Diversity and genetic struture in cajá tree (spondias mombin l.) populations in northeastern Brazil

Spondias mombin L. is a fruit tree from the American continent from the Anacardiaceae family. In Brazil it is common in different vegetation types but is more frequent in the Atlantic and Amazonian rainforests. It is economically important because of its fruits, which are widely consumed raw or processed as fruit jellies, juices and ice creams. The leaves have great importance in the pharmaceutical industry because of their antibacterial properties. In the state of Pernambuco, cajá tree is widely distributed in the Zona da Mata region and less frequently in the Agreste and Sertão areas. In this work diversity and genetic structure were studied in four populations of cajá tree from Pernambuco's Zona da Mata, Northeast Brazil, using isozymes polymorphism analyses from electrophoreses. The result showed 100% of polymorphism (P) for nine alleles and the average of alleles per locus s was 2.4. The expected heterozygosity ranged from 0.530 to 0.574 and the observed heterozygosity , from 0.572 to 0.735. It was not observed inbreeding and the average F IT was -0.175, whereas within population inbreeding (f) varied from -0.08 to- 0.37. The genetic divergence among the populations (F ST) ranged from 0.006 to 0.028 and the average was 0.026. The average of estimated gene flow (Nm) was high (5.27). The CG-IPA population, corresponding to the germplasm collection of IPA, showed more than 96% of genetic similarity with other populations; therefore, it is a good representative of the existent genetic diversity in the Zona da Mata region.

Avaliação de diploides de bananeira (Musa spp.) quanto à tolerância a salinidade

A salinidade é um fator comum de estresse abiótico que afeta a produção agrícola onde ela existe. Uma das estratégias para promover a reincorporação de áreas salinizadas e o aumento da produtividade consiste no desenvolvimento e na seleção de genótipos tolerantes, o que permitirá a identificação de parentais a serem utilizados em cruzamentos. Esta pesquisa teve como objetivo identificar genótipos diploides de bananeira tolerantes à salinidade a serem utilizados em futuros trabalhos de melhoramento genético, visando à obtenção de cultivares adaptados a solos salinos. Foram avaliados nove genótipos diploides (AA) de bananeira. Para a avaliação dos parâmetros de crescimento, foram feitas medições de área foliar, altura e contagem do número de folhas. As plantas foram tratadas durante 21 dias com 0 e 100 mM de NaCl, num delineamento experimental inteiramente casualizado, com três repetições. Aos 21 dias, foi feita a determinação do peso da matéria fresca, utilizando-se balança analítica. A obtenção do peso da matéria seca das partes limbo foliar, pseudocaule e raízes+rizoma de cada planta foi feita após secagem em estufa a 60ºC até peso constante. o genótipo Tjau Lagada, que sofreu menor redução da área foliar, possivelmente apresentará uma produção relativamente superior aos genótipos avaliados neste estudo. o genótipo 0116-01, por ter apresentado maior tolerância à salinidade, poderá ser utilizado em futuros cruzamentos, disponibilizando genes a serem incorporados em cultivares produtivas utilizadas em programas de melhoramento que visem à obtenção de cultivares adaptadas às regiões de solos salinos no Nordeste brasileiro.

Diversity and genetic structure in natural populations of Hancornia speciosa var. speciosa Gomes in northeastern Brazil

Hancornia speciosa Gomes is a fruit tree native from Brazil that belongs to Apocinaceae family, and is popularly known as Mangabeira. Its fruits are widely consumed raw or processed as fruit jam, juices and ice creams, which have made it a target of intense exploitation. The extractive activities and intense human activity on the environment of natural occurrence of H. speciosa has caused genetic erosion in the species and little is known about the ecology or genetic structure of natural populations. The objective of this research was the evaluation of the genetic diversity and genetic structure of H. speciosa var. speciosa. The genetic variability was assessed using 11 allozyme loci with a sample of 164 individuals distributed in six natural populations located in the States of Pernambuco and Alagoas, Northeastern Brazil. The results showed a high level of genetic diversity within the species (e= 0.36) seeing that the most of the genetic variability of H. speciosa var. speciosa is within its natural populations with low difference among populations ( or = 0.081). The inbreeding values within ( = -0.555) and among populations ( =-0.428) were low showing lacking of endogamy and a surplus of heterozygotes. The estimated gene flow ( m ) was high, ranging from 2.20 to 13.18, indicating to be enough to prevent the effects of genetic drift and genetic differentiation among populations. The multivariate analyses indicated that there is a relationship between genetic and geographical distances, which was confirmed by a spatial pattern analysis using Mantel test (r = 0.3598; p = 0.0920) with 1000 random permutations. The high genetic diversity index in these populations indicates potential for in situ genetic conservation.

Parasitismo de Meloidogyne enterolobii em espécies de myrtaceae

A meloidoginose da goiabeira, causada por M. enterolobii (= M. mayaguensis), é hoje considerado o principal problema fitossanitário desta cultura em todo o País, pois sua incidência resulta em acentuada queda de produtividade e, na maioria das vezes, a morte das plantas em médio prazo. Visando a seu controle, objetivou-se neste trabalho identificar fontes de resistência em goiabeira Psidium guajava L e araçazeiro Psidium sp.. Foram avaliados onze genótipos oriundos do Câmpus da Universidade Federal de Lavras (UFLA) e de regiões próximas ao município de Lavras (MG). As sementes foram coletadas de frutos fisiologicamente maduros e germinadas em piscinas hidropônicas. Ao atingirem 12 cm de altura, foram transferidas para casa de vegetação, em vasos individuais, e mantidas a temperatura entre 18-38°C. Após quinze dias, foram inoculados individualmente com suspensão de 10.000 ovos e juvenis de segundo estádio (J2) de M. enterolobii e avaliadas após 120 dias. As reações dos hospedeiros foram enquadradas nos parâmetros estabelecidos pelo fator de reprodução FR, estimado pelo quociente Pf/Pi, em que Pf representa a população final e Pi a população inicial. O delineamento utilizado foi o inteiramente casualizado, com 11 genótipos, uma espécie de nematoide e seis repetições, sendo duas plantas em cada genótipo adicionadas como controle. Quatro meses depois, foram avaliados os teores foliares de macro e micronutrientes e o grau de infecção de cada genótipo. A produção média de ovos e J2 de M. enterolobii variou de 200 a 428.146,1, e os fatores de reprodução (FR) médios variaram de 0,02(ALU1) a 42,81(A-PASTO). Os genótipos de araçazeiro (Psidium spp.) AUFLA1, AUFLA4, AUFLA5 e APASTO e os genótipos de goiabeira (P. guajava) G-ROXA e G-AMAR foram considerados suscetíveis. Os genótipos de araçazeiro ALU1, ALU2, ALU3, AROXO-C e AROXO-U mostraram-se resistentes, reforçando a necessidade de trabalhos complementares com essas espécies para uso como porta-enxerto em cultivares comerciais de goiabeira. A absorção de macro e micronutrientes é alterada quando os genótipos de Psidium spp. são inoculados com o fitonematoide M. enterolobii.

Ejaculate Characteristics Depend on Social Environment in the Horse (Equus caballus).

Sperm competition theory predicts semen characteristics to be affected by the social environment. We used the polygamous horse (Equus caballus) to experimentally study within-subject plasticity in response to different social environments. Stallions were sequentially exposed, over a period of 8 weeks each, to other stallions and then singly to mares, or vice versa (in adjacent boxes separated by grills). Ejaculates were collected to determine semen characteristics. Highest sperm numbers were found in stallions that were first exposed to other stallions and then to mares, while lowest sperm numbers were observed in stallions that had been exposed to mares but not yet to other stallions. One of three sperm velocity measures (curvilinear velocity) was consistently elevated in stallions that were first exposed to stallions and then to mares. Sperm number after exposure to mares and curvilinear sperm velocity after exposure to stallions were both positively correlated to average blood testosterone levels during the corresponding period of exposure. We conclude that ejaculate characteristics are plastic traits affected by the social environment in horses.

Identificación de características forenses avanzadas a partir del ADN: etnogeografía, patología delictiva y morfoanatomía

Resumen: El uso de la molécula de ADN ha revolucionado el mundo de la Genética forense, convirtiéndose en una herramienta poderosa para resolver un gran número de casos judiciales. El ADN permite identificar a una persona determinada con una probabilidad de error ínfima. Utilizando el análisis del ADN los ámbitos clásicos de actuación han sido: la resolución de grandes delitos, las pruebas de paternidad o de parentesco en general y por último la identificación de restos humanos que estuviesen muy dañados. Sin embargo, los investigadores policiales buscan obtener más información a partir de las moléculas de ADN extraídas de muestras biológicas (sangre, saliva, semen, células epiteliales, etc.). En este trabajo abordaremos los conocimientos que en la actualidad se tienen de algunas de las características del aspecto externo de los individuos a partir del análisis del ADN de las muestras, su interés desde el punto de vista jurídico y sus limitaciones. En concreto nos centraremos en cuatro grandes áreas de estudio: el análisis del origen geográfico de los ancestros, la genética del comportamiento, la genética médica y las predicciones de características externas tales como el color de los ojos, del cabello, de la piel, la estatura y la edad. Palabras clave: Genética forense, ADN. Origen étnico-geográfico. Patología delictiva. Características externas visibles.

A análise seminal deve ser requisitada para homens com histórico de fertilidade prévia?

OBJETIVOS: determinar se a história prévia de fertilidade pode predizer o atual status de fertilidade de um paciente masculino examinado por infertilidade do casal. MÉTODOS: estudo retrospectivo envolvendo análises seminais de 183 pacientes consecutivos subférteis avaliados entre setembro de 2002 e março de 2004. Foram excluídos do estudo os pacientes que haviam se submetido a radioterapia, quimioterapia, orquiectomia ou vasectomia. Os valores médios de todas as análises foram usados em pacientes com múltiplas análises de sêmen. Pacientes com concentração espermática superior a 20x10(6) espermatozóides/mL, motilidade superior a 50% e espermatozóides com morfologia estrita superior a 14% foram considerados normais. Os pacientes foram divididos em dois grupos, segundo o status de fertilidade: infertilidade primária (118 pacientes) e infertilidade secundária (65 pacientes). Os dados foram analisados pelos testes estatísticos chi2 e teste t de Student. RESULTADOS: não houve diferença na idade média entre os pacientes com infertilidade primária, 37,3±6,3, e infertilidade secundária, 38,1±5,9; p=0,08. No grupo de pacientes com infertilidade primária, 51,9% (61 pacientes) tiveram uma concentração espermática normal, 70,3% (83 pacientes) tiveram a motilidade espermática normal e 26,37% (31 pacientes), por sua vez, morfologia normal. No grupo de pacientes com infertilidade secundária, 53,8% (35 pacientes) tiveram concentração espermática normal, 75,4% (49 pacientes) tiveram motilidade espermática normal e 32,3% (21 pacientes), morfologia normal. Nenhuma diferença significativa foi detectada na concentração espermática (21,3x10(6)/mL versus 23,1x10(6)/mL; p=0,07), motilidade (45,2 versus 48,1%; p=0,08) e morfologia (6,1 versus 6,4%; p=0,09) entre os grupos de pacientes com infertilidade primária e secundária. CONCLUSÕES: a análise seminal deve ser solicitada mesmo em casos de fertilidade masculina prévia. Os médicos não devem presumir que um paciente possui uma análise seminal normal, baseados no fato de este possuir história de estabelecimento de uma gravidez no passado.

Criopreservação de sêmen humano: comparação entre métodos de congelação e tipos de envase

OBJETIVOS: comparar duas diferentes técnicas de congelação e dois tipos de envase do sêmen humano durante processo de criopreservação. MÉTODOS: estudo experimental, no qual foi analisada a criopreservação de 18 amostras de sêmen de 18 voluntários. Após a adição de meio crioprotetor, "Test-yolk buffer" , as amostras de sêmen foram envasadas em palhetas com capacidade de 0,25 mL ou em criotubos de 2 mL e submetidas à criopreservação por dois métodos, um lento e outro rápido, totalizando quatro tratamentos distintos: RP (congelação pelo método rápido e envasado em palheta), RT (rápido-criotubo), LP (lento-palheta) e LT (lento-criotubo). As amostras, após 24 horas, foram descongeladas em temperatura ambiente e mantidas a 37ºC. Os dados coletados foram analisados através do teste t de Student, com p<0,05, utilizando o programa de computador SPSS for Windows® versão 11.0.0. RESULTADOS: houve redução da motilidade espermática após o processo de criopreservação. A taxa de motilidade inicial foi 58,1% e as motilidades após os diferentes métodos de criopreservação foram: 19,2% (RP), 27% (RT), 21,1% (LP) e 30,3% (LT). Houve redução significativa na morfologia normal. A taxa de morfologia normal inicial foi 14,2% e as morfologias após os diferentes métodos de criopreservação foram: 12,8% (RP), 12,6% (RT), 12,6% (LP) e 12,4% (LT). CONCLUSÕES: o método de criopreservação lento com envase em criotubo esteve associado à melhor motilidade espermática após o descongelamento. Não houve diferença entre os métodos quando avaliada a morfologia espermática.

Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu)

The aim of this study was to compare different staining methods for the evaluation of sperm morphology by light microscopy and also to describe the morphometry of the entire sperm in collared peccaries (Pecari tajacu). Semen from 10 males was obtained by electroejaculation and evaluated for sperm motility, vigor, and concentration. Semen smears were prepared through three different staining methods: Bengal rose, brome-phenol blue, and eosin-nigrosin. Smears were evaluated under light microscopy and sperm morphologic alterations were determined in percentage. In addition, sperm morphometric analysis was conducted by light microscopy coupled to image analyzer software. The smears stained with Bengal Rose provide the best results for the visualization of the sperm tail, midpiece, and head. The use of eosin-nigrosin stain did not allow an adequate impregnation, and some sperm presented a few contrasts with the background. A higher incidence of bent coiled tails was verified in the use of brome-phenol blue staining (P<0.05). Through morphometric evaluation, it was observed that the tail occupies the greatest proportion (89%) of the sperm which presents a discretely elongated head. According to the results, the use of the Bengal Rose stain is recommended for the morphologic evaluation of the collared peccary sperm.

The seasonal and ovarian status effects on in vitro production of domestic cat embryos between Equator and Tropic of Capricorn

From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. Ovaries were classified according to their status and were sliced in PBS for cumulus oocyte complex (COC) releasing. Grade I COC were washed three times in H-MEM supplemented with BSA, glutamine, sodium pyruvate, cysteine, streptomycin and penicillin. Oocytes were incubated in groups of 20-30 in 400µL of DMEM supplemented with FSH, LH, estradiol, IGF-I and basic fibroblast growth factor under mineral oil for 30 or 36 hours at 38°C in humidified environment of 5% de O2, 5% CO2 and 90% N2. COC were fertilized in Ham's F-10 medium supplemented with BSA, cysteine, pyruvate and streptomycin/penicillin (culture medium) with fresh semen selected through swim up technique. Eighteen hours later, the presumptive zygotes were denuded, the percentage of cleavage was determined and every 10 zygotes were transferred to 100mL drops of culture medium for culture during three days. After 72 hours of culture the percentage of morulae formation was evaluated and these structures were transferred to drops of the same culture medium. At the eighth day of culture blastocyst formation was analyzed. During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.

Degenerative joint disease in cattle and buffaloes in the Amazon region: a retrospective study

A retrospective study of the epidemiological and clinic-pathological aspects of cattle and buffaloes with degenerative joint disease (DJD) was conducted in the state of Pará, Brazil. From 1999 to 2014, eleven cattle and 24 buffaloes were evaluated. All the treated animals with suspected DJD underwent a clinical examination of the musculoskeletal system. In seven cattle and eight buffaloes with clinical signs of the disease postmortem examination was performed. The common clinical signs observed in both species were chronic lameness, stiff gait, postural changes, audible crackles in the affected limb, prolonged recumbency, difficulty in getting up and progressive weight loss. The lesions observed at necropsy were: irregular articular surfaces, erosion of the articular cartilage and the underlying bone tissue, and proliferation of the periarticular bone tissue with formation of osteophytes. The most affected joints in cattle and buffaloes wereof the hind limb. In buffaloes, the main predisposing factor to the onset of DJD was phosphorus deficiency. In cattle, defects of the anatomical conformation of the hind limbs, chronic trauma due to the activities performed, such as semen collection, and advanced age possibly contributed to the emergence of the disease.

Production of a cloned calf from a fetal fibroblast cell line

The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5%) were successfully fused and 24 (28.9%) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8%) of which were pregnant on day 35 and 3 (16.7%) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).

Effects of pentoxifylline treatment before freezing on motility, viability and acrosome status of poor quality human spermatozoa cryopreserved by the liquid nitrogen vapor method

The objective of the present study was to investigate the effects of the direct addition of pentoxifylline (PF) to the ejaculates of men with poor sperm quality before freezing on post-thaw sperm motility, viability, acrosome integrity, and agonist-induced acrosome reaction. Semen specimens from 16 infertile men with impaired sperm count and motility (oligoasthenozoospermia) were divided into two equal aliquots: one received no treatment (control) while the other was incubated with 5 mM PF (treated). Both aliquots were cryopreserved by the liquid nitrogen vapor method. Motility was assessed according to WHO criteria. Acrosome integrity and spontaneous and calcium ionophore-induced acrosome reactions were assessed with fluorescein isothiocyanate-conjugated peanut agglutinin combined with a supra-vital dye (Hoechst-33258). Cryopreservation impaired sperm motility (percentage reduction: 87.4 (interquartile range, IQ: 70.3-92.9) vs 89.1 (IQ: 72.7-96.0%)), viability (25.9 (IQ: 22.2-29.7) vs 25.6 (IQ: 19.7-40.3%)) and acrosome integrity (18.9 (IQ: 5.4-38.9) vs 26.8 (IQ: 0.0-45.2%)) to the same extent in both treated and control aliquots. However, PF treatment before freezing improved the acrosome reaction to ionophore challenge test scores in cryopreserved spermatozoa (9.7 (IQ: 6.6-19.7) vs 4.8 (IQ: 0.5-6.8%); P = 0.002). These data show that pre-freeze treatment of poor quality human sperm with pentoxifylline did not improve post-thaw motility or viability nor did it prevent acrosomal loss during the freeze-thaw process. However, PF, as used, improved the ability of thawed spermatozoa to undergo the acrosome reaction in response to calcium ionophore. The present data indicate that treatment of poor quality human sperm with PF may enhance post-thaw sperm fertilizing ability.

Minor sperm abnormalities in young male post-pubertal patients with juvenile dermatomyositis

The objective of the present study was to identify sperm abnormalities in young male patients with juvenile dermatomyositis (JDM). In 2005, 18 male JDM patients, diagnosed according to the criteria of Bohan and Peter, were followed at the Pediatric Rheumatology Unit and Rheumatology Division, of our Institution. Of the 18 males, 11 were pre-pubertal and 7 were post-pubertal. Two of 7 post-pubertal JDM male patients were excluded: one for orchidopexy for cryptorchidism and the other for testicular ectopia in the left testis. The remaining 5 post-pubertal JDM patients were prospectively evaluated on the basis of two semen analyses, according to the World Health Organization (WHO), urologic evaluation, testicular Doppler ultrasound hormone profile. The data of the JDM patients were compared with those of 5 age-matched healthy controls. The median age 18, was similar in JDM patients and controls. All JDM patients had teratozoospermia (abnormal sperm morphology), as did 4 (80%) of the controls. One of JDM patients had previous oligoasthenoteratozoospermia treated with intravenous cyclophosphamide with normalization of the number and concentration of the sperm after 5 years. All sperm parameters (sperm concentration, total sperm count and total motile sperm count by WHO, and sperm morphology by Kruger strict criteria), testicular volumes by Prader orchidometer and ultrasound, and hormones were similar in JDM patients compared with controls. The frequency of anti-sperm antibodies was similar in both groups. All JDM patients had minor sperm abnormalities in the head, midpiece, and/or tail of spermatozoids. Serial semen analyses in larger study populations are necessary to identify the extent and duration of sperm abnormalities in male patients with idiopathic inflammatory myopathies.