Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats.

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

The applicability of fingernail lead and cadmium levels as subchronic exposure biomarkers for preschool children.

Preschool children are exposed daily to metals in their homes and at daycare centers (DCC). Metal exposure and health effects are associated even at low levels, and children comprise a group of public health concern. Nail metals have been studied for exposure biomonitoring and compared to other biological media. The aim of this study was to explore the applicability of preschool fingernail lead and cadmium as subchronic exposure biomarkers. Nail lead and cadmium levels (NLL and NCL) of 602 preschool children (age: 1-4 years) who attended 21 DCC in São Paulo, Brazil, in 2013 were analyzed. Results were compared against blood lead and cadmium levels (BLL and BCL) found in a previous study. Inductively coupled plasma mass spectrometry (ICP-MS) analyses were performed for both samples. DCC and metal contaminated sites (MCS) were georeferencing. Logistic regression tests were applied to verify associations between nail metal levels and risk factors (sex, age, maternal education, secondary smoking, DCC geographic district, vehicle flow density, relative altitude and distance between DCC and nearest MCS) (p < 0.05). BLL was stratified by exposure level (low: <5 µg.dL-1; high: >13.9 µg.dL-1; medium: ≥5 µg.dL-1 and ≤13.9 µg.dL-1) and also tested the associations in order to verify if nail lead levels are affected by exposure intensity defined by blood metals concentrations. Radius distance and relative altitude of DCC to nearest MCS were associated with high NLL and NCL. Abnormal appearance of nails was associated with high NLL and low NCL. Lead and cadmium exposure magnitude had no significant impact on NLL. NLL should only be used for initial screening, and when financial resources are scarce, especially in areas located near contaminated sites. Preschool children were co-exposed to both lead and cadmium, reinforcing the need for broader studies evaluating exposure to environmental pollutants for more than one chemical element.

Behavioral effects of Bj-PRO-7a, a proline-rich oligopeptide from Bothrops jararaca venom.

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M1R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), α-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M1R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Behavioral effects of Bj-PRO-7a, a proline-rich oligopeptide from Bothrops jararaca venom

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M1R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), α-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M1R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Frequency of enterotoxins, toxic shock syndrome toxin-1, and biofilm formation genes in Staphylococcus aureus isolates from cows with mastitis in the Northeast of Brazil.

Staphylococcus aureus is among the microorganisms more frequently associated with subclinical bovine mastitis. S. aureus may produce several virulence factors. This study aimed at determining the frequency of virulence factors such as enterotoxins, toxic shock syndrome toxin 1, and ica adhesion genes. In addition, we assessed antimicrobial drug resistance in S. aureus isolated from clinical and subclinical cases of mastitis. A total of 88 cows with clinical or subclinical mastitis were sampled, resulting in 38 S. aureus isolates, from which 25 (65.78%) carried toxin genes, including seb, sec, sed, tst, and icaD adhesion gene. These S. aureus isolates belong to 21 ribotypes and three S. aureus strains belonged to the same ribotype producing ica adhesion gene. Approximately 90% of S. aureus strains obtained in our study demonstrated multiple resistance to different antimicrobial agents. The most efficacious antimicrobial agents against the isolates were gentamicin, amoxicillin, and norfloxacin. Gentamicin was the most efficacious agent inhibiting 78.95% of the S. aureus isolates. The least efficacious were penicillin, streptomycin, and ampicillin. Our results can help in understanding the relationship between virulence factors and subclinical mastitis caused by S. aureus. Further research about diversity of S. aureus isolates and genes responsible for the pathogenicity of subclinical mastitis is essential.

Participation of TLR2 and TLR4 in Cytokines Production by Patients with Symptomatic and Asymptomatic Chronic Chagas Disease.

Chagas disease (CD), caused by the protozoan Trypanosoma cruzi, is a serious public health issue. Its evolution involves an acute stage, characterized by no specific symptoms, and the chronic stage during most individuals are asymptomatic, but about 30-40% of them become symptomatic presenting the cardiac or digestive disease. Host immune response mechanisms involved in symptomatic or asymptomatic chronic disease are not fully understood. The pro-inflammatory cytokines are crucial in host resistance. However, a fine control of this inflammatory process, by action of anti-inflammatory cytokines, is necessary to avoid tissue injury. This control was found to be responsible for no clinical manifestations in asymptomatic individuals. Toll-like receptors (TLRs) are extremely important in defining the cytokine profile released in response to a micro-organism. We found that patients with the cardiac form predominantly released the pro-inflammatory cytokines: IFN-γ, TNF-α and IL-17 with the involvement of both, TLR2 and TLR4. In contrast, patients with asymptomatic disease release predominantly the anti-inflammatory cytokines IL-10 and TGF-ß, but also with TLR2 and TLR4 participation. The mechanisms by which stimulation of the same TLRs results in release of different pattern of cytokines, depending on the patients group that is being evaluated, are discussed.

Non-invasive screening of diabetics in primary care for NAFLD and advanced fibrosis by MRI and MRE.

BACKGROUND: Current guidelines do not recommend screening for non-alcoholic fatty liver disease (NAFLD) or advanced fibrosis. Patients with type 2 diabetes mellitus (T2DM) are known to be at increased risk for NAFLD and advanced fibrosis. AIM: To assess the feasibility in diabetics in a primary care setting of screening for NAFLD and advanced fibrosis, by using non-invasive magnetic resonance imaging (MRI) to estimate the hepatic proton density fat fraction (MRI-PDFF) and magnetic resonance elastography (MRE) to estimate hepatic stiffness. METHODS: We performed a cross-sectional analysis of a prospective study that included 100 (53% men) consecutively enrolled diabetics who did not have any other aetiology of liver disease. All patients underwent a standardised research visit, laboratory tests, MRI-PDFF, and MRE. RESULTS: Mean (±s.d.) age and body mass index (BMI) was 59.7 (±11.2) years and 30.8 (±6.5) kg/m(2) , respectively. The prevalence of NAFLD (defined as MRI-PDFF ≥5%) and advanced fibrosis (defined as MRE ≥3.6 kPa) was 65% and 7.1%, respectively. One patient with advanced fibrosis had definite hepatocellular carcinoma. When compared to those without NAFLD, patients with NAFLD were younger (P = 0.028) and had higher mean BMI (P = 0.0008), waist circumference (P < 0.0001) and prevalence of metabolic syndrome (84.6% vs. 40.0%, P < 0.0001). Only 26% of those with NAFLD had elevated alanine aminotransferase. CONCLUSIONS: This proof-of-concept study demonstrates that T2DM has significant rates of both NAFLD and advanced fibrosis. Concomitant screening for NAFLD and advanced fibrosis by using MRI-proton density fat fraction and magnetic resonance elastography in T2DM is feasible and may be considered after validation in a larger cohort.

Real-time PCR for detection of Brucella ovis and Histophilus somni in ovine urine and semen / PCR em tempo real para detecção de Brucella ovis e Histophilus somni em urina e sêmen ovino

A epididimite infecciosa ovina é uma das principais enfermidades reprodutivas de carneiros. O presente estudo teve por objetivo desenvolver protocolos de PCR em tempo real para B. ovis e H. somni e avaliar sua aplicabilidade em amostras de sêmen e urina de carneiros. Delinearam-se primers e sondas espécie-específicos para cada agente. As sondas foram delineadas com o sistema TaqMan incorporando um marcador FAM para B. ovis e Cy5 para H. somni na extremidade 5' e um quencher na extremidade 3'. A PCR em tempo real para B. ovis e H. somni foi altamente sensível, uma vez que a amplificação de DNA ocorreu com até 0,2ng de DNA/reação. A especificidade dos iniciadores e sondas foi avaliada com amostras de DNA de outros agentes causadores de epididimite ovina e nenhuma amplificação inespecífica foi observada. A aplicabilidade da técnica em amostras biológicas também foi confirmada, pois não houve perda de eficácia (P>0,05) quando comparada à PCR convencional com amostras de sêmen e urina de carneiros experimentalmente infectados.

Molecular characterization of neuropathogenic Equine Herpesvirus 1 Brazilian isolates / Caracterização molecular de isolados brasileiros de herpesvírus equino 1

Este trabalho descreve a caracterização molecular de oito amostras de herpesvírus equino 1 isoladas do sistema nervoso central de equinos que morreram com sinais neurológicos no estado de Minas Gerais. As amostras de EHV-1 foram isoladas em cultivo celular, e a caracterização molecular foi feita por genotipagem e identificação do marcador neuropatogênico por meio das técnicas de PCR, restrição enzimática e sequenciamento. A caracterização molecular desses isolados pode ser a base para o desenvolvimento de novas formulações vacinais com maior eficácia para a prevenção de doença neurológica causada pelo EHV-1.(AU)

Diagnóstico etiológico de enfermidades do sistema nervoso central de equinos no Estado de Minas Gerais, Brasil / Etiologic diagnosis of diseases of the central nervous system of horses in Minas Gerais State, Brazil

O Brasil possui o quarto maior rebanho equino do mundo, e o Estado de Minas Gerais detém o maior número de equinos do país. Portanto, um diagnóstico preciso das doenças neurológicas dos equinos é prioridade no estado. Sendo assim, o objetivo deste estudo foi identificar, utilizando a Reação em Cadeia pela Polimerase (PCR), os agentes infecciosos responsáveis por enfermidades que afetam o sistema nervoso central (SNC) de equinos. De janeiro de 2009 a janeiro de 2011, foi realizado um levantamento dos casos de encefalites e encefalomielites em equinos no Estado de Minas Gerais, utilizando-se amostras de SNC de equinos que morreram com sinais neurológicos. Das 217 amostras de SNC, 47 (21,7%) foram positivas para o vírus da raiva pelo método de imunofluorescência indireta e inoculação em camundongos. Nas 170 amostras negativas para o vírus da raiva, o herpes-vírus equino-1 (EHV-1) foi diagnosticado em 20 (11,8%) e o herpes-vírus suíno-1 (SHV-1), em uma amostra por meio de PCR, e o vírus encefalite de Saint Louis (SLEV), em outra amostra, através de transcrição reversa (RT) e PCR (RT-PCR). Constatou-se que o vírus da raiva é o principal agente causador de encefalite em equinos, apesar do crescente número de casos de encefalomielite associados ao EHV-1 no Estado de Minas Gerais.(AU)

Brazil has the fourth largest equine herd in the world and the State of Minas Gerais has the largest equine population in the country. Therefore, an accurate diagnosis of cases of neurologic diseases is a priority in Minas Gerais. The aim of this study was to identify by Polymerase Chain Reaction (PCR) infectious agents associated with neurological disease in the central nervous system (CNS) of horses. A survey of encephalitis and encephalomyelitis in horses in Minas Gerais State was performed on samples of CNS from horses that died with neurological signs from January 2009 to January 2011. Forty seven CNS samples from 217 (21.7%) horses were positive for rabies virus by the indirect immunofluorescence assay and mouse inoculation. Among the 170 samples that were negative for rabies, EHV-1 was detected in 20 (11.8%) and the swine herpesvirus-1 (SHV-1) was detected in one sample by PCR, and the Saint Louis encephalitis virus (SLEV) was identified in another sample by reverse transcription (RT) and PCR (RT-PCR). Rabies virus is the most common causative agent of encephalitis in horses, despite the increasing number of cases of encephalitis associated with EHV-1 in the State of Minas Gerais.(AU)

Indirect ELISA for diagnosis of Brucella ovis infection in rams / ELISA indireto para diagnóstico da infecção por Brucella ovis em carneiros

Brucella ovis is a major cause of epididymitis in sexually mature rams, resulting in subfertility, infertility, and economic losses for the sheep industry worldwide. The aim of this study was to develop an indirect ELISA (iELISA) using recombinant proteins, namely rBoP59 and rBP26, as antigens for serological diagnosis of B. ovis infection. The BoP59 and BP26 recombinant proteins were expressed in E. coli and purified by affinity chromatography. Antigenicity was tested by Western blot and iELISA. Standardization of iELISA was performed with 500ng and 1µg BoP59 and rBP26 per well, testing serum from uninfected and experimentally infected rams. rBP26 was effective in distinguishing positive from negative rams. The rBP26 iELISA developed in this study is the first to use a completely purified rBP26 as antigen resulting in high sensitivity (100%) and specificity (90.2%), and an overall accuracy equal to 1.0...

Brucella ovis é uma das principais causas de epididimite em carneiros sexualmente maduros, resultando em subfertilidade e infertilidade e consequentes perdas econômicas para a ovinocultura em todo o mundo. O objetivo deste estudo foi desenvolver ELISA indireto (ELISAi), utilizando como antígeno proteínas recombinantes BoP59r e BP26r para diagnóstico da infecção por B. ovis. BoP59r e BP26r foram expressas em E. coli e purificadas por cromatografia de afinidade e a antigenicidade testada por Western blot e ELISAi. A padronização do ELISAi foi realizada testando 500 ng e 1 µg de BoP59r e BP26r por poço e soros de carneiros infectados e não infectados. A BP26r foi eficiente em diferenciar ovinos negativos de positivos. O ELISAi com BP26 desenvolvido neste estudo foi o primeiro a utilizar BP26 completamente purificada como antígeno, resultando em elevada sensibilidade (100%) e sensibilidade (90,2%), com acurácia igual a 1,0...

Identification and genetic characterization of a new Brazilian genotype of Toxoplasma gondii from sheep intended for human consumption.

Recent studies have demonstrated that strains of Toxoplasma gondii in Brazil are frequently different from those detected in other countries, thus making an accurate phylogenetic analysis difficult. The aim of this study was to genetically characterize T. gondii samples from sheep raised in southern Bahia and intended for human consumption, by means of PCR-RFLP and sequencing techniques. Experimental samples were obtained from 200 sheep brains purchased at butcher's shops in Itabuna, Bahia, Brazil. In total, three samples (#54, #124 and #127) were T. gondii-positive. The application of multilocus PCR-RFLP using ten molecular markers (SAG1, SAG2, SAG3, BTUB, c22-8, PK1, GRA6, L358, c-29-2 and Apico) revealed a single genotype common to all samples of this study, which differed from any other published T. gondii genotypes. An atypical allele was detected in the L358 genetic marker; this has not previously been shown in any other South American T. gondii isolates. Phylogenetic analysis on the sequences from multilocus PCR sequencing revealed that these three samples were classified into the same lineage. Extensive indel regions were detected in the Apico genetic marker. Together, our findings revealed a new Brazilian T. gondii genotype. Further research should be conducted to enrich the database of Brazilian T. gondii genotypes from different regions. This will make it possible to understand the phylogenetic relationship between isolates.

Anti-ulcerogenic and antisecretory effects of Celtis iguanaea (Jacq. ) Sargent hexane leaf extract / Efeito anti-ulcerogênico e antisecretório do extrato hexânico das folhas de Celtis iguanaea (Jacq. ) Sargent

The Celtis iguanaea (Jacq.) Sargent (Cannabaceae) is one of the native species of the Cerrado region of Brazil widely used in folk medicine to treat dyspepsia. The objective of the present study was to evaluate the gastroprotective effect of the Celtis iguanaea (Jacq.) Sargent (HE) hexane leaf extract in the lesion and gastric secretion models.Antiulcerogenic activity of the Celtis iguanaea (HE) hexane leaf extract was observed with the experimental models, such as indomethacin and pyloric ligation-induced gastric ulcers. In order to evaluate the antisecretory activity of this extract, isolated Rana catesbeiana mucosa and pyloric ligation in mice were used. The HE treatment reduced the lesion index of indomethacin and pyloric ligation-induced ulcer. This extract also reduced the gastric acid secretion and total acidity (increasing the gastric pH) in mice. The secretion of H+ was reduced in the basal values (15.58 ± 1.99 µEq H+/g/15 min) when isolated Rana catesbeiana mucosa was incubated with HE. Intraduodenal administration of HE reduced the gastric secretion produced by bethanecol or histamine. The antiulcerogenic and antisecretory efficacy of HE in this study suggest anticholinergic and antihistaminergic mechanism or interruption of intracellular events that are linked to acid secretion.

Celtis iguanaea (Jacq.) Sargent (Cannabaceae) é uma das espécies nativas do cerrado brasileiro, sendo amplamente utilizada na medicina popular para o tratamento de dispepsia. O objetivo do presente trabalho foi avaliar o efeito gastroprotetor do extrato hexânico das folhas de Celtis iguanaea (Jacq.) Sargent (EH) em modelos de secreção e de lesões gástricas. A atividade antisecretória do extrato hexânico das folhas de Celtis iguanaea (EH) foi avaliada no modelo de mucosa isolada de Rana catesbeiana e pelo modelo de ligadura pilórica em camundongos. A avaliação da atividade anti-ulcerogênica do EH foi determinada no modelo de úlceras gástricas induzidas por indometacina e ligadura pilórica. Quando a mucosa de Rana catesbeiana isolada foi incubada com o EH, a secreção de H+ foi reduzida em comparação com o valor basal (15,58 ± 1,99 µEq H+/g/15 min). O tratamento com o EH reduziu o índice de lesões na úlcera induzida por indometacina e ligadura pilórica. O EH também reduziu o volume de secreção, a acidez total, e aumentou o pH gástrico. A administração intraduodenal reduziu a secreção gástrica produzida por betanecol ou histamina. A eficácia anti-ulcerogênica e antisecretória do EH neste estudo, sugere um mecanismo anticolinérgico e antihistaminérgico ou interrupção de eventos intracelulares que estão ligados à secreção ácida.

Estudo farmacognóstico de folhas e raízes da Spiranthera odoratissima A. St. -Hil. (Rutaceae) / Pharmacognostic study of the leaves and roots of Spiranthera odoratissima A. St. -Hil. (Rutaceae)

A Spiranthera odoratissima A. St.-Hil. (manacá) é um arbusto cujas raízes são utilizadas popularmente contra dores musculares, de estômago e de cabeça, disfunções hepáticas, reumatismo, e como estimulante do apetite. As folhas são empregadas como depurativo do sangue, nas afecções renais e hepáticas. O objetivo deste trabalho foi realizar o estudo morfo-anatômico de S. odoratissima, bem como a triagem fitoquímica, a determinação dos teores de água, cinzas totais e insolúveis em ácido. O material vegetal foi coletado em Leopoldo de Bulhões/ Goiás e os estudos morfo-anatômicos e a triagem fitoquímica foram realizados utilizando as técnicas convencionais. Verificou-se que a folha é hipoestomática, com estômatos anomocíticos e a epiderme apresenta papilas na face abaxial. O mesofilo é dorsiventral e a nervura principal possui drusas de oxalato de cálcio e cavidades secretoras. O pecíolo apresenta sistema vascular em forma circular. Na raiz, em crescimento primário, observou-se cilindro vascular com organização diarca. Na prospecção fitoquímica detectaram-se fenóis, taninos, cumarinas, flavonoides, triterpernos/esteróis, antraquinonas e antocianinas nas folhas e alcaloides, cumarinas, amido e saponinas nas raízes. Esses resultados são parâmetros importantes para o controle de qualidade dessa matéria-prima vegetal.

Spiranthera odoratissima A. St.-Hil. (manacá) is a shrub whose roots are popularly used against stomach pains, headache, muscle pain, liver disorders, rheumatism and as an appetite stimulant. The leaves are used as a blood depurative and for liver and kidney disease. The purpose of this work was to perform the morphological and anatomical study, the phytochemical screening and determine the water content and the levels of total and acid of S. odoratissima. The plant material was collected in Leopoldo de Bulhões/ Goiás and the morpho-anatomical studies and the phytochemical screening were performed using conventional techniques. It was verified that the leaf is hypostomatic, with anomocytic stomata and the epidermis has papillae on the abaxial surface. The mesophyll is dorsiventral and the midrib presents druses of calcium oxalate and secretory cavities. The petiole has a circular vascular system. The root in primary growth presents vascular cylinder with diarchic organization. Phenols, tannins, coumarins, flavonoids, triterpenes/sterols, anthraquinones and anthocyanins were detected in the phytochemical screening of the leaves, and alkaloids, coumarins, saponins and starch were detected in the roots. These results are important parameters for the quality control of the vegetal raw material.

Aspectos reprodutivos e produtivos de vacas da raça Holandesa com puerpério normal ou patológico / Reproductive and productive parameters of Holstein cows with normal or pathological puerperium

Foram avaliadas 103 vacas da raça Holandesa de primeira, segunda ou terceira ordens de lactação, com o objetivo de comparar aspectos reprodutivos e produtivos dos animais com puerpério normal ou patológico. Exames ginecológicos foram realizados semanalmente, do parto ao final do puerpério, por meio de palpação transretal, ultrassonografia e vaginoscopia. As incidências de partos gemelares, partos auxiliados, retenção de placenta e infecções uterinas até o primeiro serviço foram de 11,7%, 20,4%, 38,8% e 75%, respectivamente. A involução uterina e a primeira ovulação ocorreram aos 33,5±11,1 e 43,6±21,5 dias após o parto, respectivamente, com diferenças (P<0,001) entre vacas com ou sem infecção uterina. O primeiro serviço ocorreu aos 75,9±31,0 dias após o parto e a taxa de gestação foi de 20,7%. A produção de leite diária foi de 27,9±7,3 litros por vaca até 42 dias após o parto. O prejuízo com a redução na produtividade e o descarte do leite contaminado com resíduos de antibiótico foi equivalente a 598 litros de leite por vaca com retenção de placenta e infecção uterina. A alta incidência de transtornos puerperais interferiu na fertilidade e na produção de leite, causando elevado impacto econômico para a propriedade.

The study was performed with 103 Holstein cows of first, second or third orders of lactation, and aimed to compare reproductive and productive parameters of animals with normal or pathological puerperium. Uterine and ovarian examinations were performed weekly until 42 days after parturition, by rectal palpation, vaginoscopy and ultrasonography. The incidence of twin pregnancies, assisted births, retained placenta, and uterine infection until the first service, were 11.7%, 20.4%, 38.8% and 75%, respectively. Uterine involution and detection of the first corpus luteum occurred at 33.5±11.1 and 43.6±21.5 days after parturition, respectively, and differed (P<0.01) between cows with or without uterine infection. The first artificial insemination occurred in average at 75.9±31.0 days after parturition, and the pregnancy rate was 20.7%. Average daily milk yield was 27.9±7.3 liters/cow until 42 days after parturition. Financial losses related to decreased milk production and discarding of milk contaminated with antibiotic residues corresponded to 598 liters of milk/cow presenting retained placenta and uterine infection. The high incidence of puerperal diseases negatively affected the fertility and milk production, causing large economic impact to the milk production system.

Species-specific nested PCR as a diagnostic tool for Brucella ovis infection in rams / Nested PCR espécie-específica para o diagnóstico da infecção por Brucella ovis em carneiros

The aim of the present study was to evaluate a species-specific nested PCR based on a previously described species-specific PCR for detection of B. ovis in semen and urine samples of experimentally infected rams. The performance of the species-specific nested PCR was compared with the results of a genus-specific PCR. Fourteen rams were experimentally infected with the Brucella ovis REO 198 strain and samples of semen and urine were collected every week up to 180 days post infection. Out of 83 semen samples collected, 42 (50.6%) were positive for the species-specific nested PCR, and 23 (27.7%) were positive for the genus-specific PCR. Out of 75 urine samples, 49 (65.3%) were positive for the species-specific nested PCR, whereas 11 (14.6%) were genus-specific PCR positive. Species-specific nested PCR was significantly more sensitive (P<0.001) than the genus-specific PCR in semen and urine from experimentally infected rams. In conclusion, the species-specific nested PCR developed in this study may be used as a diagnostic tool for the detection of B. ovis in semen and urine samples from suspected rams.

O presente estudo objetivou avaliar uma técnica de nested PCR espécie-específica delineada a partir de PCR espécie-específica descrita anteriormente para detecção de B. ovis em sêmen e urina de carneiros infectados experimentalmente. O desempenho da nested PCR espécie-específica foi comparado com os resultados de uma PCR gênero-específica. Quatorze carneiros foram infectados experimentalmente com Brucella ovis REO 198 e amostras de sêmen e de urina foram colhidas semanalmente até 180 dias após a infecção. De 83 amostras de sêmen, 42 (50,6%) foram positivas pela nested PCR espécie-específica, e 23 (27,7%) foram positivas pela PCR gênero-específica. De 75 amostras de urina, 49 (65,3%) foram positivas pela nested PCR espécie-específica, enquanto 11 (14,6%) foram positivas em PCR gênero-específica. A técnica de nested PCR espécie-específica foi significativamente mais sensível (P<0,001) do que a PCR gênero-específica no sêmen e na urina de carneiros infectados experimentalmente. Em conclusão, a nested PCR espécie-específica desenvolvida neste estudo pode ser utilizada como ferramenta de diagnóstico para detecção de B. ovis em sêmen e urina de carneiros suspeitos.

Anti-inflammatory and antinociceptive activities of LQFM002 - A 4-nerolidylcatechol derivative.

AIMS: The current study describes the synthesis and pharmacological evaluation of (E)-N-(3,7-dimethylocta-2,6-dienyl)-1,3-dimethyl-1H-pyrazol-5-amine (LQFM002), a compound originally designed through a molecular simplification strategy from 4-nerolidylcatechol. LQFM002 was evaluated for preservation of the PLA(2) enzyme inhibitory effects of the lead compound, 4-nerolidylcatechol, using in vitro and in vivo models. MAIN METHODS: Rota-rod, open field and pentobarbital-induced sleeping tests were used to evaluate the effects of LQFM002 on the central nervous system. A gel plate assay of PLA(2) activity, carrageenan-induced pleurisy and TNF-α levels was used to assay anti-inflammatory activity. Antinociceptive activities of LQFM002 were evaluated with acetic acid-induced writhing, formalin and hot-plate tests, while involvement of the opioid pathway in the LQFM002 antinociceptive effect was investigated with naloxone pre-treatment. KEY FINDINGS: LQFM002 inhibited PLA(2) activity, cell migration into the pleural cavity, and capillary permeability (Evan's blue concentration) and reduced TNF-α levels in pleural exudates. LQFM002 also reduced acetic acid-induced writhing and the licking time in both phases of the formalin test and increased latency in the hot-plate test. Pre-treatment with 8.25 µmol/kg naloxone (3mg/kg) reversed the analgesic effects of LQFM002 in the early phase of the formalin test. SIGNIFICANCE: LQFM002 showed anti-inflammatory activity, which possibly involved reduction of leukocyte migration and TNF-α levels. LQFM002 also demonstrated inhibition of PLA(2) activity in vitro. LQFM002 had an antinociceptive effect that involved the opioidergic system.

Monocyte-derived macrophages from Zebu (Bos taurus indicus) are more efficient to control Brucella abortus intracellular survival than macrophages from European cattle (Bos taurus taurus).

Brucellosis is one of the most important zoonotic diseases in the world. Considering its strict zoonotic nature, understanding of the pathogenesis and immunity of Brucella spp. in natural animal hosts is essential to prevent human infections. Natural resistance against brucellosis has been demonstrated in cattle, and it is associated with the ability of macrophages to prevent intracellular replication of Brucella abortus. Identification of breeds that are resistant to B. abortus may contribute for controlling and eradicating brucellosis in cattle. This study aimed to compare macrophages from Nelore (Bos taurus indicus) or Holstein (Bos taurus taurus) regarding their resistance to B. abortus infection. Macrophages from Nelore were significantly more efficient in controlling intracellular growth of B. abortus when compared to Holstein macrophages even under intralysosomal iron restricting conditions. Furthermore, Nelore macrophages had higher transcription levels of inducible nitric oxide synthase (iNOS) and TNF-α at 12h post-infection (hpi) and higher levels of IL-12 at 24 hpi when compared to Holstein macrophages. Conversely, Holstein macrophages had higher levels of IL-10 transcripts at 24 hpi. Macrohages from Nelore also generated more nitric oxide (NO) in response to B. abortus infection when compared to Holstein macrophages. In conclusion, cultured Nelore macrophages are more effective in controlling intracellular replication of B. abortus, suggesting that Nelore cattle is likely to have a higher degree of natural resistance to brucellosis than Holstein.

Diagnosis of Brucella ovis infection by serology and PCR in urine samples from naturally infected rams in the state of Piauí / Diagnóstico da infecção por Brucella ovis por sorologia e reação em cadeia pela polimerase (PCR) em amostras de urina de carneiros naturalmente infectados no estado do Piauí

A brucelose ovina causada pela Brucella ovis é uma doença reprodutiva de carneiros caracterizada por epididimite, orquite, com consequente diminuição da fertilidade e prejuízos econômicos significativos. O presente trabalho teve por objetivo avaliar a aplicabilidade da técnica de PCR como um método de diagnóstico em campo, comparado-a com a técnica de IDGA. Foram coletadas amostras de urina e soro de 90 carneiros oriundos de 31 rebanhos localizados no Estado do Piauí. Quatro das 31 (12,9%) propriedades avaliadas apresentaram animais positivos. Dezoito (20%) amostras de urina foram positivas pela PCR, enquanto o método de IDGA identificou 16 (17,8%) carneiros soropositivos. Embora os métodos tenham apresentado concordância baixa na estatística Kappa (k=0,04), não foi observada diferença estatística entre as técnicas (P>0,05) pelo teste exato de Fisher. A combinação dos dois testes aumentou significativamente a detecção de animais positivos para 34,4% (P <0,05), sugerindo que a associação de métodos de diagnóstico como a técnica de PCR em amostras de urina e sorologia por IDGA e a avaliação clínica dos animais é necessária para um diagnóstico eficiente na infecção por B. ovis.