Efeito de um quelante de cálcio, um detergente e da lecitina de soja sobre a qualidade do sêmen caprino congelado-descongelado

The aim of this paper was verifying the effect of the Equex STM Paste and EDTA addition to a Tris-egg yolk extender, on the postthaw goat sperm viability. Nine semen samples of two adult goats were collected by artificial vagina and cryopreserved. It was also objective of this study, to evaluate the utilization of a soybean lecithin based commercial extender (Bioexcell® - IMV, L'Aigle, French) for the goat semen freezing. They were formed five experimental groups: TRIS; TRIS+EDTA; TRIS+EQUEX; TRIS+EDTA+EQUEX e Bioexcell. After evaluation, the semen was diluted in the five extenders and packed in 0.25mL straws with 100 million of motile spermatozoa. The samples were cooled at 0,46°C/min to 5°C, submitted at 75min of equilibration time and frozen in liquid nitrogen vapour. The thawing was accomplished in 37°C water bath for 50s. There were no differences (P>0,05) on the means of post-thaw total and progressive sperm motility among the groups TRIS, TRIS+EQUEX and TRIS+EQUEX+EDTA. The Bioexcell group obtained the least (P<0,05) percentage of post-thawing total and progressive sperm motility. After the thermotolerance test, it was observed the greatest (P<0,05) rates of total and progressive sperm motility in the Equex STM groups (TRIS+EQUEX and TRIS+EQUEX+EDTA). Thus, it can be affirmed that the Equex addition promotes better maintenance rates in the pos-thaw sperm viability, when compared with the extenders that did not contain it.

The energetic cost of maintenance in ruminants: From classical to new concepts and prediction systems

The cost of maintenance makes up a large part of total energy costs in ruminants. Metabolizable energy (ME) requirement for maintenance (MEm) is the daily ME intake that exactly balances heat energy (HE). The net energy requirement for maintenance (NEm) is estimated subtracting MEm from the HE produced by the processing of the diet. Men cannot be directly measured experimentally and is estimated by measuring basal metabolism in fasted animals or by regression measuring the recovered energy in fed animals. MEm and NEm usually, but not always, are expressed in terms of BW0.75. However, this scaling factor is substantially empirical and its exponent is often inadequate, especially for growing animals. MEm estimated by different feeding systems (AFRC, CNCPS, CSIRO, INRA, NRC) were compared by using dairy cattle data. The comparison showed that these systems differ in the approaches used to estimate MEm and for its quantification. The CSIRO system estimated the highest MEm, mostly because it includes a correction factor to increase ME as the feeding level increases. Relative to CSIRO estimates, those of NRC, INRA, CNCPS, and AFRC were on average 0.92, 0.86, 0.84, and 0.78, respectively. MEm is affected by the previous nutritional history of the animals. This phenomenon is best predicted by dynamic models, of which several have been published in the last decades. They are based either on energy flows or on nutrient flows. Some of the different approaches used were described and discussed.

Effect of maintenance therapy with or without the use of chlorhexidine in teeth restored with composite resin in patients with diabetes mellitus

The objective of this study was to evaluate the effects of maintenance therapy with or without the use of 0.12% chlorhexidine in the periodontal tissues of patients with diabetes mellitus who had carious lesions restored with composed resin. Twenty patients were selected, all of whom had diabetes mellitus in addition to carious cervical lesions in previously treated teeth. After 90 days, improvement in plaque and gingival indices and probing depth were noticed among patients in the group that received 0.12% chlorhexidine.

Design of a labVIEW system applied to predictive maintenance

The implementation of vibration analysis techniques based on virtual instrumentation has spread increasingly in the academic and industrial branch, since the use of any software for this type of analysis brings good results at low cost. Among the existing software for programming and creation of virtual instruments, the LabVIEW was chosen for this project. This software has good interface with the method of graphical programming. In this project, it was developed a system of rotating machine condition monitoring. This monitoring system is applied in a test stand, simulating large scale applications, such as in hydroelectric, nuclear and oil exploration companies. It was initially used a test stand, where an instrumentation for data acquisition was inserted, composed of accelerometers and inductive proximity sensors. The data collection system was structured on the basis of an NI 6008 A/D converter of National Instruments. An electronic circuit command was developed through the A/D converter for a remote firing of the test stand. The equipment monitoring is performed through the data collected from the sensors. The vibration signals collected by accelerometers are processed in the time domain and frequency. Also, proximity probes were used for the axis orbit evaluation and an inductive sensor for the rotation and trigger measurement. © (2013) Trans Tech Publications, Switzerland.

Haemonchus contortus: A multiple-resistant Brazilian isolate and the costs for its characterization and maintenance for research use

The aim of this work was to determine the resistance level of Haemonchus contortus isolated from the Santa Inês flock of the Embrapa (Brazilian government's Agricultural Research Company), Southeast Livestock Unit (CPPSE), as well as to determine costs of characterizing and maintaining this isolate in host donors. Forty-two male Santa Inês lambs were experimentally infected with 4000 H. contortus infective larvae of the field isolate of CPPSE, called Embrapa2010, and divided into six treatment groups, which received triclorfon, albendazol plus cobalt sulfate, ivermectin, moxidectin, closantel and levamisole phosphate, as well as a negative control group (water). Egg per gram (EPG) counts were performed at 0, 3, 7, 10 and 14. days post treatment when the animals were slaughtered for parasite count. The data were analyzed using the RESO statistical program, considering anthelmintic resistance under 95% of efficacy. EPG and worm count presented a linear and significant relation with 94% determination coefficient. The susceptibility results obtained by RESO through both criteria (EPG and worm count) were equal, except for closantel, showing that the isolate Embrapa2010 is resistant to benzimidazoles, macrocyclic lactones and imidazothiazoles. The need of a control group did not appear to be essential since the result for susceptibility in the analyses with or without this group was the same. Suppression in egg production after treatment did not occur in the ivermectin and moxidectin groups. In the control group, the establishment percentage was just 12.5 because of the low number of third-stage larvae, resistance (innate and infection immunity) of the animals studied plus good nutrition. Drug classes presented similar efficacy between adults and immature stages. The costs for isolate characterization were calculated for 42 animals during 60. days. The total cost based on local market rates was approximately US$ 8000. The precise identification of Brazilian isolates and their establishment in host donors would be useful for laboratorial anthelmintic resistance diagnoses through in vitro tests, which has an annual cost of approximately US$ 2500 for maintenance in host donors. © 2012 Elsevier B.V.

Ultrastructure of the spermatophores and spermatozoa from the gecarcinid land crab, Cardisoma guanhumi Latreille, 1828, (Crustacea, Brachyura, Gecarcinidae) from the western Atlantic

The ultrastructure of the sperm of the gecarcinid land crab Cardisoma guanhumi is described by scanning and transmission electron microscopy. Ultrastructural sperm morphology lends support for the monophyletic origin of the Thoracotremata, and for the placement of C. guanhumi in that clade. Additionally, it further attests the low level of variability within the thoracotreme clade. With regard to ultrastructural morphology, small differences concerning the apical button, the hyaline periopercular rim, the inner acrosome zone, and the outer two lamellations in the outer acrosome zone have been found between the spermatozoa of C. guanhumi and Cardisoma carnifex, the only other species in this genus studied for spermatozoal morphology. Copyright © 2013 Taylor & Francis.

Influence of Abutment Type and Esthetic Veneering on Preload Maintenance of Abutment Screw of Implant-Supported Crowns

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Heparin and penicillamine-hypotaurine-epinephrine (PHE) solution during bovine in vitro fertilization procedures impair the quality of spermatozoa but improve normal oocyte fecundation and early embryonic development

The presence of heparin and a mixture of penicillamine, hypotaurine, and epinephrine (PHE) solution in the in vitro fertilization (IVF) media seem to be a prerequisite when bovine spermatozoa are capacitated in vitro, in order to stimulate sperm motility and acrosome reaction. The present study was designed to determine the effect of the addition of heparin and PHE during IVF on the quality and penetrability of spermatozoa into bovine oocytes and on subsequent embryo development. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes and mitochondrial function, was diminished (P < 0.05) in the presence of heparin and PHE. Oocyte penetration and normal pronuclear formation rates, as well as the percentage of zygotes presenting more than two pronuclei, was higher (P < 0.05) in the presence of heparin and PHE. No differences were observed in cleavage rates between treatment and control (P > 0.05). However, the developmental rate to the blastocyst stage was increased in the presence of heparin and PHE (P > 0.05). The quality of embryos that reached the blastocyst stage was evaluated by counting the inner cell mass (ICM) and trophectoderm (TE) cell numbers and total number of cells; the percentage of ICM and TE cells was unaffected (P > 0.05) in the presence of heparin and PHE (P < 0.05). In conclusion, this study demonstrated that while the supplementation of IVF media with heparin and PHE solution impairs spermatozoa quality, it plays an important role in sperm capacitation, improving pronuclear formation, and early embryonic development. © 2013 The Society for In Vitro Biology.

Cryoprotective effect of different glycerol concentrations on domestic cat spermatozoa

Cryopreservation of spermatozoa is a pivotal tool in assisted reproduction, and studies aiming to establish optimal freezing/thawing protocols are essential to enhance sperm survival. The objectives of the present study were to (1) compare the cryoprotective efficiency of three different glycerol concentrations (3%, 5%, and 7%) on the basis of post-thaw sperm quality and (2) investigate whether the incidence of morphologically abnormal sperm in fresh samples is related to cryodamage sensitivity. Semen was collected from six tomcats using an artificial vagina (total 18 ejaculates). Each ejaculate was diluted using Tris-egg yolk-based extender (TEY), evaluated, equally divided into three aliquots, and rediluted using TEY with and without glycerol to achieve final concentrations of 3%, 5%, and 7%. Samples were loaded into 0.25 mL straws, equilibrated for 60 minutes at 5 °C, frozen, and then thawed at 46 °C for 12 seconds. Fresh and frozen-thawed samples were evaluated for sperm motion parameters (computer-assisted sperm analysis), plasma membrane integrity (PMI; propidium iodide and carboxyfluorescein diacetate), and DNA integrity (acridine orange). Plasma and acrosomal membrane integrity were assessed by flow cytometry (propidium iodide and fluorescein isothiocyanate-conjugated pea (Pisum sativum) agglutinin) immediately after thawing. Sperm motion parameters were also evaluated at 30 and 60 minutes of postincubation. For all treatment groups, cryopreservation significantly impaired the PMI and sperm motion parameters, except for straightness and amplitude of lateral head displacement. DNA integrity showed a slight reduction (P < 0.05) when 3% glycerol was used. The percentage of total motility, progressive motility, and rapid spermatozoa were significantly lower immediately after thawing and up to 60 minutes of incubation for the 3% glycerol group when compared with 5% and 7%. No difference (P > 0.05) was found for PMI, acrosome integrity, and DNA integrity among post-thaw groups. However, higher (P < 0.05) incidence of viable cells with reacted acrosome and dead cells with intact acrosome were observed with 7% and 3% glycerol, respectively. Percentage of morphologically abnormal spermatozoa in fresh sample was positively correlated with PMI only in the 3% glycerol group and negatively correlated with sperm motility in the 5% and 7% groups. In conclusion, the final concentration of 5% glycerol offered better cryoprotective effect for ejaculated cat sperm, and the relationship found between prefreezing sperm morphology and post-thaw sperm quality showed to be dependent on final glycerol concentration. © 2013 Elsevier Inc.

The roles of marginal lagoons in the maintenance of genetic diversity in the Brazilian migratory fishes Prochilodus argenteus and P. costatus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cryopreservation of bovine spermatozoa from the epididymal tail using conventional and automated methods

Cryopreservation of sperm is important to preserve the gerrnplasm from animals of genetic value, which can die unexpectedly. This study compares conventional and automated methods of cryopreservation of spermatozoa obtained from the epididymis of bulls post-mortem. Twenty-two epididymides were obtained from a commercial slaughterhouse. Spermatozoa were collected from the tail of the epididymis using the retrograde flow technique. Thus, the samples, which were diluted in 10 ml of extender without glycerol (Botubov (R) I, Botupharma, Botucatu, SP, Brazil), were evaluated on motility, sperm vigor, structural and functional (swelling hypoosmotic test) membrane integrity, mitochondrial activity, sperm viability and ADN fragmentation. The samples were divided into two aliquots and diluted in extender with glycerol (Botubov (R) II, Botupharma, Botucatu, SP, Brazil) at a concentration of 50x10(6) motile sperm/0.5 French straws. One sample was frozen by the conventional method (4 hours at 5 degrees C, in a refrigerator and 20 min in nitrogen vapor) and the other by the automated method (Cryogen (R) Dualflex, Neovet, Uberaba, MG, Brazil). The parameters were higher in all the tests of fresh sperm samples, with the exception of the swelling hypoosmotic test, which showed no significant difference when the results were compared with sperm frozen by the conventional method. The average motility of fresh spermatozoa was 74%, and conventional and automated averages were 29 and 25%, respectively. Therefore, although cryopreservation techniques reduce sperm quality parameters, the viability of the sperm is maintained, and these methods can be used to preserve sperm.

Melatonin Protective Role in Mouse Cauda Epipidymal Spermatozoa Damage Induced by Sodium Arsenite

We evaluated the sperm parameters such as cauda epididymis weight, sperm count, sperm morphology and sperm DNA stability of adult CF-1 male mice treated daily (oral exposure) with the toxic sodium arsenite (As, 7.0 mg/kg/body weight); Melatonin (Me, 10.0 mg/kg/bw), Me (10.0 mg/kg/bw) plus As (7.0 mg/kg/bw) and Negative Control (NaCl 0.9%) to assess acute (8.3 days), chronic (33.2 days) and recovery of testicular damage (66.4 days). Arsenic decreases the number of sperm from chronic treatment (33.2 days) and this effect continued until 66.4 days of treatment. The toxic effect of As also altered the morphology of spermatozoa in all treatment periods when compared to the negative control group. However, Metalonin induced protective effects in periods of 33.2 and 66.4 days of treatment. Additionally, the stability of DNA was significantly affected by arsenic in all periods, but the chronic treatment (33.2 days) in the AsMe revealed increased stability compared to the group treated with arsenic only. Melatonin partially protects sperm toxicity caused by Arsenic, especially during periods of 33.2 and 66.4 days.

Maintenance of Manioc Diversity by Traditional Farmers in the State of Mato Grosso, Brazil: A 20-Year Comparison

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)