Distillation methods affect the chemical composition of Varronia curassavica Jacq. essential oil? / Métodos de destilação afetam a composição química do óleo essencial de Varronia curassavica Jacq?

The objective of this work was to evaluate the chemical composition of essential oil from Varronia curassavica Jacq. obtained by microwave (MI) and hydrodistillation (HD) extraction methods. The MI method tested three powers (500, 600, and 700W), three distillation times (20, 30, and 40 min.), and three water volumes (0, 25, and 50 mL per sample). The HD method tested three distillation times (100, 120, and 140 min.) and three water volumes (1.0, 1.5, and 2.0 L per 3-liter flask). The essential oils were analyzed by GC/MS-FID. The optimal condition for the essential oil extraction by the MI method was 700W for 40 min. (3.28%), regardless of the volume of water. In its turn, the best condition for essential oil extraction by the HD method was 120 min. with 1.0 L of water per flask (3.34%). The most abundant compounds for MI (700 W for 40 min. without water) were shyobunol (26.53%) and bicyclogermacrene (4.96%); and the most abundant compounds for HD (120 min. with 1.0 L of water/flask) were shyobunol (24.00%) and germacrene D-4-ol (10.23%). Methyl farnesoate (2E, 6E) and farnesyl acetate (2Z, 6E) were not detected in the essential oil extracted by HD; however, they were identified by the MI method. By increasing the distillation time and/or volume of water in HD, a reduction was observed for the content of the chemical compounds -elemene (from 1.23 to 0.97%), Ecaryophyllene (from 5.49 to 4.35%), -humulene (from 1.80 to 1.43%), alloaromadendrene (from 1.78 to 1.44%), bicyclogermacrene (from 5.63 to 4.55%), and germacrene D-4-ol (from 11.40 to 9.86%). Power, extraction time, and their interactions influenced the content of essential oil obtained by microwave extraction (MI). Within each power, the highest essential oil content was extracted at the longest distillation time (40 min.), except for 600W, where no significant difference was detected between 30 and 40 min. The optimal essential oil contents for both extraction methods were statically similar by the t-test for dependent samples. However, the MI method presents advantages, such as shorter distillation time and less energy and water consumption.

O objetivo deste trabalho foi avaliar a composição química do óleo essencial de Varronia curassavica Jacq. obtido pelos métodos de extração micro-ondas (MI) e hidrodestilação (HD). Para MI, foram testadas três potências (500, 600 e 700W), três tempos de destilação (20, 30 e 40 min.) e três volumes de água (0, 25 e 50 mL por amostra). Para HD, foram testados três tempos de destilação (100, 120 e 140 min.) e três volumes de água (1,0; 1,5 e 2,0 L por balão de 3 litros). Os óleos essenciais foram analisados por CG/EM-FID. Maiores teores de óleo essencial foram obtidos nas condições de 700 W por 40 min. (3.28%), independente do volume de água para MI, e 120 min. com 1,0 L de água por balão para HD (3,34%). Os compostos mais abundantes para MI (700W, por 40 min., sem água) foram o shyobunol (26,53%) e biciclogermacreno (4,96%) e para HD (120 min. com 1,0 L de água /balão) foram shyobunol (24,00%) e germacreno D -4 -ol (10,23%). Metil farnesoato (2E, 6E) e farnesil acetato (2Z, 6E) não foram detectados no óleo essencial extraído por HD, porém, foram detectados nas amostras extraídas por MI. Com o aumento do tempo de destilação e/ou do volume de água em HD, houve redução no conteúdo dos constituintes químicos -elemeno (de 1,23 para 0,97%), E-cariofileno (de 5,49 para 4,35%), -humuleno (1,80 para 1,43%), aloaromadendreno (de 1,78 para 1,44%), biciclogermacreno (de 5,63 para 4,55%) e germacreno D-4-ol (de 11,40 para 9,86%). A potência, o tempo de extração e suas interações influenciaram no teor de óleo essencial obtido na extração por micro-ondas (MI). Dentro de cada potência, o maior teor de óleo essencial foi obtido no tempo mais longo de extração (40 min.), exceto para 600 W, que não apresentou diferença significativa entre 30 e 40 min. Nas condições ótimas de extração, os teores de óleo essencial obtidos foram estatisticamente semelhantes pelo teste t para amostras dependentes. No entanto, a extração por micro-ondas apresenta algumas vantagens em relação a HD, como menor tempo de destilação e menor consumo de energia e água.

Deltamethrin-induced nuclear erythrocyte alteration and damage to the gills and liver of Colossoma macropomum.

Deltamethrin is one of the most commonly used pyrethroids in the world, and it has a high toxic potential, mainly on aquatic organism. Thus, the purpose of this study was to evaluate LC50 values of deltamethrin on tambaqui (Colossoma macropomum) fingerlings and to investigate genotoxic effects and histopathological responses. Fish were exposed to different concentrations of deltamethrin (0, 6.16 × 10-3; 6.44 × 10-2; 1.34 × 10-1, and 1.93 × 10-1 mg L-1) for 96 h. In addition, a genotoxicity analysis was carried out on peripheral blood erythrocytes and histopathological changes were classified by the severity degree of damage and organ functioning. The 96 h LC50 value for tambaqui was estimated at 5.56 × 10-2 mg L-1 using a static test system. Nuclear abnormalities in exposed fish included micronuclei, blebbed, notched, 8-shaped, and binucleated nuclei forms. Deltamethrin significantly induced a notched nucleus compared to other abnormalities. A histopathological examination showed hepatic lesions and gill damage. Deltamethrin was found to be highly toxic; it induced genotoxicity and caused liver and gill inflammation in tambaqui.

The effects of gelatin supplementation prior to cooling on ram semen quality and fertility.

The physical and chemical characteristics of gelatin have been used to justify its inclusion in extenders to preserve the sperm quality and improve results of cervical artificial insemination with cooled semen. The objective of this study was to evaluate the effects of gelatin supplementation in cooling extender on the quality and fertility of ram semen stored at 5°C. Semen samples (n = 24) of Santa Inês rams (n = 6) were diluted in Glycine-Yolk-Milk extender without (control) or with 1.5% of gelatin. The samples were loaded into 0.25 mL straws, cooled to 5°C and stored vertically for 48 and 72 hours. Sample quality was evaluated using straw homogeneity tests based on pH, osmolality and the proportion of spermatozoa (PS) in both upper and lower segments of straws (US and LS), analyses of sperm motility, plasma and acrosomal membrane integrity, and by fertility after artificial insemination. Differences between the US and LS of straws were found for pH and PS (%). They were significant only in the control group at both times: pH - 5.96 vs. 5.71 at 48 h and 6.13 vs. 5.89 at 72 h; PS - 21.66 vs. 78.34 at 48 h and 20.87 vs. 79.13 at 72 h. Storage in gelatin had very little, to no effect on the sperm kinetics or on the sperm membrane integrity evaluations. The addition of gelatin to the extender did not affect the pregnancy rate which ranged from 4.4 to 26.1%. We conclude that gelatin is effective in maintaining the physical and chemical homogeneity of the semen samples. Further research is needed in order to optimize the use of gelatin supplementation and elucidate any potential benefits.

Essential oils of Varronia curassavica accessions have different activity against white spot disease in freshwater fish.

The aim of this study was to evaluate the antiprotozoal activity of essential oils from Varronia curassavica accessions against different stages of Ichthyophthirius multifiliis. Essential oils from each accession were tested in vitro at the concentrations 0, 10, 25, 50, 75, 100, and 200 mg/L. The VCUR-001, VCUR-202, VCUR-509, and VCUR-601 accessions presented the major compounds α-pinene, germacrene D-4-ol, (E)-caryophyllene and epiglobulol, and sabinene, respectively. These isolated compounds were tested in vitro at a concentration proportional to that found in the essential oil which caused 100% mortality of the parasite. The concentrations of 10 and 50 mg/L of the essential oil of accession VCUR-202 provided 100% mortality of trophonts and tomonts, respectively. For the accession VCUR-509, 100% mortality of trophonts and tomonts was observed at concentrations 75 and 200 mg/L of essential oil, respectively. The same mortality was observed at concentration 200 mg/L in both stages of the parasite for the other accessions. The major compounds α-pinene, sabinene, and the (E)-caryophyllene + epiglobulol mixture caused 100% mortality of trophonts and tomonts. The in vivo assay for white spot disease control was performed in a therapeutic bath of 1 h with the essential oil of accession VCUR-202 at concentrations of 0.5 and 2.0 mg/L. A significant reduction of about 30% of trophonts on infected fish was observed, independent of the oil concentration. The V. curassavica essential oil, especially the VCUR-202 accession, is a potential source of raw material for the formulation and commercialization of bioproducts to control freshwater white spot disease in fish.

Use of cryotubes for the cryopreservation of tambaqui fish semen (Colossoma macropomum).

Tambaqui (Colossoma macropomum) is a freshwater fish of great importance to aquaculture in several South American countries. Recent studies have developed a protocol for semen cryopreservation in 0.25 and 0.5 mL straws; however, this technique has limitations for fingerling production at a large scale due to the high fecundity of tambaqui. The main objective of this study was to evaluate the feasibility of using cryotubes (1.6 and 4.5 mL) for tambaqui semen cryopreservation. Semen samples were diluted in freezing solution (5% glucose solution, 10% methylglycol, 5% egg yolk), stored in 1.6 and 4.5 mL cryotubes, frozen in liquid nitrogen vapor at -175°C and transferred to a cryogenic container at -196°C. The cryotubes were thawed in a water bath at 60°C for 70 or 90 s and the motility (total motility - TM; progressive motility - PM; curvilinear velocity - VCL; straight line velocity - VSL and average path velocity - VAP) and the viability of sperm were evaluated. There was no significant difference in sperm motility and viability post-thawing between 1.6 and 4.5m L cryotubes, except for TM (47% and 40%, respectively). Thawing for 90 s provided better results, being used in fertilization trials. Although the fertilization rate did not differ between the cryotubes (41-45%), it was significantly lower than that for fresh semen (74%). A strong positive correlation was observed between the sperm motility and fertilization rate (r=0.69-0.89). We conclude that 1.6 and 4.5 mL cryotubes have high potential for tambaqui semen cryopreservation when thawed for a minimum time of 90 s at 60°C.

Hormonal induction and semen characteristics of tambaqui Colossoma macropomum.

In the hatchery-bred tambaqui Colossoma macropomum, spontaneous semen release does not occur, and hand-stripping produces reduced semen volume. The goal of this work is to evaluate the effects of hormonal induction with carp pituitary extract (CPE) on both qualitative (visual aspect, pH, motility, viability and morphological abnormalities) and quantitative (volume, concentration and number of spermatozoa per ejaculate) traits of tambaqui semen. Eleven males were treated with CPE (induced), and 11 were left untreated as a control (non-induced). All analysed parameters except motility and percentage of viable spermatozoa presented significant differences (p < 0.05) between the induced and non-induced treatments. CPE induction resulted in a 25-fold increase in semen volume and a 10-fold increase in the number of spermatozoa collected. However, both sperm concentration and the frequency of sperm with morphological abnormalities (commonly detached heads or bent tails) were significantly lower in CPE-induced fish. The hormonal induction of tambaqui males with CPE is efficient and positively influences some qualitative and quantitative properties of semen. Additionally, semen collection via gentle abdominal massage occurs more readily in CPE-induced fish.