Eugenol as an efficient anesthetic for neotropical fish Prochilodus nigricans (Teleostei, Prochilodontidae) / [Eugenol como um anestésico eficaz para o peixe neotropical Prochilodus nigricans (Teleostei, Prochilodontidae)]

The use of anesthetics in fish farming is essential to reduce stress during management. The present study proposes to evaluate the effect of eugenol as an anesthetic for the management of curimatã (Prochilodus nigricans). Fifty specimens were used (221.34±9.0 g; 25.8 ±1.1 cm), submitted to five treatments with concentrations of 0, 25, 50, 100 and 200 mgL−1 eugenol for 10 min. The times required to reach each anesthetic stage and the recovery of each fish were recorded. To check bloodglucose levels individual blood samples were taken from the fish before immersion and after ten minutes of anesthetic exposure. Evaluation of the anesthetic effect ongills was performed by histopathological analysis. The concentration of 50 mgL−1 was within the ideal limit of three minutes recommended for surgical anesthesia induction and did not significantly increase glucose levels when compared to the control group (35.7±19.4 mg dL−1), besides not causing gill injuries. At this concentration the fish presented controlled blood glucose levels within the basal levels, besides not taking the risk to have later problems concernig mortality, because this concentration ensures the good health of the gills and good recovery of the animals.(AU)

O uso de anestésicos em pisciculturas é indispensável para reduzir o estresse durante o manejo. O presente trabalho propôs avaliar o efeito do eugenol como anestésico para manejo do curimatã (Prochilodus nigricans). Foram utilizados 50 espécimes (221,34±9,0 g; 25,8 ±1,1 cm), submetidos a cinco tratamentos com concentrações de 0, 25, 50, 100 e 200 mg L−1 de eugenol em exposição de 10 minutos. Foram registrados os tempos necessários para atingir cada estágio anestésico e a recuperação de cada peixe. Para verificação dos níveis de glicose sanguínea, foram realizadas coletas individuais de sangue dos peixes antes da imersão e após os 10 minutos de exposição ao anestésico. A avaliação do efeito do anestésico sobre as brânquias foi realizada por meio de análises histopatológicas. A concentração de 50 mg L−1 ficou dentro do limite ideal de três minutos preconizado para indução à anestesia cirúrgica e não elevou significativamente os níveis de glicose quando comparada ao grupo controle (35,7±19,4 mg dL−1), além de não provocar lesões branquiais. Nessa concentração, os peixes apresentaram níveis de glicose sanguínea controlada, dentro dos teores basais, além de não correrem o risco de apresentarem problemas posteriores em relação à mortalidade, pois essa concentração garante a boa saúde das brânquias e uma boa recuperação dos animais.(AU)

Diferentes momentos de aplicação de gonadotrofina coriônica equina em protocolo de inseminação artificial em tempo fixo para vacas de leite / Different times of application of equine chorionic gonadotropin in fixed-time artificial insemination protocols for dairy cows

Avaliaram-se diferentes momentos de aplicação da gonadotrofina coriônica equina (eCG) em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de leite. Foram utilizadas 76 fêmeas, as quais receberam, no dia zero (D0) do protocolo, dispositivos intravaginais de progesterona, sendo esses retirados no D9, e os animais foram, então, distribuídos aleatoriamente em três tratamentos: T1 - aplicação de eCG no momento da retirada dos dispositivos; T2 e T3 - aplicação de eCG 48h e 24h antes da retirada dos dispositivos, respectivamente. No D10 os animais receberam 1mg de GnRH, e a IATF foi realizada 52 horas após a retirada do implante. Não houve diferença (P>0,05) para intervalo entre a retirada do implante à ovulação (72,56±3,92h), o diâmetro do maior folículo no D9 (10,88±1,49mm), o diâmetro do folículo ovulatório (15,15±1,16mm) e do segundo maior folículo (7,49±0,52mm), a taxa de crescimento folicular (1,38±0,04mm/dia), a taxa de ovulação (96,67%), o intervalo entre diâmetro final e inicial do folículo dominante (73,49±3,84h), a área de corpo lúteo (2,27±0,43cm²), a porcentagem de CL no ovário direito (53,00%) e no esquerdo (26,33%) e a taxa de gestação (33,33%). O momento da aplicação da eCG não influenciou na eficiência do protocolo. Recomenda-se a utilização da eCG no momento da retirada do implante por otimização do manejo.(AU)

Different times of application of equine chorionic gonadotropin (eCG) in Fixed-Time Artificial Insemination (FTAI) protocols for dairy cows were evaluated. A total of 76 females were used, which received intravaginal progesterone devices on day zero (D0) of the protocol, which were withdrawn on D9, and the animals were then randomly distributed in three treatments: T1- application of eCG at the time of device withdrawal; T2 and T3 - application of eCG 48h and 24h before withdrawal of the devices, respectively. On D10 the animals received 1mg of GnRH and the FTAI was carried out 52 hours after the removal of the implant. There was no difference (P>0.05) for interval from implant removal to ovulation (72.56±3.92h), diameter of the largest follicle of D9 (10.88±1.49mm), diameters of ovulatory follicle (15.15±1.16mm) and of the second largest follicle (7.49±0.52mm), follicle growth rate (1.38±0.04mm/day), ovulation rate (96.67%), interval between final diameter and initial diameter of dominant follicle (73.49±3.84h), corpus luteum area (2.27±0.43cm²), percentage of CL in right ovary (53.00%) and in left ovary (26.33%) and pregnancy rate (33.33%). The time of the application of eCG did not influence the efficiency of the protocol. We recommend the use of eCG at the time of the removal of the implant for management optimization.(AU)

Diagnostic accuracy of thromboelastometry and its correlation with the HPLC-MS/MS quantification test

The aim of the study was to evaluate the diagnostic accuracy of thromboelastometry for assessing rivaroxaban concentrations. The accuracy of thromboelastometry was compared with the high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method, which is the gold standard for drug plasma monitoring (the reference standard). Forty-six clinically stable patients were treated with 10, 15, or 20 mg of rivaroxaban once daily (OD group) or 15 mg twice a day (BID group) (no particular indication for treatment). Patient samples were collected 2 h after the use of the medication (peak) and 2 h before the next dose (trough). The rivaroxaban plasma concentrations were determined via HPLC-MS/MS, and thromboelastometry was performed using a ROTEM® delta analyzer. There were significant prolongations in clotting time (CT) for the 10, 15, and 20 mg of rivaroxaban treatments in the OD groups. In the 15 mg BID group, the responses at the peak and trough times were similar. At the peak times, there was a positive correlation between the plasma concentration of rivaroxaban and CT (Spearman correlation rho=0.788, P<0.001) and clot formation time (rho=0.784, P<0.001), and a negative correlation for alpha angle (rho=−0.771, P<0.001), amplitude after 5 min (rho=−0.763, P<0.001), and amplitude after 10 min (rho=−0.680, P<0.001). The CT presented higher specificity and sensitivity using the cut-off determined by the receiver characteristics curve. ROTEM has potential as screening tool to measure possible bleeding risk associated with rivaroxaban plasma levels.

Óleo de peixe associado ao ácido ascórbico no diluidor para criopreservação de sêmen caprino / Fish oil associated to ascorbic acid in diluter for cryopreservation goat semen

O presente estudo teve como objetivo avaliar o efeito da inclusão de óleo de peixe associado ao ácido ascórbico no diluidor para criopreservação de sêmen caprino. Dois machos da raça Boer foram submetidos à coleta de sêmen pelo método de vagina artificial, sendo os ejaculados avaliados quanto aos aspectos físicos e morfológicos. Após avaliação, formou-se um pool, seguido do fracionamento em cinco grupos: G1 - diluidor citrato-gema e G2, G3, G4 e G5 - diluidor citrato-gema acrescido de 1,0; 2,0; 3,0 e 4,0% de óleo de peixe e 0,05% de ácido ascórbico, respectivamente. Após descongelamento, foram realizadas avaliações físicas do sêmen e os testes complementares de termorresistência lento (TTR), hiposmótico (HO), integridade acrossomal e compactação da cromatina espermática. Houve comportamento linear crescente (P<0,05) para motilidade pós-descongelamento. Não houve diferença (P>0,05) para vigor pós-descongelamento (2,00±0,24). No TTR não houve diferença (P>0,05) para motilidade e vigor espermáticos entre os tempos cinco e 180min, com médias inicial e final de 62,17±12,13 e 14,29±10,55 para motilidade e de 2,00±0,52 e 0,49±0,44 para vigor. Não houve diferença (P>0,05) para o HO, com porcentagem média de espermatozoides reativos de 23,5±5,96%. Houve comportamento linear crescente para acrossoma íntegro e decrescente para acrossoma irregular (P<0,05). Não houve diferença (P>0,05) na compactação da cromatina, com 97,06±1,17% de cromatina íntegra. A inclusão até 4% de óleo de peixe acrescido de ácido ascórbico no diluidor melhorou motilidade e integridade de acrossoma após a criopreservação.(AU)

The study aimed to evaluate the effect of fish oil inclusion associated with ascorbic acid in the thinner for goat semen cryopreservation. Two male Boers underwent semen collection through the artificial vagina method, ejaculates being then assessed for physical and morphological aspects. After evaluation, a pool was formed, followed by the split into five groups: G1 - yolk-citrate extender and G2, G3, G4 and G5 - yolk-citrate extender plus 1.0; 2.0; 3.0 and 4.0% fish oil and 0.05% ascorbic acid, respectively. After thawing, physical evaluations of semen were assessed and additional testing slow heat resistance (TTR), hiposmotic (HO), acrosome integrity and compression of sperm chromatin. There was linear increase (P<0.05) post-thaw motility. No difference was obtained for post-thaw vigor and there was no influence of the association of fish oil and ascorbic acid in TTR. Plasma membrane integrity, by hyposmotic test (HO), presented a mean of reactive spermatozoa of 23.5±5.96% (P>0.05). There was linear increase for intact acrosome and decreasing acrosome irregular (P<0.05). In the analysis of the chromatin compaction, approximately 3% of damages (P>0.05) were observed. The inclusion of 4% fish oil plus ascorbic acid in diluter improved motility and acrosome integrity after cryopreservation.(AU)

Biomarcadores histológicos em brânquias de Sciades herzbergii (Siluriformes, Ariidae) capturados no Complexo Estuarino de São Marcos, Maranhão / Histological biomarkers in gills of Sciades herzbergii (Siluriformes, Ariidae) caught in the Estuarine Complex of São Marcos, Maranhão

Neste estudo, objetivou-se avaliar a qualidade ambiental de duas áreas do complexo estuarino de São Marcos, utilizando-se as alterações branquiais em Sciades herzbergii como biomarcadores de contaminação aquática. Foram realizadas coletas no período de estiagem (setembro/dezembro de 2015) e no período chuvoso (março/junho de 2016). Foram capturados 40 exemplares de S. herzbergii em dois locais distintos do complexo estuarino: A1) Porto do Itaqui Bacanga (área potencialmente impactada); A2) Igarapé do Puca, Alcântara (área controle). Foram registradas as variáveis abióticas: salinidade, temperatura, pH e oxigênio dissolvido. No laboratório, as brânquias de cada espécime foram fixadas em formol a 10% e mantidas em álcool a 70% até a execução da técnica histológica usual. As alterações branquiais identificadas nos peixes da área potencialmente contaminada foram: deslocamento do epitélio (DE), congestão (CO), aneurisma (AN), fusão lamelar (FL), hiperplasia (HI), dilatação capilar (DC), necrose (NE) e proliferação de células epiteliais (PCE). As alterações na região controle foram: deslocamento do epitélio (DE), fusão lamelar (FL), necrose (NE) e proliferação de células epiteliais (PCE). As alterações branquiais foram mais frequentes na região portuária (84%) do que na região controle (16%). Esses resultados indicam que os peixes da área potencialmente contaminada estão apresentando respostas biológicas ao estresse local.(AU)

This study aimed to compare an environmental quality of two areas of the estuarine complex of São Marcos using gills of changes in Sciades herzbergii as biomarkers of aquatic contamination. Samples were made during the dry season (September/ December 2015) and rainy season (March/ June 2016). They were captured 40 specimens of S. herzbergii at two different points of the estuarine complex: A1) Itaqui port (potentially contaminated area); A2) Igarapé do Puca, Alcântara (control area). Abiotic variables were recorded: salinity, temperature, pH and dissolved oxygen. In the laboratory, the gills of each species were fixed in 10% formaldehyde and kept in 70% alcohol until the usual histological technique was performed. The gill alterations identified in the fish of the potentially contaminated area were: epithelial displacement (DE), congestion (CO), aneurysm (AN), lamellar fusion (FL), hyperplasia (HI), capillary dilation (DC), necrosis and epithelial cell proliferation (PCE). The changes in the control region were: epithelial displacement (DE), lamellar fusion (FL), necrosis (NE) and epithelial cell proliferation (PCE). Gill alterations were more frequent in the port region (84%) than in the control region (16%). These results indicate that fish from the potentially contaminated area are presenting biological responses to local stress.(AU)

Effects of simvastatin/ezetimibe on microparticles, endothelial progenitor cells and platelet aggregation in subjects with coronary heart disease under antiplatelet therapy

It is not known whether the addition of ezetimibe to statins adds cardiovascular protection beyond the expected changes in lipid levels. Subjects with coronary heart disease were treated with four consecutive 1-week courses of therapy (T) and evaluations. The courses were: T1, 100 mg aspirin alone; T2, 100 mg aspirin and 40 mg simvastatin/10 mg ezetimibe; T3, 40 mg simvastatin/10 mg ezetimibe, and 75 mg clopidogrel (300 mg initial loading dose); T4, 75 mg clopidogrel alone. Platelet aggregation was examined in whole blood. Endothelial microparticles (CD51), platelet microparticles (CD42/CD31), and endothelial progenitor cells (CD34/CD133; CDKDR/CD133, or CD34/KDR) were quantified by flow cytometry. Endothelial function was examined by flow-mediated dilation. Comparisons between therapies revealed differences in lipids (T2 and T3<T1 and T4 for total cholesterol, LDL-C, and triglycerides; P<0.002 for all), as well as for endothelial function (T2>T1 and T4, P=0.001). Decreased platelet aggregation was observed after aspirin (arachidonic acid, T1<T3 and T4, P=0.034) and clopidogrel (adenosine, T3 and T4<T1 and T2, P<0.0001) therapy. Simvastatin/ezetimibe diphosphate did not change platelet aggregation, the amount of circulating endothelial and platelet microparticles, or endothelial progenitor cells. Cardiovascular protection following therapy with simvastatin/ezetimibe seems restricted to lipid changes and improvement of endothelial function not affecting the release of microparticles, mobilization of endothelial progenitor cells or decreased platelet aggregation.

Differences in synthesis and absorption of cholesterol of two effective lipid-lowering therapies

Effective statin therapy is associated with a marked reduction of cardiovascular events. However, the explanation for full benefits obtained for LDL cholesterol targets by combined lipid-lowering therapy is controversial. Our study compared the effects of two equally effective lipid-lowering strategies on markers of cholesterol synthesis and absorption. A prospective, open label, randomized, parallel design study, with blinded endpoints, included 116 subjects. We compared the effects of a 12-week treatment with 40 mg rosuvastatin or the combination of 40 mg simvastatin/10 mg ezetimibe on markers of cholesterol absorption (campesterol and β-sitosterol), synthesis (desmosterol), and their ratios to cholesterol. Both therapies similarly decreased total and LDL cholesterol, triglycerides and apolipoprotein B, and increased apolipoprotein A1 (P < 0.05 vs baseline for all). Simvastatin/ezetimibe increased plasma desmosterol (P = 0.012 vs baseline), and decreased campesterol and β-sitosterol (P < 0.0001 vs baseline for both), with higher desmosterol (P = 0.007) and lower campesterol and β-sitosterol compared to rosuvastatin, (P < 0.0001, for both). In addition, rosuvastatin increased the ratios of these markers to cholesterol (P < 0.002 vs baseline for all), whereas simvastatin/ezetimibe significantly decreased the campesterol/cholesterol ratio (P = 0.008 vs baseline) and tripled the desmosterol/cholesterol ratio (P < 0.0001 vs baseline). The campesterol/cholesterol and β-sitosterol/cholesterol ratios were lower, whereas the desmosterol/cholesterol ratio was higher in patients receiving simvastatin/ezetimibe (P < 0.0001 vs rosuvastatin, for all). Pronounced differences in markers of cholesterol absorption and synthesis were observed between two equally effective lipid-lowering strategies.

Effect of passive stretching on the immobilized soleus muscle fiber morphology

The aim of the present study was to determine the effect of stretching applied every 3 days to the soleus muscle immobilized in the shortened position on muscle fiber morphology. Eighteen 16-week-old Wistar rats were used and divided into three groups of 6 animals each: a) the left soleus muscle was immobilized in the shortened position for 3 weeks; b) during immobilization, the soleus was stretched for 40 min every 3 days; c) the non-immobilized soleus was only stretched. Left and right soleus muscles were examined. One portion of the soleus was frozen for histology and muscle fiber area evaluation, while the other portion was used to identify the number and length of serial sarcomeres. Immobilized muscles (group A) showed a significant decrease in weight (44 ± 6 percent), length (19 ± 7 percent), serial sarcomere number (23 ± 15 percent), and fiber area (37 ± 31 percent) compared to the contralateral muscles (P < 0.05, paired Student t-test). The immobilized and stretched soleus (group B) showed a similar reduction but milder muscle fiber atrophy compared to the only immobilized group (22 ± 40 vs 37 ± 31 percent, respectively; P < 0.001, ANOVA test). Muscles submitted only to stretching (group C) significantly increased the length (5 ± 2 percent), serial sarcomere number (4 ± 4 percent), and fiber area (16 ± 44 percent) compared to the contralateral muscles (P < 0.05, paired Student t-test). In conclusion, stretching applied every 3 days to immobilized muscles did not prevent the muscle shortening, but reduced muscle atrophy. Stretching sessions induced hypertrophic effects in the control muscles. These results support the use of muscle stretching in sports and rehabilitation.

Effect of one stretch a week applied to the immobilized soleus muscle on rat muscle fiber morphology

We determined the effect of stretching applied once a week to the soleus muscle immobilized in the shortened position on muscle fiber morphology. Twenty-six male Wistar rats weighing 269 ± 26 g were divided into three groups. Group I, the left soleus was immobilized in the shortened position for 3 weeks; group II, the soleus was immobilized in the shortened position and stretched once a week for 3 weeks; group III, the soleus was submitted only to stretching once a week for 3 weeks. The medial part of the soleus muscle was frozen for histology and muscle fiber area evaluation and the lateral part was used for the determination of number and length of serial sarcomeres. Soleus muscle submitted only to immobilization showed a reduction in weight (44 ± 6 percent, P = 0.002), in serial sarcomere number (23 ± 15 percent) and in cross-sectional area of the fibers (37 ± 31 percent, P < 0.001) compared to the contralateral muscles. The muscle that was immobilized and stretched showed less muscle fiber atrophy than the muscles only immobilized (P < 0.05). Surprisingly, in the muscles submitted only to stretching, fiber area was decreased compared to the contralateral muscle (2548 ± 659 vs 2961 ± 806 µm², respectively, P < 0.05). In conclusion, stretching applied once a week for 40 min to the soleus muscle immobilized in the shortened position was not sufficient to prevent the reduction of muscle weight and of serial sarcomere number, but provided significant protection against muscle fiber atrophy. In contrast, stretching normal muscles once a week caused a reduction in muscle fiber area.

Alteracoes da cronaxia, da reobase e da acomodacao no musculo esqueletico desnervado submetido a eletroestimulacao / Alterations of the chronaxie, reobase and accomodation in denervated skeletal muscle submitted to electrical stimulation

O uso da eletroestimulacao (EE) no tratamento das lesoes nervosas perifericas e controverso. Este estudo avaliou a alteracao da cronaxia, da reobase e da acomodacao no musculo tibial anterior (TA) desnervado de ratos submetidos a EE. Quatro grupos de ratos forma divididos: TA desnervado com eletrodiagnostico (ED) semanal (DSN+ED); TA desnervado com EE em dias alternados (DSN+EE+ED); TA direito desnervado (DSN); e TA inervado (INV). O eletrodiagnostico do grupo DSN+EE+ED forneceu os parametros EE (20 contracoes intensas com corrente monopolar, exponencial; periodo de pulso igual a 2x o valor da cronaxia; frequencia de 20Hz; amplitude em nivel motor; e tempo ON 3,0 s e OFF 6,0 s). Apos 4 semanas, nota-se, em todos os musculos desnervados, diminuicao na reobase, caracterizando hiperexcitabilidade muscular. A reobase foi mairo no musculo so desnervado (1+0 mA), se comparada ao DSN+ED (0,5+0mA, p=0,00006) e ao DSN+EE+ED (0,58+0,19mA, P=0,0001). A cronaxia do grupo DSN+EE+ED foi melhor (4,06+2,1ms) se comparada aos grupos DNS+ED (8,4+0,93MS,P=0006) e DSN (5,31+0,53ms, p=0,45). A acomodacao diminuiu nos grupos DSN+ED (0,75+0,42mA) e DSN+EE+ED (0,75+0,6mA), mantendo-se com valores proximos a pre-desnervacao no grupo somente desnervado (DSN 1,8+0,45mA). Concluiu-se que, embora a EE e o ED tenha atrasado a recuperacao da reobase e da acomodacao, sua utilizacao conjunta favoreceu a rcuperacao da cronaxia. Esse resultado indica o efeito benefico no tratamento do musculo desnervado

A new model for the immobilization of the rat hind limb

An alternative device for the immobilization of the hind limb of the rat was developed to study the effects of chronic disuse on the soleus and tibialis anterior muscles, maintained for 3 weeks in the shortening and the stretching positions, respectively. The proposed device is made of steel mesh and cotton materials, and has some advantages when compared to cast or plaster cast: it is cheaper, lighter (12 g or 4 percent of the body weight of the rat) and the same unit can be easily adjusted and used several times in the same animal or in animals of similar size. Immobilization is also useful to restrain the movements of the hip, knee, and ankle joints. Male rats (291 ± 35 g and aged 14 ± 2 weeks) were used to develop and test the model. The soleus muscle of 18 rats was maintained in a shortened position for 21 consecutive days and lost 19 ± 7 percent of its length (P = 0.008) and 44 ± 6 percent of its weight (P = 0.002) compared to the contralateral intact muscle. No difference (P = 0.67) was found in the stretched tibialis anterior of the same hind limb when compared to the contralateral muscle. No ulcer, sore or foot swelling was observed in the animals. Immobilization was effective in producing chronic muscle disuse in the hind limbs of rats and is an acceptable alternative to the traditional methods of immobilization such as cast or plaster cast