Comparative study on the antioxidant and biological activities of carvacrol, thymol, and eugenol derivatives

Four derivatives of thymol, carvacrol, and eugenol were synthesized: 4-(hydroxymethyl)-5-isopropyl-2-methylphenol, 4,4'-methylenebis(5-isopropyl-2-methyl)phenol, 4-allyl-6-(hydroxymethyl)-2-methoxyphenol, and 4-(hydroxymethyl)-2-isopropyl-5-methylphenol. The obtained derivatives showed remarkably better antioxidative properties according to 1,1-diphenyl-2-picrylhydrazyl assay (50% inhibitory concentrations = 4-156 microg/mL) and Rancimat assay (protection factors = 1.55-5.84) when compared with parent compounds and values similar to or better than those of butylated hydroxytoluene and vitamin C. At concentrations of 10 mM carvacrol derivatives had no toxic effect on viability of Escherichia coli K-12 (determined by minimum inhibitory concentrations). Other phenol derivatives showed reduced cytotoxic effect on E. coli K-12 at concentrations of 2-5 mM on the basis of 50% lethal dose measurements. In comparison with the parent compounds, phenol derivatives showed reduced cytotoxic effect for Saccharomyces cerevisiae cells (determined by yeast colony reduction). On the other hand, the majority of synthesized compounds had dose-dependent antiproliferative effects on human uterine carcinoma cells (HeLa), which makes them potentially interesting for the adjuvant experimental cancer treatments. The 4,4'-methylenebis(5-isopropyl-2-methyl)phenol derivative of carvacrol showed lower inhibiting capacity also for the HeLa cells, which makes this particular derivative attractive as an efficient antioxidant with negligible cytotoxic effects.     

Tannin composition of barbatimão species

A comparative study of tannin composition of three species known as barbatim?o (Stryphnodendron adstringens, Stryphnodendron polyphyllum and Dimorphandra mollis) was developed by TLC, hydrolysis followed by PC, and colorimetry. Principal component analysis of the data obtained showed strong differentiation between the genus Dimorphandra and Stryphnodendron. Additionally, barks from the two Stryphnodendron sp. also presented considerable differences in the tannin structure.     

Practical diets with essential oils of plants activate the complement system and alter the intestinal morphology of Nile tilapia

The effect of the essential oils (EOs) of peppermint, Mentha piperita L., and tea tree, Melaleuca alternifolia (Maiden & Betche) Cheel, on the haematological, biochemical, and immunological parameters and intestinal morphology of Nile tilapia, Oreochromis niloticus L., was evaluated. Fish (58.09 ± 5.87 g) were fed 100 mg/kg and 250 mg/kg of each EO and sampled on days 7, 14, 30 and 60 after starting supplementation. The haematological and biochemical parameters were not altered by the supplementation of EOs compared to the control (p > .05). With regard to the immunological parameters, the activation of the complement system of fish fed 250 mg/kg peppermint and 100 mg/kg and 250 mg/kg tea tree EOs were significantly higher compared to the control after 60 days of feeding (p < .05). The complement system plays an essential role in innate immunity and contributes significantly to the acquired immune response; thus, its activation through supplementation with EOs is promising for the formulation of nutritional additives in aquaculture. Regarding intestinal morphology, fish fed 250 mg/kg tea tree EO presented higher villus size compared to all other groups (p < .05), which represents a healthier gut. These fish present a larger intestinal surface, which can result in better absorption and utilization of the nutrients. Based on the responses found in this study, both EOs were considered promising for the formulation of feed additives for Nile tilapia.     

Compound odontoma in three dogs

Three young, female dogs were operated for compound odontoma. All tumors were considered stage III with treatment consisting of partial mandibulectomy or maxillectomy. Microscopic examination of the resected tissue confirmed the diagnosis. Relatively aggressive, resective surgery resulted in prolonged tumor-free intervals.     

Aluminum tolerance in barley: Methods for screening and genetic analysis

Aluminum (Al) tolerance in roots of two cultivars of barley was studied using hematoxylin staining and root re-growth procedures. This study was performed in two F₂ segregating populations originated from crosses between the tolerant FM-404 and sensitive Harrington cultivars. The F₂ progeny analysed with hematoxylin staining revealed a segregation ratio of 3 tolerant: 1 sensitive, showing that the Al tolerance is controlled by a single gene with complete dominance for tolerance. The root re-growth measures do not confirm the 3:1 ratio. This last result can be explained due to the occurrence of genes that affect root growth rate or to the difficulties found in the evaluation of root re-growth. Barley has a complex root system, which makes it difficult to measure root re-growth after an extended period in nutrient solution. Due to the simplicity, reliability and better precision, the hematoxylin staining is the best procedure to determine the Al tolerance and its inheritance in barley. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of Metarhizium anisopliae fungus on off-host Rhipicephalus (Boophilus) microplus from tick-infested pasture under cattle grazing in Brazil

This work aimed to assess the effect of the fungus Metarhizium anisopliae on off-host Rhipicephalus (Boophilus) microplus from tick-infested Brachiaria decumbens pasture undergoing cattle grazing. For this purpose a naturally tick-infested Brachiaria decumbens pasture of 60 m × 100 m with twelve grazing Holstein Friesian-Nelore (Bos indicus) cross breed bovines was sprayed 12 times, 21 days apart with an aqueous conidial suspension of the E9 isolate of M. anisopliae fungus. Control pasture was treated with conidial suspension vehicle only. Efficacy of treatment was evaluated by tick larvae counts on the pasture and that of engorged female ticks on grazing cattle. Fungus persistence on grass stems as well as soil pasture was assessed after each spray. Efficacy of fungus against ticks was also measured by an in vitro immersion test. Whereas in vitro test showed a clear pathogenic effect of the fungus on ticks, no pathogenic effect over R. (B.) microplus ticks was detected in the field trial by spraying pasture with fungal suspension. Fungus from the suspension could be recovered from both the grass stem as well as the soil of sprayed pasture, even though the numbers obtained varied distinctly and could only be recovered shortly after spray. Pasture environments with exposure to sun and rain, seem to be very detrimental to the fungus, thus suspensions which provide fungus with protection or more resistant isolates to these should be looked for. In order to achieve high fungal efficacy against tick under field conditions, accurate laboratory assays, optimization of application strategy and knowledge of interactions between fungal strains and environmental factors are warranted.     

Biochemical parameters in the blood of gilthead sea bream (Sparus aurata Linnaeus, 1758) supplemented with commercially available β‐glucan‐based product (IMUNO‐2865®)

The aim of this study was to evaluate the effects of a promising new immune stimulant in aquaculture (IMUNO‐2865^®) on biochemical parameters in sea bream during the winter stress period. A total of 640 sea bream were fed throughout 90 days with diets containing 0 (Group 1), 1 (Group 2), 10 (Group 3) and 25 g (Group 4) of IMUNO‐2865^® kg^?1 of feed. Samples were taken each month and 90 days after supplementation. No statistical differences among treatment groups were noticed for the following biochemical parameters: glucose (GLU), aspartate aminotransferase (AST), plasma cholesterol (CHOL), lactate dehydrogenase (LDH), urea (URE) and creatinine (CREA). At the final sampling, total ammonia (NH₃) was higher in Groups 3 and 4 compared to the control and the low supplementation group (p < .05), while total proteins (TP) was significantly higher in Group 4 compared to all other groups, and in Groups 2 and 3 compared to the control group (p < .05). Blood Ca⁺⁺ levels were significantly higher after 60 days of feeding in all treatment groups compared to the control, and remained elevated in Group 4 even after 90 days following cessation of supplementation (p < .05). The results of this study described the increase of biochemical parameters in the blood of sea bream after use of IMUNO‐2865^® but future research is needed to evaluate its potentially immunostimulative effect on fish in commercial aquaculture.     

Biochemical properties of bull spermatozoa separated in iodixanol density solution

Bull spermatozoa samples contain variable portion of motile and normal morphology spermatozoa along with spermatozoa incapable of fertilization due to their pathologic changes. As semen quality is influenced by biochemical and morphological characteristics of all spermatozoa, the aim of the study was to separate spermatozoa in discontinuous iodixanol density gradient solution and to determine their cholesterol, phospholipid, triacylglycerol and lipid peroxide concentrations and creatine kinase activity. The study was performed in winter and included seven Simmental bulls aged 1.5-3.5 years. Semen samples were collected by use of artificial vagina. Upon evaluation of semen quality (volume, concentration and progressive sperm motility), the samples were centrifuged in iodixanol density solution to obtain two sperm fractions. The two fractions included sperms with progressive motility greater than 90% and less than 20%, respectively. A statistically significantly higher lipid peroxide concentration was determined in sperm fraction with <20% progressive motility. Different sperm subpopulations can be obtained by separating bull spermatozoa in different iodixanol density gradient solutions, while monitoring their biochemical properties can help assess the sperm quality.     

Effects of fasting and refeeding on the antioxidant system in cockerels and pullets

The effect of fasting and refeeding on total antioxidant status (TAS), glutathione peroxidase (GSH-Px) activity and concentration of some non-enzymatic antioxidant compounds was studied in cockerels and pullets. Blood was collected before and after 48-h fasting and 24 h after refeeding. In cockerels, fasting resulted in a significant decrease of TAS and uric acid concentration. After refeeding, the concentration of TAS remained significantly lower as compared to the control level. At the same time, blood plasma level of total lipids increased in comparison to the control and post-fasting values. In pullets, fasting resulted in a significant decrease of whole blood haemolysate GSH-Px activity and blood plasma concentrations of albumin and uric acid. Simultaneously, a significant increase in total lipids and cholesterol was obtained. In pullets, refeeding resulted in a further decrease of TAS to undetectable values, a significant decrease of blood plasma cholesterol, and a significant increase of GSH-Px in the whole blood haemolysate and in blood plasma uric acid content. The results indicate that fasting has a negative impact on the antioxidant defence system of the blood, which leads to a reduced resistance to oxidative stress in both cockerels and pullets. However, pullets seem to be more susceptible to fasting-provoked oxidative stress than cockerels.