Phagocytotic activity and gene expression of leukocytes isolated from Astyanax lacustris.

The constant intensification of aquaculture has considerable increased the stress levels of farmed fish and, consequently, the number and intensity of diseases outbreaks. Thus, studies on fish immune response, especially regarding the interaction of fish leukocytes with potential pathogens and xenobiotics are of great importance in order to develop new prophylactic and curative strategies. We isolated leukocytes from the head kidney of Astyanax lacustris-an important Neotropical fish species for aquaculture and a potential model for Neotropical aquaculture research-using a Percoll centrifugation protocol. The isolated leukocytes were incubated with lipopolysaccharide (LPS), and the expression of genes IL-1ß, IL-8, LysC, and LysG were measured. We assessed the phagocytotic activity of leukocytes using Congo red-dyed yeast, a novel and cost-effective protocol that has been developed in this study. The isolated leukocytes responded to LPS induction, exhibiting strong IL-1ß and IL-8 upregulation, two of the most important pro-inflammatory interleukins for vertebrates immune reponse. The optimal concentration of yeast for the phagocytic assay was 106 cells mL-1, resulting in acceptable phagocytic capacity (PC) but without excess of yeasts during the counting process, ensuring a high precision and accuracy of the method. To the best of our knowledge, the present study is the first to investigate the in vitro gene expression and phagocytic activity of leukocytes isolated from A. lacustris. Our findings will serve as a reference for future studies on the immunology and toxicology of Neotropical fish.

Phagocytotic activity and gene expression of leukocytes isolated from Astyanax lacustris / Atividade fagocítica e expressão gênica de leucócitos isolados de Astyanax lacustris

The constant intensification of aquaculture has considerable increased the stress levels of farmed fish and, consequently, the number and intensity of diseases outbreaks. Thus, studies on fish immune response, especially regarding the interaction of fish leukocytes with potential pathogens and xenobiotics are of great importance in order to develop new prophylactic and curative strategies. We isolated leukocytes from the head kidney of Astyanax lacustris­an important Neotropical fish species for aquaculture and a potential model for Neotropical aquaculture research­using a Percoll centrifugation protocol. The isolated leukocytes were incubated with lipopolysaccharide (LPS), and the expression of genes IL-1ß, IL-8, LysC, and LysG were measured. We assessed the phagocytotic activity of leukocytes using Congo red-dyed yeast, a novel and cost-effective protocol that has been developed in this study. The isolated leukocytes responded to LPS induction, exhibiting strong IL-1ß and IL-8 upregulation, two of the most important pro-inflammatory interleukins for vertebrates immune reponse. The optimal concentration of yeast for the phagocytic assay was 106 cells mL-1, resulting in acceptable phagocytic capacity (PC) but without excess of yeasts during the counting process, ensuring a high precision and accuracy of the method. To the best of our knowledge, the present study is the first to investigate the in vitro gene expression and phagocytic activity of leukocytes isolated from A. lacustris. Our findings will serve as a reference for future studies on the immunology and toxicology of Neotropical fish.

A constante intensificação da aquicultura tem aumentado consideravelmente os níveis de estresse dos animais cultivados e, consequentemente, o número e a intensidade dos surtos de doenças. Logo, estudos sobre a resposta imune dos peixes, especialmente relacionados com a interação dos leucócitos de peixes com potenciais patógenos e xenobióticos, são de grande importância para o desenvolvimento de novas estratégias profiláticas e curativas. No presente trabalho, nós obtivemos sucesso ao isolar leucócitos oriundos do rim cranial de Astyanax lacustris ­ uma importante espécie de peixe Neotropical para a aquicultura e um modelo em potencial para pesquisas em aquicultura Neotropical ­ usando um protocolo de centrifugação com Percoll. Os leucócitos isolados foram incubados com lipossacarídeo (LPS) e, a expressão dos genes IL-1ß, IL-8, LysC, e LysG foi avaliada. Ainda, um novo protocolo para avaliação da atividade fagocítica dos leucócitos utilizando leveduras coradas com Vermelho Congo foi estabelecido. Os leucócitos isolados responderam à indução com LPS, exibindo up regulation dos genes IL-1ß e IL-8, duas das interleucinas pró-inflamatórias mais importantes para a resposta imune de vertebrados. Além do mais, a concentração ótima de leveduras para a avaliação da fagocitose foi de 106 células mL-1, resultando em uma capacidade fagocítica (PC) aceitável, mas sem excesso de leveduras durante o processo de contagem, garantindo maior precisão e eficácia do método. Até o presente momento, o presente estudo é o primeiro a investigar a expressão gênica e atividade fagocítica de leucócitos isolados de A. lacustris através da abordagem in vitro. Ainda, nossos resultados servirão de referência para futuros estudos em imunologia e toxicologia de peixes Neotropicais.

Prospection of genomic regions divergently selected in racing line of Quarter Horses in relation to cutting line.

Selection of Quarter Horses for different purposes has led to the formation of lines, including racing and cutting horses. The objective of this study was to identify genomic regions divergently selected in racing line of Quarter Horses in relation to cutting line applying relative extended haplotype homozygosity (REHH) analysis, an extension of extended haplotype homozygosity (EHH) analysis, and the fixation index (F ST) statistic. A total of 188 horses of both sexes, born between 1985 and 2009 and registered at the Brazilian Association of Quarter Horse Breeders, including 120 of the racing line and 68 of the cutting line, were genotyped using single nucleotide polymorphism arrays. On the basis of 27 genomic regions identified as selection signatures by REHH and F ST statistics, functional annotations of genes were made in order to identify those that could have been important during formation of the racing line and that could be used subsequently for the development of selection tools. Genes involved in muscle growth (n=8), skeletal growth (n=10), muscle energy metabolism (n=15), cardiovascular system (n=14) and nervous system (n=23) were identified, including the FKTN, INSR, GYS1, CLCN1, MYLK, SYK, ANG, CNTFR and HTR2B.

Risk factors for ocular toxoplasmosis in Brazil.

The aim of this study was to investigate risk factors for ocular toxoplasmosis (OT) in patients who received medical attention at a public health service. Three hundred and forty-nine consecutive patients, treated in the Outpatient Eye Clinic of Hospital de Base, São José do Rio Preto, São Paulo state, Brazil, were enrolled in this study. After an eye examination, enzyme-linked immunosorbent assay (ELISA) was used to determine anti-Toxoplasma gondii antibodies. The results showed that 25.5% of the patients were seronegative and 74.5% were seropositive for IgG anti-T. gondii antibodies; of these 27.3% had OT and 72.7% had other ocular diseases (OOD). The presence of cats or dogs [odds ratio (OR) 2.22, 95% confidence interval (CI) 1.24-3.98, P = 0.009] and consumption of raw or undercooked meat (OR 1.77, 95% CI 1.05-2.98, P = 0.03) were associated with infection but not with the development of OT. Age (OT 48.2 ± 21.2 years vs. OOD: 69.5 ± 14.7 years, P < 0.0001) and the low level of schooling/literacy (OT vs. OOD: OR 0.414, 95% CI 0.2231-0.7692, P = 0.007) were associated with OT. The presence of dogs and cats as well as eating raw/undercooked meat increases the risk of infection, but is not associated with the development of OT.

Cerebral and ocular toxoplasmosis related with IFN-y, TNF-a, and IL-10

This study analyzed the synthesis of Interferon gamma (IFN-γ), Tumor Necrosis Factor alpha (TNF-α), and Interleukin 10 (IL-10) in chronically infected patients which developed the symptomatic disease as cerebral or ocular toxoplasmosis. Blood from 61 individuals were divided into four groups: Cerebral toxoplasmosis/AIDS patients (CT/AIDS group) (n = 15), ocular toxoplasmosis patients (OT group) (n = 23), chronic toxoplasmosis individuals (CHR group) (n = 13) and healthy individuals (HI group) (n = 10). OT, CHR, and HI groups were human immunodeficiency virus (HIV) seronegative. The diagnosis was made by laboratorial (PCR and ELISA) and clinical subjects. For cytokine determination, peripheral blood mononuclear cells (PBMC) of each patient were isolated and stimulated in vitro with T. gondii antigen. IFN-γ, TNF-α, and IL-10 activities were determined by ELISA. Patients from CT/AIDS and OT groups had low levels of IFN-γ when were compared with those from CHR group. These data suggest the low resistance to develop ocular lesions by the low ability to produce IFN-γ against the parasite. The same patients, which developed ocular or cerebral toxoplasmosis had higher TNF-α levels than CHR individuals. High TNF-α synthesis contribute to the inflammatory response and damage of the choroid and retina in OT patients and in AIDS patients caused a high inflammatory response as the TNF-α synthesis is not affected since monocytes are the major source this cytokine in response to soluble T. gondii antigens. IL-10 levels were almost similar in CT/AIDS and OT patients but low when compared with CHR individuals. The deviation to Th2 immune response including the production of anti-inflammatory cytokines, such as IL-10 may promote the parasite's survival causing the tissue immune destruction. IL-10 production in T. gondii-infected brains may support the persistence of parasites as down-regulating the intracerebral immune response. All these indicate that OT and CT/AIDS patients produced low levels of IL-10 (Th2 response) and IFN-γ (Th1 response). They produced high TNF-α suggesting a high inflammatory response triggered by the parasite.

Seleção de características morfofuncionais de cavalos da raça Mangalarga Marchador por meio da análise de componentes principais / Identification of morphofunctional traits in Mangalarga Marchador horse using principal component analysis

Dados de 14.288 animais da raça Mangalarga Marchador, nascidos de 1990 a 2005, foram utilizados para avaliar a redução da dimensionalidade do espaço multivariado para características morfofuncionais, por meio da análise de componentes principais. Foram consideradas as características: altura na cernelha, altura na garupa, comprimento da cabeça, comprimento do pescoço, comprimento do dorso, comprimento da garupa, comprimento da espádua, comprimento do corpo, largura da cabeça, largura das ancas, perímetro do tórax, perímetro da canela e a pontuação da marcha. Para tais características, obtiveram-se sete componentes principais, a partir da matriz de correlação, que apresentaram variância inferior a 0,7 (autovalor inferior a 0,7). Isso sugere sete variáveis para descarte, por apresentarem maiores coeficientes de ponderação, em valor absoluto, a partir do último componente principal. A razão para isso é que variáveis altamente correlacionadas com os componentes de menor variância representam variação praticamente insignificante. Com base nesses resultados, recomendam-se as seguintes características para serem mantidas em trabalhos futuros com esta base de dados: pontuação da marcha, altura na garupa, comprimento do dorso, comprimento da garupa, largura da cabeça e perímetro da canela.

Records from 14,288 animals of the Mangalarga Marchador breed, born from 1990 to 2005, were used to discard morphofunctional traits in a principal component analysis. The following traits were used: height at withers, height at croup, lengths of head, neck, back, croup, hip length and body, widths of head, hip width, thorax perimeter, cannon bone circumference and gait score. For the traits considered it was observed that 7 principal components showed variation lower than 0.7; suggesting that seven variables could be discarded. The reason is that when variable are highly correlated with the principal components of smaller variance, their variation is practically insignificant. Based on those results the recommendation is to maintain the following traits for future research with this database: gait score, height at croup, length of back, length of croup, width of head and cannon bone circumference.

Seleção de características morfofuncionais de cavalos da raça Mangalarga Marchador por meio da análise de componentes principais / Identification of morphofunctional traits in Mangalarga Marchador horse using principal component analysis

Dados de 14.288 animais da raça Mangalarga Marchador, nascidos de 1990 a 2005, foram utilizados para avaliar a redução da dimensionalidade do espaço multivariado para características morfofuncionais, por meio da análise de componentes principais. Foram consideradas as características: altura na cernelha, altura na garupa, comprimento da cabeça, comprimento do pescoço, comprimento do dorso, comprimento da garupa, comprimento da espádua, comprimento do corpo, largura da cabeça, largura das ancas, perímetro do tórax, perímetro da canela e a pontuação da marcha. Para tais características, obtiveram-se sete componentes principais, a partir da matriz de correlação, que apresentaram variância inferior a 0,7 (autovalor inferior a 0,7). Isso sugere sete variáveis para descarte, por apresentarem maiores coeficientes de ponderação, em valor absoluto, a partir do último componente principal. A razão para isso é que variáveis altamente correlacionadas com os componentes de menor variância representam variação praticamente insignificante. Com base nesses resultados, recomendam-se as seguintes características para serem mantidas em trabalhos futuros com esta base de dados: pontuação da marcha, altura na garupa, comprimento do dorso, comprimento da garupa, largura da cabeça e perímetro da canela.(AU)

Records from 14,288 animals of the Mangalarga Marchador breed, born from 1990 to 2005, were used to discard morphofunctional traits in a principal component analysis. The following traits were used: height at withers, height at croup, lengths of head, neck, back, croup, hip length and body, widths of head, hip width, thorax perimeter, cannon bone circumference and gait score. For the traits considered it was observed that 7 principal components showed variation lower than 0.7; suggesting that seven variables could be discarded. The reason is that when variable are highly correlated with the principal components of smaller variance, their variation is practically insignificant. Based on those results the recommendation is to maintain the following traits for future research with this database: gait score, height at croup, length of back, length of croup, width of head and cannon bone circumference.(AU)

Developmental aspects of early pregnancy in mares

In mares, the embryo mi grates to the uterus between days 5 and 6 postovulation beginning its mobility through all uterine segments, which is essential for the maternal recognition of pregnancy. During the mobility phase, the embryonic vesicle shows a linear growth rate until its fixation between days 15 and 17, when the orientation phenomenon occurs. From fixation to day 28 of pregnancy, the embryonic growth is less evident (plateau) by cross-section ultrasound examination. After this period the linear growth rate is reestablished until day 46. This plateau is attributed to the increased uterine tone that compresses the vesicle and to volume expansion, making it difficult to detect the conceptus growth only by the cross-section diameter. Around day 20, the embryo proper is visualized as an echogenic spot in the ventral aspect of the vesicle. Additionally, development of allantoic sac, embryonic heartbeat, yolk sac regression and posterior umbilical cord formation also can be visualized from days 20 to 40. An intimate interaction between uterus and conceptus is essential for the normal pregnancy development. Color- and spectral-Doppler ultrasonography can be useful for the evaluation of this interface. A gradual increase on uterine vascularity during the early pregnancy and transient changes in endometrial vascularity accompanying the vesicle location during the mobility phase have been described. Around day 38 of gestation, the formation of the endometrial cups begins and, consequently, the synthesis of the equine chorionic gonadotropin (eCG) induces the formation and development of supplementary corpora lutea, which are important to secrete progesterone and to maintain pregnancy until around day 120.

Developmental aspects of early pregnancy in mares

In mares, the embryo mi grates to the uterus between days 5 and 6 postovulation beginning its mobility through all uterine segments, which is essential for the maternal recognition of pregnancy. During the mobility phase, the embryonic vesicle shows a linear growth rate until its fixation between days 15 and 17, when the orientation phenomenon occurs. From fixation to day 28 of pregnancy, the embryonic growth is less evident (plateau) by cross-section ultrasound examination. After this period the linear growth rate is reestablished until day 46. This plateau is attributed to the increased uterine tone that compresses the vesicle and to volume expansion, making it difficult to detect the conceptus growth only by the cross-section diameter. Around day 20, the embryo proper is visualized as an echogenic spot in the ventral aspect of the vesicle. Additionally, development of allantoic sac, embryonic heartbeat, yolk sac regression and posterior umbilical cord formation also can be visualized from days 20 to 40. An intimate interaction between uterus and conceptus is essential for the normal pregnancy development. Color- and spectral-Doppler ultrasonography can be useful for the evaluation of this interface. A gradual increase on uterine vascularity during the early pregnancy and transient changes in endometrial vascularity accompanying the vesicle location during the mobility phase have been described. Around day 38 of gestation, the formation of the endometrial cups begins and, consequently, the synthesis of the equine chorionic gonadotropin (eCG) induces the formation and development of supplementary corpora lutea, which are important to secrete progesterone and to maintain pregnancy until around day 120.(AU)

Avaliação genética para peso corporal em um rebanho Nelore / Genetic evaluation for body weight in a Nellore herd

Estimaram-se as herdabilidades para os efeitos genéticos direto e materno e as correlações genéticas entre essas variáveis para os pesos ao desmame (P205), ao ano (P365) e ao sobreano (P550) em um rebanho Nelore do norte de Minas Gerais. O modelo estatístico incluiu os efeitos aditivos direto e materno, os efeitos fixos de grupo de contemporâneos (fazenda, sexo, regime alimentar, estação (seca e água) e ano de nascimento do animal) e o efeito da covariável idade da vaca ao parto (linear e quadrático). Os componentes de variância e os valores genéticos foram estimados utilizando-se o método REML. A tendência genética foi obtida utilizando-se a regressão do valor genético médio anual em relação ao ano de nascimento dos animais. As estimativas de herdabilidade do efeito aditivo direto () para P205, P365 e P550 foram 0,60, 0,69 e 0,75, respectivamente. Estes coeficientes de são de alta magnitude, indicando que o rebanho apresenta variabilidade genética aditiva relativa e, portanto, espera-se progresso genético considerável utilizando a seleção. Pela análise da tendência genética, verificou-se que houve evolução nos valores genéticos dos animais ao longo dos anos estudados.

The heritabilities for direct and maternal genetic effects and genetic correlations between these effects were estimated for weight at 205 (P205), 365 (P365), and 550 days (P550) in a Nelore herd in northern Minas Gerais. The statistical model included direct and maternal additive effects, in addition to the fixed effects of contemporary group (farm, gender, diet, season - dry and water -, and year of birth) and the covariate age of cow at calving (linear and quadratic effects). The variance components and genetic values were estimated by REML method. The genetic trend was obtained using the regression of the annual mean genetic value in relation to the year birth. The heritability estimates for the direct additive effect () for P205, P365, and P550 were equal to 0.60, 0.69, and 0.75, respectively. These coefficients showed high magnitude, indicating that the herd in question presents a great additive genetic variability and therefore it is expected a great progress using genetic selection. By the analysis of genetic trend, it was verified a development in animals genetic values over the years studied.

Avaliação genética para peso corporal em um rebanho Nelore / Genetic evaluation for body weight in a Nellore herd

Estimaram-se as herdabilidades para os efeitos genéticos direto e materno e as correlações genéticas entre essas variáveis para os pesos ao desmame (P205), ao ano (P365) e ao sobreano (P550) em um rebanho Nelore do norte de Minas Gerais. O modelo estatístico incluiu os efeitos aditivos direto e materno, os efeitos fixos de grupo de contemporâneos (fazenda, sexo, regime alimentar, estação (seca e água) e ano de nascimento do animal) e o efeito da covariável idade da vaca ao parto (linear e quadrático). Os componentes de variância e os valores genéticos foram estimados utilizando-se o método REML. A tendência genética foi obtida utilizando-se a regressão do valor genético médio anual em relação ao ano de nascimento dos animais. As estimativas de herdabilidade do efeito aditivo direto (H2O) para P205, P365 e P550 foram 0,60, 0,69 e 0,75, respectivamente. Estes coeficientes de H2O são de alta magnitude, indicando que o rebanho apresenta variabilidade genética aditiva relativa e, portanto, espera-se progresso genético considerável utilizando a seleção. Pela análise da tendência genética, verificou-se que houve evolução nos valores genéticos dos animais ao longo dos anos estudados.(AU)

The heritabilities for direct and maternal genetic effects and genetic correlations between these effects were estimated for weight at 205 (P205), 365 (P365), and 550 days (P550) in a Nelore herd in northern Minas Gerais. The statistical model included direct and maternal additive effects, in addition to the fixed effects of contemporary group (farm, gender, diet, season - dry and water -, and year of birth) and the covariate age of cow at calving (linear and quadratic effects). The variance components and genetic values were estimated by REML method. The genetic trend was obtained using the regression of the annual mean genetic value in relation to the year birth. The heritability estimates for the direct additive effect (H2O) for P205, P365, and P550 were equal to 0.60, 0.69, and 0.75, respectively. These coefficients showed high magnitude, indicating that the herd in question presents a great additive genetic variability and therefore it is expected a great progress using genetic selection. By the analysis of genetic trend, it was verified a development in animals genetic values over the years studied.(AU)

Avanços na criopreservação de embriões equinos / Advances of equine embryo cryopreservation

A aplicação de técnicas de preservação pelo frio em embriões equinos se tornou indispensável na atual estrutura imposta à indústria equina. A refrigeração, a congelação ou a vitrificação são técnicas que podem ser utilizadas com sucesso na reprodução assistida desta espécie. Entretanto, os embriões equinos têm características que os diferenciam das demais espécies, exigindo que os protocolos sejam adequados aos diferentes estádios dos embriões. Em função da escassez de literatura com criopreservação de embriões equinos, esta revisão explora tanto a refrigeração de embriões visando a seu transporte durante a estação ovulatória, quanto à congelação e vitrificação visando a sua utilização futura, seja na transferência a uma receptora de melhor qualidade ou na otimização da produção quando necessita seguir uma estação reprodutiva determinada pelo ano hípico. O sucesso da criopreservação depende de vários fatores, dos quais se destacam o diâmetro dos embriões no momento da criopreservação, pois os embriões nas fases iniciais de desenvolvimento (<300 μm) são mais resistentes ao processo de congelação ou vitrificação, bem como o tipo de crioprotetor e a metodologia empregada. Este trabalho tem como objetivo descrever diferentes aspectos relacionados com as características do embrião equino, bem como levantar as metodologias e os respectivos resultados descritos na literatura.

The use of low temperature techniques to preserve embryos became essential at the actual level of the equine industry. The cooling, freezing, and vitrification are successful techniques applied on assisted reproduction for this specie. This review explores extensively the equine embryos cryopreservation aiming their transport during the ovulatory season. Furthermore, the utilization of freezing and vitrification aiming the storage and future use of embryos, and the production optimization in cases when a breeding season is determined by racing calendar are considered. Accomplishment of these techniques depends on the embryos diameter at the cryopreservation moment. Smaller embryos at early stages of development (<300μm) are more resistant to freezing or vitrification processes due to less amount of fluid present at the blastocoele and a not completed developed embryonary capsule, allowing a better cellular dehydration and consequently better post-transfer pregnancy rates.

Avanços na criopreservação de embriões equinos / Advances of equine embryo cryopreservation

A aplicação de técnicas de preservação pelo frio em embriões equinos se tornou indispensável na atual estrutura imposta à indústria equina. A refrigeração, a congelação ou a vitrificação são técnicas que podem ser utilizadas com sucesso na reprodução assistida desta espécie. Entretanto, os embriões equinos têm características que os diferenciam das demais espécies, exigindo que os protocolos sejam adequados aos diferentes estádios dos embriões. Em função da escassez de literatura com criopreservação de embriões equinos, esta revisão explora tanto a refrigeração de embriões visando a seu transporte durante a estação ovulatória, quanto à congelação e vitrificação visando a sua utilização futura, seja na transferência a uma receptora de melhor qualidade ou na otimização da produção quando necessita seguir uma estação reprodutiva determinada pelo ano hípico. O sucesso da criopreservação depende de vários fatores, dos quais se destacam o diâmetro dos embriões no momento da criopreservação, pois os embriões nas fases iniciais de desenvolvimento (<300 μm) são mais resistentes ao processo de congelação ou vitrificação, bem como o tipo de crioprotetor e a metodologia empregada. Este trabalho tem como objetivo descrever diferentes aspectos relacionados com as características do embrião equino, bem como levantar as metodologias e os respectivos resultados descritos na literatura.(AU)

The use of low temperature techniques to preserve embryos became essential at the actual level of the equine industry. The cooling, freezing, and vitrification are successful techniques applied on assisted reproduction for this specie. This review explores extensively the equine embryos cryopreservation aiming their transport during the ovulatory season. Furthermore, the utilization of freezing and vitrification aiming the storage and future use of embryos, and the production optimization in cases when a breeding season is determined by racing calendar are considered. Accomplishment of these techniques depends on the embryos diameter at the cryopreservation moment. Smaller embryos at early stages of development (<300μm) are more resistant to freezing or vitrification processes due to less amount of fluid present at the blastocoele and a not completed developed embryonary capsule, allowing a better cellular dehydration and consequently better post-transfer pregnancy rates. (AU)

The contextual interference effect in acquisition of dart-throwing skill tested on a transfer test with extended trials.

Previous studies of contextual interference have shown that practicing several motor skills randomly (high contextual interference) facilitates retention and transfer in comparison to practicing the same tasks in a blocked order (low contextual interference). However, many studies have not supported this phenomenon in motor learning, and some researchers have questioned whether the effect can be tested by using only a few trials on the transfer test. The present study used a different methodological approach in which the number of test trials was increased to assess whether the contextual interference effect is sustained over an extended number of trials in the transfer phase. Undergraduate students (N=32) were randomly allocated to either a blocked or random group. The participants practiced 80 acquisition trials in the dart-throwing task from distances of 300 cm and 420 cm, using two different grips. The transfer test after a 10-min. interval consisted of 40 trials with a new grip at a distance of 360 cm. The results did not support the contextual interference effect since there were no significant differences between groups on transfer. These findings suggest that the contextual interference effect may not be a global learning phenomenon which can be generalized to all learning situations.

River water quality improvement by natural and constructed wetland systems in the tropical semi-arid region of northeastern Brazil.

The efficiencies of a natural Typha spp wetland (Wn) formed on a river bed and its effluent treatment in a constructed wetland (Wc, subsurface horizontal flow) were investigated in northeastern Brazil (Paraiba State). The Wc system (12 tanks with stone gravel, 4.13 m2, 0.22 m3, 20 Typha spp rhizomes, m(-2) each, with 38, 29, and 19 mm x d(-1) hydraulic loadings, and 5, 7, and 10 days HRT) was fed daily with effluent from a Wn. Wn removal presented the highest values after Typha spp were cut during the 5th week. Removal values were (1st and 2nd periods or before and after cutting): 75% and 81% BOD5; 10-53% total phosphorus; 13%-55% ammonia; 89%-91% FC; 90-96% coliphages and bacteriophages. Wc removals increased with time with best results on 10 d HRT. Removals were also higher in the 2nd period: 74%-78% BOD5; 58%-82% ammonia; 90% FC; 94-98% FS; and 92%-96% coliphages and bacteriophages. Despite the high remaining values of FC (1.4 x 10(4) CFU/100 ml) and FX (4 x 10(3) CFU/100 ml), the removals were satisfactory and HRT dependent, suggesting a gradual optimization of the system with time. The Wc exhibited good efficiency for improving water quality from polluted river.

Ovarian activity and plasma concentrations of progesterone and estradiol during pregnancy in jennies.

The objectives of this study were to determine ovarian activity (with ultrasound) and plasma concentrations of progesterone and estradiol during pregnancy in jennies. There was considerable ovarian activity during the second month of pregnancy. Secondary corpora lutea (total of 2 to 7 per jenny) were formed (mainly by luteinization) starting on Day 41.8 +/- 1.0 (range Days 38 to 46; ovulation = Day 0). The echogenicity of the primary and secondary corpora lutea gradually decreased during pregnancy. Plasma progesterone concentrations increased between Days 0 and 10 (0.9 and 19.9 ng/mL, respectively), gradually decreased to Day 30 (12.1 ng/mL), increased between Days 30 and 40 (plateau, at approximately 17 ng/mL), gradually declined from Days 110 to 160 (nadir of approximately 6 ng/mL), and remained nearly constant until increasing again just before parturition. Plasma estradiol concentrations increased gradually from Day 65, peaked (1.2 ng/mL) on Day 165 (> or = 1 ng/mL on Days 150 to 210), and decreased thereafter, with very low concentrations during the last 20 d before parturition. Ovarian function and hormone profiles were generally similar to those previously reported during pregnancy in mares.

Ultrasonographic evaluation of the conceptus from days 10 to 60 of pregnancy in jennies.

Daily ultrasound examinations were conducted from Days 10 to 60 (ovulation = Day 0) of pregnancy to monitor the conceptus in jennies (n = 12). The embryonic vesicle was first detected on Day 11.5 +/- 0.9 (mean +/- SD; range 10 to 13 d) and was mobile until movement ceased (fixation) on Day 15.5 +/- 1.4 (range, 13 to 18 d). The vesicle was spherical from Days 10 to 18 (mean growth rate, 3.2 mm/d), non spherical (irregular) with a reduced growth rate (0.5 mm/d) from Days 19 to 29, and then grew at a moderate rate (1.6 mm/d) up to Day 46. On average, detection of the embryo proper (consistently located on the ventral aspect of the yolk sac) and embryonic heartbeat were Days 20.7 +/- 1.2 and 23.5 +/- 1.3, respectively. Formation of the allantoic sac was first detected on Day 24.4 +/- 1.7 and was complete on Day 36.8 +/- 1.6. Descent of the fetus (and formation of the umbilical cord) began on Day 37.9 +/- 1.7 and was complete on Day 44.1 +/- 2.1. Crown-rump length averaged 3.7, 15.4, 22.7, 37.5 and 59.6 mm on Days 20, 30, 40, 50 and 60, respectively. In general, morphologic features and dates of occurrence were similar to those reported previously in the mare.

Morte embrionária precoce em éguas: aspectos clínicos e hormonais / Early embryonic death in mares: clinical and hormonal aspects

O presente estudo objetivou diagnosticar as causas da morte embrionária precoce em 128 éguas. Amostras de soro foram coletadas nos dias 4, 7, 10, 13, 16, 19, 21 e 30 após ovulaçäo, para dosagens hormonais e mensuraçöes do corpo lúteo. O diâmetro e as características da vesícula embrionária foram avaliados, através da ultra-sonografia, a partir do 12º dia pós-ovulaçäo. Das 128 éguas estudadas, 17 (13.28 por cento) apresentaram morte embrionária. O diâmetro do corpo lúteo, bem como as concentraçöes plasmáticas de progesterona, foi semelhante nos grupos que apresentaram morte embrionária e nos que mantiveram a gestaçäo. Os níveis de estrogeno plasmáfico foram mais elevados no grupo das fêmeas que mantiveram a gestaçäo. Os exames citológicos, microbiológicos e histopatológicos revelaram que a maioria das éguas com diagnóstico de morte embrionária eram portadoras de endometrites

Follicular dynamics in Mangalarga mares.

Ovarian follicular activity was studied by ultrasonography during 17 oestrous cycles in 9 Mangalarga mares during the second half of the ovulatory season. Sixteen oestrous cycles were considered normal and one 3-wave cycle showing a prolonged luteal phase was considered atypical. Daily ultrasonographic examinations were performed and the compiled data on follicular dynamics were studied retrospectively. One major wave of follicular growth was observed in 13 of the 16 normal cycles (81.25%), whereas 2 major waves occurred in 3 cycles (18.75%). The mean (+/- s.d.) days of emergence of the primary wave of follicular development in cycles containing one or 2 waves were Day 6.0 +/- 2.3 and Day 11.0 +/- 1.0, respectively. The secondary wave of follicular development in 2-wave cycles emerged on Day 0.0 +/- 3.6. The day of wave divergence for primary waves of follicular development in cycles which exhibited one or 2 major waves were Day 12.2 +/- 3.5 and Day 17.3 +/- 3.0, respectively. Divergence of secondary waves occurred in only one of the 3 cycles which exhibited 2 major follicular waves (Day 7). The mean (+/- s.d.) maximum diameters of the dominant follicle in the primary wave of oestrous cycles exhibiting one and 2 major waves were 39.0 +/- 3.9 mm and 34.7 +/- 2.5 mm, respectively. The mean (+/- s.d.) maximum diameter of the dominant follicle present in the secondary wave was 34.3 +/- 11.0 mm. The mean (+/- s.d.) lengths of the interovulatory intervals for cycles containing one and 2 major waves were 19.4 +/- 2.2 and 23.3 +/- 2.5 days, respectively. These data indicate that most Mangalarga mares show one major follicular wave during the oestrous cycle but a small percentage of mares show 2 major waves.

Cryopreservation of equine embryos with glycerol plus sucrose and glycerol plus 1,2-propanediol.

Six or 7-day-old equine embryos were divided into 4 groups; Group 1, n = 15, Day 7 embryos destined for immediate transfer; Group 2, n = 15, Day 6 embryos destined for deep-freezing with glycerol plus sucrose as cryoprotectant; Group 3, n = 10, Day 6 embryos destined for deep-freezing with glycerol plus 1,2-propanediol as cryoprotectant and Group 4, n = 3, fresh embryos destined for ultrastructural analysis. All the frozen/thawed embryos were transferred to recipient mares, except 3 embryos in Group 3 that were subjected to ultrastructural analysis. After thawing the cryoprotectants were removed by successive dilutions in PBS + 15% v:v fetal calf serum (FCS) containing decreasing concentrations of the cryoprotectants. Pregnancy was diagnosed ultrasonographically in 53.3%, 13.3% and 0% of the mares in Groups 1, 2 and 3 respectively. Ultrastructural analysis showed differences between frozen/thawed and fresh embryos. In the former, embryonic cells were deformed and showed dilation of the intercellular and perivitelline spaces, a decrease of desmosome number in the junctional complexes, few microvilli on the apical surface of the trophectoderm and an almost total absence of pinocytotic vesicles. Most of the mitochondria showed regions containing dilation and irregularities on the cristae, which appeared electron-dense. The results obtained with Groups 2 and 3 embryos showed that the cryoprotectants employed were not effective in protecting the embryos against damage during freezing and thawing. Indeed, the ultrastructural changes observed in the Group 3 embryos explained the absence of any established pregnancies in this group of mares.