Sanitary conditions of the Goliath grouper Epinephelus itajara in captivity on estuarine environment: case report / Condições sanitárias do mero Epinephelus itajara em cativeiro em ambiente estuarino: relato de caso

This study aimed to report the sanitary conditions through the hematological analysis of grouper E. itajara reared in captivity on estuarine conditions. Seven Goliath groupers (1,881.5±1,246.03g) were captured and kept in two tanks located on estuary. After 20 days, fish were collected for morphologic and hemato-physiologic evaluation. Two fish had clinical signs such as hemorrhagic spots and loss of scale due to agonistic behavior. Blood samples were collected, and the hematological parameters (biochemical, erythrogram and leukogram) were determined. Blood cells were characterized by their size, color and shape. Univariate statistic and principal components analysis were used to identify a hematological standard between fish with or without clinical signs. Four leukocyte types were found: lymphocyte, monocyte, neutrophil and basophil. Regardless of the clinical signs the cell morphology did not present any difference among the fish. However, there is a significant correlation between erythrocyte and lactate on fish with clinical signs. Thus, agonistics encountered among the fish is a stressing factor in captivity conditions making it necessary to have adequate management related to the size of fish and stocking density.(AU)

Este estudo avaliou as condições sanitárias do peixe mero E. itajara mantido sob cativeiro, em condições estuarinas, pelas análises hematológica e morfológica. Sete peixes (1.881,5±1.246,03g) foram capturados e mantidos em tanques localizados no estuário. Após 20 dias, os peixes foram coletados para avaliações morfológica e hematológica. Dois peixes tiveram sinais clínicos, como manchas hemorrágicas e perda de escamas devido ao comportamento agonístico entre os peixes. Amostras sanguíneas foram coletadas dos peixes anestesiados com auxílios de seringas umedecidas com EDTA 3%. Determinaram-se os parâmetros hematológicos (bioquímico, eritrograma e leucograma). Células sanguíneas foram caracterizadas por seu tamanho, cor e padrão. Estatística univariada e análises de componentes principais foram usadas para identificar um padrão hematológico entre os peixes com e sem sinais clínicos. Quatro tipos de leucócitos foram encontrados: linfócito, monócito, neutrófilo e basófilo. Os sinais clínicos não apresentaram diferença entre os peixes e a morfologia celular. Contudo, observou-se uma correlação entre os peixes com sinais clínicos e a quantidade de eritrócitos e lactato. Assim, encontros antagônicos entre os peixes são um fator estressante em condições de cativeiro, tornando-se necessário um manejo mais adequado relacionado ao tamanho dos indivíduos e à densidade de estocagem.(AU)

Efficacy of Ocimum gratissimum essential oil against the monogenean Cichlidogyrus tilapiae gill parasite of Nile tilapia / Eficácia do óleo essencial de Ocimum gratissimum contra o monogenea Cichlidogyrus tilapiae parasita de brânquias de tilápia-do-nilo

The phythotherapy is an alternative to use of chemotherapeutical agents against monogenean infection. This study evaluated the anthelmintic activity of essential oil Ocimum gratissimum against monogenean Cichlidogyrus tilapiae as well as its acute toxicity in tilapia juveniles. The mean lethal concentration (LC50) and different concentrations of the essential oil, both in vitro and in vivo assays (short and long-term baths) were assessed. The LC50 was 40.70mg.L-1 and in the in vitro assay this concentration showed 80% efficacy at the last two hours and in the in vivo assay 65.87% efficacy in long-term bath. However, it provoked morphological alterations on the gills such as hyperplasia and edema. The parasites exposure at the highest concentration (320mg.L-1) showed 100% mortality after 2h exposure in the in vitro assay, whereas in the in vivo assay, short-term baths of 5min for 3 consecutive days showed an efficacy of 87.71% without gills damage. These results demonstrate the anthelminthic activity of essential oil O. gratissimum and the safety concentration to use in Nile tilapia.(AU)

A fitoterapia é uma alternativa ao uso de agentes químicos contra infecções por parasitos monogenéticos. Este estudo avaliou a atividade anti-helmíntica do óleo essencial Ocimum gratissimum contra o monogenea Cichlidogyrus tilapiae , bem como sua toxicidade aguda e histopatologia em juvenis de tilápias. Foram avaliadas a concentração letal média (CL50) e diferentes concentrações de óleo essencial, tanto in vitro como in vivo (banho de curta e longa duração). A CL50 foi de 40,70mg.L-1; no ensaio in vitro, essa concentração apresentou 80% de eficácia, e no ensaio in vivo 65,87% de eficácia em banho de exposição crônica. No entanto, provocou alterações morfológicas nas brânquias, como hiperplasia e edema. A exposição dos parasitas na concentração mais elevada (320mg.L-1) mostrou 100% de mortalidade após duas horas de exposição no ensaio in vitro, enquanto no ensaio in vivo, em banho de curta duração de cinco minutos, durante três dias consecutivos, apresentou uma eficácia de 87,71%, sem danos às brânquias. Esses resultados demonstraram a atividade anti-helmíntica do óleo essencial de O. gratissimum e a concentração de segurança para uso na tilápia-do-nilo em banhos de curta duração.(AU)

Artificial neural networks as auxiliary tools for the improvement of bean plant architecture.

Classification using a scale of visual notes is a strategy used to select erect bean plants in order to improve bean plant architectures. Use of morphological traits associated with the phenotypic expression of bean architecture in classification procedures may enhance selection. The objective of this study was to evaluate the potential of artificial neural networks (ANNs) as auxiliary tools in the improvement of bean plant architecture. Data from 19 lines were evaluated for 22 traits, in 2007 and 2009 winter crops. Hypocotyl diameter and plant height were selected for analysis through ANNs. For classification purposes, these lines were separated into two groups, determined by the plant architecture notes. The predictive ability of ANNs was evaluated according to two scenarios to predict the plant architecture - training with 2007 data and validating in 2009 data (scenario 1), and vice versa (scenario 2). For this, ANNs were trained and validated using data from replicates of the evaluated lines for hypocotyl diameter individually, or together with the mean height of plants in the plot. In each scenario, the use of data from replicates or line means was evaluated for prediction through previously trained and validated ANNs. In both scenarios, ANNs based on hypocotyl diameter and mean height of plants were superior, since the error rates obtained were lower than those obtained using hypocotyl diameter only. Lower apparent error rates were verified in both scenarios for prediction when data on the means of the evaluated traits were submitted to better trained and validated ANNs.

Fish sperm subpopulations: Changes after cryopreservation process and relationship with fertilization success in tambaqui (Colossoma macropomum).

Fish tambaqui (Colossoma macropomum) is the native Brazilian fish with the highest agricultural production under intensive aquaculture in South America. However, the decrease in the genetic variability in fish farms has become necessary the improvement of cryopreservation process through new statistical studies of spermatozoa (like subpopulation studies). The evaluation of the kinetic data obtained with a computer-assisted sperm analysis system, applying a two-step cluster analysis, yielded in tambaqui three different subpopulations in fresh sperm: SP1, considered as a slow nonlinear subpopulation; SP2, considered as a fast nonlinear subpopulation, and finally; SP3, considered as a fast linear subpopulation. For cryopreserved sperm, the cluster analysis yielded only two sperm subpopulations: SP1', considered as a slow nonlinear subpopulation and SP2', which seemed to be an intermediate subpopulation (showing medium motility and velocity values) merged from SP2 and SP3 obtained from fresh sperm. Coefficients of correlation (r) and determination (r2) between the sperm subpopulations from fresh sperm and the fertilization rates were calculated, and SP2 and SP3 (the fast-spermatozoa subpopulations) showed a high-positive correlation with the fertilization rates (r = 0.93 and 0.79, respectively). In addition, the positive significant correlations found in curvilinear velocity (r = 0.78), straight line velocity (r = 0.57), and average velocity (r = 0.75) indicate that sperm kinetic features seem to be a key factor in the fertilization process in tambaqui, as occur in other fish species.

Diallel analysis to choose parents for black bean (Phaseolus vulgaris L.) breeding.

In this study, conducted in two different seasons, we aimed to choose parents to obtain promising segregating populations for the extraction of black bean (Phaseolus vulgaris L.) lines that are superior in terms of disease resistance, plant architecture, and grain yield. Twelve parents were arranged in two groups to compose a partial diallel in a 5 x 7 scheme. Group 1 was composed of parents with black grains and erect plant architecture, while group 2 was composed of parents that had carioca grains and were resistant to the main fungal diseases that occur in the common bean. The following traits were evaluated: severity of angular leaf spot (ALS), plant architecture (PAG), and grain yield (YIELD). The data were analyzed according to a partial diallel model using parents and F1 hybrids. In the genetic control of ALS and PAG, additive effects were predominant, while for YIELD, additive effects were predominant in one season and dominance effects were in another season, because it is a more complex trait than ALS and PAG. For YIELD, we observed an interaction between general combining ability and specific combining ability between seasons. The genes that control ALS, PAG, and YIELD were in eight of the 12 parents evaluated in the diallel. The cultivar 'BRS Estilo' is suitable to use as a parent in common bean breeding in terms of ALS, PAG and YIELD. Recurrent selection is the most recommended option for simultaneously breeding for PAG, YIELD, and resistance to angular leaf spot in bean culture.

Morphoagronomic characterization and genetic diversity of a common bean RIL mapping population derived from the cross Rudá x AND 277.

Recombinant inbred lines (RILs) are a valuable resource for building genetic linkage maps. The presence of genetic variability in the RILs is essential for detecting associations between molecular markers and loci controlling agronomic traits of interest. The main goal of this study was to quantify the genetic diversity of a common bean RIL population derived from a cross between Rudá (Mesoamerican gene pool) and AND 277 (Andean gene pool). This population was developed by the single seed descent method from 500 F2 plants until the F10 generation. Seven quantitative traits were evaluated in the field in 393 RILs, the parental lines, and five control cultivars. The plants were grown using a randomized block design with additional controls and three replicates. Significant differences were observed among the RILs for all evaluated traits (P < 0.01). A comparison of the RILs and parental lines showed significant differences (P < 0.01) for the number of days to flowering (DFL) and to harvest (DH), productivity (PROD) and mass of 100 beans (M100); however, there were no significant differences for plant architecture, degree of seed flatness, or seed shape. These results indicate the occurrence of additive x additive epistatic interactions for DFL, DH, PROD, and M100. The 393 RILs were shown to fall into 10 clusters using Tocher's method. This RIL population clearly contained genetic variability for the evaluated traits, and this variability will be crucial for future studies involving genetic mapping and quantitative trait locus identification and analysis.

Genetic potential of segregating populations of red beans conducted by the bulk method with selection.

This study evaluated the performance and estimated the genetic potential of segregating populations of red bean. Twenty populations of the second cycle of recurrent selection for red bean breeding at Universidade Federal de Viçosa were advanced to the F5 generation in bulk with selection for grain appearance. Populations plus five controls were evaluated for grain yield in three dry seasons (2004, 2005, and 2007) in a 5 x 5 lattice design with three replications, in four 4-m long rows. In the mean of the three crops, populations formed three distinct groups in which some populations had a clearly superior grain yield. Populations 288RVCI, 291RVCI, 295RVCI, 297RVCI, 300RVCI, and 303RVCI were the most promising. Over generations, segregating populations by the bulk method with selection for grain appearance may have reduced variability in grain yield. The 300RVCI population deserves further attention as it combines variability, high mean, and a high probability of generating above-standard lines.

Potential of hypocotyl diameter in family selection aiming at plant architecture improvement of common bean.

Cultivars of common bean with more erect plant architecture and greater tolerance to degree of lodging are required by producers. Thus, to evaluate the potential of hypocotyl diameter (HD) in family selection for plant architecture improvement of common bean, the HDs of 32 F2 plants were measured in 3 distinct populations, and the characteristics related to plant architecture were analyzed in their progenies. Ninety-six F2:3 families and 4 controls were evaluated in a randomized block design, with 3 replications, analyzing plant architecture grade, HD, and grain yield during the winter 2010 and drought 2011 seasons. We found that the correlation between the HD of F2 plants and traits related to plant architecture of F2:3 progenies were of low magnitude compared to the estimates for correlations considering the parents, indicating a high environmental influence on HD in bean plants. There was a predominance of additive genetic effects on the determination of hypocotyl diameter, which showed higher precision and accuracy compared to plant architecture grade. Thus, this characteristic can be used to select progenies in plant architecture improvement of common beans; however, selection must be based on the means of at least 39 plants in the plot, according to the results of repeatability analysis.

Effect of generations and environments in the analysis of a partial diallel to improve bean earliness.

The objective of this study was to select genitors based on F1 and F2 generations, evaluated in different environments, to obtain segregating populations for the identification of strains showing improved earliness, yield, and carioca-type grains. Nine bean strains were crossed in a partial diallel scheme (4 x 5), in which group 1 included 4 strains with early cycles and group 2 included 5 elite strains. The F1 and F2 generations and the genitors were assessed in Coimbra and Viçosa in randomized blocks with 3 replications. The following characteristics were evaluated: days between sowing and emergence, and grain yield. We observed an interaction between the effects of general combining ability and specific combining ability with the environments (crop, location, and generation) for both grain earliness and yield. Genetic analysis of earliness revealed a predominance of additive effects and grain yield dominance effects. The strain Goiano Precoce may be used as a genitor in breeding programs to improve earliness, while strains RP1 and VC33 can be used to increase grain yield. We observed genetic complementation between strains Goiano Precoce and RP1, BRSMG Madrepérola and BRS Estilo for earliness and between RP1 and Rosinha Precoce for grain yield.

Standardization of European eel (Anguilla anguilla) sperm motility evaluation by CASA software.

The development of powerful computer-assisted sperm analysis software has made kinetic studies of spermatozoa possible. This system has been used and validated for several species, but some technical questions have emerged regarding fish sample evaluations (i.e., frame rate, sperm dilution, chamber model, time of analysis, magnification lens, etc.). In the present study, we have evaluated the effects of different procedural and biological settings with the aim to correctly measure sperm quality parameters of the European eel. The use of different chambers did not affect the sperm motility parameters. However, regarding lens magnification, 10× was the most accurate lens, showing the least variation in the acquired data. Similarly, the frame rate setting resulted in a dramatic effect in some sperm kinetic parameters, primarily in terms of curvilinear velocity; we therefore recommend using the camera's highest available frame rate setting. Finally, the reduction in sperm motility over postactivation times suggests that sperm analysis should be performed within the first 60 seconds after activation of the European eel sperm. In conclusion, some protocol variables of sperm analysis by computer-assisted sperm analysis software can affect the measurement of eel sperm quality parameters, and should be considered before directly comparing results obtained by different laboratories. Moreover, because marine fish species show relatively similar features of sperm kinetic parameters, these results could be considered in the evaluation of the motility of sperm from other fish species.

Genetic improvement of plant architecture in the common bean.

Knowledge of genetic control of plant architecture in the common bean can help breeders define the most adequate breeding strategy to optimize gains. We examined genetic control of plant architecture in the common bean by means of partial diallel crosses. Fourteen bean lines were crossed under a partial diallel scheme, in which group 1 was composed of 8 erect plant lines and group 2 of 6 carioca-type grain lines. The F1 plants from the crosses and the 14 parents were evaluated during spring (March sowing) for the characteristics plant architecture score, hypocotyl diameter, mean plant height, and grain yield. The additive effects were found to predominate in the genetic control of plant architecture score and hypocotyl diameter. Thus, selection of more erect plants can be done in early generations. Heritability estimate in a wide sense was greater for hypocotyl diameter (0.81) than for plant architecture score (0.60), showing that the latter is a more complex characteristic than the former. Hypocotyl diameter has the potential to be used in the selection of more erect plants, either by considering selection between and within families, or by combined or phenotypic recurrent selection. The reason for this is that the diameter measurements are taken from individual plants inside the plots.

Cryopreservation of tambaqui (Colossoma macropomum) semen: extenders, cryoprotectants, dilution ratios and freezing methods.

The tambaqui is an Amazonian fish of great economic and environmental importance to Brazil and other South American countries. Several semen cryopreservation methodologies have been tested for different Brazilian fish species; however, there is little information on the use of this technique on tambaqui semen. The aim of the present study was to investigate the effect of osmolarity and activation solutions on sperm kinetics and, glucose solutions, cryoprotectants, dilution ratios, egg yolk and freezing methods on tambaqui semen freezing. The osmolarity of 230 mOsm was suitable for simultaneously yielding higher sperm motility (85%) and motility time (54 sec.) and osmolarities above 360 mOsm maintain immobile tambaqui sperm. The tambaqui semen can be successfully cryopreserved when diluted 1:9 in freezing medium composed of 5 percent glucose solution (290 mOsm) with 10 percent methylglycol and 5 percent egg yolk, and frozen directly in a dry shipper container.

Matrix metalloproteinase-9 genetic variations affect MMP-9 levels in obese children.

OBJECTIVE: Matrix metalloproteinase-9 (MMP-9) is involved in the atherosclerotic process and functional polymorphisms in the MMP-9 gene affect MMP-9 expression/activity, and are associated with cardiovascular diseases. However, no study has tested the hypothesis that functional MMP-9 polymorphisms could affect MMP-9 levels in obese children. We investigated whether three MMP-9 gene polymorphisms (C-1562T (rs3918242), 90(CA)((14-24)) (rs2234681) and Q279R (rs17576)), or haplotypes, affect MMP-9 levels in obese children. METHODS: We studied 175 healthy control children and 127 obese children. Plasma MMP-9, tissue inhibitor of MMPs (TIMP)-1 and adiponectin concentrations were measured using enzyme-linked immunosorbent assay. RESULTS: We found similar MMP-9 genotypes, allelic and haplotypes distributions in the two study groups (P>0.05). However, we found lower plasma MMP-9 concentrations in obese subjects carrying the CC or the QQ genotypes for the C-1562T and the Q279R polymorphisms, respectively, in obese children compared with children with the other genotypes, or with non-obese children with the same genotypes (all P<0.05). Moreover, we found lower MMP-9 levels and lower MMP-9/TIMP-1 ratios (which reflect net MMP-9 activity) in obese children carrying the H2 haplotype (which combines the C, H and Q alleles for the three polymorphisms, respectively) when compared with obese children carrying the other haplotypes, or with non-obese children carrying the same haplotype (P<0.05). CONCLUSIONS: Our findings show that MMP-9 genotypes and haplotypes affect MMP-9 levels in obese children and adolescents, and suggest that genetic factors may modify relevant pathogenetic mechanisms involved in the development of cardiovascular complications associated with obesity in childhood.

Distinction between plant samples according to allele dosage by semiquantitative polymerase chain reaction.

The lack of informativity of samples from heterozygotic individuals is one of the hindrances in the mapping of quantitative trait loci of outbred populations, since it is not normally possible to identify the origin of each allele. One way to include these individuals in analyses would be to genotype their endosperm, considering that a heterozygote (Aa) has AAa or Aaa endosperm, when the female genitor donates the A or a allele, respectively. We used semiquantitative polymerase chain reaction to determine allele dosages in DNA mixtures, by simulating the observed conditions for endospermic tissue. Semiquantitative polymerase chain reaction on agarose gels, along with regression analysis, allowed differentiation of the samples according to the amount of DNA. This type of information will help decrease the number of non-informative individuals in quantitative trait locus mapping of outbred populations, thereby increasing mapping accuracy.

Variabilidade genética e limite da seleção em populações de diferentes tipos de acasalamento / Genetic variability and selection limit in populations of different mating designs

Populações de cinco diferentes tipos de acasalamento, submetidas à seleção baseada no melhor preditor linear não-viesado (BLUP), foram avaliadas quanto às perdas genéticas por fixação de alelos desfavoráveis e limite da seleção, durante 50 gerações. Foram utilizados dados simulados na obtenção do genoma dos indivíduos de todas as populações. Uma característica quantitativa de herdabilidade 0,10 foi estudada em populações de seleção, com a seguinte estrutura de dados: razão sexual de 10, 20, 25 e 50 e tamanho efetivo da população de 36,36, 19,05, 15,38, e 7,84, respectivamente. Para cada razão sexual, formaram-se populações correspondentes aos tipos de acasalamento efetuados entre os reprodutores, em todas as gerações: acasalamentos preferenciais entre meios-irmãos e irmãos completos, acasalamentos preferenciais entre meios-irmãos, acasalamentos ao acaso, exclusão de acasalamentos entre irmãos completos e exclusão de acasalamentos entre meios-irmãos e irmãos completos. Valores percentuais mais baixos para locos fixados desfavoravelmente e limite da seleção mais alto foram observados na menor razão sexual (d= 10), na qual houve também melhor distinção entre os tipos de acasalamento estudados.

Efeitos da seleção individual e da seleção baseada no BLUP em populações diferentes, quanto ao tipo de acasalamento / Individual selection and selection based on BLUP effects in different populations, according to mating design

Dados simulados foram utilizados para avaliar o desempenho da seleção individual e da seleção baseada no melhor preditor linear não-viesado (BLUP), em populações que diferiam entre si, quanto ao tipo de acasalamento, no decorrer de 50 gerações. Estudou-se uma característica quantitativa com herdabilidade igual a 0,10, em populações de 600 indivíduos, que apresentaram a seguinte estrutura: valores de razão sexual (d), de 10 e 50, tamanhos efetivos de população (Ne), de 36,36 e 7,84 e intensidade de seleção dos machos (i m), de 2,23 e 2,82. Em cada valor de d, formaram-se populações correspondentes ao tipo de acasalamento, em todas as gerações: acasalamentos preferenciais de meios-irmãos e irmãos completos, acasalamentos preferenciais entre meios-irmãos, acasalamentos ao acaso, exclusão de acasalamentos entre irmãos completos e exclusão de acasalamentos de meios-irmãos e irmãos completos. As características avaliadas ao longo das gerações foram: valores fenotípicos médios, consangüinidade média, perda percentual por fixação de alelos desfavoráveis e limite da seleção. Observou-se que o BLUP, o tipo de acasalamento, excluindo acasalamentos entre irmãos e a menor razão sexual, juntos, proporcionaram os melhores resultados de valores fenotípicos, durante 45 gerações de seleção. Nessa estratégia de seleção, verificou-se também atraso no número de gerações necessárias para se atingir determinado nível de consangüinidade.

Immunocytochemistry applied to aspiration biopsy cytology. Diagnostic contribution in 100 cases of previously stained, routine specimens.

OBJECTIVE: To assess the contribution of immunocytochemistry (ICC) to aspiration biopsy cytology (ABC), in a diagnostic context, on routine, previously stained cytologic specimens. STUDY DESIGN: Among 5,221 consecutive cases of ABC, 5.3% were subjected to ICC in the clinical-morphologic context. One hundred of these cases, with a final clinical and histopathologic diagnosis, were studied to determine the contribution of this ancillary technique to the final cytologic diagnosis. All cases had histopathologic study and prospective ICC, performed on usual smears, alcohol fixed and previously stained by the Papanicolaou technique, and were subjected to an avidinbiotin-peroxidase complex method. RESULTS: ICC was contributory in 82% of cases. The contribution of ICC to ABC of lymphoid tissue, thyroid and related organs, soft tissue and miscellaneous cases was, respectively, 84% (39 cases), 88% (26), 72% (18) and 76% (17). ICC was noncontributory in 18 cases, due mainly to misleading interpretation (6%), uncharacteristic profile (5%) and inconclusive immunostain (7%). CONCLUSION: ICC could be successfully applied in routine ABC specimens since the usually investigated antigenic determinants are preserved, allowing previous morphologic study and screening of the smears. The principal contribution of ICC applied to lymph nodes, thyroid and soft tissue aspirates was, respectively, confirmation of metastatic neoplasms, differential of follicular versus C-cell proliferation and assessment of mesenchymal lineages.

Squamous-cell carcinoma of the lower lip: a retrospective study of 58 patients.

The charts of 58 patients with squamous-cell carcinomas of the lower lip, treated at the General Hospital of the University of São Paulo Medical School from January 1980 to December 1989, were retrospectively analyzed. In addition to regular demographic data, all available information was collected regarding: smoking and drinking habits; sun exposure; clinical stage; macroscopic features of the primary lesions; type of treatment; and follow-up. A meticulous pathological analysis, comprising the histologic differentiation grade, maximal tumor thickness, sun elastosis, perineural spread, vascular and muscular invasion, surgical margins, peritumoral inflammatory infiltrate, and positive lymph nodes, with or without extracapsular spread, was undertaken as well. The evaluation of the overall 5-year survival showed significant statistical differences, with prognostic implications, for the following variables: maximal tumor thickness, T-stage and positive nodes.

[Acute myeloid leukemia after treatment of Hodgkin's disease]. / Leucemia mielóide aguda após tratamento da doença de Hodgkin.

The development of acute leukemia, particularly acute myeloid leukemia, represents a serious complication in patients treated with radio and/or chemotherapy for Hodgkin's disease. It has been reported with increasing frequency in the last years. Two such cases, that occurred in 87 patients treated for Hodgkin's disease, are reported. Complete autopsy was performed in both. The patients were less than 30 years old, received combined therapy during a prolonged time (more than 12 months), with an interval superior to 44 months between the diagnosis of Hodgkin's disease and the appearance of acute myeloid leukemia. The survival time was less than 12 months. Residual Hodgkin's disease was not observed.