RESUMO
Background: Assisted reproduction techniques in birds have contributed to many species' conservation and sustainable use. One of these techniques is semen cryopreservation, which is possible following the discovery of suitable cryoprotectants. Aims: This study aimed to characterize the fresh and post-thaw ejaculates of different species of birds of prey. Methods: The following species were included in the study: red-tailed hawk (Buteo jamaicensis) n=3, golden eagle (Aquila chrysaetos) n=3, and Harris's hawk (Parabuteo unicinctus) n=3. Twenty-five ejaculates were obtained for each species. The percentage of spermatozoa motility, viability, and morphology were evaluated. Results: Evident differences were observed among the ejaculates of the three species, particularly in sperm length and between the fresh and post-thaw parameters of the same species in which the motility reduced to approximately 40% after thawing. It was demonstrated that sperm cryopreservation of the studied species was possible using the same freezing protocol. Conclusion: This study showed that sperm characteristics could influence the parameters obtained during their in vitro conservation, both in the fresh and post-thaw states.
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Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) control antral follicular growth by regulating several processes, such as the synthesis of hormones and signaling molecules, proliferation, survival, apoptosis, luteinization, and ovulation. To exert these effects, gonadotropins bind to their respective Gs protein-coupled receptors, activating the protein kinase A (PKA) pathway or recruiting Gq proteins to activate protein kinase C (PKC) signaling. Although the action mechanism of FSH and LH is clear, recently, it has been shown that both gonadotropins promote the synthesis of sphingosine-1-phosphate (S1P) in granulosa and theca cells through the activation of sphingosine kinase 1. Moreover, the inhibition of SPHKs reduces S1P synthesis, cell viability, and the proliferation of follicular cells in response to gonadotropins, and the addition of S1P to the culture medium increases the proliferation of granulosa and theca cells without apparent effects on sexual steroid synthesis. Therefore, we consider that S1P is a crucial signaling molecule that complements the canonical gonadotropin pathway to promote the proliferation and viability of granulosa and theca cells.
Assuntos
Gonadotropinas , Lisofosfolipídeos , Folículo Ovariano , Esfingosina , Esfingosina/análogos & derivados , Esfingosina/metabolismo , Esfingosina/farmacologia , Lisofosfolipídeos/metabolismo , Lisofosfolipídeos/farmacologia , Feminino , Animais , Humanos , Gonadotropinas/metabolismo , Folículo Ovariano/metabolismo , Folículo Ovariano/efeitos dos fármacos , Hormônio Luteinizante/metabolismo , Hormônio Foliculoestimulante/metabolismo , Hormônio Foliculoestimulante/farmacologia , Transdução de Sinais/efeitos dos fármacos , Células da Granulosa/metabolismo , Células da Granulosa/efeitos dos fármacosRESUMO
Introducción: Los medios de enseñanzas son los ejes vertebrales de gran parte de las acciones de enseñanzas y aprendizaje desarrolladas en cualquiera de los niveles o modalidades de la educación. Objetivos: Sistematizar y resaltar los fundamentos y ventajas de la guía de autoayuda sobre la fractura de cadera en el adulto mayor, como medio de enseñanza en la superación profesional del médico de la atención primaria. Métodos: Analítico sintético, hipotético deductivo, análisis documental y la entrevista. Resultados: Se elaboró una guía de autoayuda sobre fractura de cadera en el adulto mayor, a partir de falencias detectadas en la atención primaria de salud, que limitan el desempeño de los médicos de este nivel en actividades de promoción de salud en este grupo social. La guía de autoayuda fue diseñada y estructurada, como medio de enseñanza de estrategias de superación profesional, determinando las particularidades del contenido que se desea enseñar y las que se necesitan aprender por estos profesionales, para mejorar su desempeño en la promoción de salud en relación con el adulto mayor con fractura de cadera. Su creación fue concebida como elemento dinamizador del contenido unido a los demás componentes del proceso en busca de lograr los objetivos propuestos. Conclusiones: Se presenta una guía de la colección de autoayuda sobre fractura de cadera en el adulto mayor, diseñado, pensado y elaborado como medio de enseñanza a utilizar en la superación profesional del médico de la atención primaria de salud donde se resaltan los fundamentos teóricos que lo sustentan.
Introduction: Teaching media are the backbone of a large part of the teaching and learning actions developed at every educational level or modality. Objectives: To systematize and highlight the rationale and advantages of the self-help guide for hip fracture in the elderly, considered as a teaching tool in the professional improvement of primary health care ( PHC). Methods: Analytical - synthetic, hypothetical - deductive, documentary analysis and interview. Results: A self-help guide for hip fracture in the elderly was developed, based on deficiencies detected on PHC, which limit the performance of doctors at this level in health promotion activities for the mentioned social group. The self-help guide was designed and structured as a means of teaching professional improvement strategies by determining the content particularities to be taught and those to be learned by these professionals, in order to improve their performance in health promotion for the elderly with hip fracture. This self- help guide was conceived as a dynamic element of the content, that relates to the other components of the process in search of achieving the proposed objectives. Conclusions: We present a self-help collection guide for hip fracture in the elderly, designed, conceived, and elaborated as a teaching tool to be used in the professional improvement of the PHC, and we highlight theoretical foundations
Assuntos
Idoso , Fraturas Ósseas , Atenção Primária à Saúde , Ensino , QuadrilRESUMO
Fusarium Head Blight (FHB) is a very important fungal disease that affects small grain cereals worldwide. This disease not only causes yield loses but also crops contamination with mycotoxins such as deoxynivalenol (DON) and nivalenol (NIV). Species within the Fusarium graminearum species complex have been described as the main causal agents of this disease, however lately there have been few reports of Fusarium cerealis causing the disease in wheat and barley in different parts of the world. This study evaluated the aggressiveness of F. cerealis to durum wheat cultivars and also mycotoxin production in planta. Moreover, the mycotoxin profile of F. cerealis strains was characterized molecularly and chemically. All durum wheat cultivars showed typical FHB symptoms but the disease severity varied among them in levels up to 66%. In addition, seventeen different compounds were detected in the infected heads including DON, NIV and nivalenol-3-ß-d-glucose (NIV3G). NIV was detected in all cultivars and was the most produced mycotoxin with levels ranging from 1.04 to 6.8 mg/kg. On the other hand, the molecular analysis of F. cerealis strains showed that all of them possessed NIV genotype while the chemical assessment showed that the strains were able to produce not only this toxin in vitro but also DON, zearalenone and other twenty-one secondary metabolites. The increasing incidence of F. cerealis and the possible contamination of crops with the mycotoxins that it produces are of great concern for food security and world cereal trade since it has been reported that NIV is more toxic for humans and animals than DON.
Assuntos
Fusarium/metabolismo , Micotoxinas/metabolismo , Doenças das Plantas/microbiologia , Triticum/microbiologia , Grão Comestível/microbiologia , Contaminação de Alimentos/análise , Fusarium/genética , Genótipo , Hordeum/química , Hordeum/microbiologia , Tricotecenos/análise , Tricotecenos/metabolismo , Triticum/química , Zearalenona/análise , Zearalenona/metabolismoRESUMO
Mercury is a widespread pollutant. Mercuric ions uptake into tubular cells is supported by the Organic anion transporter 1 (Oat1) and 3 (Oat3) and its elimination into urine is through the Multidrug resistance-associated protein 2 (Mrp2). We investigated the effect of recombinant human erythropoietin (Epo) on renal function and on renal expression of Oat1, Oat3, and Mrp2 in a model of mercuric chloride (HgCl2)-induced renal damage. Four experimental groups of adult male Wistar rats were used: Control, Epo, HgCl2, and Epo + HgCl2. Epo (3000 IU/kg, b.w., i.p.) was administered 24 h before HgCl2 (4 mg/kg, b.w., i.p.). Experiments were performed 18 h after the HgCl2 dose. Parameters of renal function and structure were evaluated. The protein expression of Oat1, Oat3 and Mrp2 in renal tissue was assessed by immunoblotting techniques. Mercury levels were determined by cold vapor atomic absorption spectrometry. Pretreatment with Epo ameliorated the HgCl2-induced tubular injury as assessed by histopathology and urinary biomarkers. Immunoblotting showed that pretreatment with Epo regulated the renal expression of mercury transporters in a way to decrease mercury content in the kidney. Epo pretreatment ameliorates HgCl2-induced renal tubular injury by modulation of mercury transporters expression in the kidneys.
Assuntos
Eritropoetina/uso terapêutico , Nefropatias/tratamento farmacológico , Cloreto de Mercúrio/toxicidade , Substâncias Protetoras/uso terapêutico , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Eritropoetina/genética , Eritropoetina/farmacologia , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Nefropatias/induzido quimicamente , Nefropatias/metabolismo , Nefropatias/patologia , Masculino , Mercúrio/sangue , Mercúrio/metabolismo , Mercúrio/urina , Proteína 1 Transportadora de Ânions Orgânicos/metabolismo , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Substâncias Protetoras/farmacologia , Ratos Wistar , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêutico , Ureia/sangueRESUMO
The aim of this study was to evaluate in vitro the probiotic potential and absorption of Saccharomyces cerevisiae for the aflatoxin B1 in simulated fish intestinal tract conditions. Three yeast strains were used, two from brewery: S. cerevisiae RC1 and S. cerevisiae RC3 and one from a fish farming environment: S. cerevisiae A8L2. The selected yeasts were subjected to the following in vitro tests: homologous inhibition, self-aggregation, co-aggregation, antibacterial activity, gastrointestinal conditions tolerance and adsorption of AFB1. All S. cerevisiae strains showed good capability of self-aggregation and co-aggregation with pathogenic bacteria. All yeast strains were able to survive the gastrointestinal conditions. In acidic conditions, the factors (strain vs. time) had interaction (P=0.0317), resulting in significant variation among the strains tested in the time periods analyzed. It was observed that there was also interaction (P=0.0062) in intestinal conditions, with an increased number of cells in the 12-hour period for all strains tested. In the adsorption test, the A8L2 strain was statistically more effective (P<0.005) for both AFB1 concentrations evaluated in this study (10 and 25ng/mL). Thus, it was observed that the strains of S. cerevisiae have potential probiotic and adsorbent of AFB1.(AU)
Objetivou-se, com esta pesquisa, avaliar in vitro o potencial probiótico e adsorvente de Saccharomyces cerevisiae para aflatoxina B1 em condições simuladas do trato intestinal de peixes. Foram utilizadas três cepas de leveduras, sendo duas provenientes de cervejaria: S. cerevisiae RC1 e S. cerevisiae RC3, e uma de ambiente de piscicultura: S. cerevisiae A8L2. As leveduras selecionadas foram submetidas aos seguintes testes in vitro: inibição homóloga, autoagregação, coagregação, atividade antibacteriana, viabilidade às condições gastrointestinais e adsorção de AFB1. Todas as estirpes de S. cerevisiae mostraram boa capacidade de autoagregação e coagregação com bactérias patogênicas. Todas as estirpes de levedura foram capazes de sobreviver às condições gastrointestinais. Em condições ácidas, os fatores (cepa x tempo) tiveram interação (P=0,0317), resultando em variações significativas entre as cepas testadas nos períodos de tempo analisados. Observou-se que também houve interação (P=0,0062) em condições intestinais, havendo um aumento do número de células no período de 12h para todas as cepas avaliadas. No ensaio de adsorção, a estirpe A8L2 foi a mais eficaz estatisticamente (P<0,005), para as duas concentrações de AFB1 avaliadas neste estudo (10 e 25ng. mL-1). Dessa forma, conclui-se que as cepas de Saccharomyces cerevisiae possuem potencial probiótico e adsorvente de AFB1.(AU)
Assuntos
Animais , Saccharomyces cerevisiae , Aflatoxina B1/antagonistas & inibidores , Probióticos/uso terapêutico , Peixes/fisiologia , Intestinos/microbiologia , Técnicas In Vitro , AdsorçãoRESUMO
The aim of this study was to evaluate in vitro the probiotic potential and absorption of Saccharomyces cerevisiae for the aflatoxin B1 in simulated fish intestinal tract conditions. Three yeast strains were used, two from brewery: S. cerevisiae RC1 and S. cerevisiae RC3 and one from a fish farming environment: S. cerevisiae A8L2. The selected yeasts were subjected to the following in vitro tests: homologous inhibition, self-aggregation, co-aggregation, antibacterial activity, gastrointestinal conditions tolerance and adsorption of AFB1. All S. cerevisiae strains showed good capability of self-aggregation and co-aggregation with pathogenic bacteria. All yeast strains were able to survive the gastrointestinal conditions. In acidic conditions, the factors (strain vs. time) had interaction (P=0.0317), resulting in significant variation among the strains tested in the time periods analyzed. It was observed that there was also interaction (P=0.0062) in intestinal conditions, with an increased number of cells in the 12-hour period for all strains tested. In the adsorption test, the A8L2 strain was statistically more effective (P<0.005) for both AFB1 concentrations evaluated in this study (10 and 25ng/mL). Thus, it was observed that the strains of S. cerevisiae have potential probiotic and adsorbent of AFB1.(AU)
Objetivou-se, com esta pesquisa, avaliar in vitro o potencial probiótico e adsorvente de Saccharomyces cerevisiae para aflatoxina B1 em condições simuladas do trato intestinal de peixes. Foram utilizadas três cepas de leveduras, sendo duas provenientes de cervejaria: S. cerevisiae RC1 e S. cerevisiae RC3, e uma de ambiente de piscicultura: S. cerevisiae A8L2. As leveduras selecionadas foram submetidas aos seguintes testes in vitro: inibição homóloga, autoagregação, coagregação, atividade antibacteriana, viabilidade às condições gastrointestinais e adsorção de AFB1. Todas as estirpes de S. cerevisiae mostraram boa capacidade de autoagregação e coagregação com bactérias patogênicas. Todas as estirpes de levedura foram capazes de sobreviver às condições gastrointestinais. Em condições ácidas, os fatores (cepa x tempo) tiveram interação (P=0,0317), resultando em variações significativas entre as cepas testadas nos períodos de tempo analisados. Observou-se que também houve interação (P=0,0062) em condições intestinais, havendo um aumento do número de células no período de 12h para todas as cepas avaliadas. No ensaio de adsorção, a estirpe A8L2 foi a mais eficaz estatisticamente (P<0,005), para as duas concentrações de AFB1 avaliadas neste estudo (10 e 25ng. mL-1). Dessa forma, conclui-se que as cepas de Saccharomyces cerevisiae possuem potencial probiótico e adsorvente de AFB1.(AU)
Assuntos
Animais , Saccharomyces cerevisiae , Aflatoxina B1/antagonistas & inibidores , Probióticos/uso terapêutico , Peixes/fisiologia , Intestinos/microbiologia , Técnicas In Vitro , AdsorçãoRESUMO
AIM: To investigate the impact of inoculating peanut seeds with the biocontrol agent Trichoderma harzianum ITEM 3636 on the structure of bacterial and fungal communities from agricultural soils. METHODS AND RESULTS: Polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (PCR-DGGE) and next-generation sequencing (NGS) of amplicons (or marker gene amplification metagenomics) were performed to investigate potential changes in the structure of microbial communities from fields located in a peanut-producing area in the province of Córdoba, Argentina. Fields had history of peanut smut (caused by Thecaphora frezii) incidence. The Shannon indexes (H'), which estimate diversity, obtained from the PCR-DGGE assays did not show significant differences neither for bacterial nor for fungal communities between control and inoculation treatments. On the other hand, the number of operational taxonomic units obtained after NGS was similar between all the analysed samples. Moreover, results of alpha and beta diversity showed that there were no significant variations between the relative abundances of the most representative bacterial and fungal phyla and genera, in both fields. CONCLUSIONS: Trichoderma harzianum ITEM 3636 decreases the incidence and severity of agriculturally relevant diseases without causing significant changes in the microbial communities of agricultural soils. SIGNIFICANCE AND IMPACT OF THE STUDY: Our investigations provide information on the structure of bacterial and fungal communities in peanut-producing fields after inoculation of seeds with a biocontrol agent.
Assuntos
Agentes de Controle Biológico , Microbiologia do Solo , Trichoderma , Agricultura , Arachis , Argentina , Bactérias/genética , Bactérias/isolamento & purificação , Eletroforese em Gel de Gradiente Desnaturante , Fungos/genética , Fungos/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota , Reação em Cadeia da Polimerase , Sementes , Solo/químicaRESUMO
Fusarium head blight (FHB) caused by Fusarium graminearum species complex is a devastating disease that causes extensive yield and quality losses to wheat around the world. Fungicide application and breeding for resistance are among the most important tools to counteract FHB. Biological control is an additional tool that can be used as part of an integrated management of FHB. Bacillus velezensisRC 218, Brevibacillus sp. RC 263 and Streptomyces sp. RC 87B were selected by their potential to control FHB and deoxynivalenol production. The aim of this work was to test the tolerance of these biocontrol agents to triazole-based fungicides such as prothioconazole, tebuconazole and metconazole. Bacterial growth was evaluated in Petri dishes using the spread plating technique containing the different fungicides. Bacillus velezensisRC 218 and Streptomyces sp. RC 87B showed better tolerance to fungicides than Brevibacillus sp. RC 263. Complete growth inhibition was observed at concentrations of 20 µg ml-1 for metconazole, 40 µg ml-1 for tebuconazole and 80 µg ml-1 for prothioconazole. The results obtained indicate the possibility of using these biocontrol agents in combination with fungicides as part of an integrated management to control FHB of wheat. SIGNIFICANCE AND IMPACT OF THE STUDY: This study evaluates the possibility to use biocontrol agents (Bacillus velezensisRC 218, Brevibacillus sp. RC 263 and Streptomyces sp. RC 87B) in combination with triazole-based fungicides to control Fusarium head blight in wheat. The evaluation of biocontrol agents' growth under in vitro conditions was carried out in Petri dishes containing either prothioconazole, tebuconazole or metconazole. Viability studies demonstrated that B. velezensisRC 218 and Streptomyces sp. RC 87B were more tolerant to the fungicides evaluated. Results obtained reflect the possibility to use fungicides at low doses combined with biocontrol agents.
Assuntos
Bacillus/efeitos dos fármacos , Agentes de Controle Biológico/metabolismo , Brevibacillus/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Streptomyces/efeitos dos fármacos , Triazóis/farmacologia , Antibiose/fisiologia , Argentina , Bacillus/crescimento & desenvolvimento , Bacillus/metabolismo , Brevibacillus/crescimento & desenvolvimento , Brevibacillus/metabolismo , Fusarium/efeitos dos fármacos , Doenças das Plantas/microbiologia , Streptomyces/crescimento & desenvolvimento , Streptomyces/metabolismo , Tricotecenos/biossíntese , Triticum/microbiologiaRESUMO
The objective of this study was to evaluate if short-term dietary concentrate supplementation increased IGF-I serum concentration and resulted in a reproductive response during estrus synchronization treatment in non-lactating beef cows. Thirty non-lactating beef cows (Bos indicus × Bos taurus) were allocated to the same pastureland and fed native tropical grasses as a basal diet. Cows were synchronized using a 7-day CO-Synch plus controlled internal drug release (CIDR) protocol and received fixed time artificial insemination (FTAI). Cows were divided into two groups; the control group (n = 16) received 0.5 kg of concentrate/cow/day, whereas the supplemented group (n = 14) received 4.0 kg of concentrate/cow/day. The period of supplementation was 10 days from the day of CIDR insert to FTAI. The concentration of IGF-I increased (P < 0.05) in the supplemented group, while no significant changes were observed in the control group. Moreover, at the time of insemination, IGF-I serum concentrations were higher in supplemented cows compared with control cows (P < 0.05). Notably, metabolite and insulin concentrations did not differ (P > 0.05) between treatment groups or sampling day. The response to estrus induction, measured as estrus presentation, ovulation rate, and pregnancy rate, was similar between experimental groups (P > 0.05). In conclusion, our results indicated that supplementation with dietary concentrate for 10 days in non-lactating beef cows changed the endocrine milieu, specifically increasing IGF-I serum concentration. However, these endocrine changes did not affect response to estrous induction treatment.
Assuntos
Suplementos Nutricionais , Sincronização do Estro/efeitos dos fármacos , Inseminação Artificial/veterinária , Fator de Crescimento Insulin-Like I/análise , Criação de Animais Domésticos , Animais , Comportamento Animal , Composição Corporal , Peso Corporal , Bovinos , Preparações de Ação Retardada , Dinoprosta/farmacologia , Estro/efeitos dos fármacos , Feminino , Hormônio Liberador de Gonadotropina/fisiologia , Folículo Ovariano/fisiologia , Ovulação/efeitos dos fármacos , Gravidez , Taxa de Gravidez , Progesterona/sangue , Carne Vermelha , Fatores de TempoRESUMO
Sphingosine-1-phosphate (S1P) is a bioactive polar sphingolipid which stimulates proliferation, growth and survival in various cell types. In the ovary S1P has been shown protect the granulosa cells and oocytes from insults such as oxidative stress and radiotherapy, and S1P concentrations are greater in healthy than atretic large follicles. Hence, we postulate that S1P is fundamental in follicle development and that it is activated in ovarian granulosa cells in response to FSH and VEGF. To test this hypothesis we set out: i) to evaluate the effect of FSH and VEGF on S1P synthesis in cultured bovine granulosa cells and ii) to analyse the effect of S1P on proliferation and survival of bovine granulosa cells in vitro. Seventy five thousand bovine granulosa cells from healthy medium-sized (4-7mm) follicles were cultured in 96-well plates in McCoy's 5a medium containing 10ng/mL of insulin and 1ng/mL of LR-IGF-I at 37°C in a 5% CO2/air atmosphere at 37°C. Granulosa cell production of S1P was tested in response to treatment with FSH (0, 0.1, 1 and 10ng/mL) and VEGF (0, 0.01, 0.1, 1, 10 and 100ng/mL) and measured by HPLC. Granulosa cells produced S1P at 48 and 96h, with the maximum production observed with 1ng/mL of FSH. Likewise, 0.01ng/mL of VEGF stimulated S1P production at 48, but not 96h of culture. Further, the granulosa cell expression of sphingosine kinase-1 (SK1), responsible for S1P synthesis, was demonstrated by Western blot after 48h of culture. FSH increased the expression of phosphorylated SK1 (P<0.05) and the addition of a SK1 inhibitor reduced the constitutive and FSH-stimulated S1P synthesis (P<0.05). Sphingosine-1-phosphate had a biphasic effect on granulosa cell number after culture. At low concentration S1P (0.1µM) increased granulosa cell number after 48h of culture (P<0.05) and the proportion of cells in the G2 and M phase of the cell cycle (P<0.05), whereas higher concentrations decreased cell number (10µM; P<0.05) by an increase (P<0.05) in the proportion of cells in apoptosis (hypodiploid cells). In addition, treatment with SK-178 suppressed the FSH- and VEGF-stimulated rise of the granulosa cells number (P<0.05). Interestingly, the effect of 0.1µM S1P on granulosa cell number and their proportion in G2/M phases is similar to that observed with 1ng/mL FSH. The results of this study are the first to demonstrate sphingosine-1-phosphate (S1P) synthesis in granulosa cells under the control of FSH and VEGF. The later achieved through the regulation of sphingosine kinase 1 expression. This S1P augments the proportion of cells in the G2/M phase of the cell cycle that translates in increased granulosa cell proliferation.
Assuntos
Hormônio Foliculoestimulante/metabolismo , Células da Granulosa/metabolismo , Lisofosfolipídeos/metabolismo , Esfingosina/análogos & derivados , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Bovinos , Proliferação de Células , Feminino , Esfingosina/metabolismoRESUMO
Aspergillus section Nigri is a heterogeneous fungal group including some ochratoxin A producer species that usually contaminate raisins. The section contains the Series Carbonaria which includes the toxigenic species Aspergillus carbonarius and nontoxigenic Aspergillus ibericus that are phenotypically undistinguishable. The aim of this study was to examine the diversity of black aspergilli isolated from raisins and to develop a specific genetic marker to distinguish A. ibericus from A. carbonarius. The species most frequently found in raisins in this study were Aspergillus tubingensis (35.4%) and A. carbonarius (32.3%), followed by Aspergillus luchuensis (10.7%), Aspergillus japonicus (7.7%), Aspergillus niger (6.2%), Aspergillus welwitschiae (4.6%) and A. ibericus (3.1%). Based on inter-simple sequence repeat (ISSR) fingerprinting profiles of major Aspergillus section Nigri members, a sequence-characterized amplified region (SCAR) marker was identified. Primers were designed based on the conserved regions of the SCAR marker and were utilized in a PCR for simultaneous identification of A. carbonarius and A. ibericus. The detection level of the SCAR-PCR was found to be 0.01 ng of purified DNA. The present SCAR-PCR is rapid and less cumbersome than conventional identification techniques and could be a supplementary strategy and a reliable tool for high-throughput sample analysis.
Assuntos
Aspergillus/genética , Microbiologia de Alimentos/métodos , Marcadores Genéticos/genética , Vitis/microbiologia , Argentina , Aspergillus/isolamento & purificação , Aspergillus niger/genética , Biodiversidade , Primers do DNA/genética , DNA Fúngico/genética , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Especificidade da EspécieRESUMO
Fusarium proliferatum is a member of the Fusarium fujikuroi species complex (FFSC) involved in the maize ear rot together with Fusarium verticillioides, which is a very closely related species. Recently, different studies have detected natural fumonisin contamination in wheat kernels and most of them have shown that the main species isolated was F. proliferatum. Fusarium strains obtained from freshly harvested durum wheat samples (2008 to 2011 harvest seasons) from Argentina were characterized through a phylogenetic analysis based on translation elongation factor-1 alpha (EF-1α) and calmodulin (CaM) genes, determination of mating type alleles, and evaluation of fumonisin production capability. The strains were identified as F. proliferatum (72%), F. verticillioides (24%) and other Fusarium species. The ratio of mating type alleles (MAT-1 and MAT-2) obtained for both main populations suggests possible occurrence of sexual reproduction in the wheat fields, although this seems more frequent in F. proliferatum. Phylogenetic analysis revealed greater nucleotide variability in F. proliferatum strains than in F. verticillioides, however this was not related to origin, host or harvest year. The fumonisin-producing ability was detected in 92% of the strains isolated from durum wheat grains. These results indicate that F. proliferatum and F. verticillioides, among the fumonisin producing species, frequently contaminate durum wheat grains in Argentina, presenting a high risk for human and animal health.
Assuntos
Fumonisinas/metabolismo , Fusarium/genética , Genes Fúngicos/genética , Variação Genética , Triticum/microbiologia , Argentina , Calmodulina/genética , Fumonisinas/análise , Fusarium/química , Fusarium/classificação , Fusarium/isolamento & purificação , Genes Fúngicos Tipo Acasalamento/genética , Fator 1 de Elongação de Peptídeos/genética , FilogeniaRESUMO
Myositis ossificans (MO) is a rare disease involving heterotopic ossification in the muscle or soft tissue. Myositis ossificans traumatica (MOT) disease presents as a calcification within the injured muscle, resulting from a single or repetitive injury. There are few reports of MOT in the masticatory muscles. The case of a patient with MOT in the medial pterygoid muscle caused by a complication related to the extraction of an erupted upper third molar is reported. The major symptom was severe trismus. Despite surgical treatment, the disease relapsed. MOT can lead to serious consequences for the patient. Its aetiopathogenesis needs to be better understood, so that the most appropriate treatment is established and relapses are minimized. This will improve the quality of life of these patients.
Assuntos
Dente Serotino/cirurgia , Miosite Ossificante/etiologia , Miosite Ossificante/terapia , Músculos Pterigoides/patologia , Extração Dentária/efeitos adversos , Adulto , Biópsia , Terapia Combinada , Feminino , Humanos , Imageamento por Ressonância Magnética , Miosite Ossificante/diagnóstico , Procedimentos Cirúrgicos Bucais , Modalidades de Fisioterapia , Radiografia Panorâmica , Tomografia Computadorizada por Raios XRESUMO
Mycotoxins including aflatoxins, deoxynivalenol, fumonisins and ochratoxin A are among the main fungal secondary metabolites detected as natural contaminants in South America in different commodities such as peanuts (aflatoxins), cereals (deoxynivalenol and fumonisins) or grapes (ochratoxin A). Different strategies including crop rotation, tillage practices, fungicide application and planting less susceptible cultivars are used in order to reduce the impact of these mycotoxins in both food and feed chains. The development of fungicide resistance in many fungal pathogens as well as rising of public concern on the risks associated with pesticide use led to the search for alternative environmentally friendly methods. Biological control of plant pathogens and toxigenic fungi offers an alternative that can complement chemical control in the frame of an integrated pest management to reduce the impact of mycotoxins in the food and feed chains. The advances made in Argentina on reducing the impact of toxigenic fungi and mycotoxins in peanut, grapes and cereals using the biocontrol strategy are summarised. Native bacteria, yeasts and filamentous fungi have been selected to evaluate them as potential biocontrol agents. Field trials showed that Bacillus subtilis RC 218 and Brevibacillus sp. RC 263 were effective at reducing deoxynivalenol accumulation in wheat. The application of Clonostachys rosea isolates on wheat stubble reduced Fusarium colonisation on the stubble. Bacillus amyloliquefaciens and Microbacterium oleovorans showed good activity to control both Fusarium verticillioides growth and the accumulation of fumonisins at pre-harvest stage in maize. Control of toxigenic Aspergillus flavus and aflatoxin accumulation in peanuts was achieved using a native atoxigenic Aspergillus flavus strain based on competitive exclusion of the toxigenic strains. Kluyveromyces thermotolerans strains were used as biocontrol agents to reduce the impact of Aspergillus section Nigri and ochratoxin A accumulation in grapes.
Assuntos
Arachis/microbiologia , Agentes de Controle Biológico , Grão Comestível/microbiologia , Micotoxinas/análise , Vitis/microbiologia , Aflatoxinas/análise , Argentina , Aspergillus flavus/isolamento & purificação , Bacillus subtilis/fisiologia , Brevibacillus/fisiologia , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Fumonisinas/análise , Fusarium/isolamento & purificação , Ocratoxinas/análise , Zea mays/microbiologiaRESUMO
In the spermatozoa of some species, the ubiquitin-proteasome system detects altered proteins and tags them for elimination by the proteasome. In some species' ejaculates, a high proportion of ubiquitinated spermatozoa (i.e. those having ubiquitin bound to the altered or damaged membrane proteins) has been related to infertility. The aim of this study was to assess whether the percentage of ubiquitinated spermatozoa relates to fertility of dairy bulls and whether ubiquitination increases during protein remodelling that occurs during in vitro spermatic capacitation. Thirty-two frozen semen straws from four high-fertility (ReproMax(®) ) and four normal-fertility (Normal) Holstein-Friesian sires were evaluated. Ubiquitinated and capacitated spermatozoa were quantified by sperm ubiquitin tag immunoassay and chlortetracycline stain, respectively. Fertilizing capacity of sires was assessed by in vitro fertilization. No differences were found between Normal and ReproMax(®) sires with regard to the observed percentage of ubiquitinated spermatozoa (42.97 ± 3.69% and 49.68 ± 9.27%, respectively; p > 0.05). Additionally, no differences were found in the percentage of ubiquitinated spermatozoa as a consequence of spermatic capacitation in either Normal (42.97 ± 3.69% before capacitation vs 44.67 ± 7.5% after; p > 0.05) or ReproMax(®) sires (49.68 ± 9.27% before vs 45.05 ± 7.51% after; p > 0.05). The percentage of ubiquitinated spermatozoa in a thawed sperm samples did not correlate with its in vitro fertilizing capacity; thus, this assay does not prove useful to detect in vivo fertility differences between sires. Additionally, protein degradation occurring during remodelling of the spermatozoon plasma membrane during the capacitation process does not seem to involve the ubiquitin-proteasome system.
Assuntos
Bovinos , Fertilização/fisiologia , Preservação do Sêmen/veterinária , Espermatozoides/química , Espermatozoides/fisiologia , Animais , Criopreservação/veterinária , Fertilização in vitro/veterinária , Temperatura Alta , Masculino , Preservação do Sêmen/métodos , Capacitação Espermática , Contagem de Espermatozoides/veterinária , UbiquitinaçãoRESUMO
AIMS: The objective of this work was to design an amplified fragment length polymorphism (AFLP)-derived specific primer for the detection of Fusarium solani aetiological agent of peanut brown root rot (PBRR) in plant material and soil. METHODS AND RESULTS: Specific primers for the detection of the pathogen were designed based on an amplified region using AFLPs. The banding patterns by AFLPs showed that isolates from diseased roots were clearly distinguishable from others members of the F. solani species complex. Many bands were specific to F. solani PBRR, one of these fragments was selected and sequenced. Sequence obtained was used to develop specific PCR primers for the identification of pathogen in pure culture and in plant material and soil. Primer pair FS1/FS2 amplified a single DNA product of 175 bp. Other fungal isolates occurring in soil, included F. solani non-PBRR, were not detected by these specific primers. The assay was effective for the detection of pathogen from diseased root and infected soils. CONCLUSIONS: The designed primers for F. solani causing PBRR can be used in a PCR diagnostic protocol to rapidly and reliably detect and identify this pathogen. SIGNIFICANCE AND IMPACT OF THE STUDY: These diagnostic PCR primers will aid the detection of F. solani causing PBRR in diseased root and natural infected soils. The method developed could be a helpful tool for epidemiological studies and to avoid the spread of this serious disease in new areas.
Assuntos
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Primers do DNA/química , Fusarium/isolamento & purificação , Arachis/microbiologia , DNA Fúngico/química , Fusarium/classificação , Fusarium/genética , Microbiologia do SoloRESUMO
Rheumatoid arthritis is a systemic autoimmune disease mediated by T cells. Productive engagement of T cell receptors by major histocompatibility complex-peptide leads to proliferation, differentiation and the definition of effector functions. Altered peptide ligands (APL) generated by amino acid substitutions in the antigenic peptide have diverse effects on T cell response. We predicted a novel T cell epitope from human heat-shock protein 60, an autoantigen involved in the pathogenesis of rheumatoid arthritis. Three APLs were designed from this epitope and it was demonstrated that these peptides induce the activation of T cells through their ability to modify cell cycle phase's distribution of CD4+T cells from RA patients. Also, IL-17, TNF-α and IL-10 levels were determined in PBMC from these patients. Unlike the wild-type peptide and the other two APLs, APL2 increased the IL-10 level and suppressed IL-17 secretion in these assays. Therapeutic effect of this APL in adjuvant arthritis (AA) and collagen-induced arthritis (CIA) models was also evaluated. Clinical score, histopathology, inflammatory and regulatory cytokine concentration were monitored in the animals. APL2 efficiently inhibited the progression of AA and CIA with a significant reduction of the clinical and histopathologic score. Therapeutic effect of APL2 on CIA was similar to that obtained with MTX; the standard treatment for RA. This effect was associated with a decrease of TNF-α and IL-17 levels. These results suggest that the therapeutic effect of APL2 is mediated in part by down-regulation of inflammatory cytokines and support the potential use of APL2 as a therapeutic drug in RA patients.
Assuntos
Artrite Reumatoide/tratamento farmacológico , Chaperonina 60/química , Citocinas/efeitos dos fármacos , Modelos Animais de Doenças , Peptídeos/uso terapêutico , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Citocinas/imunologia , Citocinas/metabolismo , Epitopos de Linfócito T/química , Epitopos de Linfócito T/imunologia , Feminino , Humanos , Inflamação , Interleucina-17/imunologia , Interleucina-17/metabolismo , Ligantes , Ativação Linfocitária/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos DBA , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/imunologia , Peptídeos/metabolismo , Ratos , Ratos Endogâmicos Lew , Linfócitos T/imunologia , Resultado do TratamentoRESUMO
AIMS: The objective of this study was to evaluate the biodiversity of Aspergillus section Nigri populations from Argentinean vineyards by morphological, toxigenic and AFLP analysis. MATERIALS AND METHODS: Five hundred and thirty-eight strains were isolated from grapes during 2006/07 and 2007/08 vintages. The morphological identification and toxigenic profile for all strains isolated were performed. Eighty-eight strains were selected for characterization at species level by AFLP markers. Cluster analysis showed a clear separation into four main groups: A. carbonarius, A. tubingensis, A. niger'aggregate' and Aspergillus'uniseriate'. A. carbonarius strains constituted a homogeneous group, while a high degree of genetic diversity was found within the A. niger'aggregate' and 'A. uniseriate' clusters. The A. tubingensis cluster was the most prevalent group and was clearly separated from A. niger'aggregate'. Ten strains showed 45% homology with A. tubingensis FRR 5720 ex-type strain and were considered as 'atypical' or a closely related species. AFLP results indicate that no genotypical differences can be established between ochratoxigenic and nonochratoxigenic strains. CONCLUSIONS: Aspergillus section Nigri populations on grapes were represented mainly by four groups. A. tubingensis species were separated from A. niger'aggregate' group and some of their strains produced OTA. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new data on molecular characterization of Aspergillus section Nigri populations in Argentina.
Assuntos
Aspergillus/classificação , Ocratoxinas/biossíntese , Vitis/microbiologia , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Argentina , Aspergillus/genética , Aspergillus/isolamento & purificação , Aspergillus/metabolismo , BiodiversidadeRESUMO
Vascular complications after head and neck surgical procedures are rare but can life-threatening. A pseudoaneurysm may appear weeks or months after the initial injury and may be difficult to diagnose. The authors report the case of a 43-year-old man involved in a motor vehicle accident presenting with multiple facial fractures, which after surgical treatment presented episodes of bleeding. After an angiography was performed, a pseudoaneurysm of the facial artery was detected. The patient later underwent endovascular embolization and had a satisfactory resolution.