Effect of monensin and soybean oil on rumen ciliate protozoa and correlation between protozoa with ruminal fermentation and digestive parameters

Four lactating dairy cows with ruminal cannula and 155 DIM were allotted to a 4 × 4 Latin square and fed twice daily corn silage and concentrate (55:45, %DM) to evaluate the effect of monensin and soybean oil in the diet of lactating cows on the counting of ciliate protozoa beyond establishing correlations between number of protozoa with some parameters of digestion and ruminal fermentation. The treatments consisted of the inclusion of 33 ppm of sodium monensin or soybean oil at 4% DM, as following: control diet, without oil or monensin - CT; diet with monensin - MN; diet with soybean oil - OL; diet with the combination of 33 ppm of monensin and 4% of soybean oil - OM. Occurrence of 11 ciliate genera was observed, being Entodinium the predominant in all treatments. No significant soybean oil × monensin interaction was observed on protozoa number and genera. The genera Entodinium, Dasytricha, Eremoplastron and Isotricha reduced in the diets with oil, while monensin reduced the counting of Dasytricha, Eremoplastron and Epidinium. The total number of ciliate protozoa and cellulolytics protozoa was reduced by soybean oil and monensin effects, showing a defaunatory additive effect when combined the oil and the monensin. The ciliate cellulolytics was reduced by linoleic acid intake and was related positively with NDF ruminal digestibility and ruminal ammonia. The total number of protozoa was correlated with the molar proportion of propionate in the ruminal liquid, suggesting that monensin and soybean oil, due their defaunatory effect, can reduce the loss of methane in the rumen.Quatro vacas lactantes canuladas no rúmen, com 155 dias em lactação, foram dispostas em um quadrado latino 4 × 4 e alimentadas, duas vezes ao dia, com silagem de milho e concentrado (relação 55:45, base matéria seca), para se avaliarem os efeitos da monensina e do óleo de soja na dieta de vacas lactantes sobre a contagem de protozoários ciliados, além de estabelecer correlações entre os protozoários com alguns parâmetros da digestão e da fermentação ruminal. Os tratamentos consistiram da inclusão de 33 ppm de monensina sódica ou de 4% de óleo de soja na dieta, assim representados: dieta controle, sem óleo ou monensina - CT; dieta com monensina – MN; dieta com óleo – OL; combinação de 33 ppm de monensina e 4% de óleo de soja - OM. Verificou-se a ocorrência de 11 gêneros de ciliados, sendo Entodinium o predominante em todos os tratamentos. Não foi observada interação significativa óleo de soja × monensina sobre a contagem de protozoários e gêneros. Os gêneros Entodinium, Dasytricha, Eremoplastron e Isotricha foram reduzidos nas dietas com óleo, enquanto a monensina diminuiu a contagem de Dasytricha, Eremoplastron e Epidinium. O número total de protozoários e de ciliados celulolíticos foi reduzido pelos efeitos de óleo de soja e monensina, indicando efeito aditivo defaunatório quando combinados o óleo e a monensina. Os ciliados celulolíticos foram reduzidos pelo consumo de ácido linoléico e positivamente relacionados à digestibilidade ruminal da FDN e amônia ruminal. O número total de protozoários foi correlacionado à proporção de propionato no líquido ruminal, indicando que monensina e óleo de soja, pelo seu efeito defaunatório, podem reduzir a perda de metano no rúmen

Trypanosoma cruzi high infectivity in vitro is related to cardiac lesions during long-term infection in Beagledogs

Trypanosoma cruzi is a hemoflagelate parasite associated with heart dysfunctions causing serious problems in Central and South America. Beagle dogs develop the symptoms of Chagas disease in humans, and could be an important experimental model for better understanding the immunopathogenic mechanisms involved in the chagasic infection. In the present study we investigated the relation among biological factors inherent to the parasite (trypomastigote polymorphism and in vitro infectivity) and immunoglobulin production, inflammation, and fibrosis in the heart of Beagle dogs infected with either T. cruzi Y or Berenice-78 strains. In vitro infectivity of Vero cells as well as the extension of cardiac lesions in infected Beagle was higher for Y strain when compared to Berenice-78 strain. These data suggested that in vitro infectivity assays may correlate with pathogenicity in vivo. In fact, animals infected with Y strain, which shows prevalence of slender forms and high infectivity in vitro, presented cardiomegaly, inflammation, and fibrosis in heart area. Concerning the immunoglobulin production, no statistically significant difference was observed for IgA, IgM or IgG levels among T. cruzi infected animals. However, IgA together IgM levels have shown to be a good marker for the acute phase of Chagas disease

Epilepsy in neurodegenerative diseases: related drugs and molecular pathways

Epilepsy is a chronic disease of the central nervous system characterized by an electrical imbalance in neurons. It is the second most prevalent neurological disease, with 50 million people affected around the world, and 30% of all epilepsies do not respond to available treatments. Currently, the main hypothesis about the molecular processes that trigger epileptic seizures and promote the neurotoxic effects that lead to cell death focuses on the exacerbation of the glutamate pathway and the massive influx of Ca2+ into neurons by different factors. However, other mechanisms have been proposed, and most of them have also been described in other neurodegenerative diseases, such as Alzheimers disease, Parkinsons disease, Huntingtons disease, or multiple sclerosis. Interestingly, and mainly because of these common molecular links and the lack of effective treatments for these diseases, some antiseizure drugs have been investigated to evaluate their therapeutic potential in these pathologies. Therefore, in this review, we thoroughly investigate the common molecular pathways between epilepsy and the major neurodegenerative diseases, examine the incidence of epilepsy in these populations, and explore the use of current and innovative antiseizure drugs in the treatment of refractory epilepsy and other neurodegenerative diseases.Acknowledges the support of the Spanish Ministry of Science, Innovation and Universities under the grant Juan de la Cierva (FJC2018-036012-I). Authors acknowledge the support of the Instituto de Salud Carlos III (ISCIII) Acción Estratégica en Salud, integrated into the Spanish National R+D+I Plan and financed by ISCIII Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER “Una manera de hacer Europa”) grant PI17/01474 awarded to M.B. Boada and grant PI19/00335 awarded to M.M.; M.E. acknowledges the support of the Spanish Ministry of Economy and Competitiveness under the project SAF2017- 84283-R, and CIBERNED under project CB06/05/0024. E.B.S. acknowledges the support of the Portuguese Science and Technology Foundation (FCT) for the strategic fund (UIDB/04469/2020). A.R. acknowledges the support of CIBERNED (Instituto de Salud Carlos III (ISCIII)), the EU/EFPIA Innovative Medicines Initiative Joint Undertaking, ADAPTED Grant Nº 115975, from EXIT project, EU Euronanomed3 Program JCT2017 Grant Nº AC17/00100, from PREADAPT project. Joint Program for Neurodegenerative Diseases (JPND) Grant No. AC19/00097, and from grants PI13/02434, PI16/01861 BA19/00020, and PI19/01301. Acción Estratégica en Salud, integrated in the Spanish National RCDCI Plan and financed by Instituto de Salud Carlos III (ISCIII)- Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER—“Una manera de Hacer Europa”), by Fundación bancaria “La Caixa” and Grífols SA (GR@ACE project)info:eu-repo/semantics/publishedVersio

Differential expression of proteins in genetically distinct Trypanosoma cruzi samples (TcI and TcII DTUs) isolated from chronic Chagas disease cardiac patients.

Background Trypanosoma cruzi, a hemoflagellate protozoan parasite and the etiological agent of Chagas disease (CD), exhibits great genetic and biological diversity. Infected individuals may present clinical manifestations with different levels of severity. Several hypotheses have been proposed to attempt to correlate the diversity of clinical signs and symptoms to the genetic variability of T. cruzi. This work aimed to investigate the differential expression of proteins from two distinct genetic groups of T. cruzi (discrete typing units TcI and TcII), isolated from chronically infected individuals displaying the cardiac form of CD. For this purpose, epimastigote forms of the two isolates were cultured in vitro and the cells recovered for protein extraction. Comparative two-dimensional (2D) gel electrophoreses were performed and differentially expressed spots selected for identification by mass spectrometry, followed by database searching and protein categorization. Results The 2D electrophoretic profiles revealed the complex composition of the T. cruzi extracted proteome. Protein spots were distributed along the entire pH and molecular mass ranges attesting for the integrity of the protein preparations. In total, 46 differentially expressed proteins were identified present in 40 distinct spots found in the comparative gel analyses. Of these, 16 displayed upregulation in the gel from TcI-typed parasites and 24 appeared overexpressed in the gel from TcII-typed parasites. Functional characterization of differentially expressed proteins revealed major alterations associated with stress response, lipid and amino acid metabolism in parasites of the TcII isolate, whilst those proteins upregulated in the TcI sample were primarily linked to central metabolic pathways. Conclusions The comparative 2D-gel electrophoresis allowed detection of major differences in protein expression between two T. cruzi isolates, belonging to the TcI and TcII genotypes. Our findings suggest that patients displaying the cardiac form of the disease harbor parasites capable of exhibiting distinct proteomic profiles. This should be of relevance to disease prognosis and treatment

Epilepsy in Neurodegenerative Diseases: Related Drugs and Molecular Pathways

Epilepsy is a chronic disease of the central nervous system characterized by an electrical imbalance in neurons. It is the second most prevalent neurological disease, with 50 million people affected around the world, and 30% of all epilepsies do not respond to available treatments. Currently, the main hypothesis about the molecular processes that trigger epileptic seizures and promote the neurotoxic effects that lead to cell death focuses on the exacerbation of the glutamate pathway and the massive influx of Ca2+ into neurons by different factors. However, other mechanisms have been proposed, and most of them have also been described in other neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, or multiple sclerosis. Interestingly, and mainly because of these common molecular links and the lack of effective treatments for these diseases, some antiseizure drugs have been investigated to evaluate their therapeutic potential in these pathologies. Therefore, in this review, we thoroughly investigate the common molecular pathways between epilepsy and the major neurodegenerative diseases, examine the incidence of epilepsy in these populations, and explore the use of current and innovative antiseizure drugs in the treatment of refractory epilepsy and other neurodegenerative diseases. Keywords: Alzheimer's disease; Huntington's disease; Parkinson's disease; epilepsy; multiple sclerosis; neurodegenerative diseases

Soroprevalência da doença de Chagas em escolares de dois municípios do Vale do Jequitinhonha, Minas Gerais, Brasil, seis anos após a implantação da vigilância epidemiológica

Six years after the beginning of the epidemiological surveillance of Chagas disease in Berilo and José Gonçalves de Minas, Jequitinhonha Valley, MG, Brazil, a serological inquiry was performed to observe whether the transmission of this endemy was occurring in this area. A randomized sample of 1,412 children seven to 14 years old, was screened. Six asymptomatic children were found to be positive, leading to 0.4% of prevalence. Hemoculture confirmed infection in five out of the six positive cases. Additional epidemiological investigation revealed important antecedents, such as disease reports in relatives and predisposing ecological and housing conditions. Our results demonstrated similar seroprevalence (0.4%) in schoolchildren, ranging from seven to 14 years old, and that were observed six years ago (0.2%) for children 0-9 year-old. Thus, considering the constant presence of Panstrogylus megistus in the peridomicile these findings emphasize the need of continuous improved epidemiological surveillance of Chagas disease in this region.Seis anos após o início da vigilância epidemiológica para doença de Chagas em Berilo e José Gonçalves de Minas, Vale do Jequitinhonha, MG, Brasil, foi realizado um inquérito sorológico para verificar se a transmissão desta endemia estava ocorrendo naquela área. Uma amostra aleatória de 1.412 crianças, de 7 a 14 anos, foi avaliada. Foram encontradas seis crianças positivas assintomáticas, totalizando uma prevalência de 0,4%. Hemocultura confirmou a infecção em cinco dos seis casos positivos. Uma investigação epidemiológica adicional, revelou importantes antecedentes, tais como: casos da doença em parentes e condições de moradia e ecológicas predisponentes. Nossos resultados demonstram uma soroprevalência similar (0,4%) em escolares de 7 a 14 anos àquela observada há seis anos (0,2%) em crianças de 0-9 anos. Dessa forma, considerando a presença constante de Panstrogylus megistus estes achados reforçam a necessidade de uma vigilância epidemiológica contínua e aperfeiçoada para Doença de Chagas naquela região

Análise quantitativa das lesões cardíacas na cardiomiopatia chagásica crônica canina

Lesions observed in chronic chagasic cardiopathy frequently produce electrocardiographic alterations and affect cardiac function. Through a computerized morphometrical analysis we quantified the areas occupied by cardiac muscle, connective and adipose tissues in the right atrium of dogs experimentally infected with Trypanosoma cruzi. All of the infected dogs showed chronic myocarditis with variable reduction levels of cardiac muscle, fibrosis and adipose tissue replacement. In the atrial myocardium of dogs infected with Be78 and Be62 cardiac muscle represented 34 and 50%, fibrosis 28 and 32% and adipose tissue 38 and 18%, respectively. The fibrosis observed was both diffuse and focal and mostly intrafascicular, either partially or completely interrupting the path of muscle bundles. Such histological alterations probably contributed to the appearance of electrocardiographic disturbances verified in 10 out 11 dogs which are also common in human chronic chagasic cardiopathy. Fibrosis was the most important microscopic occurrence found since it produces rearrangements of collagen fibers in relation to myocardiocytes which causes changes in anatomical physiognomy and mechanical behavior of the myocardium. These abnormalities can contribute to the appearance of cardiac malfunction, arrythmias and congestive cardiac insufficiency as observed in two of the analyzed dogs. Strain Be78 caused destruction of atrial cardiac muscle higher than that induced by strain Be62.As lesões observadas na cardiopatia chagásica crônica frequentemente produzem alterações eletrocardiográficas e afetam a função cardíaca. Através de uma análise morfométrica computadorizada nós quantificamos as áreas ocupadas por músculo cardíaco, tecido conjuntivo fibroso e tecido adiposo no átrio direito de cães experimentalmente infectados pelo Trypanosoma cruzi. Todos os cães infectados apresentaram miocardite crônica fibrosante com graus variáveis de redução de músculo cardíaco, fibrose e substituição por tecido adiposo. No miocárdio atrial dos cães infectados pelas cepas Be78 e Be62 foram observados 34 e 50% de músculo cardíaco, 28 e 32% de fibrose e, 38 e 18% de tecido adiposo, respectivamente. A fibrose observada era tanto difusa quanto focal e, principalmente intrafascicular interrompendo total ou parcialmente o percurso dos feixes musculares. Tais alterações histológicas provavelmente contribuiram para o surgimento dos distúrbios eletrocardiográficos verificados em 10 dos 11 cães estudados e que são comuns na cardiopatia chagásica crônica humana. De todos os achados microscópicos encontrados, a fibrose foi a mais importante por produzir rearranjos na fibras colágenas em relação aos miocardiócitos, modificando a fisionomia anatômica e o comportamento mecânico do miocárdio. Tais anormalidades estruturais podem contribuir para o surgimento à disfunção cardíaca, arritmias e à insuficiência cardíaca congestiva como verificado em dois cães analisados. A cepa Be78 produziu uma destruição de músculo cardíaco atrial estatisticamente superior à induzida pela cepa Be62

Trypanosoma cruzi : sensitivity of the polymerase chain reaction for detecting the parasite in the blood of mice infected with different clonal genotypes.

The polymerase chain reaction showed high sensitivity for detecting Trypanosoma cruzi in the blood of mice, independent of clonal genotype (19, 20?T. cruzi I; 32, 39?T. cruzi II) or phase of the infection (acute or chronic)

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