Canine sperm vitrification with nonpermeable cryoprotectants and coconut water extender

This study was aimed to assess the efficiency of coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants for canine semen vitrification, using a simple method that yields a high survival rate of spermatozoa for clinical use. Twelve ejaculates from 12 adult normozoospermic dogs were collected separately by digital manipulation and only the second semen fraction was used in this study. After evaluation of volume, concentration, viability, total and progressive motility, velocity parameters and morphology, semen was diluted with a coconut water extender (50% (v/v(volume per volume)) coconut water, 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% and 0.25M sucrose until final concentration of 100x106 spermatozoa/ml. After equilibration at 5ºC for 60 minutes, semen was vitrified by "direct dropping method" into liquid nitrogen in spheres with a volume of 30 µl. After a week of storage the spheres were devitrified as three of them were dropped into 0.5 mL of CaniPlus AI medium (Minitüb, Germany), which was previously warmed in a water bath at 42ºC for 2 minutes and evaluated about the above mentioned parameters. It was found that vitrification resulted in a lower percentage of viable sperms, normal morphology, total and progressive motilities (p0.05) compared to fresh semen samples. In conclusion, our results demonstrate that vitrification with coconut water extender with addition of 1% soy lecithin and 0.25M sucrose as cryoprotectants, has an excellent potential for routine canine sperm cryopreservation.(AU)

Refrigeração de sêmen canino em meio diluente à base de água de coco (Cocos nucifera) natural / Canine semen refrigeration in a natural coconut water (Cocos nucifera)-based extender / Refrigeración de semen canino en un medio diluyente natural a base de agua de coco (Cocos nucifera)

A inseminação artificial em cadelas contribui para o melhoramento genético da espécie, previne algumas doenças sexualmente transmissíveis a partir da cópula e possibilita a reprodução de animais que não poderiam copular de forma natural, seja por motivos anatômicos, geográficos ou comportamentais. Todavia, nem sempre é possível utilizar o sêmen fresco, sendo assim necessário um diluente para resfriar e mantê-lo viável por determinado período. Diante disso, o objetivo com este trabalho foi analisar o sêmen canino diluído em água de coco e refrigerado, à 5 ºC em diferentes tempos. Foram utilizados cinco cães da raça Hounds do Brasil, realizando três colheitas de sêmen de cada animal, com intervalos de sete dias. Os ejaculados foram mantidos a temperatura de 37ºC e realizado análises macroscópicas (volume, cor, aspecto e odor) e microscópicas (motilidade, vigor, concentração e morfologia espermática). Em seguida, os ejaculados foram diluídos em água de coco natural a uma concentração de 200 milhões de espermatozoides/mL, e mantidos à temperatura de 5 °C, por até 72 horas. Nos intervalos de seis, doze, vinte e quatro, trinta e seis, quarenta e oito, e setenta e duas horas, as amostras foram avaliadas quanto a motilidade e vigor espermático. Os ejaculados frescos apresentaram em média volume de 6,2 mL, cor branca, aspecto aquoso a leitoso, odor "sui generis", motilidade espermática de 89,5 %, vigor espermático 4,3, concentração média de 418 x106espermatozoides/mL e 7,3% de alterações patológicas. Após o início do resfriamento à 5 ºC, os valores de motilidade e vigor diminuíram com o passar do tempo, sendo os menores valores encontrados após 48 e 72 horas. O diluente a água de coco in natura mostrou-se eficiente para refrigeração de sêmen canino, à 5 ºC, conservando-o por um período de até 36h após a colheita, conforme preconizado pelo Colégio Brasileiro de Reprodução Animal.(AU)

improvement of the species, prevents some sexually transmitted diseases through copulation and allows the reproduction of animals that could not copulate naturally, either for anatomical, geographic or behavioral reasons. However, it is not always possible to use fresh semen, thus requiring a diluent to cool and keep it viable for a certain period. Therefore, the objective of this work was to analyze canine semen diluted in coconut water and refrigerated at 5 ºC at different times. Five Brazilian Hounds were used, performing three semen collections from each animal, with intervals of seven days. The ejaculates were kept at a temperature of 37 ºC and macroscopic (volume, color, appearance and odor) and microscopic (motility, vigor, concentration and sperm morphology) analyzes were performed. Then, the ejaculates were diluted in natural coconut water at a concentration of 200 million sperm/mL, and kept at a temperature of 5 °C for up to 72 hours. At intervals of six, twelve, twenty-four, thirty-six, forty-eight, and seventy-two hours, samples were evaluated for sperm motility and vigor. Fresh ejaculates had an average volume of 6.2 mL, white color, watery to milky appearance, "sui generis" odor, 89.5% sperm motility, 4.3 sperm vigor, average concentration of 418 x106 spermatozoa/mL and 7.3%pathological changes. After the beginning of cooling at 5 °C, the values of motility and vigor decreased over time, with the lowest values found after 48 and 72 hours. The in natura coconut water extender proved to be efficient for cooling canine semen at5 ºC, keeping it for a period of up to 36 hours after harvest, as recommended by the Brazilian College of Animal Reproduction.(AU)

Artificial insemination in bitches contributes to the genetic La inseminación artificial en perras contribuye a la mejora genética de la especie, previene algunas enfermedades de transmisión sexual a través de la cópula y permite la reproducción de animales que no podrían copular de forma natural, ya sea por razones anatómicas, geográficas o de comportamiento. Sin embargo, no siempre es posible utilizar semen fresco, por lo que se requiere un diluyente para enfriarlo y mantenerlo viable durante un cierto período. Por tanto, el objetivo de este trabajo fue analizar semen canino diluido en agua de coco y refrigerado a 5 ºC en diferentes tiempos. Se utilizaron cinco sabuesos brasileños, realizándose tres colectas de semen de cada animal, con intervalos de siete días. Los eyaculados se mantuvieron a una temperatura de 37 ºC y se realizaron análisis macroscópicos (volumen, color, apariencia y olor) y microscópicos (motilidad, vigor, concentración y morfología espermática). Luego, los eyaculados se diluyeron en agua de coco natural a una concentración de 200 millones de espermatozoides/mL y se mantuvieron a una temperatura de 5 °C hasta por 72 horas. A intervalos de seis, doce, veinticuatro, treinta y seis, cuarenta y ocho y setenta y dos horas, se evaluó la motilidad y el vigor de los espermatozoides en las muestras. Los eyaculados frescos tuvieron un volumen promedio de 6,2 mL, color blanco, apariencia acuosa a lechosa, olor "sui generis", motilidad espermática de 89,5%, vigor espermático de 4,3, concentración promedio de 418 x106 espermatozoides/mL y cambios patológicos de 7,3%. Después del inicio del enfriamiento a 5 °C, los valores de motilidad y vigor disminuyeron con el tiempo, encontrándose los valores más bajos a las 48 y 72 horas. El diluyente de agua de coco in natura demostró ser eficaz para enfriar el semen canino a5 ºC, manteniéndolo por un período de hasta 36 horas después de la cosecha, según lo recomendado por el Colegio Brasileño de Reproducción Animal.(AU)

Diferentes concentrações antibióticas adicionadas ao diluente de sêmen suíno / Different antibiotic concentrations added to the swine semen extender

Devido a contaminação bacteriana, utiliza-se antibióticos nos diluentes de sêmen para inibir o crescimento bacteriano durante a sua estocagem. Assim, este trabalho teve por objetivo determinar o limite de toxicidade de diferentes concentrações antibióticas da penicilina/estreptomicina adicionadas ao meio diluente do sêmen e investigar a influência sobre a motilidade espermática. Para isso, o sêmen de cinco reprodutores híbridos comerciais foi coletado uma vez por semana, durante nove semanas, através da técnica da mão enluvada. Cada ejaculado foi distribuído de forma igual entre todos os tratamentos, que consistiram em diferentes concentrações de solução de antibióticos, sendo: T1 = 0,0g/L (controle); T2 = 4,2g/L; e T3 = 12,6g/L. O sêmen diluído em água de coco em pó (ACP-103@) foi conservado por cinco dias e analisado nos dias: D0 (dia da coleta), D2 e D4 (último dia de conservação) quanto ao vigor e à motilidade espermática. Em D0, o sêmen conservado em diluente acrescido de 12,6g de solução antibiótica apresentou menor valor de vigor espermático em relação aos demais tratamentos (p<0,05). Já em D2 e D4, os tratamentos contendo antibióticos apresentaram resultados de vigor aquém dos obtidos no grupo controle (p<0,05). Em relação à motilidade espermática, os maiores percentuais de células móveis foram observados nas amostras de sêmen conservadas em ACP 0,0g/L (controle) quando comparados aos demais tratamentos (p<0,05). Conclui-se que as concentrações antibióticas utilizadas no estudo (4,2 e 12,6 g/L) mostraram efeitos tóxicos logo após a diluição do sêmen suíno, afetando de forma negativa parâmetros seminais durante a conservação a 17 °C.

Due to bacterial contamination, antibiotics are used in semen extenders to inhibit bacterial growth during storage. Thus, the present study aimed to determine the toxicity limit of different antibiotic concentrations of penicillin/streptomycin added to the semen extender and investigate the influence on sperm motility. For this purpose, semen from five commercial hybrid breeders was collected once a week for 9 weeks, using the gloved hand technique. Each ejaculate was distributed equally among all treatments, which consisted of different concentrations of antibiotic solution: T1 = 0.0g/L (control); T2 = 4.2g/L; T3 = 12.6g/L. The semen diluted in powdered coconut water (ACP-103@) was kept for 5 days and analyzed on the following days: D0 (collection day), D2, and D4 (last conservation day) for sperm vigor and motility. In D0, the semen preserved in the diluent added with 12.6 g of antibiotic solution showed a lower sperm vigor value compared to the other treatments (p<0.05). In D2 and D4, treatments containing antibiotics presented vigor results below those obtained in the control group (p<0.05). Regarding sperm motility, higher percentages of mobile cells were observed in semen samples conserved in ACP 0.0 g/L (control) when compared to the other treatments (p<0.05). It is concluded that the antibiotic concentrations used in the study (4.2 and 12.6 g/L) showed toxic effects soon after the swine semen, negatively affecting seminal parameters during storage at 17 °C.

Sustained deficit irrigation on yield and fruit water quality of dwarf green coconut / Efeito da irrigação com déficit hídrico sustentado sobre a produtividade e qualidade da água dos frutos do coqueiro anão verde

This study evaluated the effects of sustained deficit irrigation (SDI) on fruit yield and fruit water quality of dwarf green coconut trees. The experiment was carried out in a commercial orchard located in Camocim, Ceará, Brazil. Four years old coconut trees were irrigated during 29 months, using micro-sprinklers, at irrigation depths equivalent to 55%, 77%, 100% and 131% of crop evapotranspiration (ETc). Green coconut fruits were harvested six months after flower aperture and evaluated for number of fruits per plant, volume of coconut water per fruit and total soluble solids of the coconut water. SDI reduced coconut fruit yield, fruit water volume and coconut water yield. Conversely, SDI increased total soluble solids of the coconut water and irrigation water productivity in terms of fruits and coconut water. Deficit irrigation showed no economic advantage over full irrigation due to the small reduction in irrigation costs compared to the substantial reduction in gross revenue.

Este estudo teve como objetivo avaliar os efeitos da irrigação com déficit sustentado (SDI) sobre a produção e a qualidade da água de frutos do coqueiro Anão Verde. O experimento foi realizado em um pomar comercial localizado em Camocim, Ceará, Brasil. Plantas com quatro anos de idade foram irrigadas por microaspersão, durante 29 meses, com lâminas de irrigação equivalentes a 55%, 77%, 100% e 131% da evapotranspiração da cultura (ETc). Os frutos foram colhidos seis meses após a abertura das flores e avaliados quanto ao número de frutos por planta, volume de água de coco por fruto e sólidos solúveis totais da água de coco. A SDI reduziu a produtividade de frutos, o volume de água do fruto e a produção de água de coco por planta. Por outro lado, a SDI aumentou os sólidos solúveis totais da água de coco e a produtividade da água de irrigação em termos de frutas e água de coco. A SDI não apresentou vantagem econômica sobre a irrigação plena devido à pequena redução nos custos de irrigação em comparação à redução substancial da receita bruta.

Growth of black pepper plantlets under different substrates and irrigation levels

ABSTRACT Optimization of the use of water and the possibility of using residues as substrate in the black pepper production chain can yield economic and environmental benefits. The objective of this study was to evaluate the biometric and physiological traits of black pepper plantlets (Piper nigrum L.), cv. Bragantina, at different irrigation levels and soilless substrates. Plantlets were grown for 120 days on three substrates [biosolid + crushed coconut fiber (3:1, by volume); biosolid + granitic rock powder (3:1, by volume); and biosolid + crushed coconut fiber + granitic rock powder (2:1:1, by volume)] and at five irrigation levels [36 % (very low); 55 % (low); 85 % (moderate); 100 % (well-irrigated); and 126 % of the reference irrigation level (over-irrigated)]. The biometric traits evaluated increased as the irrigation levels increased, with the exception of root dry mass, which was not influenced by this factor, nor water use efficiency, which decreased as the irrigation levels increased. In general, plants grown with a mixture of the three wastes (biosolid + crushed coconut fiber + granitic rock powder) exhibited higher values for biometric variables and chlorophyll content when they were moderately, well or over-irrigated, in comparison with the other two substrates. It was possible to achieve the plantlet commercial standard using this three waste mixture and the moderate irrigation level.

Assessment of coconut water added to Numida meleagris egg yolk as cryoprotectant for goat semen

Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl (Numida meleagris) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen (Numida meleagris) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality.(AU)

Method for the physical characterization of substrates using centrifugation

The characterization of substrate properties supports the correct choice of components and directs management decisions in container cultivation. There are several systems of analysis, however, the routine procedures to determine the water retention curve (WRC) are time consuming and inaccurate. This study proposed a reproducible, less time-consuming, and easy to perform method for physical characterization of substrates. We used Sphagnum peat, coconut powder, carbonized rice husk, expanded vermiculite, and expanded clay in isolation. The WRC was obtained at tensions 0, 10, 50, and 100 hPa using the method of Büchner funnel, the European standard (CEN-EN13041), and by the centrifugation method of samples. In centrifugation, tensions were simulated by combining the angular velocity, sample height, and radius measured from the sample outermost point to the central point of the centrifuge. Three rotations were applied (230; 520 and 730 rpm), testing the centrifugation time for each tension and then the volumetric water content was compared between the methods. The results showed a positive correlation (p < 0.001) between the centrifugation and standard methods, considering that the centrifugation time required for moisture stabilization varied according to the material analyzed and the tension applied. However, the time required to obtain a report was reduced by half. Sample centrifugation is a promising method, ensuring that the tensile force actually acts on the entire sample to determine the water retention curve of the substrate also reducing the duration of the analyses.

Growth of black pepper plantlets under different substrates and irrigation levels

Optimization of the use of water and the possibility of using residues as substrate in the black pepper production chain can yield economic and environmental benefits. The objective of this study was to evaluate the biometric and physiological traits of black pepper plantlets (Piper nigrum L.), cv. Bragantina, at different irrigation levels and soilless substrates. Plantlets were grown for 120 days on three substrates [biosolid + crushed coconut fiber (3:1, by volume); biosolid + granitic rock powder (3:1, by volume); and biosolid + crushed coconut fiber + granitic rock powder (2:1:1, by volume)] and at five irrigation levels [36 % (very low); 55 % (low); 85 % (moderate); 100 % (well-irrigated); and 126 % of the reference irrigation level (over-irrigated)]. The biometric traits evaluated increased as the irrigation levels increased, with the exception of root dry mass, which was not influenced by this factor, nor water use efficiency, which decreased as the irrigation levels increased. In general, plants grown with a mixture of the three wastes (biosolid + crushed coconut fiber + granitic rock powder) exhibited higher values for biometric variables and chlorophyll content when they were moderately, well or over-irrigated, in comparison with the other two substrates. It was possible to achieve the plantlet commercial standard using this three waste mixture and the moderate irrigation level.

Growth of black pepper plantlets under different substrates and irrigation levels

Optimization of the use of water and the possibility of using residues as substrate in the black pepper production chain can yield economic and environmental benefits. The objective of this study was to evaluate the biometric and physiological traits of black pepper plantlets (Piper nigrum L.), cv. Bragantina, at different irrigation levels and soilless substrates. Plantlets were grown for 120 days on three substrates [biosolid + crushed coconut fiber (3:1, by volume); biosolid + granitic rock powder (3:1, by volume); and biosolid + crushed coconut fiber + granitic rock powder (2:1:1, by volume)] and at five irrigation levels [36 % (very low); 55 % (low); 85 % (moderate); 100 % (well-irrigated); and 126 % of the reference irrigation level (over-irrigated)]. The biometric traits evaluated increased as the irrigation levels increased, with the exception of root dry mass, which was not influenced by this factor, nor water use efficiency, which decreased as the irrigation levels increased. In general, plants grown with a mixture of the three wastes (biosolid + crushed coconut fiber + granitic rock powder) exhibited higher values for biometric variables and chlorophyll content when they were moderately, well or over-irrigated, in comparison with the other two substrates. It was possible to achieve the plantlet commercial standard using this three waste mixture and the moderate irrigation level.(AU)

Heterologous in vitro fertilization and embryo production for assessment of jaguar (Panthera onca Linnaeus, 1758) frozen-thawed semen in different extenders

Heterologous in vitro fertilization (IVF) is an important tool for assessing fertility of endangered mammals such as the jaguar, considering difficult access to females for artificial insemination and to obtain homologous oocytes. We aimed to evaluate the fertility of jaguar sperm cryopreserved with different extenders, using domestic cat oocytes to assess the development of hybrid embryos. Semen from four captive jaguars was obtained by electroejaculation. Samples were cryopreserved in powdered coconut water (ACP-117c) or Tris extender containing 20% egg yolk and 6% glycerol. Thawed spermatozoa were resuspended (2.0 × 106 spermatozoa/mL) in IVF medium and co-incubated with cat oocytes matured in vitro for 18 h. Presumptive zygotes were cultured for 7 days. After 48 h, cleavage rate was evaluated, and non-cleaved structures were stained for IVF evaluation. On days 5 and 7, the rate of morula and blastocyst formation was assessed. Data were analyzed using the Fisher exact test (p < 0.05). No difference was observed between ACP-117c and Tris extenders, respectively, for oocytes with 2nd polar body (2/51, 3.9 ± 2.9% vs. 2/56, 3.6 ± 3.1%), pronuclear structures (5/51, 9.8 ± 4.7% vs. 8/56, 14.3 ± 8.0%), and total IVF rates (7/36, 19.4 ± 5.0% vs. 10/37, 27.0 ± 13.8%). All the samples fertilized the oocytes, with 22.9 ± 3.2% (16/70) and 16.7 ± 3.6% (12/72) cleavage of mature oocytes for ACP-117c and Tris extenders, respectively. Morula rates of 4.3 ± 2.3% (3/70) and 5.6 ± 2.2% (4/72) were observed for ACP-117c and Tris, respectively. Only the Tris extender demonstrated blastocyst production (2/12, 16.7 ± 1.5% blastocyst/cleavage). We demonstrated that jaguar ejaculates cryopreserved using ACP-117c and Tris were suitable for IVF techniques, with blastocyst production by ejaculates cryopreserved in Tris. This is a first report of embryos produced in vitro using jaguar sperm and domestic cat oocytes through IVF.(AU)

Effect of adding anethole to the cryopreservation medium (powdered coconut water) on morphology, kinetics, and oxidative stress of buck sperm / Efeito da adição de anethole ao meio de criopreserção (água de coco em pó) na morfologia, cinética e estresse oxidativo do sêmen de bode

The quality of post-thawing goat sperm is critical to the success of artificial insemination protocols and may be influenced by extenders, cryoprotectants, and antioxidant substances. Therefore, the objective of this study was to evaluate the effects of the antioxidant anethole on goat sperm diluted in preservation medium based on powdered coconut water (ACP-101c) and frozen. For that, each ejaculate was submitted to the following treatments: ACP-101c (control); control plus supplementation with 30, 300, or 2000 µg/ mL anethole. The samples were thawed and evaluated for morphology, kinetics, membrane integrity, and reactive oxygen species (ROS). The addition of anethole increased morphological abnormalities (P < 0.05), however, it did not affect sperm kinetics. Flow cytometry analysis showed that sperm cells cryopreserved with 300 µg/mL anethole had lower acrosome integrity than those cryopreserved in other treatments. Evaluation of oxidative stress revealed that cells stored in the presence of 2000 µg/mL anethole had small amounts of ROS when compared to those preserved in the control medium alone or supplemented with 300 µg/mL anethole (P < 0.05). After cryopreservation of sperm with 2000 µg/mL anethole, the highest percentage of viable sperm without ROS was observed (P < 0.05). In conclusion, despite reducing ROS levels, the supplementation of anethole in ACP-101c did not affect sperm kinetics or membrane integrity post-thawing, however, it did cause morphological damage to sperm.(AU)

A qualidade do espermatozoide caprino pós-descongelação é crítica para o sucesso dos protocolos de inseminação artificial e pode ser influenciada por extensores, crioprotetores e substâncias antioxidantes. Portanto, o objetivo deste estudo foi avaliar os efeitos do antioxidante anetole sobre espermatozoides caprinos diluídos em meio de conservação à base de água de coco em pó (ACP-101c) e congelados. Para tanto, cada ejaculado foi submetido aos seguintes tratamentos: ACP-101c (controle); controle mais suplementação com 30, 300 ou 2000 µg / mL de anetole. As amostras foram descongeladas e avaliadas quanto à morfologia, cinética, integridade de membranas e espécies reativas de oxigênio. A adição de anetole aumentou as anormalidades morfológicas (P < 0,05), no entanto, não afetou a cinética dos espermatozoides. A análise da citometria de fluxo mostrou que as células de esperma criopreservadas com 300 µg / mL anethole tinham integridade acrosma menor do que aquelas criopreservadas em outros tratamentos. A avaliação do estresse oxidativo revelou que as células armazenadas na presença de 2000 µg / mL anethole apresentaram pequenas quantidades de ROS quando comparadas às preservadas em meio de controle isoladamente ou suplementadas com 300 µg / mL anethole (P < 0,05). Após a criopreservação de espermatozoides com 2000 µg / mL anethole, observou-se a maior porcentagem de espermatozoides viáveis sem ROS (P < 0,05). A população com espermatozoides viáveis sem ROS foi maior quando utilizado 2.000 µg / mL (P < 0,05). Em conclusão, apesar de reduzir os níveis de ROS, a suplementação de anetole em ACP-101c não afetou a cinética espermática e a integridade da membrana pós-descongelação, entretanto, causou danos morfológicos nos espermatozoides.(AU)

Sêmen ovino e caprino imunossexado e diluído em meio de conservação à base de água de coco em pó (ACP101/102c) / Ram and goat semen immunosexed and diluted in powdered coconut water-based preservation medium (ACP101/102c)

Sperm sexing aims to separate sperm populations in carriers of the "X" or "Y" chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c. Ejaculates from five rams and five goats were collected with the aid of an artificial vagina; they were evaluated and submitted to the immunosexing protocol, according to the manufacturer's recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with "Y" chromosomes (HY; HY Biotechnology, Rio de Janeiro, RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P < 0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species. The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species.(AU)

Limonoid detection and profile in callus culture of sweet orange

Plant tissue culture has emerged as an important tool to produce bioactive compounds from various plant species, including the sustainable production of limonoids that are receiving considerable attention due to the benefits associated with human health such as anticancer activities. The purpose of the present study was to evaluate the capacity of limonoids aglycone production from callus culture from sweet orange cv. Pera (Citrus sinensis) seeds and identify the compounds produced in this cell line. Callus induction occurred in Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic (2,4-D), malt extract, agar and coconut water. For the analysis and identification of the limonoids, CG-MS-EI ion-positive mode and UPLC-QTOF-ESI were used operating in positive and negative mode. An intense peak corresponding to limonin appeared in the callus extracts at a retention time of 58.1 min. in CG-MS-EI and four major limonoids aglycone by positive ion mode UPLC-QTOF-ESI: limonin, nomilin, deacetylnomilin, and nomilinic acid. The culture medium was efficient at the bioproduction of limonoids aglycone in callus cultures of C. sinensis seeds. Therefore, data obtained from UPLC-QTOF-ESI proved its importance at identifying new compounds that benefit human health, and may assist future work in the identification of known or new limonoids in Citrus species and related genera.

Limonoid detection and profile in callus culture of sweet orange

Plant tissue culture has emerged as an important tool to produce bioactive compounds from various plant species, including the sustainable production of limonoids that are receiving considerable attention due to the benefits associated with human health such as anticancer activities. The purpose of the present study was to evaluate the capacity of limonoids aglycone production from callus culture from sweet orange cv. Pera (Citrus sinensis) seeds and identify the compounds produced in this cell line. Callus induction occurred in Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic (2,4-D), malt extract, agar and coconut water. For the analysis and identification of the limonoids, CG-MS-EI ion-positive mode and UPLC-QTOF-ESI were used operating in positive and negative mode. An intense peak corresponding to limonin appeared in the callus extracts at a retention time of 58.1 min. in CG-MS-EI and four major limonoids aglycone by positive ion mode UPLC-QTOF-ESI: limonin, nomilin, deacetylnomilin, and nomilinic acid. The culture medium was efficient at the bioproduction of limonoids aglycone in callus cultures of C. sinensis seeds. Therefore, data obtained from UPLC-QTOF-ESI proved its importance at identifying new compounds that benefit human health, and may assist future work in the identification of known or new limonoids in Citrus species and related genera.(AU)

Parâmetros microscópicos de açúcar de coco comercializados na cidade de São Paulo, Brasil: identidade e qualidade / Microscopic parameters of coconut sugar marketed in the city of São Paulo, Brazil: identity and quality

O açúcar de coco vem se tornando popular no Brasil e no mundo devido às alegações nutricionais de ser um adoçante natural e mais saudável em relação a outros açúcares. Sua produção ocorre de forma artesanal a partir da extração da seiva do floema da inflorescência da palmeira. Este trabalho teve por objetivo analisar microscopicamente a identidade e a presença de matérias estranhas em amostras de açúcar de coco, bem como verificar sua conformidade em relação às legislações sanitárias brasileiras. Foram analisadas 13 amostras comerciais de açúcar de coco da cidade de São Paulo quanto aos ensaios de pesquisa e identificação de elementos histológicos e pesquisa de matérias estranhas. A identificação de elementos histológicos vegetais revelou a presença de amidos morfologicamente semelhantes ao padrão de Triticum sp. (trigo), Manihot sp. (mandioca) e Oryza sativa (arroz), além de amidos alterados. Também foram encontrados elementos histológicos vegetais compatíveis com padrão de Saccharum officinarum (cana-de-açúcar) e de Cocos nucifera (coco). Todas as amostras analisadas continham matérias estranhas com predomínio de fragmentos de insetos em 100% delas. Os resultados do estudo indicam falhas nas Boas Práticas de produção do açúcar de coco e a necessidade da adequação deste produto em relação às normas vigentes.

Coconut sugar has become popular in Brazil and worldwide due to the nutritional claims of being a natural and healthier sweetener compared to other sugars. Its production takes places in an artisanal way from the extraction of phloem sap from the inflorescence of the palm. The aim of this work was to microscopically analyze the identity and the presence of foreign matter in samples of coconut sugar, as well as to verify its compliance with Brazilian health legislation. Thirteen commercial samples of coconut sugar from the city of São Paulo were analyzed for the research and identification of histological elements and foreign matter. The identification of vegetable histological elements revealed the presence of starches morphologically similar to the Triticum sp. (wheat), Manihot sp. (cassava) and Oryza sativa (rice) pattern, besides altered starches. Vegetable histological elements compatible with the Saccharum officinarum (sugar cane) and Cocos nucifera (coconut) pattern were also found. All samples analyzed contained foreign matter with predominance of insect fragments in 100% of the samples. The results of the study indicate flaws in the Good Practices for the production of coconut sugar and the need to adapt this product to current standards.

Parâmetros microscópicos de açúcar de coco comercializados na cidade de São Paulo, Brasil: identidade e qualidade / Microscopic parameters of coconut sugar marketed in the city of São Paulo, Brazil: identity and quality

O açúcar de coco vem se tornando popular no Brasil e no mundo devido às alegações nutricionais de ser um adoçante natural e mais saudável em relação a outros açúcares. Sua produção ocorre de forma artesanal a partir da extração da seiva do floema da inflorescência da palmeira. Este trabalho teve por objetivo analisar microscopicamente a identidade e a presença de matérias estranhas em amostras de açúcar de coco, bem como verificar sua conformidade em relação às legislações sanitárias brasileiras. Foram analisadas 13 amostras comerciais de açúcar de coco da cidade de São Paulo quanto aos ensaios de pesquisa e identificação de elementos histológicos e pesquisa de matérias estranhas. A identificação de elementos histológicos vegetais revelou a presença de amidos morfologicamente semelhantes ao padrão de Triticum sp. (trigo), Manihot sp. (mandioca) e Oryza sativa (arroz), além de amidos alterados. Também foram encontrados elementos histológicos vegetais compatíveis com padrão de Saccharum officinarum (cana-de-açúcar) e de Cocos nucifera (coco). Todas as amostras analisadas continham matérias estranhas com predomínio de fragmentos de insetos em 100% delas. Os resultados do estudo indicam falhas nas Boas Práticas de produção do açúcar de coco e a necessidade da adequação deste produto em relação às normas vigentes.(AU)

Coconut sugar has become popular in Brazil and worldwide due to the nutritional claims of being a natural and healthier sweetener compared to other sugars. Its production takes places in an artisanal way from the extraction of phloem sap from the inflorescence of the palm. The aim of this work was to microscopically analyze the identity and the presence of foreign matter in samples of coconut sugar, as well as to verify its compliance with Brazilian health legislation. Thirteen commercial samples of coconut sugar from the city of São Paulo were analyzed for the research and identification of histological elements and foreign matter. The identification of vegetable histological elements revealed the presence of starches morphologically similar to the Triticum sp. (wheat), Manihot sp. (cassava) and Oryza sativa (rice) pattern, besides altered starches. Vegetable histological elements compatible with the Saccharum officinarum (sugar cane) and Cocos nucifera (coconut) pattern were also found. All samples analyzed contained foreign matter with predominance of insect fragments in 100% of the samples. The results of the study indicate flaws in the Good Practices for the production of coconut sugar and the need to adapt this product to current standards.(AU)

Coconut Cake in Diets for Quail in the Laying Phase

Currently, agro-industrial by-products have increasingly been used in animal feeding, as they constitute an alternative source of nutrients for the animal diet and a way to simultaneously reduce environmental pollution. The objective of this study was to examine increasing levels of inclusion of coconut cake in Japanese quail diets in the laying phase on their production performance and egg quality. A total of 360 Japanese quails were allotted to eight treatments with nine replicates and eight birds per experimental unit, in a randomized-block design. Five diets were formulated: a diet without inclusion of the by-product; and diets containing 3, 6, 9, and 12% coconut cake. The experiment lasted 63 days, with evaluations occurring at every 21 days. The following variables were analyzed: feed intake, laying rate, feed conversion, egg weight, specific gravity, Haugh unit, yolk, albumen and shell percentage, shell thickness, and shell weight. The treatments elicited a positive linear response from laying rate, whereas feed conversion per egg mass decreased linearly. In terms of egg-quality traits, shell percentage was influenced, increasing linearly. Coconut cake inclusion at 12% in the diet of Japanese quail in the laying improved feed conversion per egg mass and increased egg-laying rate and eggshell percentage.(AU)

Coconut Cake in Diets for Quail in the Laying Phase

Currently, agro-industrial by-products have increasingly been used in animal feeding, as they constitute an alternative source of nutrients for the animal diet and a way to simultaneously reduce environmental pollution. The objective of this study was to examine increasing levels of inclusion of coconut cake in Japanese quail diets in the laying phase on their production performance and egg quality. A total of 360 Japanese quails were allotted to eight treatments with nine replicates and eight birds per experimental unit, in a randomized-block design. Five diets were formulated: a diet without inclusion of the by-product; and diets containing 3, 6, 9, and 12% coconut cake. The experiment lasted 63 days, with evaluations occurring at every 21 days. The following variables were analyzed: feed intake, laying rate, feed conversion, egg weight, specific gravity, Haugh unit, yolk, albumen and shell percentage, shell thickness, and shell weight. The treatments elicited a positive linear response from laying rate, whereas feed conversion per egg mass decreased linearly. In terms of egg-quality traits, shell percentage was influenced, increasing linearly. Coconut cake inclusion at 12% in the diet of Japanese quail in the laying improved feed conversion per egg mass and increased egg-laying rate and eggshell percentage.

Biotecnologias como ferramentas para o desenvolvimento do Nordeste do Brasil / Biotechnologies as tools for the development of northeast Brazil

A biotecnologia tem sido um ramo de estudo diferencial para diversos setores da sociedade, apresentando, através de bioprodutos e bioprocessos, melhorias para o avanço e desenvolvimento da região Nordeste do Brasil. Vale pontuar que um importante meio que traz anualmente acréscimos inovadores à área biotecnológica é o setor acadêmico, que através de cursos stricto sensu a nível de mestrado e doutorado, promovem pesquisas relevantes para vários setores como economia, agroindústria, saúde, dentre outros. Exemplos disso são: o curso de Doutorado em Biotecnologia da RENORBIO) e o Programa Profissional de Pós-Graduação em Biotecnologia em Saúde Humana e Animal (PPGBiotec). O presente artigo se sobre o percurso trilhado pelos cursos stricto sensu mencionados, bem como ressalta a relevância da Biotecnologia para a região Nordeste do Brasil, em seus diferentes campos de investigação, com ênfase nos Recursos Naturais, Agropecuária e Saúde. Exemplificamos as biotecnologias utilizando a água de coco que vêm sendo trabalhadas desde os anos 1980’s e sua evolução até os dias atuais. Com base em toda a potencialidade da Região Nordeste para a geração de bioprodutos e bioprocessos, ressaltamos que os mesmos só serão úteis se realmente forem tratados como inovação tecnológica, gerarem nota fiscal e impactarem positivamente para o bem estar da sociedade.

Biotechnology has been a branch of differential study for various sectors of society, presenting, through bioproducts and bioprocesses, improvements for the advancement and development of the Northeast region of Brazil. It is worth noting that an important means that annually brings innovative additions to the biotechnological area is the academic sector, which through stricto sensu courses at the master's and doctoral level, promote relevant research for various sectors such as economics, agribusiness, health, among others. Examples of this are: the Doctorate course in Biotechnology (RENORBIO) and the Professional Graduate Program in Biotechnology in Human and Animal Health (PPGBiotec). This article is about the path taken by the stricto sensu courses mentioned, as well as emphasizing the relevance of Biotechnology for the Northeast region of Brazil, in its different research fields, with emphasis on Natural Resources, Agriculture and Health. We exemplify biotechnologies using coconut water that has been worked since the 1980's and its evolution to the present day. Based on all the potential of the Northeast Region for the generation of bioproducts and bioprocesses, we emphasize that they will only be useful if they are really treated as technological innovation, generate invoices and have a positive impact on society's well-being.

Biotecnologias como ferramentas para o desenvolvimento do Nordeste do Brasil / Biotechnologies as tools for the development of northeast Brazil

A biotecnologia tem sido um ramo de estudo diferencial para diversos setores da sociedade, apresentando, através de bioprodutos e bioprocessos, melhorias para o avanço e desenvolvimento da região Nordeste do Brasil. Vale pontuar que um importante meio que traz anualmente acréscimos inovadores à área biotecnológica é o setor acadêmico, que através de cursos stricto sensu a nível de mestrado e doutorado, promovem pesquisas relevantes para vários setores como economia, agroindústria, saúde, dentre outros. Exemplos disso são: o curso de Doutorado em Biotecnologia da RENORBIO) e o Programa Profissional de Pós-Graduação em Biotecnologia em Saúde Humana e Animal (PPGBiotec). O presente artigo se sobre o percurso trilhado pelos cursos stricto sensu mencionados, bem como ressalta a relevância da Biotecnologia para a região Nordeste do Brasil, em seus diferentes campos de investigação, com ênfase nos Recursos Naturais, Agropecuária e Saúde. Exemplificamos as biotecnologias utilizando a água de coco que vêm sendo trabalhadas desde os anos 1980s e sua evolução até os dias atuais. Com base em toda a potencialidade da Região Nordeste para a geração de bioprodutos e bioprocessos, ressaltamos que os mesmos só serão úteis se realmente forem tratados como inovação tecnológica, gerarem nota fiscal e impactarem positivamente para o bem estar da sociedade.(AU)

Biotechnology has been a branch of differential study for various sectors of society, presenting, through bioproducts and bioprocesses, improvements for the advancement and development of the Northeast region of Brazil. It is worth noting that an important means that annually brings innovative additions to the biotechnological area is the academic sector, which through stricto sensu courses at the master's and doctoral level, promote relevant research for various sectors such as economics, agribusiness, health, among others. Examples of this are: the Doctorate course in Biotechnology (RENORBIO) and the Professional Graduate Program in Biotechnology in Human and Animal Health (PPGBiotec). This article is about the path taken by the stricto sensu courses mentioned, as well as emphasizing the relevance of Biotechnology for the Northeast region of Brazil, in its different research fields, with emphasis on Natural Resources, Agriculture and Health. We exemplify biotechnologies using coconut water that has been worked since the 1980's and its evolution to the present day. Based on all the potential of the Northeast Region for the generation of bioproducts and bioprocesses, we emphasize that they will only be useful if they are really treated as technological innovation, generate invoices and have a positive impact on society's well-being.(AU)