogen Philasterides dicentrarchi was evaluated in turbot by measuring the production of specific antibodies and duration of protection. Four groups of turbot were vaccinated twice, on days 0 and 30, with phosphate-buffered saline, mineral oil adjuvant, antigen or antigen plus adjuvant. Specific serum antibodies were determined on day 0 and 1 month after the first and the second vaccinations. Protection was evaluated 1 month after the first vaccination and 1 and 5 months after the second vaccination. Serum antibody titres, measured by enzyme-linked immunosorbent assay, and protection, assessed by challenges, increased significantly 1 month after the second vaccination in the group injected with antigen plus adjuvant and the protection lasted for at least a further 5 months in this group. The relative protection was 77% and 66% 1 and 5 months after the second vaccination, respectively. Administration of antigen or adjuvant separately had no effect on antibody response or protection. The results indicate that emulsion containing antigen plus adjuvant induced durable protection against P. dicentrarchi after the administration of the two vaccinations, and that this preparation can be used as a vaccine against the pathogen.     

Non-specific responses of turbot (Scophthalmus maximus L.) adherent cells to microsporidian spores

We investigated non-specific responses of turbot spleen- and pronephros-resident adherent cells to spores of fish microsporidians, and the effects of the glucocorticoid dexamethasone (DX) on these responses. On average, 65% of adherent cells from the spleen and pronephros showed esterase activity (as characteristic of macrophages); 32% showed peroxidase activity (as characteristic of neutrophils), and 19% of peroxidase-positive cells were capable of phagocytosing microsporidian spores. A significantly higher proportion of adherent cells showed phagocytic activity when viable spores were the target than when non-viable spores were the target. Microsporidian spores stimulated adherent cells to produce reactive oxygen and nitrogen intermediates (ROIs and RNIs), though less effectively than the other stimulants tested. Adherent cells exposed to viable spores produced significantly less intracellular superoxide than adherent cells exposed to non-viable spores. Daily injection of fish with DX over 6 days significantly inhibited both phagocytosis of microsporidian spores and spore-induced ROI production, and similar effects were observed when adherent cells were exposed to DX in vitro.     

Potential biomedical reuse of vegetative residuals from mycorrhized grapevines subjected to warming

Grapevine leaves are widely discarded in open fields despite their known antioxidant properties. We tested the cytotoxicity of leaf extracts from three clones (CL-260, CL-1048, CL-8) of Vitis vinifera L. cv. Tempranillo against four human cancer cell lines: colon, HT-29; breast, MCF-7; lung HTB-54; and lymphoblastic leukemia, CCRF-CEM. Grapevines were cultivated at either ambient (24/14 °C) or elevated (28/18 °C) day/night temperatures, and inoculated (+M) or not (-M) with arbuscular mycorrhizal fungi (AMF). Cytotoxicity was analysed by MTT assays. Elevated air temperatures enhanced the cytotoxicity of leaf extracts from CL-260 against HT-29, CCRF-CEM and HTB-54 and that from CL-8 against MCF-7. Mycorrhization improved the cytotoxicity of leaf extracts from CL-1048 against HT-29, CCRF-CEM, HTB-54 and MCF-7. The cytotoxic activities of CL-260 against HTB-54 and CL-1048 against HT-29 were correlated, respectively, with total phenols and total antioxidant capacity. We conclude that the predicted increase in air temperature for the future climate and the mycorrhizal association of grapevines may enhance the cytotoxicity of leaves, which strengthens the potential application of these agricultural residuals for biomedicine. However, the clonal diversity in the response to AMF and air temperature highlights the importance of choosing the most adequate clone for a concrete environmental scenario.     

Design, synthesis, and biological evaluation of phosphoramide derivatives as urease inhibitors

The design, synthesis, and biological evaluation of phosphoramide derivatives as urease inhibitors to reduce the loss of ammonia has been carried out. Forty phosphorus derivatives were synthesized and their inhibitory activities evaluated against that of jack bean urease. In addition, in vivo assays have been carried out. All of the compounds were characterized by IR, (1)H NMR, MS, and elemental microanalysis. In some cases, detailed molecular modeling studies were carried out, and these highlighted the interaction between the enzyme active center and the compounds and also the characteristics related to their activity as urease inhibitors. According to the IC(50) values for in vitro inhibitory activity, 12 compounds showed values below 1 microM and 8 of them represent improvements of activity in comparison to the commercial urease inhibitor N-n-butylthiophosphorictriamide (NBPT) (100 nM) (AGROTAIN). On the basis of the activity results and the conclusions of the molecular modeling study, a structural model for new potential inhibitors has been defined.     

Effect of clofibrate and hydrocortisone on intestinal trichinellosis in mice

These investigations show the immunosuppressive effects of Clofibrate (CPIB) and hydrocortisone on the intestinal cycle of Trichinella nelsoni. They demonstrate an effect of CPIB similar to that of hydrocortisone in the establishment of the muscular and intestinal phase of this parasite. A higher retention rate of intestinal adult worms was observed in hydrocortisone-treated mice. These facts are consistent with the idea that CPIB can act on diverse immune cells, involving both thymus-dependent and thymus-independent antigens.