Progression of micturition dysfunction associated with the development of heart failure in rats: Model of overactive bladder.

Heart failure (HF) has a strong association with the development of lower urinary tract symptoms, especially overactive bladder (OAB); although this condition remains poorly investigated. In this study, we assess the aortocaval fistula (ACF) model as a novel experimental model of micturition dysfunction, associated with HF, focused on the molecular and functional studies to evaluate the autonomic nervous system and urinary bladder remodeling. Male rats were submitted to ACF for HF induction. Echocardiography, cystometric, histomorphometry and molecular analysis, as well as concentration-response curves to carbachol and ATP and frequency-response curves to electrical field stimulation (EFS) were evaluated in Sham and HF (4- and 12-weeksendpoint) groups. Compared to SHAM, HF groups exhibited progressive increases in the left ventricle (LV) mass and fractional shortening which indicates cardiac dysfunction, although HF was characterized only after 12 weeks by the reduced ejection fraction. For micturition function, HF groups presented increased non-voiding contractions (NVC) and decreased bladder capacity; however, when comparing HF groups, these urinary parameters were significantly impaired over the weeks (12-weeks). The contractile responses induced by CCh, ATP and EFS were greater in detrusor muscle (DSM) from HF rats. mRNA expression for muscarinic receptors (M2 and M3) was higher in DSM only after 12 weeks of ACF, in addition to MMP9 and TGF-beta. Histomorphometric revealed increased urothelium thickness in both HF groups, whereas DSM thickness occurred only after 12 weeks. Thus, the ACF model induced cardiac dyfunction with progressive micturition dysfunction over the weeks, characterized by increased DSM contractile mechanisms as well as extracellular matrix remodeling in the urinary bladder, representing a useful tool to evaluate the OAB associated with HF.

Five cases of nosocomial and community-acquired Legionnaires' disease in São Paulo, Brazil.

Legionella sp has been emerging over the last decade as an important cause of pneumonia both hospital and community-acquired. Following an outbreak in a Renal-Transplant Unit stocked serum was tested for antibodies against Legionella pneumophila serogroup 1, and 5 cases of Legionnaires' Disease were reviewed. Two of the cases were nosocomial and three cases were community-acquired. Clinical and laboratorial aspects were similar to those expected for other causes of pneumonia, however jaundice was encountered in two cases. This study suggests that the real incidence of pneumonia caused by Legionella sp is being underestimated and the authors emphasize the importance of considering Legionnaires' Disease when empirically treating community-acquired pneumonia.

Legionnaires' disease in the renal transplant unit of "Hospital das Clinicas, FMUSP". During a five year period (1988-1993).

Several reports have related Legionella pneumophila with pneumonia in renal transplant patients, however this association has not been systematically documented in Brazil. Therefore this paper reports the incidence, by serological assays, of Legionella pneumophila serogroup 1 in these patients during a five year period. For this purpose sera from blood samples of 70 hospitalized patients with pneumonia from the Renal Transplant Unit of Hospital das Clinicas, FMUSP collected at the acute and convalescent phase of infection were submitted to indirect immunofluorescence assay (IFA) to demonstrate anti-Legionella pneumophila serogroup 1 antibodies. Of these 70 patients studied during the period of 1988 to 1993, 18 (25.71%) had significant rises in specific antibody titers for Legionella pneumophila serogroup 1. Incidence was interrupted following Hospital water decontamination procedures, with recurrence of infections after treatment interruption. In this study, the high susceptibility (25.71%) of immunodepressed renal transplant patients to Legionella pneumophila serogroup 1 nosocomial infections is documented. The importance of the implementation and maintenance of water decontamination measures for prophylaxis of the infection is also clearly evident.

Evaluation of a rapid screening assay for bacterial identification (Dot-ELISA) in fecal samples from children.

With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA.

[Uremia, hydroelectrolytic and acid-base changes in kidney failure of chronic hepatopathies]. / Estudo da uremia, alterações hidroelectrolíticas e ácido-básicas na insuficiência renal de hepatopatias crônicas.

The author presents 16 patients with chronic liver diseases that developed renal failure. The author emphasizes the picture characteristics as prolonged evolution of non-oliguric renal failure tending to the rise of serum sodium in 2 cases and fall of serum potassium and less seriousness of metabolic acidosis compensated by respiratory alkalosis, and a further patient with metabolic alkalosis and paradoxal acidury. The collection of these elements perhaps explain the evolution of the diseases in the patients without dialysing treatment measures. The clinic pictures should be considered as part of the alterations of terminal hepatic failure.

[Determination of lactic acid in blood by enzymatic-spectrophotometry method in normal subjects]. / Determinação do ácido láctico no sangue pelo método enzimático-espectrofotométrico em indivíduos normalis.

The authors carried out duplicate dosages of lactic acid in 20 normal patients (11 females and 9 males) ages ranged from 17 to 48 years. The spectrophotometric method was employed, and the values obtained from 0.69 to 4.21 mMol/L with a mean value of 1.5095 +/- 0.975. Some of the difficulties of the method and how to avoid them are discussed.

[Association of hepatosplenic schistosomiasis mansoni and histiocytic lymphoma]. / Associação da esquistossomose mansônica hepatoesplênica e linfoma histiocítico.

A case of one patient with hepatosplenic Schistosomiasis mansoni in association with histiocytic lymphoma is presented. The authors discuss the concomitance of the two pathologic processes, by comparing with other cases of the literature and speculating about its significance, there is the possibility of the schistosomiasis being related as the causing factor of the malignant disease.

[Two cases of severe cutaneous vasculitis with thrombocytopenia associated with hepatic cirrhosis: autoimmune and infective-inflammatory component with endothelial lesion and restrictive pulmonary pattern in one case]. / Dois casos de vasculite cutânea grave com plaquetopenia associada à cirrose hepática: componente autoimune e infecto-inflamatório com lesão endotelial e padrão restritivo pulmonar em um caso.

Two clinical cases of female patients with hepatic cirrhosis and autoimmune multisystemic involvement with infectious intercurrent are reported. Case 1 presented infective endocarditis and erysipelas on the left thigh. In the course of the clinical picture a cutaneous vasculitis developed in the same place together with autoimmune thrombocytopenia, leukopenia and pulmonary restrictive picture with inflammatory pattern. There are also elevate immune complexes and complement consumption. Case 2 presented erysipelas on the left thigh cutaneous vasculitis and complement consumption. In Case 1 the infective endocarditis was treated with antibiotic therapy during 4 weeks followed by 1 mg/kg corticoid (Prednisone) with thrombocytopenia and leukopenia reversion. Case 2 presented an improvement with antibiotic only. The relation between chronic liver diseases and systemic autoimmune phenomena is commented, special attention being paid to the cutaneous, hematological and pulmonary affection.

[Clinical, biochemical and histopathological development of post-transfusional non-A, non-B hepatitis from the acute picture to chronicity during 13.3 years]. / Evolução clínico-bioquímica e histopatologica de hepatite não-A, não-B pós-transfusional, do quadro agudo até a cronicidade, com duração de 13.3 anos.

This case was of a 45 year old female patient with a post-transfusion non-A non-B hepatitis which was accompanied since an acute phase to hepatic cirrhosis during a period of 159.7 months or 13.3 years. Four hepatic biopsies were carried out and they divided the follow-up into 5 evolutive periods. The biopsies revealed a progressive histologic from chronic persistent hepatitis to an active chronic hepatitis and cirrhosis. The aminotransferases followed a floating course in the whole period, with ALT greater than AST starting from the 3rd period. The 3rd period (from 5th to 8th year) was of least activity of the aminotransferases, and the 4th and 5th periods (from 8th to 13th year) showed the highest activity of ALT. The 2nd period (from 3rd to 5th year) showed the least portion of gamma globulin and the highest of albumin in comparison with the others. There was no connection between the levels of aminotransferases and the values of gamma globulin and albumin in the follow up process. The treatment employed in the 5th evolutive period (prednisone and colchicine) did not present any biochemical improvement.

Prebiótico, probiótico e simbiótico para larvas de Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae) / Prebiotic, probiotic and symbiotic for Trichogaster leeri larvae (Bleeker, 1852, Perciformes, Osphronemidae)

Avaliou-se a suplementação de prebiótico (mananoligossacarídeo - 2g/kg), probiótico (Bacillus subtilis, Cohn, 1872, Bacillales, Bacillaceae - 2g/kg) e simbiótico (mananoligossacarídeo - 2g/kg + B. subtilis - 2g/kg), em rações para larvas de Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae), durante a transição alimentar, sobre o crescimento, a sobrevivência, a morfometria intestinal e a resistência ao estresse por exposição aérea. Para isso, 420 larvas (0,45±0,05mg) foram distribuídas em 20 aquários (3,5L), em delineamento inteiramente ao acaso, com cinco tratamentos e quatro repetições, durante 28 dias, a saber: 12 dias de alimento vivo (AV) + oito dias de coalimentação (AV + ração controle) + oito dias de ração controle; 12 dias de AV + oito dias de coalimentação (AV + ração prebiótico) + oito dias de ração prebiótico; 12 dias de AV + oito dias de coalimentação (AV + ração probiótico) + oito dias de ração probiótico; 12 dias de AV + oito dias de coalimentação (AV + ração simbiótico) + oito dias de ração simbiótico; 28 dias de AV. Larvas que receberam apenas AV apresentaram resultados de desempenho zootécnico significativamente superiores às larvas dos demais tratamentos, exceto para taxa de crescimento específico em comprimento e sobrevivência, que foram semelhantes aos resultados obtidos por larvas que receberam rações suplementadas com simbiótico. Larvas que receberam a ração controle apresentaram resultados significativamente inferiores para altura das vilosidades intestinais e taxa de resistência ao estresse comparando-se aos demais tratamentos. Os resultados deste estudo indicam que a suplementação com prebiótico, probiótico e simbiótico exerce efeito positivo sobre a sobrevivência, a morfometria intestinal e a resistência ao estresse em larvas de T. leeri, no entanto somente a suplementação com simbiótico resulta em melhora no crescimento.(AU)

The prebiotic (mannan oligosaccharide - 2g/kg), probiotic (Bacillus subtilis Cohn, 1872, Bacillales, Bacillaceae - 2g/kg) and symbiotic (mannan oligosaccharide - 2g/kg + Bacillus subtilis - 2g/kg) supplementation in diets during food transition in Trichogaster leeri (Bleeker, 1852, Perciformes, Osphronemidae) larvae on growth, survival, intestinal morphology and stress resistance to air exposure was evaluated. For this, 420 larvae (0.45±0.05mg) were distributed in 20 aquaria (3.5L) in a completely randomized design with five treatments and four replications for 28 days, namely: 12 days of live food (LF) + eight days of co-feeding (LF + control diet) + eight days of control diet; 12 days of LF + eight days of co-feeding (LF + prebiotic feed) + eight days of prebiotic feed; 12 days of LF + eight days of co-feeding (LF + probiotic feed) + eight days of probiotic feed; 12 days of LF + eight days of co-feeding (LF + symbiotic feed) + eight days symbiotic feed; 28 days of LF. Larvae fed only LF showed growth performance significantly higher than the larvae of other treatments, except for specific growth rate in length and survival that were similar to the results obtained by larvae fed diets symbiotically supplemented. Larvae fed the control diet showed significantly lower results for height of intestinal villi and stress resistance rate comparing to other treatments. The results of this study indicate that prebiotic, probiotic and symbiotic supplementation has a positive effect on survival, intestinal morphology and resistance to stress in larvae of T. leeri, however, only symbiotic supplementation results in improved growth.(AU)

Estratégia de coalimentação na sobrevivência e no crescimento de larvas de Betta splendens durante a transição alimentar / Co-feeding strategy on survival and growth of Betta splendens larvae during weaning

Conduziu-se um experimento com o objetivo de avaliar a influência do período de coalimentação na sobrevivência e no crescimento de larvas de Betta splendens. As larvas foram alimentadas com náuplios de Artemia durante sete dias e, posteriormente, submetidas aos seguintes tratamentos: jejum contínuo; ração contínua; quatro dias de coalimentação + 14 dias de ração; oito dias de coalimentação + 10 dias de ração; 12 dias de coalimentação + seis dias de ração; náuplios de Artemia durante todo o período experimental. Os valores das variáveis de desempenho das larvas do tratamento 12 dias de coalimentação + seis dias de ração e do tratamento náuplios de Artemia foram superiores aos demais tratamentos, exceto para sobrevivência, em que não houve diferença significativa com o tratamento oito dias de coalimentação + 10 dias de ração. Considerando-se apenas o tempo de oferta de Artemia (em dias), obteve-se a equação de regressão para as variáveis analisadas. A sobrevivência e a taxa de crescimento específico (TCE) apresentaram efeito quadrático, e as demais variáveis apresentaram efeito linear. O ponto de máxima para sobrevivência foi de 21,7 dias (88,92%) e para TCE foi de 26,2 dias (23,47% dia-1). O período de co-feeding influencia no crescimento e na sobrevivência de larvas de Betta splendens. Após o período de 19 dias de oferta de alimento vivo, com 12 dias de coalimentação, as larvas estão aptas a aproveitar de maneira eficiente o alimento inerte sem prejuízos ao crescimento e à sobrevivência.

An experiment was carried out to evaluate the influence of the period of co-feeding on survival and growth of Betta splendens larvae. The following strategies for co-feeding were used: continuous fasting, 18 days of inert diet, 4 days of co-feeding + 14 days of inert diet; 8 days of co-feeding + 10 days of inert diet; 12 days of co-feeding + 6 days of inert diet, nauplii of Artemia throughout the experiment period. The values of performance variables for larvae with 12 days of treatment for co-feeding + 6 days of inert diet and the nauplii treatment with Artemia were higher than the other treatments, except for survival, which did not differ significantly from the treatment with 8 days of co-feeding + 10 days of inert diet. Considering only the supply time for Artemia (in days) the regression equation was obtained for the variables analyzed. The survival and specific growth rate (SGR) showed a quadratic effect and other variables showed a linear effect. The point of maximum survival was 21.7 days (88.92%) and for SGR it was 26.2 days (23.47 % day-1). The co-feeding period influences the survival and growth of the Betta splendens larvae. After the period of 19 days of supply of live food, with 12 days of co-feeding, larvae can be fed exclusively on inert diet, without negative effects on growth and survival.

Níveis de carnitina na ração no desempenho corporal de tricogáster léri (Trichogaster leeri bleeker, 1852) / Carnitine levels in feed of body performance of the gouramis (Trichogaster leeri bleeker, 1852)

Foram avaliados os efeitos de carnitina na dieta de Trichogaster leeri, com peso inicial de 0,5±0,2g, distribuídos aleatoriamente e individualmente em aquários plásticos (1000mL), com renovação total da água a cada 48 horas. Cinco dietas isocalóricas, contendo 28 por cento de proteína bruta e 3100kcal de energia bruta, foram elaboradas com a suplementação de T1 = 0mg, T2 = 300mg, T3 = 600mg, T4 = 900mg e T5 = 1200mg de l-carnitina/kg de ração. As dietas foram administradas três vezes ao dia, ad libitum, durante 45 dias. Foram avaliados comprimento total e padrão, altura, peso final e tamanho de cauda, além do ganho de peso, conversão alimentar e fator de condição. Em todas as características analisadas, o tratamento de 900mg de carnitina/kg foi o mais eficiente no desempenho zootécnico para o T. leeri, e, após cálculos dos pontos máximos, sugere-se a concentração de 1.000mg/kg de ração. Observou-se diferença significativa para tamanho de cauda, altura do corpo, peso final e fator de condição.

The effects of carnitine were evaluated in diets fed to Trichogaster leeri, initial weight 0.5±0.2g and individually randomized into plastic containers (1000mL), with total renewal of water every 48 hours. Five isocaloric diets containing 28 percent crude protein and 3100 kcalof gross energy were made with the supplementation of: T1= 0 mg; T2= 300mg;T3= 600mg; T4= 900mg; and T5= 1200mg of l-carnitina/kg feed. The diets were administered three times daily ad libitum for 45 days. Total and standard length, height, weight and size of the tail end, in addition to weight gain, feed conversion ratio and condition factor were evaluated. In all parameters examined the treatment of 900 mg of carnitine/kg was the most efficient on the performance for the T. leeri, and after calculating the maximum points, a concentration of 1.000mg/kg feed is suggested. There was significant difference in tail length, body height, final weight and condition factors.

Fecundidade da manjuba Curimatella lepidura (CURIMATIDAE) na represa de Três Marias, rio São Francisco, MG / Fecundity of manjuba Curimatella lepidura (CURIMATIDAE) in the Três Marias reservoir, São Francisco river, MG

Foram determinados os tamanhos dos ovócitos e as fecundidades residual e relativa ao peso de corpo, ao comprimento padrão, ao peso do ovário e ao peso de corpo do líquido dos ovócitos vitelogênicos, e a fecundidade absoluta de 230 fêmeas de manjubas (Curimatellalepidura) coletadas no reservatório de Três Marias, rio São Francisco, MG durante três anos de coleta. Foi utilizado o método gravimétrico para determinar a quantidade de ovócitos vitelogênicos, que foram separados dos avitelogênicos, por diferença de peso. O diâmetro médio dos ovócitos foi de 711±44µm. O valor médio da fecundidade absoluta foi de 60.994,9±27.142,7 ovócitos por fêmea no estádio 2C de maturação (maduro). A fecundidade média relativa ao peso corporal e ao peso do ovário foi de 1.294,1±437,5 e 8.293,8±1.961,9 ovócitos por grama, respectivamente. Não houve diferença significativa (P<0,05) entre as fecundidades absolutas médias nos três anos de pesquisa. Foram encontrados elevados coeficientes de correlação, e os modelos lineares e potenciais foram os mais adequados para expressar as fecundidades absolutas e relativas, ou seja, quanto maior o grau de maturidade, maior a fecundidade dos peixes.

The sizes of vitellogenic oocytes and fecundity of 230 females of manjubas (Curimatella lepidura) from the Três Marias reservoir, river São Francisco, MG, was determined throughout three years of monthly captures. Gravimetric method was used to determine the quantities of vitellogenic oocytes, which were separated from no vitellogenics by weight difference and using water flow. The average diameter of vitellogenic oocytes was 711±44µm. The mean absolute fecundity was 60994.9±27142.7 oocytes per female in stage 2C of maturation (mature). The average fecundity relative to body weight and ovarian weight was 1294.1±437.5 and 8293.8±1961.9 oocytes per gram, respectively. There was no significant difference (P<0.05) between fecundity during the three years of research. High correlation coefficients were found, and the linear and potential models were the most appropriate to express the absolute and relative fecundities, i.e., the higher the maturity degree, higher the fecundity.

Efeito da granulometria do milho no desempenho de juvenis de pacu, Piaractus mesopotamicus (Holmberg, 1887) / Effect of corn granulometry on performance of juvenile pacu, Piaractus mesopotamicus Holmberg, 1887

Avaliou-se o efeito de diferentes granulometrias do milho da ração no desempenho de juvenis de pacu, Piaractus mesopotamicus. Foram utilizados 120 juvenis com peso inicial de 8,42±0,89, comprimento total de 7,03±0,20cm, comprimento padrão de 6,32±0,13cm e altura de 3,21±0,11cm, distribuídos em 20 aquários (300L), na densidade de seis peixes por unidade experimental (aquário), em delineamento inteiramente ao acaso, com cinco tratamentos constituídos por diferentes granulometrias - 850, 710, 500, 300, 150µm - do milho na composição da ração. Cada tratamento tinha quatro repetições. O tratamento que proporcionou melhor desempenho dos animais foi o com granulometria do milho de 150µm, destacando-se a conversão alimentar de 1,38, enquanto no tratamento com maior granulometria a conversão alimentar foi de 1,61. Assim, recomenda-se usar a granulometria de 150µm.

The effect of different particle size of corn ration on the performance of juvenile pacu, Piaractus mesopotamicus, was evaluated. A total of 120 juveniles with an initial weight of 8.42±0.89g, total length 7.03±0.20cm, standard length, 6.32±0.13cm, and height 3.21±0.11cm, were distributed into 20 tanks (300L), at a density of six fish per experimental unit (tank), in a completely randomized design with five treatments consisting of different particle sizes 850, 710, 500, 300, 150µm of corn in the ration composition. Each treatment had four replicates. Among all treatments, the best animal performance was achieved with the ration with corn particle size of 150µm, reaching a feed conversion ratio of 1.38, whereas for larger feed grain size, it was 1.61. Thus, the particle size of 150µm is recommended.

Randomized trial with itraconazole, ketoconazole and sulfadiazine in paracoccidioidomycosis.

Forty-two patients with active paracoccidioidomycosis were randomized to receive itraconazole (50-100 mg d(-1)), ketoconazole (200-400 mg d(-1)) or sulfadiazine (100-150 mg kg d(-1) up to 6 g d(-1)) for 4-6 months, followed by slow release sulfa until negativity of serological tests. All 14 patients in itraconazole and sulfadiazine groups and 13 in the ketoconazole group showed an adequate clinical response to the chemotherapy. One patient in the latter group showed treatment failure according to clinical and mycological criteria. The test of the hypothesis that the drugs reduced antibody levels up to ten months of treatment showed a p value equal to 0.0001 for itraconazole, 0.017 for ketoconazole and 0.0012 for sulfadiazine; this reduction was similar for the three groups. In this first randomized study for the treatment of paracoccidioidomycosis we could not show superiority of any one regimen over the others in the clinical and serological responses of patients with the moderately severe form of the disease.

Determinação da razão ótima de espermatozóides por ovócitos de piabanha Brycon insignis (pisces - characidae) / Determination of the optimum ratio of spermatozoa per oocyte of the piabanha Brycon insignis

Determinou-se a razão ótima de espermatozóides por ovócitos da piabanha Brycon insignis, utilizando-se dois machos e duas fêmeas da espécie, submetidos ao procedimento de desova induzida. Os gametas foram extrusados manualmente 200 horas-grau após a aplicação do extrato bruto de hipófise. Os ovócitos foram misturados e deste pool retiraram-se amostras com 2g de ovócitos (701 ovócitos/g). O sêmen do pool foi diluído em solução de NaCL 1,2 por cento de tal forma que, após a adição de 1ml do sêmen diluído aos ovócitos, fossem obtidas as seguintes razões espermatozóides por ovócitos em cada tratamento: T1=86.662, T2=173.324, T3=259.986, T4=346.648 e T5=433.310. A taxa de fertilização do sêmen não diluído usado como controle foi de 65,3 por cento. Após ativação espermática com NaHCO3 1 por cento e fecundação, os ovos foram transferidos para as incubadoras e nelas foram observadas as seguintes percentagens de fertilização em relação à do grupo-controle: T1=35,7 por cento, T2=53,1 por cento, T3=79,1 por cento, T4=93,4 por cento e T5=87,8 por cento. A percentagem de fertilização (em relação ao controle) aumentou de forma linear, segundo a equação de regressão: Y=15,55+0,0002297X (P<0,01; R²=0,98), até a proporção de 314.481 espermatozóides por ovócitos, após o que a taxa de fertilização se estabilizou no limite de 88 por cento

The optimum spermatozoa:oocyte ratio of piabanha Brycon insignis was studied. Two males and two females were induced to spawn, and the gametes were stripped after 200 hourgrades starting from the application of carp pituitary gland. Oocytes from two females were mixed, and samples of 2g (701 oocytes/g) were collected from the obtained pool and placed into plastic cups. The semen from the two males, after mixed to compose a pool, was diluted in NaCl solution (1.2 percent) so that, after the addition of 1ml of diluted sperm into the oocytes, the following spermatozoa:oocyte ratios (tri-replicated) were obtained: T1=86,662, T2=173,324, T3=259,986, T4=346,648 and T5=433,310. After the activation using 1 percent NaHCO3 and fertilization, the eggs were transferred to incubators, and observed the percentages of fertilization relative to the control. T1=35.7 percent, T2=53.1 percent, T3=79.1 percent, T4=93.4 percent and T5=87.8 percent. Undiluted semen was used as "control", which showed 65 percent of fertilization. The percentage of fertilization (relative to the control) increased linearly according to the regression Y= 15.55 + 0.0002297X (P<0.01; R²=0.98), until the proportion of 314,481 spermatozoa per oocyte, and, from this point, the fertilization rate maintained at 88 percent