RESUMO Cercopithifilaria bainae é um nematoide pertencente à família Onchocercidae, que parasita o tecido subcutâneo de cães. Sua transmissão ocorre pelo carrapato Rhipicephalus sanguineus, e sua distribuição geográfica se sobrepõe ao espalhamento desse vetor. O presente estudo relata a detecção de microfilaremia por C. bainae em um cão macho de oito anos que apresentava anorexia, hipertermia, incoordenação motora, midríase e nódulo no testículo esquerdo e infecção concomitante por Ehrlichia sp. A coleta de sangue foi realizada, e o material analisado por meio dos exames de microscopia, PCR e sequenciamento de DNA. Microfilárias medindo 150±5,5μm de comprimento e 7±1,8μm de largura foram recuperadas. A sequência de DNA obtida mostrou 98% de identidade com sequências de C. bainae disponíveis no Genbank. Este é o primeiro relato de microfilaremia de C. bainae em um cão na região Centro-Oeste do Brasil.

ABSTRACT Cercopithifilaria bainae is a nematode belonging to the family Onchocercidae that parasitizes the subcutaneous tissue of dogs. Its transmission occurs through the tick Rhipicephalus sanguineus and its geographical distribution overlaps that of this vector. The present study reports the detection of microfilaremia by C. bainae in an eight-year-old male dog that presented anorexia, hyperthermia, motor incoordination, mydriasis, a nodule in the left testicle and concomitant infection by Ehrlichia sp. Blood samples were analyzed using microscopy, PCR and DNA sequencing. Microfilariae measuring 150±5.5μm in length and 7±1.8μm in width were retrieved. The DNA sequence exhibited 98% identity with C. bainae sequences available in Genbank. This is the first report of microfilaremia by C. bainae in a dog in the central western region of Brazil.

NifA is the transcriptional activator of the nif genes in Proteobacteria. It is usually regulated by nitrogen and oxygen, allowing biological nitrogen fixation to occur under appropriate conditions. NifA proteins have a typical three-domain structure, including a regulatory N-terminal GAF domain, which is involved in control by fixed nitrogen and not strictly required for activity, a catalytic AAA+ central domain, which catalyzes open complex formation, and a C-terminal domain involved in DNA-binding. In Herbaspirillum seropedicae, a β-proteobacterium capable of colonizing Graminae of agricultural importance, NifA regulation by ammonium involves its N-terminal GAF domain and the signal transduction protein GlnK. When the GAF domain is removed, the protein can still activate nif genes transcription; however, ammonium regulation is lost. In this work, we generated eight constructs resulting in point mutations in H. seropedicae NifA and analyzed their effect on nifH transcription in Escherichia coli and H. seropedicae. Mutations K22V, T160E, M161V, L172R, and A215D resulted in inactive proteins. Mutations Q216I and S220I produced partially active proteins with activity control similar to wild-type NifA. However, mutation G25E, located in the GAF domain, resulted in an active protein that did not require GlnK for activity and was partially sensitive to ammonium. This suggested that G25E may affect the negative interaction between the N-terminal GAF domain and the catalytic central domain under high ammonium concentrations, thus rendering the protein constitutively active, or that G25E could lead to a conformational change comparable with that when GlnK interacts with the GAF domain.

RESUMO Agentes químicos têm sido usados para auxiliar na inibição da formação do biofilme e impedir desenvolvimento da doença periodontal (DP). O objetivo deste estudo foi avaliar radiograficamente a ação do extrato de própolis na progressão da DP induzida em ratos. Foram utilizados 48 ratos Wistar, divididos em 4 grupos (n=12): Controle, Pincel, Própolis e Clorexidina. Os grupos foram subdivididos para análise aos 7 e 21 dias. A progressão da DP foi avaliada radiograficamente pela distância entre a junção cemento-esmalte e a crista óssea alveolar na face mesial do 1º molar inferior. A perda óssea foi significativamente menor nos animais do grupo própolis em ambos os períodos (p ≤ 0,01). Concluiu-se por análise radiográfica que a aplicação tópica do extrato de própolis interfere reduzindo a progressão da doença periodontal induzida por ligadura em ratos, demonstrando a importância desse composto como substância auxiliar no tratamento periodontal.

ABSTRACT Chemical agents have been used to assist on inhibiting biofilm formation and on preventing the development of periodontal disease (PD). The aim of this study was to radiographically evaluate the action of propolis extract on the progression of PD induced in rats. 48 Wistar rats were divided into 4 groups (n = 12): Control, Brush, Propolis and Chlorhexidine. The groups were subdivided for analysis at 7 and 21 days. The progression of the periodontal disease was radiographically assessed by the distance between the cement-enamel junction and the alveolar bone crest mesial of the 1st molar. Bone loss was significantly lower in the Propolis group in both periods (p ≤ 0.01). By radiographic analysis, it is concluded that the topical application of propolis extract interferes by reducing the progression of ligature-induced periodontal disease in rats, demonstrating the importance of this compound as an auxiliary substance in periodontal treatment.

Objetivou-se avaliar a viabilidade econômica da produção de leite de vacas Holandês x Gir, pela inclusão de níveis crescentes de óleo de girassol (0,0; 1,5; 3,0 e 4,5%) na matéria seca de dietas baseadas em cana-de-açúcar. Os dados utilizados no estudo foram obtidos de experimento realizado com 12 vacas Holandês x Gir, multíparas, com produção média de 18,2kg/dia de leite, distribuídas em três quadrados latinos 4 x 4, conforme a produção de leite, a ordem de lactação e o peso corpóreo. As dietas foram isoproteicas, fornecidas ad libitum na forma de mistura total (relação volumoso:concentrado de 60:40, base matéria seca), e os consumos das vacas determinados diariamente. As produções individuais diárias de leite foram registradas, e sua composição foi analisada quanto aos teores de gordura, proteína e ácidos graxos. A viabilidade econômica foi realizada considerando-se os preços dos ingredientes fornecidos e o consumo diário da dieta de cada tratamento. O benefício econômico foi obtido pela diferença entre a receita, com a venda do leite, e o custo com alimentação dos animais. O pagamento por qualidade foi estimado pela média de oito sistemas utilizados por empresas do ramo de laticínios, com base nos teores de gordura, proteína e no volume de leite. A dieta sem inclusão de OG foi a que proporcionou maior benefício econômico. A inclusão de OG nas dietas baseadas em cana-de-açúcar não foi viável, economicamente, devido aos gastos adicionais com OG e às penalizações resultantes da redução dos teores de proteína e de gordura do leite.

The aim of this study was to evaluate the economic viability of the milk yield of Holstein x Gir cows by the addition of growing levels of sunflower oil (SO) (0.0, 1.5, 3.0 and 4.5%) in the dry matter of sugar-cane based diets. The data utilized in the study were obtained from an experiment conducted with 12 multiparous Holstein x Gir cows, with average yield of 18.2kg/day of milk, distributed into three 4 x 4 Latin squares according to milk yield, lactation order and body weight. The diets were isoprotein, a total mixture fed at libitum (ratio roughage concentrate of 60:40, dry matter basis) and cow consumption was determined daily. The individual milk yields were recorded daily and their composition analyzed regarding fat, protein and fatty acids content. The economic viability was carried out by considering the prices of the feedstuffs fed and the daily consumption in the diet for each treatment. The economic benefit was obtained by the difference between the receipt with the milk sale and the cost with the feeding of the animals. The pay for quality was estimated on the basis of the mean of eight systems utilized by businesses in the dairy branch on the basis of the fat and protein contents and by milk volume. The diet without the addition of SO was the one which provided the greatest economic benefit. The addition of SO in the sugar-cane based diets was not economically viable due to the additional expenditures with SO and the penalizations resulting from the reduction of the milk protein and fat content.

O objetivo deste estudo foi avaliar o perfil de ácidos graxos (AG), os índices de qualidade nutricional e a estabilidade oxidativa (EO) de manteigas produzidas do leite de vacas alimentadas com dietas à base de cana-de-açúcar contendo níveis crescentes de óleo de girassol (OG): 0 (controle); 1,5; 3,0 e 4,5% da matéria seca (MS). O perfil de AG das manteigas foi analisado por cromatografia gasosa, e a EO foi determinada utilizando-se o equipamento Rancimat®, modelo 743, operado a 120ºC e fluxo de ar de 20L/h. As concentrações dos AG rumênico (CLA cis-9, trans-11), vacênico (C18:1 trans-11) e oleico (C18:1 cis-9) na gordura das manteigas foram aumentadas em 867, 687 e 148%, respectivamente, à medida que se aumentou de 0 para 4,5% o nível de OG na dieta. Por outro lado, as concentrações dos AG saturados de cadeia média foram linearmente reduzidas (P<0,0001) na gordura das manteigas, em razão do incremento de OG nas dietas. Quanto aos índices de qualidade nutricional, houve redução linear (P<0,0001) no índice de aterogenicidade e no de trombogenicidade e aumento da relação entre AG hipo e hipercolesterolêmicos, em resposta ao aumento do nível de OG na dieta. Consistente com o incremento (P<0,0001) nas concentrações totais dos AG mono e poli-insaturados, a EO da gordura das manteigas foi linearmente reduzida (P<0,0001) em razão do incremento de OG nas dietas. Concluiu-se que a suplementação com OG melhorou a qualidade nutricional das manteigas produzidas do leite de vacas Holandês x Gir devido a mudanças positivas no perfil de AG da gordura. Entretanto, tais mudanças na composição dos AG da gordura foram acompanhadas de redução da EO das manteigas, associada à menor vida de prateleira.

The objective of this study was to evaluate the fatty acid (FA) profile, nutritional quality and oxidative stability (OE) indexes of butter obtained from milk of cows fed sugar cane-based diets containing increasing levels of sunflower oil (SO): 0 (Control); 1.5; 3.0 and 4.5% of diet DM. The butter FA profile was analyzed by gas chromatography and OE was determined using the Rancimat® equipment (model 743) operated at 120ºC and air flow of 20 L/h. The concentrations of rumenic acid (cis-9 trans-11 CLA), vaccenic acid (trans-11 C18:1) and oleic acid (cis-9 C18:1) in butter fat were increased by 867, 687 and 148%, respectively, as the dietary SO level increased from 0 to 4.5%. In contrast, the concentrations of medium chain saturated FA were linearly reduced (P<0.05) in butter fat from cows fed increasing levels of SO. Regarding the butter nutritional quality, a linear decrease (P<0.0001) in atherogenicity and thrombogenicity indexes and a linear increase (P<0.0001) in the hypocholesterolemic/ hypercholesterolemic ratio were observed compared to control, 1.5, 3.0 and 4.5% SO, respectively. Consistent with the increased (P<0.0001) concentration of mono and polyunsaturated FA, the OE of butter fat was linearly reduced (P<0.0001) as the dietary SO level increased. It was concluded that diet supplementation with SO improved the nutritional quality of butter fat of Holstein x Gir dairy cows as a result of positive changes in milk FA profile. However, these changes in milk FA composition were accompanied by a reduction in the SO of butter, which in turn is associated with a shorter shelf life.

Avaliaram-se o consumo, metabólitos sanguíneos e a produção e composição do leite de 16 vacas Holandês x Gir, manejadas em pastagem de Brachiaria brizantha cv. Marandu, suplementadas com 6kg/vaca/dia (base matéria natural) de concentrado contendo 0; 1,3; 2,6 e 3,9kg/vaca/dia de grão de soja tostado (GST). Foi utilizado o delineamento com quatro quadrados latinos (QL) 4 x 4, sendo cada fase do QL constituída de 10 dias de período de adaptação à dieta e de cinco para coleta de amostras. A suplementação da dieta com GST resultou em redução linear (P<0,05) nos consumos de matéria seca e de fibra insolúvel em detergente neutro do pasto e total. A concentração de glicose não foi afetada (P>0,05), porém a de ácidos graxos não esterificados aumentou com a inclusão do GST na dieta (P<0,05). Não houve efeito (P>0,05) da adição do GST sobre a produção e composição do leite, exceto para o teor (P=0,10) e produção (P=0,08) de gordura no leite.

Feed intake, blood metabolites, and milk yield and composition of Holstein x Gir cows grazing Brachiaria brizantha cv. Marandu receiving 6kg/cow/day (natural matter based) of concentrate containing 0, 1.3, 2.6, 3.9kg/cow/day of roasted soybean seeds (RSS) were evaluated. The experimental design was four 4 x 4 latin square (LS) in wich each phase consisted of 10 days for diet adaptation period and 5 days for samples collection. Diet supplementation with RSS decreased linearly (P<0.05) the intake of pasture, total diet dry matter and neutral detergent fiber. The plasma glucose concentration was unaffected (P>0.05), but nonesterified fatty acids increased with RSS supplementation (P<0.05). There was no effect (P>0.05) of RSS supplement on milk production and composition, except the linear reduction on the content (P=0.10) and yield (P=0.08) milk fat, which can present positive financial implication to farmers, knowing that most of the Brazilian dairy use payment programs by milk composition.

Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ54 co-factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH4Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.

O objetivo deste estudo foi identificar a atividade inibitória de óleos de copaíba sobre o crescimento dos micro-organismos: Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Citrobacter freundi, Staphylococcus aureus, Actinobacillus pleuropneumoniae e Haemophilus parasuis. Foi realizado um teste de difusão em ágar com duas soluções a 10% de óleos de copaíba obtidos de duas diferentes espécies de copaíba (Copaifera officinalis e C. langsdorffii) e um controle negativo com tween 80 e água. Os isolados clínicos de H. parasuis e A. pleuropneumoniae foram incubados em microaerofilia. Os resultados mostraram três espécies Gram-negativas inibidas por ambas as soluções de óleo de copaíba: E. coli, P. aeruginosa e S. flexneri. Na inibição de P. aeruginosa o óleo de C. officinalis foi superior ao de C. langsdorffii (P<0,05). Todas as cepas de S. aureus tiveram seu crescimento inibido pelas soluções no ensaio, sem diferença estatística entre os halos. Estes resultados sugerem que o óleo de copaíba pode ser uma fonte potencial de compostos inibitórios para ser utilizada como antimicrobianos no tratamento de infecções humanas e animais e conservação de alimentos.

We evaluated genetic variants of apolipoprotein E (APOE HhaI) and their association with serum lipids in colorectal cancer (CRC), together with eating habits and personal history. Eight-seven adults with CRC and 73 controls were studied. APOE*2 (rs7412) and APOE*4 (rs429358) were identified by polymerase chain reaction-restriction fragment length polymorphism. APOE gene polymorphisms were similar in both groups, but the ε4/ε4 genotype (6%) was present only in controls. The patients had reduced levels (mean ± SD) of total cholesterol and low-density lipoprotein cholesterol fraction (180.4 ± 49.5 and 116.1 ± 43.1 mg/dL, respectively) compared to controls (204.2 ± 55.6, P = 0.135 and 134.7 ± 50.8 mg/dL; P = 0.330, respectively) indicating that they were not statistically significant after the Bonferroni correction. The APOE*4 allele was associated with lower levels of total cholesterol, low- and high-density lipoprotein cholesterol fraction and increased levels of very low-density lipoprotein cholesterol fraction and triglycerides only among patients (P = 0.014). There was a positive correlation between the altered lipid profile and increased body mass indexes in both groups (P < 0.010). Moreover, a higher rate of hypertension and overweight was observed in controls (P < 0.002). In conclusion, the presence of the ε4/ε4 genotype only in controls may be due to a protective effect against CRC. Lower lipid profile values among patients, even those on lipid-rich diets associated with the APOE*4 allele, suggest alterations in the lipid synthesis and metabolism pathways in CRC.

Apolipoprotein E (apoE - e2, e3, e4 alleles) plays a role in the regulation of lipid metabolism, with the e4 considered to be a risk factor for coronary artery disease (CAD). We aimed to evaluate the apoE polymorphisms in Brazilians with CAD and their influence on the lipid profile and other risk factors (hypertension, diabetes mellitus, smoking). Two hundred individuals were examined: 100 patients with atherosclerosis confirmed by coronary angiography and 100 controls. Blood samples were drawn to determine apoE polymorphisms and lipid profile. As expected, the e3 allele was prevalent in the CAD (0.87) and non-CAD groups (0.81; P = 0.099), followed by the e4 allele (0.09 and 0.14, respectively; P = 0.158). The e3/3 (76 and 78%) and e3/4 (16 and 23%) were the most common genotypes for patients and controls, respectively. The lipid profile was altered in patients compared to controls (P < 0.05), independently of the e4 allele. However, in the controls this allele was prevalent in individuals with elevated LDL-cholesterol levels only (odds ratio = 2.531; 95% CI = 1.028-6.232). The frequency of risk factors was higher in the CAD group (P < 0.05), but their association with the lipid profile was not demonstrable in e4 carriers. In conclusion, the e4 allele is not associated with CAD or lipid profile in patients with atherosclerosis. However, its frequency in the non-CAD group is associated with increased levels of LDL-cholesterol, suggesting an independent effect of the e4 allele on lipid profile when the low frequency of other risk factors in this group is taken into account.