Cobia Rachycentron canadum L. reared in low‐salinity water: does dietary sodium chloride affect growth and osmoregulation?

The effects of NaCl supplementation (0.0%, 2.5%, 5.0%, 7.5% and 10.0% dry weight of a basal diet) on growth, gill histological alterations and osmoregulation of juvenile cobia reared in low‐salinity water (5 g L^?1) were assessed. At the end of the experiment, gills were sampled for Na⁺, K⁺‐ATPase activity determination and histological evaluation. In all treatments, no mortality was observed. Results showed that dietary NaCl supplementation did not alter growth. At the highest supplementations (7.5% and 10.0%), juvenile cobia showed higher feed intake and feed conversion ratio. Na⁺, K⁺‐ATPase activity was higher in fish fed the diet without salt supplementation than in those fed with NaCl‐supplemented diets. The number of chloride cells significantly increased with increasing dietary salt level, being 2.5‐fold higher in fish fed with 10.0% NaCl supplementation (41 cells mm^?2) than in those from the non‐supplemented fed group (16 cells mm^?2). These findings indicate that dietary salt supplementation stimulated chloride cell proliferation paralleled with a reduction in the gill Na⁺, K⁺‐ATPase activity, suggesting a possible decrease in energy consumption associated with osmoregulation. However, the suggested energy sparing did not have a significant impact on juvenile cobia growth.     

Genital lesions and distribution of amastigotes in bitches naturally infected with Leishmania chagasi

Recent reports indicate that Leishmania chagasi has tropism to the male canine genital system, which is associated with shedding of the organism in the semen, supporting the hypothesis of venereal transmission. The aim of this study was to describe the lesions and assess parasite load in the genital system of bitches with canine visceral leishmaniasis (CanL). Symptomatic (n=5) and asymptomatic (n=5) bitches seropositive for CanL were randomly selected at the Center for Zoonosis Control (Belo Horizonte, State of Minas Gerais, Brazil). Five serologically negative, healthy, adult bitches also from the CZC were used as controls. Samples from genital organs (vulva, vagina, cervix, uterine body, uterine horns, uterine tubes, and ovaries), liver, and spleen were histologically evaluated and processed for immunodetection of Leishmania sp., and PCR. The most significant histological change was a mild to moderate vulvar dermatitis, characterized by a histio-plasma-lymphocytic infiltrate. This change was detected in all asymptomatic, four symptomatic, and three uninfected control bitches. In one symptomatic and one asymptomatic bitch intracytoplasmic amastigotes were observed within macrophages in the inflammatory infiltrate. Samples from all the segments of the genital tract were positive in at least one infected animal, in the absence of detectable amastigotes in the tissue. These findings support the notion that L. chagasi does not have genital tropism in the bitch, which is in contrast to our previous findings in naturally infected male intact dogs.     

Mixed infection by Libyostrongylus douglassii and L. dentatus (Nematoda: Trichostrongylidae) in Struthio camelus (Ratites: Struthioniformes) from Brazil with further morphological characterization of adults

The genus Libyostrongylus includes three species, L. douglassii, L. dentatus and L. magnus that occur as parasites in the proventriculus of Struthio camelus. We confirmed a mixed infection by L. douglassii and L. dentatus in farmed ostriches from the southeast of Brazil for the first time, and provided new information on some morphological characters that differentiate these species. Adult nematodes collected from the proventriculus of ostriches were observed by light and scanning electron microscopy. Morphologic characterization and morphometric analysis of the nematodes enabled the distinction of both species and corroborated results of prior studies. Specimens of L. dentatus have a buccal capsule with a prominent esophageal tooth. Furthermore, males and females of L. dentatus were larger (4954 and 9347 microm) than those of L. douglassii (3411 and 4229 microm), but measurements for most characters in both species were smaller then those previously reported. Besides, the cephalic structure based on scanning electron microscopy differs, and L. dentatus has thick lips with round papillae, whereas L. douglassii has fine lips with lengthened papillae. The confirmation of both species in South America strongly suggests that the mixed infection may be common in farmed ostriches.     

A panel of protein candidates for comprehensive study of Caprine Arthritis Encephalitis (CAE) infection

The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE.     

Use of trichostatin A alters the expression of HDAC3 and KAT2 and improves in vitro development of bovine embryos cloned using less methylated mesenchymal stem cells

The aim of this work was to investigate the methylation and hydroxymethylation status of mesenchymal stem cells (MSC) from amniotic fluid (MSC‐AF), adipose tissue (MSC‐AT) and fibroblasts (FIB‐control) and to verify the effect of trichostatin A (TSA) on gene expression and development of cloned bovine embryos produced using these cells. Characterization of MSC from two animals (BOV1 and BOV2) was performed by flow cytometry, immunophenotyping and analysis of cellular differentiation genes expression. The cells were used in the nuclear transfer in the absence or presence of 50 nM TSA for 20 hr in embryo culture. Expression of HDAC1, HDAC3 and KAT2A genes was measured in embryos by qRT‐PCR. Methylation results showed difference between animals, with MSC from BOV2 demonstrating lower methylation rate than BOV1. Meanwhile, MSC‐AF were less hydroxymethylated for both animals. MSC‐AF from BOV2 produced 44.92 ± 8.88% of blastocysts when embryos were exposed to TSA and similar to embryo rate of MSC‐AT also treated with TSA (37.96 ± 15.80%). However, when methylation was lower in FIB compared to MSC, as found in BOV1, the use of TSA was not sufficient to increase embryo production. MSC‐AF embryos expressed less HDAC3 when treated with TSA, and expression of KAT2A was higher in embryos produced with all MSC and treated with TSA than embryos produced with FIB. The use of MSC less methylated and more hydroxymethylated in combination with embryo incubation with TSA can induce lower expression of HDAC3 and higher expression of KAT2A in the embryos and consequently improve bovine embryo production.     

Relation of Toxicity in Corn Seeds Treated with Zinc and Salicylic Acid

The beneficial effect of corn seed treatment with zinc (Zn) is directly related to the source used. The excess of this micronutrient causes seedling stress and reduces growth. Thus, assuming that the use of exogenous phytohormones can minimize such effects, we evaluated different doses and sources of Zn for the treatment of maize seeds with or without salicylic acid. The experiment took place in the laboratory, and two factorial experiments, 2 × 4 + 1, were performed in a randomized design. The seeds were treated with either ZnO or ZnSO₄ at doses of 0.5, 1, 2, and 3 g.kg^?1 seed with four replications, differing only by the addition of 4.14 mg L^?1 salicylic acid. Treating seeds with Zn and salicylic acid did not affect germination. ZnO led to a greater increase in dry mass in corn seedlings as compared with zinc sulfate, especially at higher doses (2 and 3 g kg^?1 seed). Seed treatment with sulfate reduces root and shoot length, and salicylic acid did not attenuate this toxic effect. Dry mass is not affected when oxide is used. Salicylic acid reduces the accumulation of zinc in the treatment of corn seeds, regardless of the source used.     

Biogenic amine profile in unripe Arabica coffee beans processed according to dry and wet methods

Immature coffee fruit processing contributes to a high amount of defective beans, which determines a significant amount of low-quality coffee sold in the Brazilian internal market. Unripe bean processing was tested, taking the levels of bioactive amines as criteria for evaluating the extent of fermentation and establishing the differences between processing methods. The beans were processed by the dry method after being mechanically depulped immediately after harvest or after a 12 h resting period in a dry pile or immersed in water. Seven bioactive amines were quantified: putrescine, spermine, spermidine, serotonin, cadaverine, histamine, and tyramine, with global amounts ranging from 71.8 to 80.3 mg/kg. The levels of spermine and spermidine were lower in the unripe depulped coffee than in the natural coffee. The specific conditions of dry and wet processing also influenced cadaverine levels, and histamine was reduced in unripe depulped coffee. A resting period of 12 h does not induce significant alteration on the beans and can be improved if performed in water. These results confirm that peeling immature coffee can decrease fermentation processes while providing more uniform drying, thus reducing the number of defects and potentially increasing beverage quality.     

Production of triacetylhydroxytyrosol from olive mill waste waters for use as stabilized bioantioxidant

A hydroxytyrosol triacetyl derivative was very efficiently produced as a highly pure stabilized antioxidant compound by a short treatment of olive mill waste water (OMWW) organic extracts, rich in hydroxytyrosol, with an acetylating mixture composed of HClO4-SiO2 and Ac2O (Chakborti and Gulhane reaction), in mild and safe conditions. A successive single step of middle pressure liquid chromatography (MPLC) purification of the reaction product was performed, with an overall yield of 35.6%. (This process, including both the Chakborti and Gulhane reaction and the MPLC purification, is protected by an international patent under PCT/IT2005/000781.) The o-diphenol triacetyl derivative was also produced by direct reaction of hydroxytyrosol, previously purified by MPLC, with HClO4-SiO2 and Ac2O, with an overall yield of 29.5%. A further procedure for the production of the hydroxytyrosol triacetyl derivative was consistent with the direct treatment of raw OMWW with the acetylating agent and a single step of MPLC purification, with an overall yield of 27.6%. The purified natural triacetylhydroxytyrosol confirmed the same strong protective effects against the oxidative stress in human cells as the corresponding synthetic compound, likely because of the biochemical activation of the acetyl derivative into the active parent hydroxytyrosol by esterases. We therefore propose the utilization of OMWW for recovering hydroxytyrosol as a natural antioxidant in a chemically stabilized form, with a good yield, which can be potentially used as a nontoxic functional component in nutritional, pharmaceutical, and cosmetic preparations.