A polymerase chain reaction for the detection of Brucella canis in semen of naturally infected dogs

The objective was to evaluate a PCR assay for the detection of Brucella canis in canine semen, comparing its performance with that of bacterial isolation, serological tests and PCR assay of blood. Fifty-two male dogs were examined clinically to detect reproductive abnormalities and their serum was tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR assays were performed on blood and semen samples. The findings of the semen PCR were compared (Kappa coefficient and McNemar test) to those of blood PCR, culture of blood and semen, RSAT, and 2ME-RSAT. Nucleic acid extracts from semen collected from dogs not infected with B. canis were spiked with decreasing amounts of B. canis RM6/66 DNA and the resulting samples subjected to PCR. In addition, semen samples of non-infected dogs were spiked with decreasing amounts of B. canis CFU and the resulting suspensions were used for DNA extraction and amplification. of the 52 dogs that were examined, the following tests were positive: RSAT, 16 (30.7%); 2ME-RSAT, 5 (9.6%); blood culture, 14 (26.9%); semen culture, 11 (21.1%); blood PCR, 18 (34.6%); semen PCR, 18 (34.6%). The PCR assay detected as few as 3.8 fg of B. canis DNA experimentally diluted in 444.9 ng of canine DNA (extracted from semen samples of noninfected dogs). In addition, the PCR assay amplified B. canis genetic sequences from semen samples containing as little as 1.0 x 10(0) cfu/mL. We concluded that PCR assay of semen was a good candidate as a confirmatory test for the diagnosis of brucellosis in dogs; its diagnostic performance was similar to blood culture or blood PCR. Furthermore, the PCR assay of semen was more sensitive than the 2ME-RSAT or semen culture. Examination of semen by PCR should be included for diagnosis of brucellosis prior to natural mating or AI; in that regard, some dogs that were negative on serological and microbiological examinations as well as blood PCR were positive on PCR of semen. (c) 2007 Elsevier B.V. All rights reserved.

Dinâmica do peso e da condição corporal e eficiência reprodutiva de ovelhas da raça Santa Inês e mestiças Santa Inês-Suffolk submetidas a dois sistemas de alimentação em intervalos entre partos de 8 meses

O trabalho teve como objetivo comparar a eficiência produtiva e reprodutiva de ovelhas em dois sistemas de alimentação. Noventa e oito matrizes da raça Santa Inês e trinta e nove mestiças Suffolk (3/4 Suffolk + 1/4 Santa Inês) foram divididas em dois tratamentos: tratamento 1 (T1) - mantidas em pastagem, com suplementação de silagem de capim elefante na seca invernal, e tratamento 2 (T2) -mantidas em pastagem, com suplementação de silagem de capim elefante na seca invernal e de concentrado três semanas antes e durante a estação de monta, três semanas antes do parto e durante a lactação. Verificou-se diferença (P<0,05) entre peso inicial (PI) (52,5kg e 54,33kg), e peso final (PF) (53,38kg e 55,76kg) para T1 e T2, respectivamente. Houve efeito de genótipo (P<0,05), sendo PI 51,46 kg para a raça Santa Inês (SI) e 55,38kg para as mestiças Santa Inês-Suffolk (SF). Para PF, observou-se 52,36kg para a Santa Inês e 56,78kg na mestiça Santa Inês-Suffolk. O peso pré-parto (PPP) diferiu (P<0,05) apenas entre as estações, tendo sido de 65,23kg na estação reprodutiva I (2 a 4/2002), 58,15kg na estação reprodutiva II (10 a 12/2002) e 59,73kg na estação reprodutiva III (6 a 8/2003). No peso pós-parto (PPART), também ocorreram diferenças (P<0,05) entre a raça Santa Inês (53,59kg) e a mestiça Santa Inês-Suffolk (57,05kg); no peso aos 30 dias de lactação (P30d) a Santa Inês registrou 52,94kg e a mestiça Santa Inês-Suffolk 55,45kg. O peso aos 70 dias (P70d) de lactação foi para a Santa Inês de 50,83kg e de 53,22 kg para a mestiça Santa Inês-Suffolk; e o peso aos 100dias (P100d) de lactação foi de 51,55kg e de 53,61kg para a Santa Inês e para a mestiça Santa Inês-Suffolk, respectivamente. A condição corporal inicial (CCI) foi maior (P<0,05) para o T2 2,47 do que para o T1 2,16. Na condição corporal final (CCF), 2,19 e 2,6, respectivamente para T1 e T2, mas os tratamentos não diferiram na condição corporal pré-parto (CCPP). A CCI 2,4 para a mestiça Santa Inês-Suffolk foi (P<0,05) em relação a Santa Inês 2,22. A CCF da mestiça Santa Inês-Suffolk de 2,49 também foi maior (P<0,05) que da Santa Inês que obteve 2,3, mas não diferiram na CCPP. em relação as três estações reprodutivas, apenas na estação I a CCI 2,55, CCF 2,8 e a CCPP 3,03 foram maiores (P<0,05). Já as estações reprodutivas II= 2,47 e III= 2,1 diferiram somente na CCPP que foi menor na estação III. Na estação I e T1 ambos os genótipos obtiveram 72,5% de fertilidade e o T2 apresentou para as mestiças Santa Inês-Suffolk 77% e para a Santa Inês 88%. Na estação II o T1 obteve para as mestiças Santa Inês-Suffolk 42% e para as Santa Inês 38% e o T2 resultou em 56 e 50% para as mestiças Santa Inês-Suffolk e Santa Inês respectivamente. Na estação III a fertilidade do T1 foi para as mestiças 60% e para as Santa Inês 54% e o T2 87% e 76% para as mestiças e Santa Inês respectivamente. Encontrou-se diferença na prolificidade, entre os tratamentos, sendo na estação I a prolificidade foi de 1,20 e 1,55, na estação II foi 0,90 e 1,03 e na estação III obteve-se 1,11 e 1,14, respectivamente para o T1 e T2. Concluiu-se que a suplementação melhorou o desempenho reprodutivo das ovelhas.

Bioestimulação da atividade reprodutiva de coelhas lactantes em regime de monta natural

Esta pesquisa avaliou o efeito da separação mãe-ninhada sobre o desempenho reprodutivo de coelhas lactantes e ponderal das ninhadas até aos 70 dias de idade. Examinaram-se 415 exposições ao macho de 157 fêmeas lactantes distribuídas em dois grupos: controle, com livre acesso ao ninho, e tratado, cujo acesso bloqueou-se entre o sétimo e o nono dias pós-parto, durante as 39 horas que precederam a cobrição. Das 415 exposições ao macho, 387 resultaram em cobrições e 264 em partos. As fêmeas tratadas, de quinta lactação em diante, apresentaram receptividade e taxa de parição, respectivamente, 6,7% e 10,8% maiores em comparação às fêmeas do grupo controle. Não se constatou efeito da interação tratamento x ordem de lactação, ou do tratamento, com respeito ao número de láparos/ninhada à desmama e nos 70 dias de idade. em ninhadas de primíparas, a taxa de mortalidade da separação até 70 dias de idade foi maior no grupo tratado, em relação ao controle (0,248±0,047 vs. 0,120±0,047). O peso total da ninhada à desmama foi inferior no grupo tratado, do que no controle (5.753±93 g vs. 6.222±89 g) em todas as ordens. Todavia, não se observou diferença no peso total ou médio da ninhada nos 70 dias de idade. O tratamento também reduziu o peso individual à desmama (912,4±4,9 g vs. 962,7±4,5 g) em todas as ordens de lactação, entretanto, elevou o ganho de peso médio diário pós-desmama (32,25±0,19 g/d vs. 31,44±0,17 g/d), proporcionando a ocorrência de ganho compensatório neste período. Pode-se recomendar a separação temporária mãe-ninhada para melhorar o desempenho reprodutivo de coelhas lactantes multíparas; mas deve ser evitada para as primíparas, devido à elevação da taxa de mortalidade das ninhadas.

Melhoramento e estrutura populacional em bubalinos da raça Mediterrâneo no Brasil

O objetivo deste trabalho foi avaliar a estrutura populacional e os progressos genéticos, fenotípicos e ambientais de características de desenvolvimento ponderal, em bubalinos da raça Mediterrâneo. Foram utilizadas informações dos pesos ajustados aos 205, 365 e 550 dias de idade, de 6.243 bubalinos nascidos no período de 1974 a 2003, provenientes de quatro fazendas. As estimativas dos coeficientes de herdabilidade foram altas para os pesos ajustados nas três idades. Os rebanhos apresentaram ganho genético positivo quanto a essas três características, porém, aquém do possível, provavelmente em razão da seleção baseada somente no fenótipo, realizada pelos produtores. Para aumento nos ganhos genéticos, são necessários a redução do intervalo de geração, o aumento do tamanho efetivo, o uso de reprodutores avaliados com base nos valores genéticos preditos pelo BLUP e o controle dos acasalamentos de animais aparentados.

ASSESSING THE SUITABILITY of STERILE INSECT TECHNIQUE APPLIED TO AEDES AEGYPTI

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Neuroendocrine and reproductive aspects of adult male rats exposed neonatally to an antiestrogen

The present study was designed to investigate the effects of a single dose of an estrogen antagonist-clomiphene-during neonatal life, on later neuroendocrine system and reproductive performance. Immediately after birth, male pups received clomiphene citrate (s.c.). At adulthood, although testosterone levels and wet weights of reproductive organs were not altered, the treatment induced an increased number of spermatozoa and a delay in the transit time in the cauda epididymis. Additionally, there was impairment of sexual behavior evidenced by a delay in the latencies to the first mount and first intromission. Treated rats also showed decreased dopaminergic and serotonergic neurotransmissions in the hypothalamus and decreased dopaminergic neurotransmission in the striatum. The decreased dopaminergic activity could be related to the lower sexual motivation observed. These results indicate the necessity of preventing exposure to drugs that may impair sexual differentiation, which can compromise later mating success as well as the capacity to generate descendants. (c) 2006 Elsevier B.V. All rights reserved.

Sexual behavior, neuroendocrine, and neurochemical aspects in male rats exposed prenatally to stress

The present study was designed to examine some short- and long-term effects of maternal restraint stress-during the period of sexual brain differentiation-on reproductive and endocrine systems, sexual behavior, and brain neurotransmitters in male rat descendants. Pregnant rats were exposed to restraint stress for 1 h/day from gestational days (GDs) 18 to 22. Prenatal stress did not influence the wet weight of sexual organs and the quantity of germ cells in adult male pups; however, these animals showed reduced testosterone levels, delayed latency to the first mount and first intromission, and also decreased number of ejaculations. Additionally, there was an increase in the dopamine and serotonin levels in the striatum. Our results indicate that prenatal stress had a long-term effect on neurotransmitter levels and sexual behavior. In this sense, reproductive problems caused by injuries during the fetal period can compromise the later success of mating as well as the capacity to generate descendants. (C) 2004 Elsevier B.V. All rights reserved.

Impairment of adult male reproductive function in rats exposed to ethanol since puberty

The objective of this work was to evaluate reproductive function in adult male rats exposed to ethanol since puberty. Male Wistar rats, 50 days old, received a liquid diet with 36% of the daily calories derived from ethanol or an isocaloric control diet for 55 days. The ethanol treatment impaired sexual behavior and only 22% of these rats reached ejaculation. The fertility of ethanol-treated animals was significantly reduced, mainly after natural mating. Serum testosterone levels, daily sperm production and sperm count in the epididymis were also significantly diminished after ethanol treatment, associated with an acceleration of the sperm transit time in the cauda epididymidis, decrease in sperm motility and increased percentage of abnormal shaped sperm cells. The results showed that chronic consumption of ethanol beginning at puberty impairs the reproductive function of adult male rats. (c) 2006 Elsevier B.V. All rights reserved.

Reproductive and sexual behavior changes in male rats exposed perinatally to picrotoxin

Previous studies in rats suggested that picrotoxin, a GABA(A) receptor antagonist, may cause long-term changes in male reproductive physiology and behavior in rats exposed during prenatal and postnatal periods. The present study has further examined this phenomenon. Wistar rat dams were dosed subcutaneously with 0.75 mg/kg picrotoxin in saline, or vehicle alone, during the perinatal period (day 19 of gestation, immediately after parturition, and once a day during the first 5 days of lactation). Birth weight and sexual maturation of pups were unchanged; however, plasma testosterone levels and sexual behavior was altered in male offspring. Although fertile, these males showed altered mating behavior in terms of a decrease in the mean number of mounts during a 30-min observation period with normal females. Some showed homosexual behavior when castrated and pretreated with exogenous estrogen. These findings suggest that perinatal exposure to picrotoxin alters sexual dimorphism in the developing rat brain, manifesting as altered reproductive performance and sexual behavior of males. (c) 2004 Elsevier B.V. All rights reserved.

Ejaculatory dysfunction in streptozotocin-induced diabetic rats: the role of testosterone

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Investigation of the Effects of a1-Adrenoceptor Antagonism and L-Type Calcium Channel Blockade on Ejaculation and Vas Deferens and Seminal Vesicle Contractility In Vitro

Introduction. Premature ejaculation is one of the most common male sexual dysfunctions. Current pharmacological treatments involve reduction in penile sensitivity by local anesthetics or increase of ejaculatory threshold by selective serotonin reuptake inhibitors. a1-Adrenoceptors (a1-ARs) and L-type calcium channels are expressed in the smooth muscles of the male reproductive tract, and their activations play an important role in the physiological events involved in the seminal emission phase of ejaculation.Aim. To evaluate if the inhibition of the contractility of the vas deferens and seminal vesicle by alpha(1)-AR antagonism or the L-type calcium channel blockade can delay ejaculation.Methods. The effects of the alpha(1)-AR antagonist tamsulosin and of the L-type calcium channel blockers, nifedipine and (S)-(+)-niguldipine, on contractions induced by norepinephrine in the rat vas deferens and seminal vesicles in vitro and on the ejaculation latency of male rats in behavioral mating tests were evaluated.Main Outcome Measure. Tension development of vas deferens and seminal vesicles in response to norepinephrine in vitro and behavioral mating parameters were quantified.Results. Tension development of vas deferens and seminal vesicle to alpha(1)-AR activation was significantly inhibited by tamsulosin, nifedipine, and (S)-(+)-niguldipine. Tamsulosin displayed insurmountable antagonism of contractions induced by norepinephrine in the rat vas deferens and seminal vesicle. Ejaculation latency of male rats was not modified by tamsulosin, nifedipine, or (S)-(+)-niguldipine; however, both the number and weight of the seminal plugs recovered from female rats mated with male rats treated with tamsulosin were significantly reduced.Conclusion. Seminal emission impairment by inhibition of vas deferens or seminal vesicle contractility by L-type calcium channel blockade or alpha(1)-AR antagonism is not able to delay the ejaculation. de Almeida Kiguti LR and Pupo AS. Investigation of the effects of alpha(1)-adrenoceptor antagonism and L-type calcium channel blockade on ejaculation and vas deferens and seminal vesicle contractility in vitro. J Sex Med 2012; 9: 159-168.

Female preferences based on male nutritional chemical traits

Selection favors females that attend to reliable information about male genetic quality and fitness. Male nutritional condition can be a significant sign of mate quality since poor nutrition can be related to reduced sperm quality, low sperm quantity, sexually transmitted diseases, and parasites. We tested whether female Nile tilapia, Oreochromis niloticus, preferred the chemical cues of males that were well fed with high-protein diet over those fed with low-protein diet. Females do not only discriminate between males but also show a preference for well-fed males, discriminating between the odors with respect to nutritional state, suggesting that they were responding to a food-specific chemical cue. It is therefore likely that nutritional condition is related to the production of pheromones in males. Our results suggest that information about male nutritional state can be conveyed in chemical cues and that females attend to these cues during mate choice.

Female pintado catfish choose well-fed males

This study tested the preference of pintado catfish females (Pseudoplatystoma coruscans) for associating with well-fed or food-deprived males in simultaneous choice tests. Females were tested under three different treatments in which: (1) females could choose on the basis of multiple cues from the well-fed or food-deprived males (visual plus chemical); (2) only chemical cues were presented; (3) only visual cues were presented. Females chose well-fed males when chemical cues were presented (either visual + chemical or only chemical cues). When only visual cues were presented, females spent an equal amount of time in the choice apparatus compartments. Chemical signals, either directly through control of pheromone production, or indirectly by excreted metabolites, are likely to be an important source of information about nutritional condition in mating choice.

Characterization and transferability of microsatellite markers of the cultivated peanut (Arachis hypogaea)

Background: the genus Arachis includes Arachis hypogaea (cultivated peanut) and wild species that are used in peanut breeding or as forage. Molecular markers have been employed in several studies of this genus, but microsatellite markers have only been used in few investigations. Microsatellites are very informative and are useful to assess genetic variability, analyze mating systems and in genetic mapping. The objectives of this study were to develop A. hypogaea microsatellite loci and to evaluate the transferability of these markers to other Arachis species.Results: Thirteen loci were isolated and characterized using 16 accessions of A. hypogaea. The level of variation found in A. hypogaea using microsatellites was higher than with other markers. Cross-transferability of the markers was also high. Sequencing of the fragments amplified using the primer pair AhII from 17 wild Arachis species showed that almost all wild species had similar repeated sequence to the one observed in A. hypogaea. Sequence data suggested that there is no correlation between taxonomic relationship of a wild species to A. hypogaea and the number of repeats found in its microsatellite loci.Conclusion: These results show that microsatellite primer pairs from A. hypogaea have multiple uses. A higher level of variation among A. hypogaea accessions can be detected using microsatellite markers in comparison to other markers, such as RFLP, RAPD and AFLP. The microsatellite primers of A. hypogaea showed a very high rate of transferability to other species of the genus. These primer pairs provide important tools to evaluate the genetic variability and to assess the mating system in Arachis species.

Heterologous microsatellite primer pairs informative for the whole genus Arachis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)