ABSTRACT Researchers from different areas of knowledge have used the Cox proportional-hazards model, due to its simplicity and easy interpretation when studying situations in which the response variable is the time until the occurrence of an event of interest. However, the traditional Cox proportional-hazards model is not suitable for modeling data sets that violate the assumption of proportionality of the risks (or failure rates) and the effects of covariates over time are not detected. The generalized time-dependent logistic (GTDL) model has been used as an alternative in the modeling of survival data, taking into account the assumption of non-proportionality of the risks. In the literature, we found a wide and relevant production in inferential procedures, but no contribution in diagnostic methods or techniques. In this paper, Cox-Snell, modified Cox-Snell, martingale, deviance, randomized quantiles, NMSP (normally-transformed modified survival probabilities) and NRSP (normally-transformed randomized survival probabilities) residuals are proposed to assess the suitability of the GTDL model to the data. A Monte Carlo simulation study is conducted in order to investigate the empirical distribution of these residuals. In summary, the obtained simulation results indicate the adequacy, for the GTDL model, of the randomized quantile and NRSP residuals, regardless of the proportion of censorship in the data. The GTDL library is built and made available in the R programming language. Finally, the methodology studied is applied to a set of real data, available in the literature, involving patients diagnosed with advanced-stage lung cancer. Codes for installing and using the GTDL library are shown in the Supplementary Material (https://github.com/carrascojalmar/GTDL-Material-Suplementar).

RESUMO Pesquisadores de diferentes áreas do conhecimento têm utilizado o modelo de riscos proporcionais de Cox devido à sua simplicidade e fácil interpretação ao estudar situações em que a variável resposta é o tempo até a ocorrência de um evento de interesse. No entanto, o modelo tradicional de Cox não é adequado para descrever conjuntos de dados que violam a suposição de proporcionalidade dos riscos (ou taxas de falha) e os efeitos das covariáveis ao longo do tempo não são detectados. O modelo logístico generalizado dependente do tempo (GTDL, do inglês generalized time-dependent logistic) tem sido utilizado como alternativa na modelagem de dados de sobrevivência, levando em consideração a suposição de não proporcionalidade dos riscos. Na literatura é encontrada uma ampla e relevante produção em procedimentos inferenciais, mas nenhuma contribuição em métodos ou técnicas de diagnóstico. Neste artigo, os resíduos de Cox-Snell, modificados de Cox-Snell, martingale, deviance, quantílicos aleatorizados, NMSP (do inglês normally-transformed modified survival probabilities) e NRSP (do inglês normally-transformed randomized survival probabilities) são propostos para avaliar a adequabilidade do modelo GTDL aos dados. Um estudo de simulação via Monte Carlo é conduzido com o proposito de investigar a distribuição empírica desses resíduos. Em suma, os resultados de simulação obtidos indicam a adequação, para o modelo GTDL, dos resíduos quantílicos aleatorizados e NRSP, independentemente da proporção de censura nos dados. A biblioteca GTDL é construída e disponibilizada na linguagem de programação R. Finalmente, a metodologia estudada é aplicada a um conjunto de dados reais, disponível na literatura, envolvendo pacientes diagnosticados com câncer de pulmão em estágio avançado. Os códigos de instalação e uso da biblioteca GTDL são exibidos no Material Suplementar (https://github.com/carrascojalmar/GTDL-Material-Suplementar).

RESUMO Avaliaram-se os níveis de farinha de vagem de Samanea saman (0, 10, 15, 20 e 25%) em substituição ao milho na matéria seca da dieta sobre o consumo de MS, PB e EM, a excreção urinária de creatinina e de derivados de purina totais e a síntese de proteína microbiana em cordeiros. Foram utilizados 25 cordeiros Bergamácia, não castrados, peso corporal inicial de 24±5kg e idade média de 120 dias. O delineamento experimental foi inteiramente ao acaso, com cinco tratamentos e cinco repetições. O experimento durou 84 dias e as coletas de urina spot e de 24h foram realizadas no último dia do experimento. O consumo não foi afetado (P>0,05), o volume de urina e a excreção diária de creatinina foram influenciados (P<0,05) por S. Saman substituindo o milho, observando-se efeito linear crescente e variação cúbica (P<0,05), respectivamente. Os componentes cúbicos e quadráticos foram significativos (P<0,05) para a excreção de derivados de purina totais, purinas absorvidas e síntese microbiana. A variação cúbica, com picos em 5% e 18% de substituição para a excreção urinária de creatinina e de derivados de purina, indica que os níveis utilizados de S. saman afetaram a atividade renal de cordeiros. Substituição do milho por farelo de vagem de S. saman até 10% aumenta a eficiência de síntese de proteína bruta microbiana.

ABSTRACT Evaluated the levels of rain tree (Samanea saman) pod meal (RTPM) (0, 10, 15, 20 and 25%) replacing maize in the dry matter of the diet on intake of DM, CP and ME, creatinine and total purine derivatives excretion in urine and microbial protein synthesis in lambs. Twenty-five uncastrated Bergamasca lambs were used, with an initial body weight of 24±5kg and an average age of 120 days. The experimental design was completely randomized, with five treatments and five replications. The trial lasted 84 days and the 24h and spot urine collections were performed in the last day of the experiment. The intake was not affected (P>0.05), the urine volume and daily creatinine excretion were influenced (P<0.05) by RTPM replacing maize, observing increasing linear effect and cubic variation (P<0.05), respectively. The cubic and quadratic components were significant (P<0.05) for the excretion of total purine derivatives, absorbed purines, and microbial synthesis. The cubic variation, with peaks at 5% and 18% replacement for urinary excretion of creatinine and purine derivatives, indicates that the levels used of RTPM affected the renal activity of lambs. Substitution of maize by RTPM up to 10% enhances efficiency of rumen microbial crude protein synthesis.

This work investigates the synergistic use of Theobroma cacao (Cocoa), and Elaeis guineensis (Palm oil) as corrosion inhibitors for carbon steel in 0.5 mol.L-1 hydrochloric acid solution, in the absence and presence of different mixture concentrations of the studied compounds. The efficiency of corrosion inhibitors was studied by gravimetric tests and the corrosion resistance by electrochemical tests, such as electrochemical impedance spectroscopy (EIS) and polarization curves. The chemical composition of the cocoa bark powder and palm kernel cake powder was evaluated by Fourier transform infrared spectroscopy (FTIR). The results of the gravimetric tests showed an inhibition efficiency of 79% for the cocoa bark powder and 76% for palm kernel cake powder, however, the mixture of the compounds presented an inhibition superior to 94%, showing the great synergism between the studied species. These results corroborate the values obtained from EIS.

Abstract Oil pollution has significantly contributed to coral reef decline in the last five decades and a major oil spill reached Brazilian tropical coast in August 2019. Here, we report the first evidence of direct crude oil contact from that spill on reef coral species, and evaluate the effects of this disaster on coral vitality and benthic assemblage structure on the largest coastal marine protected area (MPA Costa dos Corais) in Brazil. We compared benthic cover in reefs with and without oil and monitored Siderastrea stellata colonies 90 days after oil contact. Oil stains between 0.5 and 150 cm were found in two of the 17 reef sites investigated. Multivariate analyses did not detect significant differences between oiled and non-oiled reefs and there was no evidence of S. stellata health deterioration. These results indicate minimal acute effects on coral vitality and intertidal reef benthic assemblage structure. Future studies should investigate oil effects on specific aspects of coral biology as growth, reproduction, bleaching susceptibility and metagenomics which can deteriorate over longer time frames, and we recommend long-term coral reef monitoring to support a robust assessment and mitigation of chronic oil impacts.

Constitutional genomic imbalances are known to cause malformations, disabilities, neurodevelopmental delay, and dysmorphia and can lead to dysfunctions in the cell cycle. In extremely rare genetic conditions such as small supernumerary marker chromosomes (sSMC), it is important to understand the cellular consequences of this extra marker, as well the factors that contribute to their maintenance or elimination through successive cell cycles and phenotypic impact. The study of chromosomal mosaicism provides a natural model to characterize the effect of aneuploidy on genome stability and compare cells with the same genetic background and environment exposure, but differing in the presence of sSMC. Here, we report the functional characterization of different cell lines from two familial patients with mosaic sSMC derived from chromosome 12. We performed studies of proliferation dynamics, stability, and variability of these cells using fluorescent in situ hybridization (FISH), sister chromatid exchanges (SCE), and conventional staining. We also quantified the telomere-related genomic instability of sSMC cells using 3D telomeric profile analysis by quantitative-FISH. sSMC cells exhibited differences in the cell cycle dynamics compared to normal cells. First, the sSMC cells exhibited lower proliferation index and higher frequency of SCE than normal cells, associated with a higher level of chromosomal instability. Second, sSMC cells exhibited more telomeric-related genomic instability. Lastly, the differences of sSMC cells distribution among tissues could explain different phenotypic repercussions observed in patients. These results will help in our understanding of the sSMC stability, maintenance during cell cycle, and the cell cycle variables involved in the different phenotypic manifestations.

ABSTRACT Phytogenic additives are organic molecules that also improve ruminal fermentation, turning the supplemented individuals into more productive animals, without damage the population welfare. The objective of this study was to evaluate the effects of mesquite (Prosopis juliflora [Sw.] DC.) extract as phytogenic additive for sheep finished on pasture in the Brazilian Northeast semiarid region. Twenty-four intact lambs (Santa Ines × Dorper F1 crossbred) were used, with an initial body weight of 23.00 ± 1.83 kg. The experiment was carried out in a complete randomized design with four treatments and six replicates. Treatments consisted of four diets: Pasture and no supplementation; grazing pasture and phytogenic additive; pasture, supplementation with Tifton 85 hay, and concentrate; and pasture, phytogenic additive, and supplementation with Tifton 85 hay and concentrate. Prior to supplementation, animals received the phytogenic additive according to treatment. There were nonsignificant differences for nutrient intake and behavior patterns (P > 0.05). However, additive intake derived from mesquite provided an increase (P < 0.05) in digestibility (14.40% total digestible nutrients), N balance (27.12% retained N:ingested N) and performance (8.82% final body weight, 21.81% total weight gain, and 30.81% average daily gain) compared to animals consuming only pasture in rainy period. Thus, the use of mesquite extract as phytogenic additive is recommended for sheep finished on pasture in the Brazilian semiarid region.

RESUMO Mycoplasma bovis é um agente altamente contagioso relacionado a várias patologias em bovinos. A detecção de anticorpos reativos a M. bovis por Ensaio de Imunoadsorção Enzimática Indireto (iELISA) identifica se houve exposição ao microrganismo. O presente estudo teve como objetivo otimizar um iELISA de antígeno celular total de M. bovis, aplicando-o a amostras de soro bovino, bem como avaliar fatores de risco. Amostras de soro foram obtidas de 400 vacas de 17 rebanhos da Região Sudeste do Brasil. Na otimização do iELISA foram obtidos: 2μg/mL de antígeno, diluição dos soros 1:300 e do conjugado 1:15000. A frequência de animais reativos foi de 62,3% (249/400) e de 100% (17/17) para os rebanhos. Os fatores de risco foram: rebanhos com mais de 100 animais (OR= 3,1; IC= 95%); raça Holandesa (OR= 72,5;IC= 95%); vacas (OR= 29,7;IC= 95%); sistema intensivo (OR= 3,3; C= 95%); produção de pequenos ruminantes (OR= 4,4;IC=95%); produção de leite acima de 500L (OR= 2,9;IC= 95%); sem quarentena (OR= 1,5;IC= 95%); ordenha mecânica (OR= 5,5;IC= 95%) e casos de mastite (OR= 5,5;IC= 95%). O iELISA proposto foi capaz de detectar anticorpos reativos a M. bovis no soro bovino. O conhecimento desses fatores de risco pode auxiliar na implementação de medidas profiláticas.

ABSTRACT Mycoplasma bovis is a highly contagious agent associated with several pathologies in cattle. The detection of reactive antibodies to M. bovis by Indirect Enzyme-Linked Immunosorbent Assay (iELISA) identifies if there was an exposure to the microorganism. The current study aimed to optimize an iELISA from M. bovis total cell antigen, applying it to bovine serum samples, and to evaluate risk factors. Serum samples were obtained from 400 cows from 17 herds from Southeast Brazil. In the optimization of iELISA, the following was established: 2 μg/mL of antigen, sera dilution 1:300, and conjugate dilution 1:15000. The frequency was 62.3% (249/400) of reactive animals and 100% (17/17) of reactive herds. Risk factors were: herds with more than 100 animals (OR= 3.1; CI= 95%); Holstein breed (OR= 72.5; CI= 95%); cows (OR= 29.7; CI= 95%); intensive breeding system (OR= 3.3; CI= 95%); associated small ruminant production (OR= 4.4; CI= 95%); milk production above 500L (OR= 2.9; CI= 95%); no quarantine (OR= 1.5; CI= 95%); mechanical milking (OR= 5.5; CI= 95%) and cases of mastitis (OR= 5.5; CI= 95%). The proposed iELISA was able to detect antibodies reactive to M. bovis in bovine serum. Knowledge of these risk factors can assist in the implementation of prophylactic measures.

RESUMO A emergência de cepas de Staphylococcus aureus resistentes à meticilina associadas à pecuária (LA-MRSA) e o papel potencial dos suínos na evolução dessas cepas têm levado ao aumento do interesse na pesquisa desses microrganismos. No entanto, isso tem contribuído para a falta de estudos sobre o isolamento e a caracterização de cepas de S. aureus sensíveis à meticilina (MSSA). Neste estudo, foi analisada a prevalência de S. aureus em suínos nas fases de creche e terminação. Os perfis de suscetibilidade aos antibióticos, a tolerância a metais pesados e a produção de biofilme dos isolados foram avaliados por meio de técnicas fenotípicas e genotípicas. Um total de 1.250 colônias sugestivas de Staphylococcus spp. foi isolado de 128 suínos, das quais 63,6% (n = 795) pertenciam a esse gênero microbiano. Sessenta e sete colônias isoladas de 34 animais (26,5%) foram confirmadas como S. aureus (8,4%). Nenhuma cepa resistente ao cobre, ao zinco ou à meticilina foi detectada; entretanto, todas as cepas apresentaram perfil de resistência a pelo menos três classes diferentes de antimicrobianos e 21 produziam biofilme. Esses dados são preocupantes, pois indicam a necessidade de maior vigilância no uso de antimicrobianos, bem como reforçam a importância de estudos com cepas de MSSA.

ABSTRACT The emergence of livestock-associated methicillin-resistant Staphylococcus aureus strains (LA-MRSA) and the potential role of pigs in the evolution of these strains has led to increased interest in research of these microorganisms. However, this has contributed to a lack of research in the isolation and characterization of methicillin-susceptible S. aureus strains (MSSA). In this study, the prevalence of S. aureus in pigs in the nursery and finishing stages were analyzed. The susceptibility profiles to antibiotics, tolerance to heavy metals, and biofilm production of the isolates were evaluated using phenotypic and genotypic techniques. A total of 1,250 colonies suggestive of Staphylococcus spp. were isolated from 128 pigs, of which 63.6% (n = 795) belonged to this microbial genus. Sixty-seven colonies isolated from 34 animals (26.5%) were confirmed as S. aureus (8.4%). No strains resistant to copper, zinc, or methicillin were detected; however, all strains presented a resistance profile to at least three different classes of antimicrobials and 21 produced biofilms. These data are of concern, as they indicate the need for increased surveillance in the use of antimicrobials as well as reinforce the importance of studies on MSSA strains.

Abstract This study inventoried and characterized the richness of galling insects based on gall morphotypes and their host plants in two Cerrado sensu stricto areas of Caetité municipality in Bahia State, Brazil, to aid the identification of galling insects and their host plants, as well as to contribute to the knowledge and conservation of local biodiversity. The survey was conducted in the Moita dos Porcos archaeological site and João Barroca Farm site, adopting the random walking methodology for sampling, during 12 months. We recorded 98 gall morphotypes on 42 plant species belonging to 36 genera and 22 families. Leguminosae, Malpighiaceae and Myrtaceae demonstrated the greatest richness of galls, and the Copaifera langsdorffii was the super-host species, with 16 gall morphotypes. Most galls occur on leaves, and may be globoid, glabrous, grouped, and are usually unilocular, and brown. The galling insects identified belonged to Coleoptera, Diptera, Lepidoptera, and Thysanoptera. Eight plant host species and eight gall morphotypes were recorded for the first time in Cerrado areas in Brazil. The areas surveyed demonstrated high richness of gall morphotypes and host plants, evidencing the importance of studying and preserving different areas of the same biome.

RESUMO A cotonicultura tem forte fator de impacto na economia nacional, e o estado de Mato Grosso se destaca por ser o maior produtor de algodão herbáceo e deter o maior rebanho bovino do país, condições essas que estimulam o uso do caroço, da torta e do farelo de algodão na alimentação animal. Considerando que o gossipol está presente nos subprodutos do algodão e que seus efeitos sobre a reprodução podem reduzir a fertilidade dos animais, objetivou-se, com essa pesquisa, avaliar o efeito da ingestão de dietas com diferentes teores de gossipol livre por dia sobre a morfometria testicular e a qualidade seminal de touros da raça Nelore. Foram utilizados 28 touros, distribuídos aleatoriamente em seis tratamentos: T0, 0 grama de gossipol livre/touro/dia ; T1, 1,08 grama de gossipol livre/touro/dia; T2, 2,07 gramas de gossipol livre/touro/dia; T3, 3,24 gramas de gossipol livre/touro/dia; T4, 3,82 gramas de gossipol livre/touro/dia e T5, 5,08 gramas de gossipol livre/touro/dia. Os animais de cada tratamento foram mantidos confinados em área média de 100m2, dotada de bebedouro, cochos para mistura mineral e para volumoso/concentrado. O consumo de 3,24 gramas de gossipol livre/touro/dia alterou a qualidade espermática e a morfometria testicular de touros.

ABSTRACT The cotton industry has a strong impact factor in the Brazilian economy and the state of Mato Grosso stands out for being the largest upland-type cotton producer and also holds the largest cattle herd in the country, conditions that stimulate the use of cottonseed pie and cottonseed meal in animal feed. Whereas gossypol is present in cotton by-products and their effects on reproduction can reduce the animal fertility, this research evaluated the effect of diets with different free gossypol contents in the testicular morphometry and semen quality of Nelore bulls. 28 bulls were randomly distributed in six treatments: T0, receiving no free gossypol; T1, receiving 1.08g / bull / day; T2, receiving 2.07g / bull / day; T3, receiving 3.24g / bull / day; T4, receiving 3.82g / bull / day and T5, receiving 5.08g / bull / day of free gossypol respectively. The animals in each treatment were kept confined in an averaged area of 100m2, having fresh water, troughs for mineral mixture and roughage / concentrate. The consumption of 3.24g of free gossypol / bull / day altered the bulls sperm quality and testicular morphometry.

RESUMO Objetivou-se avaliar a seletividade aparente de ovinos em pastos de capim-marandu (Urochloa brizantha cv. Marandu) com quatro alturas no início do diferimento (15, 25, 35 e 45cm). O período de diferimento foi de 92 dias e iniciou em 21/03/2014. As avaliações ocorreram no início (primeira semana), meio (45° dia) e fim (92° dia) do período de pastejo, no inverno (21/06/2014 a 21/09/2014). Os pastos diferidos com 15 e 25cm apresentaram menor massa de forragem (MF), mas maior percentual de folha viva (FV) na MF do que os pastos diferidos com 35 e 45cm. O percentual de colmo vivo na MF e o índice de seletividade aparente (ISA) da FV foram superiores no pasto diferido com 45cm. O percentual de colmo morto (CM) na simulação de pastejo (SP) e o ISA desse componente morfológico foram menores no pasto diferido com 15cm, em comparação ao diferido com 45cm. A MF e os percentuais de FV na MF e na amostra de SP se reduziram, enquanto os percentuais de CM na MF e na amostra de SP aumentaram com o período de pastejo. O capim-marandu com 15cm no início do período de diferimento melhora a morfologia do pasto diferido. O pastejo seletivo é dificultado no decorrer do período de pastejo.

ABSTRACT The objective of this study was to evaluate the apparent selectivity of sheep in marandu palisadegrass (Urochloa brizantha cv. Marandu) pastures with four heights at the beginning of the deferment period (15, 25, 35 and 45cm). The deferment period was 92 days and started on 03/21/2014. Evaluations occurred in the beginning (first week), middle (45th day) and end (92nd day) of the grazing period, in winter (06/21/2014 to 09/21/2014). Deferred pastures with 15 and 25cm presented lower forage mass (FM), but higher live leaf (LL) percentage in FM than deferred pastures with 35 and 45cm. The live stem percentage in the FM and the apparent selectivity index (ASI) of the LL were superior in the deferred pasture with 45cm. The dead stem (DS) percentage in the grazing simulation (GS) and the ASI of this morphological component were lower in the pasture with 15cm, compared to the deferred pasture with 45cm. The FM and the LL percentages in FM and in the GS sample decreased, while the DS percentages in FM and in GS sample increased with the grazing period. Marandu palisadegrass with 15cm at beginning of the deferment period improves the morphology of the deferred pasture. Selective grazing is difficult during the grazing period.

ABSTRACT In this work, we developed and validated a HPLC-PDA method for the quantification of hibalactone in Hydrocotyle umbellata L., Araliaceae, subterraneous parts extracts and optimized its ultrasound-assisted extraction. Chromatographic separations were carried out with an isocratic mobile phase of acetonitrile/methanol/water (10:65:25), a flow of 0.8 ml min−1, detection at 290 nm and C18 column (250 × 4.6 mm, 5 µm). The method validation parameters were determined according to Brazilian legislation. The optimization of the hibalactone ultrasound-assisted extraction was performed using Box–Behnken design and response surface methodology. The HPLC method for hibalactone quantification proved to be selective, linear, precise, accurate and robust, being useful for the analysis of hibalactone in H. umbellata subterraneous parts extracts. The optimal ultrasound-assisted extraction conditions were obtained with solid-to-liquid ratio of 1:5 g ml−1, ethanolic strength of 70% (v/v) and temperature of 65 °C. The results can provide support of the quality control and standardization of raw materials from H. umbellata.

Abstract: Candida haemulonii complex (C. haemulonii, C. haemulonii var. vulnera and C. duobushaemulonii) consists of emergent multidrug-resistant pathogens that cause bloodstream and deep-seated infections. However, little is known about their virulence factors. Herein, we evaluated the presence of extracellular serine peptidases in this fungal complex. Serine peptidase activity was measured by spectrophotometry using chromogenic peptide substrates to the S1 family. Chymotrypsin-, trypsin- and elastase-like activities were detected in all fungal isolates. Since higher chymotrypsin- and trypsin-like activities were observed from the cleavage of N-succinyl-Ala-Ala-Pro-Phe-pNa and N-benzoyl-Phe-Val-Arg-pNa, respectively, these substrates were selected for further experiments. Overall, pHs 7.0 and 9.0 were those in which higher chymotrypsin- and trypsin-like activities were observed, respectively, displaying higher hydrolytic activities at 37-45°C. Additionally, the serine peptidases produced by C. haemulonii complex were inhibited by PMSF and AEBSF in a typically concentration-dependent manner. Although the Michaelis constant (Km) values obtained for chymotrypsin-like peptidases were similar, greater differences were observed for trypsin-like enzymes secreted by the different fungal isolates. This is the first time that peptidases belonging to the S1 family are described in the C. haemulonii species complex. Thus, these data open the doors for more detailed studies into potential roles of these peptidases in fungal virulence.

RESUMO A presente pesquisa foi realizada com o objetivo de avaliar os níveis de colesterol, extrato etéreo, perfil de ácidos graxos e o índice aterogênico e trombogênico da gordura intramuscular de suínos machos imunocastrados em relação às fêmeas e aos machos castrados cirurgicamente. Utilizaram-se 45 animais, com peso inicial de 25,2±2,8kg, distribuídos em delineamento inteiramente ao acaso, com três tratamentos (machos castrados, machos e fêmeas imunocastrados) e três repetições de cinco animais cada. Os animais foram abatidos a 90,3±2,7kg. As amostras de Longissimus dorsi foram coletadas, liofilizadas e analisadas quanto ao perfil de ácidos gordos, colesterol e extrato etéreo. Não foram observadas diferenças (P>0,05) para nenhuma das variáveis analisadas entre a gordura intramuscular de machos castrados cirurgicamente, machos imunocastrados e fêmeas. A imunocastração não alterou os níveis intramusculares de extrato etéreo, colesterol, composição de ácidos graxos ou o índice aterogênico e trombogênico do Longissimus dorsi.