Acute murine-betacoronavirus infection impairs testicular steroidogenesis and the quality of sperm production.

Although several testicular alterations promoted by coronavirus infection have been demonstrated, the extent, causes, and players of testicular pathogenesis are not totally understood. The present study aimed to investigate the short-term effects on male fertility of intranasally administered murine hepatitis virus strain 3 (MHV-3), a member of the genus Betacoronavirus, which causes a severe systemic acute infection. This mouse model might be used as a in vivo prototype for investigating the impact of betacoronavirus on the endocrine and exocrine testicular functions with the advantage to be performed in a biosafety level 2 condition. Herein, we performed virological, histopathological, and molecular studies regarding the testicular spermatogenesis and the spermatic quality analyses in an MHV-3-infected C57BL/6 mice. The main outcomes showed that MHV-3 infects mouse testis and induces a testicular inflammatory state, impairing the steroidogenic pathway. The infection led to several alterations in the testicular parenchyma, such as: seminiferous epithelium sloughing, retention of residual bodies, germ cell apoptosis, alterations in intercellular junction proteins, and worse spermatogenic parameters. Moreover, the levels of plasmatic testosterone as well as the quality of sperm production reduced. Therefore, the present data suggest that the viral/inflammatory impairment of the steroidogenic pathway and the consequent imbalance of androgen levels is critical in testicular pathology, disturbing the SC barrier function and the germ cell differentiation. Our study is important for comprehending the effects of beta coronavirus infections on testis function in order to develop treatments that could prevent virus-mediated male infertility.

Manutenção do ritmo sinusal após retorno à prática esportiva em paciente com fibrilação atrial tratado com descondicionamento / Maintenance of sinus rhythm after return to sports practice in a patient with atrial fibrillation treated with deconditioning

O exercício físico reduz risco de doenças cardiovasculares, metabólicas e está relacionada a aumento do bem-estar e longevidade. Alguns estudos sugerem redução do risco de arritmias, como a fibrilação atrial, em até 19% em homens e 9% em mulheres. Contudo, observa-se prevalência superior, de até 3,5 vezes, de desenvolvimento de fibrilação atrial em praticantes de exercícios de muito alta intensidade ou competitiva, tornando essa a arritmia mais frequente no atleta. Os indivíduos sob maior risco são homens, praticantes de endurance de alta intensidade e com alto volume. A presença de fibrilação atrial, ainda que paroxística, impacta negativamente sobre a capacidade funcional e performance nesses indivíduos, seja pela redução do débito cardíaco pela perda da contração atrial ou pela perda de controle da frequência cardíaca pelo sistema nervoso autônomo. A evidência atual disponível para guiar o manejo desses pacientes se baseia em estudos randomizados em pacientes portadores de FA não atletas, estudos menores em atletas ou em consenso de especialista e o tratamento se baseia nos pilares de controle de ritmo farmacológico ou intervencionista por meio de ablação por radiofrequência ou destrainamento. Esse relato de caso tem como objetivo apresentar a evolução de paciente atleta com fibrilação atrial tratado de forma não invasiva, com destreinamento, de forma a revisar e ampliar o conhecimento sobre o tema. RELATO DE CASO: Paciente, sexo masculino, 42 anos, sem comorbidades prévias, pratica esporte competitivo desde os 15 anos nas modalidades: mountain bike, triátlon e maratona. Procura atendimento médico, sem queixas, solicitando liberação para realização de prova triathlon ironman. Ao exame físico apresentava ausculta cardíaca com frequência de 50 bpm e ritmo irregular, sem outras alterações. O eletrocardiograma apresentava ritmo de fibrilação atrial com baixa resposta ventricular. Foi realizada investigação de cardiopatia estrutural com ecocardiograma que revelou átrio esquerdo aumentado, com volume indexado de 40ml/ m2. Função biventricular preservada. Sem outros achados. Em teste ergométrico, apresentava resposta adequada da frequência cardíaca ao esforço, com excelente capacidade funcional e sem alterações sugestivas de isquemia. Holter evidenciava apenas presença de fibrilação atrial. Foi optado então por tratamento com destreinamento por 3 meses. Após, paciente realizou holter que apresentava ritmo sinusal. Foi mantido seguimento por um ano, com repetição de holter 24h que se manteve em ritmo sinusal.

Avaliação da prática esportiva em atleta jovem com estenose aórtica subvalvar corrigida / Evaluation of sports practice in a young athlete with corrected subvalvular aortic stenosis

Paciente 18 anos, sexo masculino, sem outras comorbidades, ex-atleta amador praticante de Rugby, interrompeu a prática esportiva por orientação médica após o diagnóstico de estenose subvalvar aórtica grave, com gradiente médio de 81 mmHg e máximo de 158 mmHg em membrana subvalvar. Foi realizada correção cirúrgica com ressecção da membrana subaórtica associada à ressecção muscular da via de saída do ventrículo esquerdo e plastia da valva tricúspide. Após recuperação cirúrgica paciente, mantém-se assintomático e retorna periodicamente para avaliação quanto a prática de atividade física e esportiva competitiva. Três anos após cirurgia, em avaliação complementar, ao ecocardiograma, observava-se persistência de prega ventrículo-infundibular e resquício de membrana, determinando estreitamento da via de saída do ventrículo esquerdo (VE), com aceleração de fluxo gerando gradiente sistólico máximo entre VE e aorta de 42 mmHg e médio de 24 mmHg. Refluxo aórtico discreto a moderado. Função sistólica biventricular preservada, hipertrofia ventricular esquerda discreta. Foi solicitado também ecocardiograma com estresse físico que não mostrou sinais de isquemia miocárdica. Membrana subaórtica gerou obstrução importante com repercussão hemodinâmica ao esforço físico. Por fim, realizou teste ergométrico, que apresentou Infradesnivelamento de segmento ST de 2,5mm horizontam em MC5, sem dor, com comportamento adequado da pressão e da frequência cardíaca durante o esforço e capacidade funcional estimada de 13 METs. Após exames complementares, foi realizado reunião com especialistas em cardiologia do esporte e liberado para a prática de atividade física moderada, com cálculo de frequência máxima até 60% da FC reserva e resistiva com até 50% da força máxima. A estenose subvalvar aórtica é uma cardiopatia congênita relativamente frequente que resulta de uma anormalidade subjacente na arquitetura da via de saída do ventrículo esquerdo (VSVE), com fluxo turbilhonado levando à fibrose progressiva dessa região. Obstrução significativa da VSVE leva à hipertrofia ventricular e função hiperdinâmica. É a segunda causa mais comum de estenose aórtica na população pediátrica e muitos pacientes têm regurgitação aórtica associada. A correção cirúrgica geralmente é indicada a partir da 2ª década de vida e 20 até 30% dos pacientes evoluem com recorrência da obstrução. Em virtude da prevalência dessa condição em indivíduos jovens, além de outras condições de risco, a avaliação médica por especialista antes da prática esportiva competitiva torna-se essencial.

Insuficiência cardíaca de fração de ejeção reduzida após infarto agudo do miocárdio em atleta de endurance: é possível continuar no esporte? / Heart failure with reduced ejection fraction after acute myocardial infarction in an endurance athlete: is it possible to continue in sport?

Insuficiência cardíaca é das principais doenças resultando em redução da capacidade física, sendo a doença arterial coronariana sua principal causa, especialmente em atletas acima de 35 anos. Será que insuficiência cardíaca com fração de ejeção reduzida após infarto agudo miocárdio teria espaço no esporte? Paciente masculino, 56 anos, com antecedente de hipertensão arterial sistêmica, previamente atleta de corrida de rua desde 38 anos de idade (meia maratona com pace médio de 3:50 min/km) procurou atendimento em pronto socorro em março de 2016 por angina típica em repouso. Ficou em observação e liberado após analgesia. Por manter dispneia iniciou acompanhamento com cardiologista e, 2 meses após episódio de dor torácica, realizou cintilografia de perfusão miocárdica com tecnécio sestamibi associado a teste ergométrico, evidenciando hipocaptação persistente extensa em parede anterosseptal, inferosseptal e ápice de ventrículo esquerdo (VE), sem hipocaptação transitória e com fração de ejeção de VE (FEVE) de 30%. Realizada estratificação invasiva mostrando lesão de 99% em segmento médio de artéria descendente anterior com aspecto de recanalização e fluxo TIMI um com acinesia de parede anterior discinesia de parede apical. Paciente encaminhado ao serviço terciário e optado por realizar cintilografia miocárdica com tálio-201 e estresse com teste ergométrico que resultou em ausência de isquemia e de viabilidade miocárdica. Portanto, paciente foi mantido em tratamento clínico e reabilitação cardiovascular. Em acompanhamento ambulatorial após seis anos do evento, atualmente praticando corrida dois a três vezes por semana (10 a 12 km com pace 6:15 min/km), com ecocardiograma transtorácico com aumento biatrial e discreto de VE, FEVE 38% com acinesia do segmento médio do septo e do segmento apical de todas as paredes, insuficiência mitral discreta a moderada por remodelamento ventricular. Teste ergométrico realizado com protocolo de Ellestad, interrompido após 12'45" por exaustão e atingindo 98% da frequência cardíaca máxima preconizada pela idade, sem alterações sugestivas de isquemia, extrassístoles ventriculares isoladas e raras e consumo máximo estimado de oxigênio de 56 ml/kg/min, em uso de betabloqueador. Apesar do paciente apresentar excelente aptidão cardiorrespiratória, foi orientado manter atividades de leve a moderada intensidade (70% da reserva cronotrópica) devido a FEVE reduzida. Portanto, há espaço para esses pacientes no esporte não competitivo: após reabilitação cardiovascular, estratificação de risco e contínuo acompanhamento médico e multidisciplinar.

Activation of C-C motif chemokine receptor 2 modulates testicular macrophages number, steroidogenesis, and spermatogenesis progression.

The monocyte chemoattractant protein 1 (MCP-1) belongs to the CC chemokine family and acts in the recruitment of C-C motif chemokine receptor 2 (CCR2)-positive immune cell types to inflammation sites. In testis, the MCP-1/CCR2 axis has been associated with the macrophage population's functional regulation, which presents significant functions supporting germ cell development. In this context, herein, we aimed to investigate the role of the chemokine receptor CCR2 in mice testicular environment and its impact on male sperm production. Using adult transgenic mice strain that had the CCR2 gene replaced by a red fluorescent protein gene, we showed a stage-dependent expression of CCR2 in type B spermatogonia and early primary spermatocytes. Several parameters related to sperm production were reduced in the absence of CCR2 protein, such as Sertoli cell efficiency, meiotic index, and overall yield of spermatogenesis. Daily sperm production decreased by almost 40%, and several damages in the seminiferous tubules were observed. Significant reduction in the expression of important genes related to the Sertoli cell function (Cnx43, Vim, Ocln, Spna2) and meiosis initiation (Stra8, Pcna, Prdm9, Msh5) occurred in comparison to controls. Also, the number of macrophages significantly decreased in the absence of CCR2 protein, along with a disturbance in Leydig cell steroidogenic activity. In summary, our results show that the non-activation of the MCP-1/CCR2 axis disturbs the testicular homeostasis, interfering in macrophage population, meiosis initiation, blood-testis barrier function, and androgen synthesis, leading to the malfunction of seminiferous tubules, decreased testosterone levels, defective sperm production, and lower fertility index.

Insights into differentiation and function of the transition region between the seminiferous tubule and rete testis.

Seminiferous tubules physically connect to the rete testis through short segments called the transition region (TR). During fetal development, this specialized junction is considered the initial site where testis cords begin to form and to grow in length well beyond birth and into adulthood and form convoluted tubular cores. Mitotic activity of the Sertoli cell, the somatic cell of the epithelium, ceases before puberty, but modified Sertoli cells in the TR remain immature and capable of proliferation. This review presents what is known about this specialized region of the testis, with an emphasis on the morphological, molecular and physiological features, which support the hypothesis that this short region of epithelial transition serves as a specialized niche for undifferentiated Sertoli cells and spermatogonial stem cells. Also, the region is populated by an elevated number of immune cells, suggesting an important activity in monitoring and responding to any leakage of autoantigens, as sperm enter the rete testis. Several structure/function characteristics of the transition region are discussed and compared across species.

Características morfológicas do estômago de jaguatirica (Leopardus pardalis - Linnaeus, 1758) / Morphological characteristics of the stomach of the ocelot (Leopardus pardalis - Linnaeus, 1758)

Leopardus pardalis é uma das espécies de felinos neotropicais mais estudadas em seus aspectos ecológicos, porém informações de questões morfofisiológicas do trato digestório não são encontradas na literatura. Visando contribuir com tais informações, o objetivo deste artigo foi caracterizar a morfologia do estômago dessa espécie. Nesta pesquisa, foram utilizados cinco exemplares adultos, coletados após óbito por atropelamento ou doados pelo IBAMA ao Laboratório de Zoologia e Morfologia Animal da Universidade do Estado de Mato Grosso - Alta Floresta. Os animais foram fixados com solução aquosa de formaldeído a 10%, dissecados por meio de técnicas anatômicas. Além dos estudos anatômicos, também foram coletados tecidos para microscopia. As análises macroscópicas revelaram que o estômago era do tipo unicavitário, com presença de pequena e grande curvatura, localizado no antímero esquerdo e constituído pelas regiões características do órgão. Em seus aspectos microscópicos, mostrou-se formado pelas quatro túnicas usuais do tubo digestório: túnica mucosa, submucosa, muscular e serosa, cada uma com as particularidades características de cada região, principalmente na túnica mucosa. O estômago de L. pardalis revelou uma morfologia semelhante às espécies domésticas e selvagens, com particularidades histomorfológicas na região pilórica. Esse padrão pode estar relacionado a adaptações evolutivas no processo digestivo dessa espécie.(AU)

The species Leopardus pardalis is one of the species of neotropical felines more studied in its ecological aspects, however, information of morphophysiological questions of the digestive tract are not found in the literature. Aiming to contribute with such information, the objective of this work was to characterize the morphology of the stomach of this species. Five adult specimens, collected after death by trampling, or donated by IBAMA to the Laboratory of Zoology and Animal Morphology of the University of the State of Mato Grosso - Alta Floresta, were used in this research. The animals were fixed with aqueous solution of formaldehyde 10%, dissected through the basic techniques and instruments of anatomy, and later the material was collected for microscopy. The macroscopic analysis revealed that the stomach was of the unicavitary type, with small and great curvature, located in the left antimer and constituted by the characteristic regions of the organ. In its microscopic aspects it was formed by the four usual tunics of the digestive tube: tunica mucosa, submucosa, muscular and serosa, each one with the peculiarities characteristic of each region, mainly in the tunica mucosa. The stomach of L. pardalis revealed similar morphology to the domestic and wild species, with small histomorphological peculiarities in the pyloric region, which may be related to evolutionary adaptations in the digestive process of this species.(AU)

Characterization of spermatogonial cells and niche in the scorpion mud turtle (Kinosternon scorpioides).

Undifferentiated spermatogonia (Aund) or spermatogonial stem cells (SSCs) are committed to the establishment and maintenance of spermatogenesis and fertility throughout a male's life and are located in a highly specialized microenvironment called niche that regulates their fate. Although several studies have been developed on SSCs in mammalian testis, little is known about other vertebrate classes. The present study is the first to perform a more detailed investigation on the spermatogonial cells and their niche in a reptilian species. Thus, we characterized Aund/SSCs and evaluated the existence of SSCs niche in the Kinosternon scorpioides, a freshwater turtle found from Mexico to northern and central South America. Our results showed that, in this species, Aund/SSCs exhibited a nuclear morphological pattern similar to those described for other mammalian species already investigated. However, in comparison to other spermatogonial cell types, Aund/SSCs presented the largest nuclear volume in this turtle. Similar to some mammalian and fish species investigated, both GFRA1 and CSF1 receptors were expressed in Aund/SSCs in K. scorpioides. Also, as K. scorpioides Aund/SSCs were preferentially located near blood vessels, it can be suggested that this niche characteristic is a well conserved feature during evolution. Besides being valuable for comparative reproductive biology, our findings represent an important step towards the understanding of SSCs biology and the development of valuable systems/tools for SSCs culture and cryopreservation in turtles. Moreover, we expect that the above-mentioned results will be useful for reproductive biotechnologies as well as for governmental programs aiming at reptilian species conservation.

Duration of spermatogenesis and identification of spermatogonial stem cell markers in a Neotropical catfish, Jundiá (Rhamdia quelen).

Spermatogenesis is a process driven by stem cell, where germ cell cycle is under the control of a specific genotype species. Considering that Jundiá (Rhamdia quelen) is a Neotropical catfish with great economical importance and useful experimental model, little information is available on basic aspects of its reproductive biology, especially on spermatogenesis. As a result, this study aimed to characterize the male germ cells, estimate the duration of spermatogenesis and evaluate the expression of selected stem cell genes in Jundiá testis. Similar to other fish species, our results showed a remarkable decrease of germ cell nuclear volume during Jundiá spermatogenesis, particularly from type A undifferentiated to late type B spermatogonia and from diplotene to late spermatids. Using a S-phase marker, bromodeoxyuridine (BrdU), the combined duration of meiotic and spermiogenic phases in this species was estimated in approximately 7 days. This is considered very short when compared to mammals, where spermatogenesis last from 30 to 74 days. Selected stem cell genes were partially sequenced and characterized in Jundiá testis. Expression analysis showed higher plzf and pou5f3 mRNA levels in the cell fractions enriched by type A undifferentiated spermatogonia. These results were further confirmed by in situ hybridization that showed strong signal of plzf and pou5f3 mRNA in type A undifferentiated spermatogonia. Altogether, these information will expand our knowledge of the reproductive biology of this species, contributing to improve its production and management, and also for biotechnological applications, such as germ cell transplantation.

Oral administration of Simbioflora® (synbiotic) attenuates intestinal damage in a mouse model of 5-fluorouracil-induced mucositis.

The use of probiotics to prevent or treat mucosal inflammation has been studied; however, the combined effect of probiotics and prebiotics is unclear. The aim of this study was to test whether oral administration of a synbiotic (Simbioflora®) preparation containing Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium lactis plus fructooligosaccharide could help control mucosal inflammation in experimental mucositis induced by 5-fluorouracil (5-FU). Male BALB/c mice were randomly divided into six groups: control (CTL), control + prebiotic (CTL+P), control + synbiotic (CTL+S), mucositis (MUC), mucositis + prebiotic (MUC+P), and mucositis + synbiotic (MUC+S). Mice from the CTL+S, MUC+S, CTL+P, and MUC+P groups received synbiotic or prebiotic daily by oral gavage for 13 days. Mice in the CTL and MUC groups received the same volume of saline. On day 11, mice in the MUC, MUC+P, and MUC+S groups received an intraperitoneal injection of 300 mg/kg 5-FU to induce mucositis. After 72 h, all mice were euthanised. Intestinal permeability, intestinal histology, and biochemical parameters were analysed. Group MUC showed a greater weight loss and increased intestinal permeability (0.020 counts per min [cpm]/g) compared to the CTL group (0.01 cpm/g) P<0.05. Both treatments attenuated weight loss compared to the MUC group. Nonetheless, the synbiotic caused a greater reduction in intestinal permeability (0.012 cpm/g) compared to the MUC (0.020 cpm/g) and MUC+P (0.016 cpm/g) groups P<0.05. Mice in groups MUC+P and MUC+S displayed significant recovery of lesions and maintenance of the mucus layer. There were no differences in the short-chain fatty acid concentrations in the faeces between the MUC and CTL groups (P>0.05). Increased acetate and propionate concentrations were evidenced in the faeces of the MUC+P and MUC+S groups. Only the synbiotic treatment increased the butyrate concentration (P<0.05). The results indicate that administration of synbiotic can decrease mucosal damage caused by mucositis.

Spermatogenesis in a neotropical marsupial species, Philander frenatus (Olfers, 1818).

Despite the singular morphology of the male genital system and the different reproductive strategies of marsupials, little emphasis has been given to the testis morphology and spermatogenic kinetics in this mammalian order. The present study aimed to investigate the testis function and the duration of spermatogenesis in the southeastern four-eyed opossum, Philander frenatus. Testes of six adult males were routinely processed for histological and stereological analyses. In order to determine the duration of spermatogenesis, intratesticular injections of tritiated thymidine were performed 1h, 13days and 21days before the sacrifice. Based on the development of the acrosomic system, ten stages of the seminiferous epithelium cycle were characterized. The mean body and testis weights for the P. frenatus were respectively 326±20g and 0.4±0.05g, providing a gonadosomatic index of 0.3±0.02%. The most advanced germ cell types labeled at 1h, 13days and 21days after thymidine injections were, respectively, preleptotene spermatocytes at stage IV, pachytene spermatocytes at stage IV and diplotene spermatocytes at stage IX. Based on the stages frequencies and the most advanced labeled germ cells, each spermatogenic cycle and the entire spermatogenic process lasted respectively 13.5±0.5 and 60.9±2.4days. When compared to the vast majority of eutherian mammals already investigated, these data indicate that the Philander frenatus presents a relatively long duration of spermatogenesis.

Antimicrobial effects of Piper hispidum extract, fractions and chalcones against Candida albicans and Staphylococcus aureus.

Three chalcones, 2'-hydroxy-4,4',6'-trimethoxychalcone, 2'-hydroxy-4,4',6'-tetramethoxychalcone, and 3,2'-dihydroxy-4,4',6'-trimethoxychalcone, were isolated from the leaves of Piper hispidum in a bioguided fractionation of crude extract. The antimicrobial activity of crude extract of P. hispidum leaves was determined against bacteria Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus and yeasts Candida albicans, C. parapsilosis and C. tropicalis. Fractions and chalcones were tested against C. albicans and S. aureus. The checkerboard assay was performed to assess synergic interactions between extract and antifungal drugs, and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay was used to evaluate anti-biofilm effects of extract. The extract was active against yeasts, S. aureus and B. subtilis with MIC values between 15.6 and 62.5µg/mL. Synergistic effects of extract associated with fluconazole and nystatin were observed against C. albicans, with fractional inhibitory concentration indices of 0.37 and 0.24, respectively. The extract was also effective against C. albicans and S. aureus biofilm cells at concentrations of 62.5 and 200µg/mL, respectively. Thus, P. hispidum may be a possible source of bioactive substances with antimicrobial properties.

Sertoli cells are capable of proliferation into adulthood in the transition region between the seminiferous tubules and the rete testis in Wistar rats.

Sertoli cells (SCs) play a crucial role in testis differentiation, development and function, determining the magnitude of sperm production in sexually mature animals. For over 40 years, it has been considered that these key testis somatic cells stop dividing during early pre-pubertal phase, between around 10 to 20 days after birth respectively in mice and rats, being after that under physiological conditions a stable and terminally differentiated population. However, evidences from the literature are challenging this dogma. In the present study, using several important functional markers (Ki-67, BrdU, p27, GATA-4, Androgen Receptor), we investigated the SC differentiation status in 36 days old and adult Wistar rats, focusing mainly in the transition region (TR) between the seminiferous tubules (ST) and the rete testis. Our results showed that SCs in TR remain undifferentiated for a longer period and, although at a lesser degree, even in adult rats proliferating SCs were observed in this region. Therefore, these findings suggest that, different from the other ST regions investigated, SCs residing in the TR exhibit a distinct functional phenotype. These undifferentiated SCs may compose a subpopulation of SC progenitors that reside in a specific microenvironment capable of growing the ST length if needed from this particular testis region. Moreover, our findings demonstrate an important aspect of testis function in mammals and opens new venues for other experimental approaches to the investigation of SC physiology, spermatogenesis progression and testis growth. Besides that, the TR may represent an important site for pathophysiological investigations and cellular interactions in the testis.

Molecular typing and differences in biofilm formation and antibiotic susceptibilities among Prototheca strains isolated in Italy and Brazil.

Bovine mastitis caused by Prototheca is a serious and complex problem that accounts for high economic losses in the dairy industry. The main objective of this study was to identify and characterize at genetic level different Prototheca strains and provide the most complete data about protothecal antibiotic resistance. The study involves 46 isolates from Italian (13 strains) and Brazilian (33 strains) mastitic milk. These strains were identified by multiplex PCR and single strand conformation polymorphism analysis and characterized by randomly amplified polymorphic DNA (RAPD)-PCR. Moreover, biofilm production and antibiotic susceptibility were evaluated. Forty-two strains resulted as Prototheca zopfii genotype 2, whereas 4 isolates could belong to a potential new Prototheca species. The RAPD-PCR, performed with 3 primers (M13, OPA-4, and OPA-18), showed a notable heterogeneity among isolates and grouped the strains according to the species and geographical origin. Biofilm production was species-dependent and P. zopfii genotype 2 strains were classified as strong biofilm producers. In vitro antibiotic susceptibility tests indicated that Prototheca strains were susceptible to antibacterial drugs belonging to aminoglycosides group; the highest activity against Prototheca strains was observed in the case of colistin sulfate, gentamicin, and netilmicin (100% of susceptible strains). It is interesting to note that all the Italian P. zopfii genotype 2 strains showed lower minimum inhibitory concentration values than the Brazilian ones. Nisin showed more efficacy than lysozyme and potassium sorbate, inhibiting 31% of the strains. Results obtained in this study confirmed that RAPD-PCR is a rapid, inexpensive, and highly discriminating tool for Prototheca strains characterization and could give a good scientific contribution for better understanding the protothecal mastitis in dairy herd.

Successful xenogeneic germ cell transplantation from Jundia catfish (Rhamdia quelen) into adult Nile tilapia (Oreochromis niloticus) testes.

Fish germ cell transplantation presents several important potential applications for aquaculture, including the preservation of germplasm from endangered fish species with high genetic and commercial values. Using this technique in studies developed in our laboratory with adult male Nile tilapias (Oreochromis niloticus), all the necessary procedures were successfully established, allowing the production of functional sperm and healthy progeny approximately 2months after allogeneic transplantation. In the present study, we evaluated the viability of the adult Nile tilapia testis to generate sperm after xenogeneic transplant of germ cells from sexually mature Jundia catfish (Rhamdia quelen) that belong to a different taxonomic order. Therefore, in order to investigate at different time-periods post-transplantation, the presence and development of donor PKH26 labeled catfish germ cells were followed in the tilapia seminiferous tubules. From 7 to 20days post-transplantation, only PKH26 labeled spermatogonia were observed, whereas spermatocytes at different stages of development were found at 70days. Germ cell transplantation success and progression of spermatogenesis were indicated by the presence of labeled PKH26 spermatids and sperm on days 90 and 120 post-transplantation, respectively. Confirming the presence of the catfish genetic material in the tilapia testis, all recipient tilapias evaluated (n=8) showed the genetic markers evaluated. Therefore, we demonstrated for the first time that the adult Nile tilapia testis offers the functional conditions for development of spermatogenesis with sperm production from a fish species belonging to a different order, which provides an important new venue for aquaculture advancement.

Antidermatophytic activity of hydroalcoholic extracts from Rosmarinus officinalis and Tetradenia riparia.

Rosmarinus officinalis and Tetradenia riparia are used in folk medicine for the treatment of disease, including infectious diseases and skin disorders. The purpose of this study was to investigate the antifungal activity of hydroalcoholic extracts from R. officinalis and T. riparia against strains of Trichophyton rubrum, T. mentagrophytes and Microsporum gypseum. Hydroalcoholic extracts prepared with dried leaves from R. officinalis, Psidium guajava and T. riparia were assayed against dermatophyte species by the microdilution technique and by microscopy. R. officinalis and T. riparia were the most active against dermatophytes, as determined from the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC), and were investigated further. Fluorescence microscopy and scanning electron microscopy were used to investigate inhibition of hyphal growth by the two extracts, and showed a strong inhibition and an irregular growth pattern. Both extracts showed good action against dermatophytes, inhibiting fungal growth and causing alterations in their hyphae. Therefore, R. officinalis and T. riparia are potential sources of new compounds for the development of antifungal drugs.

Integrated Environmental Assessment of streams in the Sinos River basin in the state of Rio Grande do Sul, Brazil.

The deterioration of environmental quality in the Sinos River basin is directly associated with the impacts of intense industrialization and urbanization. An integrated environmental assessment (IEA) was conducted in July and September of 2012, in areas along the sources of the EstânciaVelha/Portão, Pampa and Schmidt streams using physical, chemical and biological methods. The water in the three sampling sites was not proper for human consumption, presented a low toxic contamination index (TCI) and mesotrophic characteristics. One site was included in Class 4, and two, in Class 3, according to current legislation. The rapid assessment protocol (RAP) indicated a natural environmental condition for habitat diversity and environmental impact in the three sites. The Tradescantia pallida (Rose) D.R. Hunt var. purpurea Boom biomarker showed water genotoxicity in two of the sites. The integrated diagnosis of water quality in these streams is fundamentally important to ensure the sustainable management of water resources and their multiple uses, as well to estimate their contribution to pollution in this river basin.

Glycosylflavonoids from Cecropia pachystachya Trécul are quorum sensing inhibitors.

The Cecropia genus is widely distributed in Latin America including at least 60 species, and some of them are commonly used in traditional medicine for the treatment of several diseases. We used Cecropia pachystachya Trécul to search for quorum sensing (QS) inhibitors compounds and found that the aqueous extract of C. pachystachya leaves is a promising source of substances with this activity. Using as biosensor Chromobacterium violaceum ATCC 31532 and Escherichia coli pSB403, the compounds chlorogenic acid (2), isoorientin (3), orientin (4), isovitexin (6), vitexin (7), and rutin (9) were identified as QS inhibitors. None of these compounds inhibited the growth of neither the used biosensors nor the microorganisms Staphylococcus aureus ATCC 23591, Escherichia coli ATCC 25922 and Saccharomyces cerevisiae, used here as growth inhibition controls. Along with the rutin, here we presented for the first time the QS-inhibition potential of the C-glycosyl flavonoids. The prospective of this evidence lead to the use of these compounds as antipathogenic drugs or antifoulants.

Viabilidade econômica da terceira ordenha em sistemas de produção de leite com ordenhadeira tipo circuito aberto / Economic viability of the third milking in milk production systems using open circuit type milking mechanics

Objetivou-se analisar a viabilidade econômica da terceira ordenha em sistemas de produção de leite que utilizam ordenhadeira mecânica, do tipo circuito aberto, visando fornecer aos técnicos e pecuaristas informações que os auxiliem no processo de tomada de decisões. Especificamente pretendeu-se ainda: a) estimar o custo de uma ordenha; b) estimar, por meio de simulação, o custo da terceira ordenha; c) desenvolver uma equação matemática que permita estimar a quantidade mínima de leite produzido em duas ordenhas, a partir da qual será viável economicamente a realização da terceira ordenha. Os dados foram coletados em três sistemas de produção de leite, em uma rotina de duas ordenhas diárias, sendo realizadas três coletas de dados em cada um, perfazendo um total de nove. Considerando-se os dados médios, seria viável a realização da terceira ordenha se a produtividade média diária das vacas em lactação em duas ordenhas fosse igual ou superior a 38,13kg de leite. A equação matemática desenvolvida pode auxiliar o técnico e o pecuarista a estimarem a quantidade mínima de leite produzido por uma vaca em duas ordenhas, a partir da qual será viável economicamente a realização da terceira ordenha, com precisão e considerável rapidez.

This study aims to analyze the economic viability of the third milking in milk production systems using open circuit milking mechanics, aiming to provide technicians and farmers with information to help in the decision-making process. Specifically intend to a) estimate the cost of a milking; (b) estimate, by means of simulation, the cost of the third milking; (c) develop a mathematical equation that allows estimating the minimum amount of milk produced in two milkings, from which it will be economically feasible to hold the third milking. The data were collected in three milk production systems, in a routine of two milkings per day, with three collections of data in each, a total of nine collections. Considering the average data, it would be feasible to hold the third milking if the productivity of daily average lactating cows is equal or greater than 38.13 kg of milk. The mathematical equation developed may help the technician and the farmer to estimate the minimum quantity of milk produced by a cow in two milkings, from which it will be economically feasible to hold the third milking with precision and considerable speed.

Germ cell transplantation as a potential biotechnological approach to fish reproduction.

Although the use of germ cell transplantation has been relatively well established in mammals, the technique has only been adapted for use in fish after entering the 2000s. During the last decade, several different approaches have been developed for germ cell transplantation in fish using recipients of various ages and life stages, such as blastula-stage embryos, newly hatched larvae and sexually mature specimens. As germ cells can develop into live organisms through maturation and fertilization processes, germ cell transplantation in fish has opened up new avenues of research in reproductive biotechnology and aquaculture. For instance, the use of xenotransplantation in fish has lead to advances in the conservation of endangered species and the production of commercially valuable fish using surrogated recipients. Further, this could also facilitate the engineering of transgenic fish. However, as is the case with mammals, knowledge regarding the basic biology and physiology of germline stem cells in fish remains incomplete, imposing a considerable limitation on the application of germ cell transplantation in fish. Furthering our understanding of germline stem cells would contribute significantly to advances regarding germ cell transplantation in fish.