Human cornea conservation in coconut water solution / Conservação de córneas humanas em solução de água de coco

ABSTRACT Purpose: The aim of this study was to evaluate the physical and chemical characteristics of coconut water and to analyze the use of coconut water solution for the conservation of human corneas. Methods: This was an experimental and controlled study performed at the Eye Bank of the General Hospital of Fortaleza. The coconut water-based solution was prepared at the Goat Seed Technology Laboratory of the Department of Veterinary Medicine of the State University of Ceará. Discarded corneas from the Eye Bank were divided into two groups for sequential experiments: G1, coconut water-based solution (experimental group), and G2, conservative treatment with OPTISOL GS® (control group). The osmolality of corneas in G1 was analyzed sequentially at 275, 300, 325, 345, 365, and 400 mOsm/L. The viability of the corneas was determined by specular microscopy and biomicroscopy on the first, third, and seventh days. Results: Corneas preserved in a solution of 365 and 345 mOsm/L had a transparency of 8 mm until the third day and had diffuse edema in the periphery, central folds, and partial epithelium loss until the seventh day. The 365-mOsm/L solution was associated with the worst results during follow-up. Corneas placed in Optisol-GS retained their original aspects. Conclusions: Coconut water-based preservative partially maintained corneal transparency and epithelial integrity, especially during the first three days of follow-up. The coconut water-based solutions used were not effective for use as preservatives in a human eye bank.(AU)

RESUMO Objetivos: As características físico-químicas e o baixo custo da água de coco foram fundamentais para o este estudo. Analisar o uso de solução a base de água de coco como meio de conservação de córneas humanas em banco de olhos. Métodos: Estudo experimental e controlado realizado no Banco de Olhos do Hospital Geral de Fortaleza. Utilizou-se solução à base de água de coco preparada no laboratório de Tecnologia de Sêmen de Caprinos do Departamento de Medicina Veterinária da Universidade Estadual do Ceará. Foram usadas córneas de descartes divididas em dois grupos: G1 (Conservante com água de coco) - grupo experimental e G2 (grupo Conservante com OPTISOL GS®) grupo controle, em experimentos sequenciais. A osmolaridade do G1 foi analisada sequencialmente com 275, 300, 325, 345, 365 e 400 mOsm/L. A viabilidade das córneas foram realizadas por microscopia especular e biomicroscopia nos 1º, 3º e 7º dias. Resultados: As córneas em solução de 365 e 345 mOsm/L apresentavam transparência nos 8mm centrais até o 3º dia, com edema em toda periferia, dobras centrais e edema 2+, com perda parcial do epitélio até 7º dia, sendo o de maior osmolaridade com melhor transparência durante o seguimento. Grupo com 275, 300 e 400 mOsm/L, córnea opaca, edema difuso, perda total do epitélio no 3º dia. As córneas em Optisol mantiveram seus aspectos. Conclusões: O conservante à base de água de coco manteve em parte a transparência corneana e a integridade epitelial, especialmente nos primeiros 3 dias de seguimento. A solução conservante com água de coco nas formulações utilizadas não se mostrou eficaz para o uso em banco de olhos humanos.(AU)

Effect of refrigeration at -1°C on spermatozoa quality of domestic cats / Efeito da refrigeração a -1°C na qualidade de espermatozoides de gatos domésticos

The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P<0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.(AU)

O objetivo deste trabalho foi avaliar a qualidade espermática de gatos domésticos obtidos por eletroejaculação e recuperação da cauda do epidídimo após a refrigeração a -1°C e a 4°C por 24 e 48 horas. Vinte e nove gatos adultos (2 a 6kg) foram utilizados. A colheita de espermatozoides foi realizada por eletroejaculação (EEJ) e, após 48 horas, os gatos foram orquiectomizados, e as amostras espermáticas foram obtidas a partir do ducto deferente e da cauda do epidídimo (EPD). As amostras foram diluídas em ACP-117® e as características espermáticas foram avaliadas em três momentos distintos: fresco, 24 e 48 horas após a refrigeração. Para ser possível comparar as duas temperaturas de refrigeração, a primeira etapa foi analisar se havia diferença entre as técnicas de colheita. Após isto, dois experimentos foram conduzidos: no primeiro, espermatozoides de 14 gatos foram utilizados e a refrigeração foi realizada a -1°C; e no segundo, amostras de 15 gatos foram utilizados e os espermatozoides foram refrigerados a 4°C. A cinética espermática foi avaliada por análise computadorizada (CASA), a concentração por câmara de Neubauer, a morfologia espermática foi avaliada pela coloração de Karras modificada, e a integridade da membrana foi avaliada por eosina nigrosina. Os resultados obtidos foram analisados no software R, versão 3.2.5, utilizando o teste de Mann-Whitney para variáveis com distribuições anormais, considerando significância ao nível de 5%. No ejaculado, maiores valores de defeitos morfológicos totais foram observados após 24 e 48 horas de refrigeração a 4°C (P<0,022) em comparação com refrigeração a -1°C, usando o teste de Friedman. Para quantificar a diminuição na qualidade espermática, as reduções dos parâmetros foram calculadas entre os pontos de tempo (F-24h/F-48h/24h-48h). Na EPD, uma maior redução na qualidade espermática foi detectada após 24 horas de refrigeração a 4°C, tanto na motilidade e na cinética espermática quanto nos índices de movimento e velocidade, em comparação com a refrigeração a -1°C. Com base nos resultados, pode concluir-se que a refrigeração dos espermatozoides felino a -1°C, até 48 horas, foi eficaz na manutenção da qualidade espermático colhidos por EEJ e EPD, e pode ser uma alternativa para a criopreservação de espermatozoides em felinos domésticos.(AU)

Use of the ACP® and BTS extenders for cooling at 15°C white-lipped peccary (Tayassu pecari) semen / Uso dos diluidores ACP® e BTS para refrigeração a 15°C de sêmen de queixada (Tayassu pecari)

Knowledge about reproduction of white-lipped peccary is of great importance to assist with the conservation of this species and enable its rational use in captivity. This study aimed to evaluate the effect of ACP-103®, ACP-116® and BTS semen extenders on sperm viability during cooling of Tayassu pecari semen. Five ejaculates from four adult males were chilled. The animals were submitted to the protocols of sedation and anesthesia for semen collection by the electroejaculation method. After collection, the semen was macro- and microscopically assessed and diluted to reach 35x106 spermatozoa/mL in each of the three different extenders tested. The fresh-extended semen was packed in a BotuFLEX® thermal box to keep samples at 15°C for 24 hours. After cooling, the following semen parameters were analyzed: sperm motility, functional and structural integrity of sperm membranes, mitochondrial activity, chromatin condensation, and the thermoresistance test was performed. The parameters sperm motility, structural and functional integrity of sperm membranes, mitochondrial activity, and chromatin condensation were preserved after use of the extenders tested, and were similar to those of in natura semen (p>0.05). Curvilinear velocity (VCL) (p<0.05) was the only parameter with reduced values after cooling regardless of the extender used. The percentage of sperm with normal morphology was greater in samples cooled using the BTS extender (p<0.05). The ACP-103®, ACP-116® and BTS extenders can be used for the cooling and preservation of white-lipped peccary semen at 15°C for 24 hours.(AU)

Para auxiliar na conservação da espécie e permitir o uso racional do queixada em cativeiro é de grande importância o conhecimento sobre a reprodução da espécie. Objetivou-se avaliar o efeito dos diluidores de sêmen ACP-103®, ACP-116® e BTS na viabilidade espermática durante a refrigeração do sêmen do Tayassu pecari. Foram refrigerados cinco ejaculados provenientes de quatro machos adultos. Os animais foram contidos com auxílio de puçá e submetidos ao protocolo de sedação e anestesia para realização da coleta de sêmen pelo método da eletroejaculação. Depois da coleta, o sêmen foi avaliado macro e microscopicamente e diluído para atingir 35x106 espermatozoides/mL em cada um dos três diferentes diluidores testados. O sêmen diluído foi acondicionado em caixa térmica BotuFLEX® para manter as amostras a 15°C por um período de 24 horas. Depois da refrigeração, os espermatozoides foram avaliados quanto aos parâmetros de movimento espermático, integridade funcional e estrutural das membranas espermáticas, atividade mitocondrial, condensação da cromatina e teste de termorresistência. Os diluidores testados preservaram as características cinéticas, a integridade estrutural e funcional das membranas espermáticas, a atividade mitocondrial e a condensação da cromatina semelhante ao sêmen in natura (P>0,05). O único parâmetro que reduziu com o processo de refrigeração independente do diluidor utilizado foi a Velocidade Curvilinear (VCL) (P<0,05). Foi observado aumento do percentual de espermatozoides morfologicamente normais nas amostras refrigeradas em BTS (P<0,05). Os diluidores ACP-103®, ACP-116® e BTS podem refrigerar e conservar o sêmen de queixada a 15°C por 24 horas.(AU)

Physicochemical coconut water assessment and of the microbiological quality of its sugar extracted of five coconut ecotypes at the marc delorme station, côte d’ivoire

The objective of this study was to produce sugar from coconut water of immature nuts aged 8 to 9 months from West African Tall (WAT), Malayan Yellow Dwarf (MYD), Equatorial Guinea Green Dwarf (EGD) ecotypes and two hybrids PB113+ and PB121+ cultivated at Marc Delorme research station, southern Côte d'Ivoire. 12 coconut trees were randomly selected per cultivar on experimental fields. The coconut water from immature nuts of ranks 19 and 20 have undergone the production of 3 different sugars. Syrup and brown sugar were produced by hot and white sugar by freeze-drying. The volume, the Dry Metter content, pH and Brix degree of coconut water and microbiological characteristics of sugars of the latter were assessed. Pearson’s correlation at 5% level was tested. The results shown that 2 immature nuts from PB121+ and PB113+ as against 3 from EGD and WAT or 4 from MYD were necessary to obtain one liter of coconut water suggesting that nuts from hybrids are bigger. The samples of coconut water extracted from MYD and EGD nuts contained high soluble solids rates varying from 6.32 to 6.98%. Their sugar content was higher. Thus, EGD produced 66.18 g/liter white sugar by freeze-dried, 61.64 g/liter of semi crystalline brown sugar and 64.84 g/liter sugar syrup. Likewise, from immature water of MYD nuts, extracted sugar yields were 62.62 g/liter for white sugar obtained by freeze-dried, 59.95 g/liter for syrup sugar and 55.90 g/liter for semi crystalline brown sugar. Significant positive correlations exists between immature coconut water’s soluble solids content and the sugar yield (r = + 0.93). Sugar obtained offers satisfactory microbiological quality thus, can be integrated in the food practices of the consumers.

Reduction of the DNA damages, Hepatoprotective Effect and Antioxidant Potential of the Coconut Water, ascorbic and Caffeic Acids in Oxidative Stress Mediated by Ethanol

ABSTRACT Hepatic disorders such as steatosis and alcoholic steatohepatitis are common diseases that affect thousands of people around the globe. This study aims to identify the main phenol compounds using a new HPLC-ESI+-MS/MS method, to evaluate some oxidative stress parameters and the hepatoprotective action of green dwarf coconut water, caffeic and ascorbic acids on the liver and serum of rats treated with ethanol. The results showed five polyphenols in the lyophilized coconut water spiked with standards: chlorogenic acid (0.18 µM), caffeic acid (1.1 µM), methyl caffeate (0.03 µM), quercetin (0.08 µM) and ferulic acid (0.02 µM) isomers. In the animals, the activity of the serum γ-glutamyltranspeptidase (γ-GT) was reduced to 1.8 I.U/L in the coconut water group, 3.6 I.U/L in the ascorbic acid group and 2.9 I.U/L in the caffeic acid groups, when compared with the ethanol group (5.1 I.U/L, p<0.05). Still in liver, the DNA analysis demonstrated a decrease of oxidized bases compared to ethanol group of 36.2% and 48.0% for pretreated and post treated coconut water group respectively, 42.5% for the caffeic acid group, and 34.5% for the ascorbic acid group. The ascorbic acid was efficient in inhibiting the thiobarbituric acid reactive substances (TBARS) in the liver by 16.5% in comparison with the ethanol group. These data indicate that the green dwarf coconut water, caffeic and ascorbic acids have antioxidant, hepatoprotective and reduced DNA damage properties, thus decreasing the oxidative stress induced by ethanol metabolism.

Histocytoarchitectural Changes of Wistar Rat’s Kidney Exposed to Young Coconut Water Before and After Induction with Carbon Tetrachloride.

Background: Young coconut water (YCW) has been used by individuals to boast immunity and that of the experimental animals. In this study an attempt was made to investigate the protective effect of YCW against the Carbon tetrachloride (CCl4) induced renal toxicity in rats. Methods: A total of 20 male adult wistar rats which were not previously subjected to any experiment were divided into four groups. Each group has five rats. Group 1 (Normal control) received basal diet, olive oil and water. Group 2 (Positive control) received basal diet, olive oil and YCW (100ml/kg). Group 3 (Negative control) received basal diet, water and CCl4 diluted with water and olive oil. Group 4 (Experimental group) received basal diet and YCW (100ml/kg) and then intoxicated with CCl4 diluted with water and olive oil. After day 7, the rats were sacrificed and their kidneys were collected and processed histologically following standard protocols. Results: Group 1 (Normal control) displayed normal histocyto-archetature of the kidney. Group 2 (Positive control) revealed hyperplasia with mild inflammatory response. Group 3 (Negative control) showed hypercellularity, mild cystic spaces, necrosis, and loose glomerular membrane indicative of high inflammatory response. Group 4 (Experimental group) revealed moderate cellular activities in line with moderate inflammatory response. Conclusion: The administration of YCW on the rat before intoxication with CCl4 suppresses the deleterious effect of CCl4 on the experimental group.

The Effects of Different Media, Sucrose Concentrations and Natural Additives on Plantlet Growth of Phalaenopsis Hybrid 'Pink'

ABSTRACT The effects of various sucrose concentrations as carbon source and natural additives in different media on plantlet growth of Phalaenopsis hybrid 'Pink' were studied. Plantlets were cultured on two media (Murashige and Skoog [MS] and Vacin and Went [VW]) supplemented with 0, 10, 20, 30 and 40 g L-1 sucrose either with 0, 10 and 20% (v/v) coconut water (CW) or carrot juice (CJ) as natural additives. After four months of culture, the combination of sucrose and CW supplemented with both media affected plantlet growth where most of the plantlets showed slow growth and survival frequency (0-80%) with increasing concentrations of CW in all sucrose concentrations. However, plantlet growth on both media containing only 20 g L-1 sucrose without CW was optimal in terms of root number, root length, leaf number, leaf length, leaf width, fresh weight, dry weight and plant height. The combination of sucrose and CJ supplemented with MS medium resulted in overall good plantlet growth with 100% survival frequency. The combination of sucrose (20 g L-1) and CJ (10%) supplemented with MS medium increased root length, leaf length, leaf width and plant height. Plantlet growth was also optimal in the combination of 20 g L-1 sucrose and 10% CJ supplemented with VW medium. The results of this study indicate that Phalaenopsis hybrid 'Pink' cultured on the combination of sucrose (20 g L-1) and CJ (10%) supplemented with either MS or VW media can be used for plantlet growth of this species.

Fibroblast Viability after Storage at 20 °C in Milk, Hank's Balanced Salt Solution and Coconut Water

Abstract The objective of this study was to evaluate the effectiveness of various storage media at 20 °C in maintaining the viability of human periodontal ligament fibroblasts (HPLF) over time. HPLF were maintained at 20 °C in skim milk (SM), whole milk (WM), freshly prepared Hank's balanced salt solution (HBSS), Save-A-Tooth(r), natural coconut water (NCW), coconut water industrialized (ICW) and tap water (negative control) for 3, 6, 24, 48, 72, 96 and 120 h. Cells maintained in Minimal Essential Medium (MEM-37) at 37 °C served as a positive control. Cell viability was determined by MTT assay. Statistical analysis was performed by Kruskal-Wallis test and Scheffe test (α = 5%). From 24 h, NCW was significantly better in maintaining cell viability than all other tested storage media (p<0.05). SM and WM were significantly better than HBSS for up to 72 h. Save-A-Tooth(r) and ICW were the worst conservation storage media. In conclusion, the effectiveness of the tested storage media to maintain the viability of the periodontal ligament cells was as follows, in a descending order: NCW > MEM-37> SM and IM> HBSS> ICW > Save-A-Tooth(r)> tap water.

Resumo O objetivo deste estudo foi avaliar a efetividade de vários meios de conservação a 20 °C em manter a viabilidade de fibroblastos do ligamento periodontal humano (FLPH) ao longo do tempo. FLPH foram conservados a 20 °C em leite desnatado (LD), leite integral (LI), solução salina balanceada de Hank (HBSS) recém preparada, Save-A-Tooth(r) (Save), água de coco natural (ACN), água de coco industrializada (ACI) e água de torneira (água - controle negativo) por 3, 6, 24, 48, 72, 96 e 120 h. Células conservadas em Meio Essencial Mínimo (MEM-37) a 37 °C serviram como controle-positivo. A viabilidade celular foi determinada pelo ensaio MTT. A análise estatística dos dados foi realizada pelos testes Kruskal-Wallis e Scheffé (α=5%). A partir de 24 h, ACN foi significantemente melhor em manter a viabilidade celular do que todos os outros meios testados (p<0,05). LD e LI foram significantemente melhores do que a HBSS por até 72 h. Save e ACI foram os piores meios de conservação. Concluindo, a efetividade dos meios de conservação testados em manter a viabilidade das células do ligamento periodontal foi a seguinte em ordem decrescente: ACN > MEM-37 > LD e LI > HBSS > ACI > Save > água.

Recovery of sperm after epididymal refrigeration from domestic cats using ACP-117c and Tris extenders / Recuperação de espermatozoides após refrigeração do epidídimo de gatos domésticos utilizando os diluidores ACP-117c e Tris

We aimed to compare fresh sperm and sperm cooled to 4ºC that had been recovered from the epididymides of cats using powdered coconut water (ACP-117c) and Tris extenders. Sixty epididymides were divided into 6 groups: 10 fresh epididymides were recovered using Tris (T0h); 10 were kept at 4°C/2h and recovered using Tris (T2h); 10 were kept at 4°C/4h and recovered using Tris (T4h); 10 fresh were recovered using ACP-117c (A0h); 10 were kept at 4°C/2h and recovered using ACP-117c (A2h), and 10 were kept at 4°C/4h and recovered using ACP-117c (A4h). The testis-epididymis complexes (TEC) control were not cooled. The others were cooled at 4°C for 2 or 4h. The epididymis was separated and the sperm was recovered by the modified flotation method. Sperm kinetic parameters were evaluated by a computer-system analysis, and vigor, viability, concentration, membrane function and morphology of the sperm were assessed under a light microscope. The progressive motility with ACP-117c declined after 2h of cooling, but did not differ between fresh and 4h. The vigor and membrane function were higher in A4h than A0h. The vigor at T2h and T4h were decreased compared to T0h. T0h was higher than A0h for vigor and sperm membrane function. However, after 4h of cooling, ACP-117c maintained a higher percentage of living cells. Feline epididymal sperm quality can be maintained to the degree necessary for artificial breeding programs following cooling and ACP-117c may be successfully used to recover cat sperm that have been cooled for up to 4h.(AU)

Objetivou-se comparar a qualidade de espermatozoides recuperados a fresco e após refrigeração a 4ºC do epidídimo de gatos domésticos utilizando-se os diluidores ACP-117c e Tris. Sessenta epidídimos foram distribuídos em seis grupos: 10 epidídimos a fresco com o Tris (T0h), 10 a 4°C/2h e recuperados com Tris (T2h), 10 a 4°C/4h e recuperados com Tris (T4h), 10 epidídimos a fresco com o ACP-117c (A0h), 10 a 4 °C/2h e recuperados com ACP-117c (A2h), 10 a 4°C/4h e recuperados com ACP-117c (A4h). Os complexos testículo-epidídimo (CTE) do controle não foram refrigerados. Os outros foram refrigerados a 4°C durante duas e quatro horas. Os epidídimos foram separados das demais estruturas, e os espermatozoides recuperados pela técnica de flutuação modificada. Os parâmetros cinéticos foram avaliados em um sistema computadorizado, e o vigor, a viabilidade, a concentração, a funcionalidade de membrana e a morfologia celular foram avaliados em microscopia de luz. A motilidade progressiva com ACP-117c declinou após duas horas de refrigeração, mas não diferiu entre a recuperação a fresco e após refrigeração por quatro horas. Vigor e integridade funcional da membrana celular foram significativamente superiores no grupo A4h em comparação ao A0h. O vigor espermático em T2h e T4h reduziu significativamente em comparação com T0h. T0h foi significativamente superior ao A0h quanto aos parâmetros de vigor e integridade funcional da membrana espermática, entretanto, após quatro horas de refrigeração, o ACP-117c apresentou um maior percentual de células vivas. Os espermatozoides epididimários de felinos domésticos conseguem manter a qualidade necessária para serem utilizados em programas de reprodução artificial após serem refrigerados e recuperados por meio da técnica de flutuação modificada, e o diluidor ACP-117c pode ser utilizado com sucesso para recuperação de células espermáticas refrigeradas de gatos por até quatro horas.(AU)

Evaluation of coconut water neutralized by different agents on the viability of human fibroblasts: an in vitro study / Avaliação da água de coco neutralizada por diferentes agentes na viabilidade de fibroblastos humanos: estudo in vitro

Objective: This study evaluated four types of pH adjustment of the coconut water (CW) on viability of human fibroblasts (HFF). Material and method: Natural and industrialized CW were adjusted to pH 7.0 using: (1) Sodium Hidroxide (NaOH), (2) Sodium bicarbonate (NaHCO3), (3) Triethanolamine (C6H15NO3), (4) 2-Amino-2-Methil-1-Propanol (C4H11NO). Fibroblasts were plated at 2×104/ well in 96 well plates and maintained in the CW solutions for 2 h and 4 h. Positive control was represented by HFF maintained in DMEM and the negative control by tap water. Cell viability was analyzed by MTT formazan method. Data were analyzed by 3-way ANOVA followed by Tukey's and Dunnet's test. Result: There are no significant effect on the cell viability regarding type of CW, period of evaluation, and the interactions between CW and period of evaluation, CW and pH adjustment method, pH adjustment method and period of evaluation (p>0.05). Conclusion The product used for CW pH adjustment did not influenced HFF viability, thought there are a tendency of better performance in natural CW.

Objetivo: Avaliar a eficácia de quatro tipos de substâncias usadas para ajuste do pH da água de coco (AC) sobre a viabilidade de fibroblastos humanos (HFF). Material e método: O pH da AC natural e industrializada foi ajustado para pH 7,0 utilizando: (1) Hidróxido de Sódio (NaOH), (2) bicarbonato de sódio (NaHCO3), (3) Trietanolamina (C6H15NO3), (4) 2-Amino-2- Methil-1-propanol (C4H11NO). Células HFF foram plaqueadas em 2×104 células/poço em placas de 96 poços e mantidas nas diferentes soluções de AC acima durante 2 h e 4 h. Controle positivo foi representado por HFF mantidas em DMEM e o controle negativo por água da torneira. A viabilidade celular foi avaliada pelo método de MTT Formazan. Os dados foram analisados por 3-way ANOVA seguido pelo teste de Tukey e Dunnett. Resultado: A viabilidade celular não é influenciada pelo período de avaliação, e as interações entre AC e período de avaliação, AC e método de ajuste de pH, método de ajuste de pH e período de avaliação (p> 0,05). Conclusão: O produto utilizado para ajuste do pH não interfere na viabilidade de FH, embora, haja uma tendência de melhor desempenho em AC natural.

Aloe vera na criopreservação do sêmen de tambaqui (Colossoma macropomum) / Aloe vera in the cryopreservation of tambaqui (Colossoma macropomum) sperm

This study aimed to evaluate the extract of Aloe vera (AV) associated or not with 10% Dimethylsulfoxide (DMSO) in cryopreservation of tambaqui semen. For the formation of the pools (n= 14), 30 males were hormonally induced twice. Each pool had the objective motility, curvilinear velocity, straight-line velocity, average path velocity and morphology analyzed before and after cryopreservation of semen. The means for cryopreservation were constituted of Powder Coconut Water-104 diluent added DMSO and/or AV (5 or 10%). After cryopreservation, motility, velocities and morphology were reduced significantly when compared to fresh semen. For sperm motility the best treatment was that using only DMSO (20,86±8,31) and DMSO + 5% AV (15.71±9.77). For the velocities, the worse treatment was DMSO+10% AV. Treatment with only the addition of DMSO had a significantly higher effect than others on percentage of morphologically normal sperm. The mean correlation found was between motilityand the rate of morphologically normal sperm (r = 0.687). In conclusion, the addition of AV does not provide greater protection for spermatozoa during cryopreservation.

Short-term preservation of Pecari tajacu ovarian preantral follicles using phosphate buffered saline (PBS) or powdered coconut water (ACP(r)) media / Preservação de folículos ovarianos pré-antrais de Pecari tajacu por curtos períodos utilizando meios à base de solução salina fosfatada tamponada (PBS) ou água de coco em pó (ACP(r))

We compare protocols for the short-term preservation of collared peccarie's ovarian preantral follicles (PFs) by using phosphate buffered saline- (PBS) or powdered coconut water- (ACP(r)) based medium. For morphology analysis each pair of ovaries collected from six females was divided into nine fragments. One fragment was destined for morphology analysis (histology and transmission electron microscopy - TEM), constituting the control group and the other fragments were placed in tubes with PBS or ACP(r), packed in 5 L Styrofoam boxes, stored for 4h, 12h, 24h, and 36h, and then analyzed. For viability analysis a pair of ovaries from two additional females was divided into nine fragments; one fragment was immediately destined for viability analysis (Trypan blue test) and the other fragments were stored as previously described, until 24h and then analyzed. After 4h storage in ACP(r) medium, the follicular integrity was similar to control (87.8% vs 94.4%, respectively); however, ultrastructural analyses revealed swollen mitochondria as the first signals of PF degeneration. It was observed that ACP(r) (66.7%) was more efficient than PBS (49.4%) to preserve the morphological integrity after 36h storage (P<0.05); however, no differences were observed on follicular viability (P>0.05). In conclusion, the use of the ACP(r) is recommended for the short-term preservation of Pecari tajacu preantral follicles...

Compararam-se protocolos para a preservação por curtos períodos de folículos ovarianos pré-antrais (PFs) de catetos, utilizando meios à base de solução salina tamponada (PBS) ou água de coco em pó (ACP(r)). Para a análise morfológica, cada par de ovários coletados de seis fêmeas foi dividido em nove fragmentos. Um fragmento foi destinado para a análise da morfologia (histologia e microscopia eletrônica de transmissão - MET), constituindo o grupo controle, e os demais fragmentos foram colocados em tubos contendo PBS ou ACP(r), acondicionados em caixas térmicas de poliestireno expandido de 5L, armazenados durante quatro, 12, 24 e 36 horas, e, então, analisados. Para a análise da viabilidade, pares de ovários de duas fêmeas adicionais foram divididos em nove fragmentos; um deles foi imediatamente destinado à análise da viabilidade (teste com azul de Trypan), os outros fragmentos foram armazenados como descrito previamente até 24h e, então, foram analisados. Após quatro horas de armazenamento em meio ACP(r), a integridade folicular foi similar ao grupo controle (87,8% vs. 94,4%, respectivamente); contudo, a análise ultraestrutural revelou mitocôndrias edemaciadas como os primeiros sinais de degeneração dos PFs. Foi observado que o ACP(r) (66,7%) foi mais eficiente do que o PBS (49.4%) em preservar a integridade morfológica após 36h (p<0,05); entretanto, nenhuma diferença foi observada para a viabilidade folicular (P>0,05). Em conclusão, o uso da ACP(r) é recomendado para a preservação por curtos períodos de folículos pré-antrais de Pecari tajacu...

EFECTO INDUCTOR DEL AGUA DE COCO SOBRE LA GERMINACIÓN DE SEMILLAS Y BROTAMIENTO DE LOS CORMOS DE LA HIERBA DE LA EQUIS Dracontium grayumianum / Inductive Effect of Coconut Water on Germination of Seeds and Sprouting of Corms of Dracontium grayumianum

El objetivo de este estudio fue determinar el efecto promotor de agua de coco, ácido giberélico, de la estratificación fría y escarificación mecánica, sobre la germinación de semillas de Dracontium grayumianum, y el efecto de ácido giberélico y agua de coco sobre el brotamiento de cormos de la misma especie. Las semillas no sometidas a los tratamientos inductores fueron incapaces de germinar, pero la inmersión en agua de coco tuvo efectos notables, produciendo un porcentaje de germinación del 50%, superior al efecto logrado con los demás tratamientos. El endospermo líquido del coco también tuvo efecto favorable sobre el brotamiento de cormos bajo condiciones de vivero, al igual que el tratamiento con una solución de ácido giberélico. Este es el primer reporte del uso de agua de coco como agente promotor de la germinación de semillas con alto nivel de latencia, lo que coloca este recurso como una alternativa adicional, altamente eficaz y de bajo costo, para ser utilizado en estrategias de propagación vegetal de especies con semillas de latencia profunda.

The objective of this study was to determine the promotional effect of coconut water, gibberellic acid, cold stratification and mechanical scarification on seed germination of Dracontium grayumianum, and the effect of gibberellic acid and coconut water on the sprouting of corms of the same species. The seeds without the inductive treatment were unable to germinate, but the immersion in coconut water had significant effects, producing a germination rate of 50%, higher than the effect achieved with other treatments. The liquid endosperm of coconut also had favorable effect on the sprouting of corms under nursery conditions, like the treatment with gibberellic acid solution. This is the first report of the use of coconut water as a promoter of seed germination with high latency, which places this resource as an additional alternative, highly efficient, and cost-effective, for use in plant propagation strategies of species with seeds of deep dormancy.

Conservação do sêmen e liquefação do coágulo seminal de macaco-prego (Cebus apella) em água de coco em pó (ACP-118®), em diferentes temperaturas / Semen conservation and seminal clot liquefaction of capuchin monkey (Cebus apella) in powdered coconut water extender (PCW) at different temperatures

O objetivo do estudo foi avaliar a água de coco em pó (ACP) na conservação do sêmen e liquefação do coágulo seminal de Cebus apella. O sêmen de seis machos adultos foi coletado por eletroejaculação (EEJ), diluído em solução à base de ACP-118® e submetido à incubação em banho-maria a 33, 35 e 37°C, por 24 horas. Avaliou-se a integridade espermática por meio da coloração eosina-nigrosina a cada uma hora durante as seis horas iniciais e após 24 horas de incubação. Os volumes médios e as concentrações espermáticas das frações coagulada e líquida foram de 0,20±0,02 e 0,20±0,10mL; 1,1±0,3x10(8) e 1,3±0,9x10(7) espermatozoides mL-1, respectivamente. Somente em uma amostra da fração líquida foram verificados espermatozoides com motilidade (20 por cento) e vigor (4), perdurando por 40 minutos. A maior parte do coágulo liquefez em ACP-118® após 12 horas de incubação. O melhor tratamento observado foi sob 33°C, por manter até 47±12,8 por cento de espermatozoides vivos após 24 horas. Conclui-se que o diluente à base de ACP é eficiente na liquefação do coágulo seminal e na manutenção da integridade espermática até 24 horas após a EEJ, nas temperaturas de 33, 35 e 37°C.

The aim of this study was to evaluate the powdered coconut water (PCW) in the semen conservation and seminal clot liquefaction. The semen of six adult male Cebus apella was collected by electroejaculation (EEJ), diluted in ACP-118® extender and stayed in water bath at 33, 35 and 37°C for 24 hours. The sperm integrity was evaluated by eosin-nigrosine staining every one hour during the six initial hours and after 24 hours of incubation. The average volumes and sperm concentrations of clotted and liquid fractions were 0.20±0.02 and 0.20±0.10mL, 1.1±0.3x108 and 1.3±0.9x107 sperm mL-1, respectively. Immediately after collection, only in a sample of liquid fraction was observed 20 percent motility and vigor 4, which stopped after 40 minutes. Most of the clot was liquefied in ACP-118® after 12 hours of incubation. The best observed treatment was 33°C, because it kept 47±12.8 percent of sperm integrity after 24 hours. It was concluded that the PCW extender is effective in the liquefaction of seminal clot and maintenance of sperm viability 24 hours after the EEJ at 33, 35 and 37°C.

Constituents and antioxidant activity of two varieties of coconut water (Cocos nucifera L) / Constituintes químicos e atividade antioxidante de duas variedades de água de coco (Cocos nucifera L)

Uma análise dos componentes da água-de-coco (Cocos nucifera L.) de duas variedades da fruta (verde e amarelo) por hidrodestilação e extração com solvente, mostrou a presença de álcoois, cetonas, tióis, ácidos carboxílicos, fenóis, e ésteres. Significativa atividade antioxidante foi observada, usando o método DPPH, para as amostras obtidas por hidrodestilação e extração de éter de petróleo para ambas as variedades do coco.

An analysis of the constituents of coconut (Cocos nucifera L.) water from two fruit varieties (green and yellow) by hydrodistillation and solvent extraction showed the presence of alcohols, ketones, thiols, carboxylic acids, phenols, and esters. Substantial antioxidant activity was observed, using the DPPH assay, for the samples obtained by hydrodistillation and petroleum ether extraction of both coconut varieties.

Coconut water medium in the laboratory diagnosis of cholera

Coconut water, which is readily available for the preparation of low cost culture media specially by peripheral laboratories of this country, have been found to be a satisfactory basic nutritional requirement for the culture of V. cholera. Colonies formed on Coconut Water Agar were characteristic and distinct from the colonies of commonsals. In a comparative study with MEA and TCBS, no significant difference were observed in the recovery percentage of V. cholerae from rectal swabs pre-enriched in Coconut Water Broth. Preliminary enrichment of the fecal samples in coconut Water Broth for six hours at 37 C and inoculating the broth inoculum on Coconut Water Agar seemed to be the cultural procedure of choice in its use in the laboratory diagnosis of cholerae.(Summary)