RESUMO A brucelose é uma importante zoonose de proporções mundiais. Os testes sorológicos utilizados no Brasil, antígeno acidificado tamponado (RBT), soroaglutinação lenta/2-mercaptoetanol (2-ME), polarização fluorescente (FPA) e fixação de complemento (FC), são baseados no antígeno lipopolissacarídeo liso (S-LPS) da Brucella abortus. O objetivo deste estudo foi avaliar uma proteína BP26 recombinante utilizada como antígeno em um teste rápido imunocromatográfico de fluxo lateral (rBP26-LFIA) para o diagnóstico sorológico da brucelose bovina. As características de desempenho analítico foram avaliadas em amostras de soro bovino positivas e negativas previamente caracterizadas pelos testes RBT e 2-ME. As estimativas da sensibilidade e especificidade analíticas foram de 73,91% e 97,14%, respectivamente. Soros bovinos reagentes à Neospora, Trypanossoma vivax e Leptospira foram utilizados para avaliação de especificidade, em razão de essas doenças serem causas frequentes de aborto em bovinos, além de ser possível reação cruzada induzida por vacinas comerciais contra Leptospira, em testes sorológicos para a brucelose bovina que utilizam S-LPS como antígeno. Como conclusão, o teste rBP26-LFIA, com sua atual padronização, apresentou um bom desempenho analítico. Contudo, uma futura avaliação do desempenho diagnóstico do teste rBP26-LFIA com amostras provenientes de regiões com prevalência conhecidas é necessária para sua recomendação de uso no programa brasileiro de controle e erradicação da brucelose bovina. mundiais Brasil RBT, , (RBT) lenta/2mercaptoetanol lenta2mercaptoetanol lentamercaptoetanol lenta/2 mercaptoetanol lenta 2 2ME, 2ME ME (2-ME) FPA (FPA FC, FC (FC) SLPS S LPS (S-LPS abortus BP BP2 rBP26LFIA rBPLFIA rBP26 LFIA rBP (rBP26-LFIA 2ME. ME. 2-ME 7391 73 91 73,91 9714 97 14 97,14% respectivamente Neospora conclusão rBP26LFIA, LFIA, padronização Contudo (RBT 2mercaptoetanol lenta2 lenta/ (2-ME (FC rBP2 739 7 9 73,9 971 1 97,14 73, 97,1 97,

ABSTRACT Brucellosis is an important bacterial disease of global distribution with zoonotic potencial. Serological tests used in Brazil for diagnosis of bovine brucellosis, including the Rose Bengal test (RBT), 2-mercaptoethanol (2-ME), fluorescent polarization (FPA), and complement fixation (FC), are based on the smooth lipopolysaccharide antigen (S-LPS) of Brucella abortus. The aim of this study was to evaluate a recombinant BP26 protein used as antigen in a rapid lateral flow immunochromatographic assay (rBP26-LFIA) for serological diagnosis of bovine brucellosis. Analytical performance of rBP26-LFIA was evaluated in positive and negative bovine serum samples previously characterized by RBT and 2-ME. Estimates of analytical sensitivity and specificity were 73.91% and 97.14%, respectively. Bovine sera reactive to Neospora, Trypanosoma vivax or Leptospira were used to assess specificity because these diseases are commonly associated with abortion in cattle. In addition to a possible cross-reaction induced by commercial vaccines against Leptospira in serological tests for bovine brucellosis using S-LPS as an antigen. In conclusion, rBP26-LFIA, with its current standardization, had good analytical performance. However, a future evaluation of diagnostic performance by rBP26-LFIA with samples from regions with known prevalence is necessary for its recommendation for use in the Brazilian program for the control and eradication of bovine brucellosis. potencial RBT, , (RBT) 2mercaptoethanol mercaptoethanol 2 2ME, 2ME ME (2-ME) FPA, FPA (FPA) FC, FC (FC) SLPS S LPS (S-LPS abortus BP BP2 rBP26LFIA rBPLFIA rBP26 LFIA rBP (rBP26-LFIA 2ME. ME. 2-ME 7391 73 91 73.91 9714 97 14 97.14% respectively Neospora cattle crossreaction cross reaction conclusion rBP26LFIA, LFIA, standardization However (RBT (2-ME (FPA (FC rBP2 739 7 9 73.9 971 1 97.14 73. 97.1 97.

RESUMO Este estudo apresenta uma descrição macroscópica do plexo braquial na preguiça-real, Choloepus didactylus, e na preguiça-comum, Bradypus variegatus, a fim de identificar os pontos de bloqueio anestésico para realização de procedimentos cirúrgicos. Seis C. didactylus e nove B. variegatus tiveram seus membros torácicos dissecados, expondo-se o plexo braquial, e posteriormente localizados os pontos de bloqueio. Em geral, a formação do plexo braquial em C. didactylus envolveu nervos C5-T1, enquanto o plexo braquial em alguns indivíduos da espécie envolveu C4-T2. Em B. variegatus, o plexo braquial foi constituído por C6-T2. Em ambas as espécies, as ramificações deram origem a três troncos que se uniram para formar um tronco comum do plexo braquial. Na técnica de bloqueio do plexo braquial, foram realizados quatro acessos: supraclavicular, infraclavicular, axilar e distal. Os resultados deste estudo contribuem para procedimentos cirúrgicos e anestésicos e pesquisas sobre anatomia comparativa entre animais silvestres. preguiçareal, preguiçareal preguiça real, real preguiça-real preguiçacomum, preguiçacomum comum, preguiça-comum C B dissecados expondose expondo geral C5T1, C5T1 CT C5 T1, T1 T C5-T1 C4T2. C4T2 C4 T2. T2 C4-T2 C6T2. C6T2 C6 C6-T2 espécies acessos supraclavicular infraclavicular distal silvestres C5T C5-T C4T C4-T C6T C6-T

ABSTRACT This study presents a macroscopic description of the brachial plexus in the royal sloth, Choloepus didactylus, and the common sloth, Bradypus variegatus, to identify the points for anesthetic block to perform surgical procedures. Six C. didactylus and nine B. variegatus had their forelimbs dissected, exposing the brachial plexus, and the points for blockage were located subsequently. In general, the formation of the brachial plexus in C. didactylus involved C5-T1 nerves, whereas the brachial plexus in a few individuals of the species involved C4-T2. In B. variegatus, the brachial plexus was contributed by C6-T2. In both species, the ramifications gave rise to three trunks that joined together to form a common brachial plexus trunk. In the brachial plexus block technique, four accesses were performed: supraclavicular, infraclavicular, axillary, and distal. The results of this study contribute to surgical and anesthetic procedures and research on comparative anatomy among wild animals. sloth C B dissected subsequently general C5T1 CT C5 T1 T C5-T nerves C4T2. C4T2 C4 T2. T2 C4-T2 C6T2. C6T2 C6 C6-T2 trunk technique performed supraclavicular infraclavicular axillary distal animals C5T C4T C4-T C6T C6-T

Abstract Pterodon pubescens (Benth.) Benth., Fabaceae, fruits have been investigated for their anti-inflammatory and antinociceptive activities, and have demonstrated effectiveness in inflammatory conditions. A physiochemical and microbiological stability study was conducted to investigate two nanoemulsion-based delivery systems of two different hydrophilic surfactants (polyethylene glycol-40H castor oil or polyethylene glycol-40 castor oil). The nanoemulsions, containing P. pubescens oil, lecithin, hydrophilic surfactant and water, were analyzed for droplet size distribution, polydispersity index, pH, consistency index, stability against centrifugal force, and active content/vouacapan derivatives. The physicochemical characteristics were followed for 365 days. The nanoemulsion system was evaluated for anti-inflammatory activity by using with a peritonitis model, immediately after preparation and after 365 days of storage at 25 °C. The stability study demonstrated that proper storage (25 °C) preserved the characteristics of the nanoemulsion containing 7.5% polyethylene glycol-40H castor oil, 5% lecithin, and 5% P. pubescens oil. Further, it ensured a shelf life of 365 days as a phytotherapeutic formulation. In the peritonitis assay induced by carrageenan, nanoemulsion prepared with polyethylene glycol-40H castor oil (125 mg/kg) reduced leukocyte migration, even after 365 days of storage (25 °C), highlighting its potential for the treatment of inflammatory diseases. However, further studies are needed to confirm its clinical effectiveness.

RESUMO Hyptis pectinata, conhecida popularmente como Sambacaitá é uma erva medicinal nativa utilizada pela população local para o tratamento de enfermidades, cujas propriedades farmacológicas, antimicrobianas e medicinais foram relatadas em diversos estudos. O óleo essencial das folhas é rico em compostos terpênicos responsáveis pelas suas propriedades. Devido à existência de fatores que podem influenciar a composição química dos óleos essenciais, o objetivo deste trabalho foi avaliar a influência do armazenamento do óleo essencial Hyptis pectinata durante 360 dias em duas temperaturas diferentes. O óleo essencial foi destilado a partir de folhas secas de plantas de sete meses de idade, utilizando o método de hidrodestilação e a composição química foi determinada usando CG-EM e CG-DIC. Os efeitos do armazenamento no óleo essencial Hyptis pectinata foram testados ao longo de um ano (0, 15, 30, 60, 90, 120, 150, 180, 240, 300 e 360 dias), utilizando duas temperaturas: ambiente (±32ºC) e freezer (-20ºC). Os compostos predominantes no óleo essencial foram o β-cariofileno, óxido de cariofileno e germancreno D e houve variação na concentração dos constituintes químicos do óleo essencial, dependendo da temperatura de armazenamento, bem como ao longo dos 360 dias. O armazenamento do óleo essencial, à temperatura ambiente ao longo de um ano resultou em maiores concentrações de β-elemeno, α-copaeno, germacreno D, óxido de cariofileno e (E, E) -α-farneseno e menores teores de α-humuleno e β-cariofileno, quando comparado com os resultados obtidos a partir de armazenamento do óleo essencial no freezer. Estes resultados indicam que o óleo essencial deve ser estocado no freezer, o que propicia maior estabilidade na concentração dos constituintes químicos.

ABSTRACT Hyptis pectinata, popularly known as “sambacaitá”, is a native medicinal herb used by local people for treating disease. Its pharmacological, antimicrobial, and medicinal properties have been reported in several studies. The essential oil of the leaves is rich in terpene compounds responsible for its properties. Because there are factors that can influence the chemical composition of essential oils, the objective of this study was to evaluate the influence of storage on Hyptis pectinata essential oil for 360 days at two different temperatures. The essential oil was distilled from dried leaves of seven-month-old plants using the hydrodistillation method, and the chemical composition was determined using GC-MS and GC/FID. The effects of storage on the essential oil Hyptis pectinata were tested over the course of a year (0, 15, 30, 60, 90, 120, 150, 180, 240, 300, and 360 days), using two temperatures: room (±32ºC) and freezer (-20ºC). The predominant compounds in the essential oil are β–caryophyllene, caryophyllene oxide, and germancrene D. The concentration of the chemical constituents of the essential oil varied depending on the storage temperature, and over the 360 days. The storage of the essential oil at room temperature over one year resulted in higher concentrations of β-elemene, α-copaene, germacrene D, caryophyllene oxide, and (E,E)-α-farnesene and lower content of α-humulene and β-caryophyllene, compared to the results obtained from storing the essential oil in a freezer. These results indicate that the essential oil should be stored in a freezer, which provides greater stability to the concentration of the chemical constituents.

The presence of the single nucleotide polymorphisms in exon 1 of the mannose-binding lectin 2 (MBL2) gene was evaluated in a sample of 159 patients undergoing coronary artery bypass surgery (71 patients undergoing valve replacement surgery and 300 control subjects) to investigate a possible association between polymorphisms and heart disease with Chlamydia infection. The identification of the alleles B and D was performed using real time polymerase chain reaction (PCR) and of the allele C was accomplished through PCR assays followed by digestion with the restriction enzyme. The comparative analysis of allelic and genotypic frequencies between the three groups did not reveal any significant difference, even when related to previous Chlamydia infection. Variations in the MBL plasma levels were influenced by the presence of polymorphisms, being significantly higher in the group of cardiac patients, but without representing a risk for the disease. The results showed that despite MBL2 gene polymorphisms being associated with the protein plasma levels, the polymorphisms were not enough to predict the development of heart disease, regardless of infection with both species of Chlamydia.

Este estudo descreve o efeito da esfingosina 1-fosfato (S1P) no desenvolvimento de folículos pré-antrais, portanto da ativação e viabilidade de folículos caprinos cultivados in vitro. Fragmentos de ovários foram cultivados por um ou sete dias em meio essencial mínimo com diferentes concentrações de S1P (0, 1, 10, 50, 100 ou 200ng/mL). Os fragmentos de ovário foram processados para análise histológica em microscopia óptica, microscopia eletrônica e microscopia de fluorescência. O tratamento usando 1ng/mL de S1P foi capaz de manter a porcentagem de folículos normais durante o período de cultivo de sete dias. Ao final do período de cultivo, houve uma redução significativa (p<0,05) na porcentagem de folículos primordiais em todos os grupos tratados com S1P, comparados com os grupos controle (FC e CC), seguida por um aumento do número de folículos ativados (intermediários, primários e secundários). Adicionalmente, na cultura por sete dias com meio suplementado com S1P (1ng/mL), houve preservação da ultraestrutura das organelas e manteve-se a viabilidade dos folículos pré-antrais avaliados por microscopia de fluorescência. Em conclusão, após sete dias de cultura, o meio suplementado com 1ng/mL de S1P ativa o desenvolvimento de folículos pré-antrais de caprino, cultivados in situ e mantém as viabilidades oocitária e folicular.

This study describes the effect of sphingosine 1-phosphate (S1P) for development of preantral follicle, therefore the activation and follicular viability of caprine follicles cultured in vitro. Ovarian fragments were cultured for 1 or 7 days in Minimum Essential Medium with different S1P concentrations (0, 1, 10, 50, 100 or 200ng/mL). All ovarian fragments were processed for histological analysis in optical microscopy, transmission electron microscopy and fluorescence analysis. The treatment using 1ng/mL of S1P was able to maintain the percentage of normal follicles with the progression of the culture from day 1 to 7. At end of the 7-day culture period there was a significant reduction (P<0.05) in the percentage of primordial follicles in all groups treated with S1P, compared with fresh control (FC) and Control Culture (CC), which was followed by an increase of activated follicles (intermediary, primary and secondary). In addition, the culture for 7 days with media supplemented with S1P with 1ng/mL preserved the ultrastructure of organelles and kept the preantral follicular viability when evaluated by fluorescence microscopy. In conclusion, after 7 days of culture, the 1ng/mL of S1P activates the development of preantral caprine follicles, cultured in situ and maintains the oocitary and follicular viability.

O objetivo desse estudo foi investigar se o TGF-β afeta a sobrevivência, ativação e crescimento de folículos primordiais caprinos inclusos no córtex ovariano após o cultivo in vitro. Ovários de cabras foram coletados em abatedouro e fragmentos de tecido ovariano foram cultivados por um e sete dias em meio essencial mínimo alfa (α-MEM+) sozinho ou suplementado com TGF-β (1, 5, 10 ou 50ng/mL). Fragmentos ovarianos não cultivados e cultivados foram processados para análise histológica e ultraestrutural. Os resultados mostraram que, comparado ao controle fresco, houve diminuição no percentual de folículos morfologicamente normais em todos os tratamentos somente após sete dias de cultivo. O TGF-β não afetou a ativação folicular independente da concentração testada, contudo, o diâmetro folicular foi superior (P<0.05) no tratamento com 10ng/mL de TGF-β quando comparado ao controle fresco e aos demais tratamentos. Além disso, essa mesma concentração manteve a ultraestrutura normal dos folículos após sete dias de cultivo. Em conclusão, o TGF-β apresentou efeito adicional no crescimento folicular e na manutenção da integridade ultraestrutural de folículos pré-antrais caprinos inclusos no tecido ovariano quando utilizado na concentração de 10ng/mL durante sete dias de cultivo.

The objectives of this study were to investigate whether TGF-β affect the survival, activation and further growth of goat primordial follicles enclosed in ovarian cortex after in vitro culture. Goat ovaries were collected from an abattoir and pieces of ovarian tissues were cultured for one or seven days in a supplemented alpha Minimum Essential Medium, alone or containing TGF-β (1, 5, 10 or 50ng/mL). Ovarian tissues from the fresh control as well as those cultured were processed for histological and ultrastructural studies. The results showed that when compared with fresh control, there was decrease in the percentages of histologically normal follicles in all treatments only after seven days culture. TGF-β did not affect the activation of preantral follicles regardless of its concentration, however, larger follicles diameter (P<0.05) was observed using 10ng/mL TGF-β than in the fresh control and other treatments. Moreover, this concentration maintained the normal ultrastructure after seven days of culture. In conclusion, TGF-β showed additional effect on the follicle growth and the maintenance of ultrastructural integrity of goat preantral follicles enclosed in ovarian tissue when used at 10ng/mL during seven days of culture.

The objective of this study was to determine the effects of GDF-9, IGF-I, and GH alone or combined on preantral follicle survival, activation and development after 1 and 7 days of in vitro culture. Either fresh (non-cultured) or cultured ovarian tissue was processed for histological and fluorescence analysis. For all media tested, the percent of normal follicles was greater when compared to minimum essential medium supplemented (MEM+) alone, except when ovarian tissue was cultured with GDF-9/IGF-I or GDF-9/GH (P < 0.05). Fluorescence analysis showed that the percent of viable follicles after 7 days of culture was similar for non-cultured tissue and for all treatments tested. The percent of primordial follicles was reduced (P < 0.05) and there was a significant and concomitant increase in the percent of intermediate and primary follicles in all treatments tested after 7 days of culture when compared to non-cultured tissue. After 7 days of culture, the highest percent of intermediate follicles was observed with IGF-I/GH (61.3%), and the highest percent of primary follicles was achieved with IGF-I (57.7%). After 7 days of culture in MEM+ containing GDF-9, IGF-I and GH alone or in all associations, a significant increase in follicular diameter was observed when compared to MEM+ alone and non-cultured tissue. In conclusion, GDF-9, IGF-I and GH alone or in combination maintain preantral follicle survival and promote primordial follicle activation. Nevertheless, the data showed that IGF-I/GH and IGF-I alone are efficient in promoting the transition from primordial to intermediate follicles and from intermediate to primary follicles, respectively.

Two different methods to detect Fasciola hepatica in faeces using kappa index were compared with four tamises technics and sequencial fitration in two tamises using Flukefinder® apparatus. High accordance between the two methods was observed. The values showed high sensibility and specificity.

The influence of apolipoprotein E alleles and genotypes on plasma lipid levels was determined in 185 individuals of mixed ethnicity living in Ouro Preto, Brazil. DNA was obtained from blood samples and the genotypes were determined by an RFLP-PCR procedure. The *3 allele was the most frequent (72%), followed by *4 (20%) and *2 (8%); *4 frequency was higher and *2 frequency was lower in the dyslipidemic group than in the normal control group. The *2 carriers presented lower LDL and total cholesterol levels compared to the *3 and *4 carriers. All six expected genotypes were observed in the individuals genotyped: E2/2 (2.1%), E4/4 (2.7%), E2/4 (3.7%), E2/3 (8.0%), E3/3 (53.3%), E3/4 (29.9%); no difference in genotype frequencies was found between the normal and dyslipidemic groups. Compared with *2, the presence of *3 increases more than two times the risk for dyslipidemia (OR = 2.31; P = 0.025; 95% CI = 1.06-5.06) and the presence of *4 increases it three times (OR = 3.31; P = 0.006; 95% CI = 1.36-8.04). The only significant effect of genotype was an increased risk for dyslipidemia in the *4 genotype carriers (E3/4 + E4/4) compared with the *2 genotype carriers (E2/2 + E2/3) with OR = 3.69 (95% CI = 1.25-10.88). The present study indicates that in the Ouro Preto admixed population the presence of APOE *2 can confer a protective effect, whereas the presence of APOE *4 implies an enhanced risk for dyslipidemia.

Bezerros traçadores foram utilizados para avaliar a contaminação sazonal das pastagens por helmintos gastrintestinais e pulmonares em uma propriedade de exploração leiteira na região Campo das Vertentes, Minas Gerais. Os animais se infectaram durante todos os meses do ano. As maiores cargas parasitárias foram recuperadas no período chuvoso (setembro - abril), e o pico foi observado em abril-maio, final do período chuvoso. As espécies recuperadas foram Cooperia punctata, C. spatulata, Haemonchus contortus, H. similis, Oesophagostomum radiatum, Trichostrongylus axei, T. colubriformis, Trichuris discolor, T. globulosa, Dictyocaulus viviparus e Agriostomum vryburgi. Para avaliar a dinâmica das infecções, foi acompanhado um grupo de vacas com seus bezerros lactantes no período de janeiro de 1999 a fevereiro de 2000. Os valores da contagem de ovos por grama de fezes (OPG) dos bezerros apresentaram dois picos, em maio de 1999 e fevereiro de 2000, enquanto o pico da contagem de OPG das vacas ocorreu em julho e agosto de 2000. Nas coproculturas, os gêneros de maior ocorrência foram Cooperia e Haemonchus nos bezerros, e Haemonchus e Trichostrongylus nas vacas.

Tracer calves used to monitor contamination of pasture with pulmonary and gastrointestinal nematode helminths on a dairy farm were infected throughout the year. The highest parasite burdens were observed during the rainy season (September - April). However, the highest peak occurred in April and May 1999, the end of the rainy season. Helminth species recovered from the calves were Cooperia punctata, C. spatulata, Haemonchus contortus, H. similis, Oesophagostomum radiatum, Trichostrongylus axei, T. colubriformis, Trichuris discolor, T. globulosa, Dictyocaulus viviparus and Agriostomum vryburgi. A group of lactating cows and their accompanying calves were followed during the period from January 1999 to February 2000 to evaluate the dynamics of their infection. Parasite eggs per gram of feces (EPG) exhibited two peaks in the calves, May 1999 and February 2000; while EPG peaks in cows occurred in July and August 2000. Cooperia and Haemonchus were the most prevalent genera in fecal cultures from calves, whereas in cow fecal cultures, the highest prevalence was for Haemonchus and Trichostrongylus.