RESUMO
BACKGROUND: Bulls are of great importance in the productive chain and for this reason they should have a good semen quality. There is no doubt that sperm morphology is very important to bull fertility, although little is known about how exactly the abnormal morphologies may affect sperm functions. OBJECTIVES: To detail the morphological description of the aplastic midpiece defect (AMD), as well as to understand its consequences for male fertility based on membrane and acrosome status, mitochondrial membrane potential and DNA integrity parameters. MATERIALS AND METHODS: The bulls were divided into two groups: control, consisting of satisfactory potential breeders (n = 3); and AMD, consisting of unsatisfactory potential breeders with a high percentage of AMD (n = 3). Bulls were evaluated by the breeding soundness evaluation; five ejaculates were collected from each animal and analyzed by flow cytometry. RESULTS: Spermatozoa from AMD group exhibited lower sperm motility and vigor (p < 0.05). In addition, it also exhibited lower mitochondrial membrane potential (p < 0.05), a higher percentage of spermatozoa with DNA fragmentation (p < 0.05), lower acrosome and plasma membrane integrity (p < 0.05), and higher lipid bilayer sperm membrane disorganization (p < 0.05) in comparison with control bulls. DISCUSSION: These findings may be due to oxidative stress and a reduction of the energy production capacity in addition to an alteration in the structural composition of the sperm cell. Moreover, semen with a high percentage of AMD may also be undergoing apoptosis. CONCLUSION: Bulls with a high percentage of AMD in their semen are not suitable for reproduction. Furthermore, it suggests there is a putative genetic basis for this sperm defect.
Assuntos
Bovinos , Fertilidade , Espermatozoides/anormalidades , Animais , Masculino , Peça Intermédia do Espermatozoide/patologia , Espermatozoides/fisiologiaRESUMO
Anastrepha obliqua (Macquart) is an agricultural pest of wide geographic distribution infesting a vast list of host species in America. However, little is known about the past geographic distribution of this species. In this study, we investigated the potential past and current distribution of species in South America. In this sense, the MaxEnt algorithm was used to model the ecological niche of the species in the past (Last Interglcial Maximum; Last Glacial Maximum) and current periods. The results suggested that under the current climatic conditions, A. obliqua showed high environmental suitability to become established in most South American countries, especially in Brazil. The lowest suitability indices were observed in Chile, Argentina, and Uruguay. The past analysis for Last Glacial Maximum revealed that there was no significant change in the distribution potential of the species when compared to the current model; however, in the Last Interglacial Maximum period, there was a large reduction in the areas of suitability for the species when compared to the current and Last Glacial Maximum distribution models. The analysis also revealed vast areas of refuges for the species mainly on the coast of Brazil, as well as Venezuela, Bolivia, Guyana, and Surinam. The results presented here may be useful for future phylogeographical studies in order to test if the refuge areas concentrate greater genetic diversity for this species. In addition, our study provides important information for understanding the current dynamics of A. obliqua, which may be useful for control programs in places where this species can become a pest.
Assuntos
Distribuição Animal , Tephritidae , Animais , Ecossistema , Modelos Teóricos , Filogeografia , América do SulRESUMO
We assessed the transferability of 120 EST-derived Eucalyptus microsatellite primers to Campomanesia adamantium and C. pubescens. Both species are berry trees native to the Brazilian Cerrado, and population genetic information is poor. Twelve markers were used to analyze the genetic variability of four sampled populations. Regarding DNA extraction, we sampled leaf tissues from two populations of each species (80 individuals). Of the 120 primers evaluated, 87 did not amplify any PCR products, and 21 rendered nonspecific amplification. Twelve primers were successfully transferred, providing a low combined probability of genetic identity for both species (5.718 x 10(-10) for C. adamantium; 1.182 x 10(-11) for C. pubescens) and a high probability of paternity exclusion (0.99939 for C. adamantium; 0.99982 for C. pubescens). The average number of alleles in the polymorphic loci was 6.8 for C. adamantium and 7.8 for C. pubescens, ranging from 2 to 16 alleles per locus. The observed heterozygosity values for C. adamantium and C. pubescens were 0.504 and 0.503, respectively, and the expected heterozygosity values for C. adamantium and C. pubescens were 0.517 and 0.579, respectively. The populations exhibited structured genetic variability with qP values of 0.105 for C. adamantium and 0.249 for C. pubescens. Thus, we concluded that these 12 microsatellite markers, transferred from Eucalyptus, were efficient for population genetic studies of C. adamantium and C. pubescens.
Assuntos
Variação Genética , Repetições de Microssatélites , Myrtaceae/genética , Alelos , DNA de Plantas , Etiquetas de Sequências Expressas , Genética PopulacionalRESUMO
Microsatellite markers are a useful tool for ecological monitoring of natural and managed populations. A technical limitation is the necessity for investment in the development of primers. Heterologous primers can provide an alternative to searching for new loci. In bees, these markers have been used in populational and intracolonial genetic analyses. The genus Melipona has the largest number of species among bee genera, about 70, occurring throughout the Neotropical region. However, only five species of the genus Melipona have specific microsatellite markers. Given the great diversity of this genus, this number is not representative. We analyzed the transferability of 49 microsatellite loci to four other species of the genus Melipona (M. scutellaris, M. mondury, M. mandacaia, and M. quadrifasciata). Four individuals of each species, from different localities, were used in amplification tests. Primer pairs described for five Melipona species and for Trigona carbonaria were tested. Among the 49 loci, 22 gave amplification products for all four species, while three gave nonspecific bands and five showed no amplification products. The remaining loci varied in the pattern of amplification, according to the species examined. The number of alleles ranged from 1 to 6. The results demonstrate the possibility of using these heterologous markers in other Melipona species, increasing the number of loci that can be analyzed and contributing to further genetic analyses of intra- and intercolonial structure, which is required for conservation measure planning, genetic improvement and resolution of taxonomic problems.
Assuntos
Abelhas/genética , Mordeduras e Picadas/genética , Primers do DNA/metabolismo , Repetições de Microssatélites/genética , Animais , Loci Gênicos/genética , Reação em Cadeia da Polimerase , Especificidade da EspécieRESUMO
Arbekacin is an aminoglycoside used in Japan for treating infections caused by gentamicin and oxacillin-resistant S. aureus (ORSA). The objective of this study was to determine the in vitro antimicrobial activity of arbekacin against 454 clinical isolates of ORSA. The isolates were consecutively collected between January and July, 2000, from patients hospitalized in 8 Brazilian medical centers. The antimicrobial susceptibility testing was performed by disk diffusion method according to NCCLS recommendations. The vast majority of the isolates, 453 strains (99.8%), were considered susceptible to arbekacin based on the criteria proposed by the Requirements for Antibiotic Products of Japan. Only 1 isolate (0.2%) was classified as resistant. On the other hand, high rates of resistance were demonstrated for other aminoglycosides, such as gentamicin (97.6% resistance) and amikacin (97.0% resistance). Resistance rate was also high for ciprofloxacin (98.0%). All isolates were considered susceptible to vancomycin. The excellent in vitro antimicrobial activity of arbekacin demonstrated in this study indicates that this antimicrobial agent may play an important role in the treatment of severe ORSA infections, especially those that show poor clinical response with vancomycin monotherapy. Since the aminoglycosides should not be used as monotherapy to treat Gram positive infections, further studies evaluating in vitro and in vivo synergistic activity of arbekacin combinations are necessary to clarify the clinical role of this aminoglycoside.
Assuntos
Aminoglicosídeos , Antibacterianos/farmacologia , Dibecacina/análogos & derivados , Dibecacina/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Brasil , Resistência Microbiana a Medicamentos , Hospitais , Humanos , Testes de Sensibilidade Microbiana , Oxacilina/farmacologia , Resistência às Penicilinas , Penicilinas/farmacologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/isolamento & purificaçãoAssuntos
População Negra , Sobrevivência de Enxerto , Transplante de Rim/fisiologia , População Branca , Adulto , Brasil , Feminino , Seguimentos , Rejeição de Enxerto/epidemiologia , Teste de Histocompatibilidade , Humanos , Transplante de Rim/imunologia , Masculino , Estudos Retrospectivos , Fatores de TempoRESUMO
The antimicrobial susceptibility of 239 coagulase-negative staphylococci (CNS) isolates consecutively collected from blood culture in patients admitted in a 600-bed teaching hospital was evaluated. The isolates were identified to the species level by conventional methods and the MicroScan Positive Combo Panel type 6 system, and their susceptibility to vancomycin, teicoplanin, and oxacillin were tested by agar dilution, disk diffusion, and MicroScan-WalkAway system. The species distribution was as follows: Staphylococcus epidermidis 120 (50.2%), S. hominis 29 (12.1%), S. haemolyticus 24 (10.0%), S. cohnii 14 (5.9%), and isolates from other CNS species 52 (21.8%). The percentage of resistance to oxacillin was 74.5% by agar dilution. The highest percentages of oxacillin resistance were found among S. haemolyticus (95.8%) and S. epidermidis (80.8%). Teicoplanin resistance (MIC > or = 32 micrograms/mL) was detected in five S. haemolyticus isolates, whereas intermediate resistance (MIC = 16 micrograms/mL) was detected in nine strains. These isolates with reduced susceptibility to teicoplanin were resistant to oxacillin, but remained susceptible to vancomycin (MIC < or = 4 micrograms/mL). Two isolates, one S. haemolyticus and one S. epidermidis, showed a vancomycin MIC of 8 micrograms/mL, and both MicroScan and disk diffusion methods classified these isolates as susceptible. Our results showed that glycopeptide resistance is emerging among CNS isolates in our institution and the disk diffusion method may not detect isolates with decreased susceptibility to these antimicrobial agents.
Assuntos
Antibacterianos/farmacologia , Infecções Estafilocócicas/microbiologia , Staphylococcus/efeitos dos fármacos , Teicoplanina/farmacologia , Vancomicina/farmacologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Bacteriemia/microbiologia , Técnicas de Tipagem Bacteriana , Sangue/microbiologia , Pré-Escolar , Coagulase/metabolismo , Resistência Microbiana a Medicamentos , Eletroforese em Gel de Campo Pulsado , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Oxacilina/farmacologia , Penicilinas/farmacologia , Staphylococcus/classificação , Staphylococcus/enzimologia , Staphylococcus/isolamento & purificaçãoRESUMO
Production of industrial enzymes including cellulases requires minimum cost with the downstream processing. The objective of this work was to analyze the precipitation of cellulases by ammonium sulfate in the presence of hydroxypropyl(methylcellulose) as a co-precipitant through the use of statistical experimental design. The model generated with the experimental results showed that high protein recovery can be achieved at high levels of temperature, aging times, and rate of salt-solution addition, and at a low mixing level. The results also allowed the observation that activity recovery was improved at high levels of temperature, rate of salt addition and mixing level, and a low level of aging time.
RESUMO
A simple and inexpensive shaker/Erlenmeyer flask system for large-scale cultivation of insect cells is described and compared to a commercial spinner system. On the basis of maximum cell density, average population doubling time and overproduction of recombinant protein, a better result was obtained with a simpler and less expensive bioreactor consisting of Erlenmeyer flasks and an ordinary shaker waterbath. Routinely, about 90 mg of pure poly(ADP-ribose) polymerase catalytic domain was obtained for a total of 3 x 10(9) infected cells in three liters of culture.
Assuntos
Baculoviridae , Insetos/citologia , Poli Adenosina Difosfato Ribose , Polinucleotídeo Adenililtransferase/isolamento & purificação , Proteínas Recombinantes/isolamento & purificação , Animais , Técnicas In VitroRESUMO
A simple and inexpensive shaker/Erlenmeyer flask system for largescale cultivation of insect cells is described and compared to a commercial spinner system. On the basis of maximum cell density, average population doubling time and overproduction of recombinant protein, a better result was obtained with a simpler and less expensive biorector consisting of Erlenmeyer flasks and an ordinary shaker waterbath. Routinely, about 90 mg of pure poly(ADP-ribose) polymerase catalytic domain was obtained for a total of 3 x 10(9) infected cells in three liters of culture.
Assuntos
Animais , ADP Ribose Transferases , Baculoviridae , Técnicas In Vitro , Insetos/citologia , Poli Adenosina Difosfato Ribose , Polinucleotídeo Adenililtransferase/isolamento & purificação , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
Two antibodies, one monoclonal and one polyclonal, were produced and used to identify Taenia solium eggs, using the enzyme-linked immuno-electrotransfer blot technique (EITB). Different life-stages of Taenia solium and T. saginata, including eggs from gravid proglottids recovered, post-treatment, from patients infected with the tapeworms, and eggs of Diphyllobothrium pacificum and Hymenolepis nana from other patients were tested with these antibodies. The monoclonal antibody only recognized the eggs and immature proglottids of T. solium. The polyclonal antibody, however, not only reacted with the eggs, cysticerci and immature proglottids of T. solium but also with the eggs and immature proglottids of T. saginata. The sensitivity and specificity of the EITB were both 100% using the monoclonal antibody but only 78% and 60%, respectively, using the polyclonal antibody. Diagnostic bands for T. solium eggs corresponded to proteins of 22.5 kDa using the polyclonal antibody and 22.5-37 kDa using the monoclonal antibody. Species-specific fluorescence was obtained with an anti-T. solium monoclonal antibody which bound to egg-derived oncospheres of T. solium but not to those of T. saginata.