Associations of occupational physical activity and active transport with depressive symptoms in a middle-income country.

OBJECTIVE: This study investigated the associations of occupational physical activity and active transport with depressive symptoms, stratified by income range, among Brazilian working adults. STUDY DESIGN: This cross-sectional study drew on data from the 2019 Brazilian National Health Survey. METHODS: We used the Patient Health Questionnaire (PHQ-9) to assess depressive symptoms in working adults aged 18-65 years, with physical activities (occupational physical activity and active transport) self-reported through a specially developed questionnaire. Crude and adjusted logistic regression models were fitted. RESULTS: Individuals at the highest level of occupational physical activity (odds ratio [OR] 1.35; 95% confidence interval [95% CI] 1.19-1.53) and at the moderate level of active transport to work (OR 1.66; 95% CI 1.24-2.22) returned increased odds of depressive symptoms as compared with those who were inactive in these domains. Stratified by income group, light active transport to work (OR 0.57; 95% CI 0.34-0.95) in the lower income group (Range 1) was associated with lower odds of depressive symptoms. On the other hand, individuals at the highest level of active transport to work returned higher odds of depressive symptoms in the low- and intermediate-income groups, Ranges 3 and 4 (OR 1.92; 95% CI 1.22-3.00 and OR 2.91; 95% CI 1.71-4.95, respectively). CONCLUSION: Our results suggest that occupational physical activity and active transport may be a risk factor for depressive symptoms. They also point to differences in this relationship by income range. Further studies are needed to pursue the analysis of how specific domains of physical activity contribute to depressive symptoms.

Alpha lipoic acid controls degeneration and ensures follicular development in ovine ovarian tissue cultured in vitro.

This study aims to evaluate the effects of adding alpha lipoic acid (ALA) to the in vitro ovarian tissue culture medium, either fresh or after vitrification/warming. For this purpose, 10 ovaries from five adult sheep were used. Each pair of ovaries gave rise to 16 fragments and were randomly distributed into two groups: fresh (n = 8) and vitrified (n = 8). Two fresh fragments were fixed immediately and considered the control, while another six were cultured in vitro for 14 days in the absence; presence of a constant (100 µM/0-14 day) or dynamic (50 µM/day 0-7 and 100 µM/day 8-14) concentration of ALA. As for the vitrified fragments, two were fixed and the other six were cultured in vitro under the same conditions described for the fresh group. All the fragments were subjected to morphological evaluation, follicular development and stromal density (classical histology), DNA fragmentation (TUNEL), senescence (Sudan Black), fibrosis (Masson's Trichome), and endoplasmic reticulum stress (immunofluorescence). Measurements of the antioxidant capacity against the free radicals 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and estradiol (E2) levels in the culture medium was performed. The results showed that in the absence of ALA, in vitro culture of vitrified ovarian fragments showed a significant reduction (P < 0.05) in follicular morphology and increased the presence of senescence and tissue fibrosis (P < 0.05). Dynamic ALA maintained E2 levels unchanged (P > 0.05) until the end of vitrified ovarian tissue culture and controlled the levels of ABTS and DPPH radicals in fresh or vitrified cultures. Therefore, it is concluded that ALA should be added to the vitrified ovarian tissue in vitro culture medium to reduce the damage that leads to loss of ovarian function. To ensure steroidogenesis during in vitro culture, ALA should be added dynamically (different concentrations throughout culture).

Anethole supplementation during in vitro maturation increases in vitro goat embryo production in a concentration-dependent manner.

During in vitro maturation (IVM) cumulus-oocyte complexes (COCs) are exposed to conditions that can trigger oxidative stress, thus, reducing oocyte maturation and viability. Aiming to mitigate these detrimental conditions, the effects of IVM medium supplementation with anethole have been tested. Anethole, also known as trans-anethole (1-methoxy-4 [1-propenyl]-benzene), is a naturally occurring phenylpropanoid with various pharmacological properties, including antioxidant effects. However, no study has examined anethole effect on goat COCs during IVM. Thus, the aim of this study was to evaluate the effects of different anethole concentrations on oocyte maturation, oxidative stress, and in vitro development of caprine embryos after parthenogenetic activation. Goat COCs were selected and randomly distributed into the following treatments: TCM-199+ medium (control), or TCM-199+ medium supplemented with 30 µg/mL (AN30); 300 µg/mL (AN300) or 2000 µg/mL (AN2000) of anethole. After IVM, part of the COCs was chosen for oocyte viability and chromatin configuration, intracellular reactive oxygen species levels, and mitochondrial membrane potential assessment. Another part of COCs was parthenogenetically activated, and presumptive zygotes were cultured for 7 days. Results demonstrated that anethole at 30 µg/mL increased oocyte maturation and cleavage rates when compared to the other treatments (P < 0.05), as well as oocyte viability and in vitro embryo production when compared to the control treatment (P < 0.05). Additionally, treatment with anethole at 2000 µg/mL decreased oocyte nuclear maturation and cleavage rates when compared to other treatments (P < 0.05) and embryo production if compared to control and AN30 treatments (P < 0.05). Moreover, anethole at 2000 µg/mL increased mitochondrial membrane potential when compared to the other treatments (P < 0.05). In conclusion, anethole exerts a concentration-dependent effect during goat COCs IVM. For a more desirable outcome of oocyte viability and maturation, and in vitro embryo production, the use of anethole at 30 µg/mL is recommended.

Effects of maize genotypes, nitrogen rates and sources in yield, nutritional status, and fumonisins incidence.

The maize yield, nutritional status, and grain fumonisins concentration were evaluated in different genotypes, doses, and nitrogen sources (N) in two years and three locations. Two experiments were carried out in each area and year in an experimental design of a subdivided plot with four replications. One experiment involved a 4x2 factorial treatment: four nitrogen (N) doses (0, 80, 160, and 240 kg ha-1) in coverage and having urea as a source of N and two genotypes. Another experiment involved a 4x2 factorial treatment: four N sources: urea, urea covered with polymer, ammonium nitrate, and ammonium nitrate + urea (UAN), at a dose of 160 kg ha-1, in two genotypes. The genotype generally influenced maize yield more than N doses and sources, mainly due to the bushy stunt/corn stunt tolerance of AG7098 PRO2 and AG8677 PRO2. The N doses linearly increased the N leaf content. However, the N sources did not affect the N leaf content. The N doses and sources had no significant effect on the content of fumonisins, which was affected only by the genotypes in Sete Lagoas in 2016 (N doses experiment) and 2017 (N sources experiment). The hybrids, P3630H and AG8677PRO2 (Sete Lagoas, 2016, N doses experiment and 2017, N sources experiment, respectively) exceeded the Brazilian legislation for Maximum Tolerance Limit for fumonisins in corn grains, which is 5,000 µg kg-1. The best result was obtained with AG7098 PRO2, with yields (above 10,000 kg ha-1) and fumonisins consistently below 5,000 µg kg-1. Therefore, the selection of corn hybrids is a strategy to reduce the occurrence of fumonisins in the grains.

Definition of protocols for cryopreservation and three-dimensional in vitro culture of prepubertal goat testicular tissue after histomorphological, ultrastructural, and functional analysis.

This study aims to define the best method (slow freezing or vitrification) and fragment size (1, 5, or 9 mm³) for prepubertal goat testis cryopreservation, as well as to evaluate testicular morphological integrity after cryopreservation and in vitro culture (IVC). Initially (experiment I), 1, 5, or 9 mm³ testis fragments were cryopreserved by slow freezing using a Mr. Frosty container with 20% Dimethylsulfoxide (DMSO) or vitrified using the Ovarian Tissue Cryosystem (OTC) device, (Equilibration solution - ES: 10% DMSO and 10% ethylene glycol - EG; Vitrification solution - VS: 20% DMSO and 20% EG) and then subjected to morphological analysis, type I and III collagen quantification and gene expression (Oct4, C-kit, Bax, and Bcl-2). Subsequently, (experiment II), fresh or cryopreserved by slow freezing testis fragments were cultured in vitro and submitted to morphological analysis by scanning electron microscopy. The data from the experiment I revealed fewer morphological alterations in 1 and 5 mm³ fragments after vitrification and slow freezing, respectively. The percentage of type I collagen fibers in 5 and 9 mm³ frozen was higher than in fresh or vitrified fragments. For type III collagen, fresh or frozen fragments of 1 and 5 mm3 showed a higher percentage than fragments of 9 mm3. Gene expression for Oct4 and C-kit after slow freezing or vitrification in the 5 mm3 fragments was lower than that observed in the fresh fragments. The Bax:Bcl-2 ratio in the 1 and 9 mm³ fragments was lower than in the 5 mm³ fragments for fresh fragments or after freezing. In experiment II, fragments cultured in vitro, previously frozen or not, showed more morphological alterations than fresh or frozen fragments. We concluded that slow freezing of 5 mm³ fragments was the best protocol for cryopreserving prepubertal goat testis and although the results of IVC are encouraging, it still needs improvement to restore testicular function after cryopreservation.

Bose-Einstein condensation of photons in microcavity plasmas.

Bose-Einstein condensation of a finite number of photons propagating inside a plasma-filled microcavity is investigated. The nonzero chemical potential is provided by the electrons, which induces a finite photon mass and allows condensation to occur. We derive an equation that models the evolution of the photon-mode occupancies, with Compton scattering taken into account as the mechanism of thermalization. The kinetic evolution of the photon spectrum is solved numerically, and we find evidence of condensation down to nanosecond timescales for typical microplasma conditions, n_{e}∼10^{14}-10^{15}cm^{-3}. The critical temperature scales almost linearly with the number of photons, and we find high condensate fractions at microcavity-plasma temperatures, for experimentally achievable cavity lengths (100-500µm) and photon numbers (10^{10}-10^{12}).

Effect of carvacrol antioxidant capacity on oocyte maturation and embryo production in cattle.

Carvacrol (C10H14O), an efficient phenolic antioxidant substance for several cell types, may become a useful antioxidant for female germ cells and embryo culture. This study investigates the effects of carvacrol supplementation on bovine oocytes in in vitro maturation (IVM) and embryo production. In total, 1222 cumulus-oocyte complexes were cultured in TCM-199+ alone (control treatment) or supplemented with carvacrol at the concentrations of 3 µM (Carv-3), 12.5 µM (Carv-12.5), or 25 µM (Carv-25). After IVM, the oocytes were subjected to in vitro fertilization and embryo production, and the spent medium post-IVM was used for evaluating the levels of reactive oxygen species and the antioxidant capacity (2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2'-azinobis-3-ethyl-benzothiozoline-6-sulphonic acid quantification). A greater (P < 0.05) antioxidant potential was observed in the spent medium of all carvacrol-treated groups compared with the control medium. Moreover, the addition of carvacrol to the maturation medium did not affect (P > 0.05) blastocyst production on days 7 and 10 of culture; however, the total number of cells per blastocyst was reduced (P < 0.05) in two carvacrol-treated groups (Carv-3 and Carv-25). In conclusion, carvacrol demonstrated a high antioxidant capacity in the spent medium after oocyte maturation; however, although embryo production was not affected, in general, carvacrol addition to IVM medium reduced the total number of cells per blastocyst. Therefore, due to the high antioxidant capacity of carvacrol, new experiments are warranted to investigate the beneficial effects of lower concentrations of carvacrol on embryo production in cattle and other species.

How omnivory affects the survival and choices of earwig Doru luteipes (Scudder) (Dermaptera: Forficulidae)? / Como a onivoria afeta a sobrevivência e escolhas da tesourinha Doru luteipes (Scudder) (Dermaptera: Forficulidae)?

Abstract Doru luteipes (Scudder, 1876) is an omnivorous predator that finds different food resources in the corn plant: eggs of Spodoptera frugiperda (J.E. Smith, 1797), uredospores of Puccinia polysora (Underw, 1897), and pollen. Knowing the survival and food preferences of this predator is essential to define its relevance as a biological control agent. We hypothesize that the foraging behavior and predatory capacity of D. luteipes may be affected when several food resources, especially eggs of S. frugiperda, uredospores of P. polysora, and pollen are concurrently in the same plant. The survival of D. luteipes in the nymph stage and their preference among food resources, often available in corn plants, were determined. To verify the survival of D. luteipes, newly hatched nymphs were fed exclusively with 1- uredospores of P. polysora, 2- eggs of S. frugiperda, 3- corn pollen, 4- a combination of uredospores + eggs, and 5- artificial diet (control). In another experiment, nymphs and adults of D. luteipes with 24 and 48 hours of fasting were individually released in the center of a container with four diets: 1- uredospores of P. polysora, 2- eggs of S. frugiperda, 3- corn pollen, 4- artificial diet, and maintained for 10 minutes, to evaluate the food choice and feeding time. The exclusive feeding with S. frugiperda eggs caused low nymph survival (8%), but the combination of P. polysora uredospores + S. frugiperda eggs allowed 58.3% survival. D. luteipes preferred feeding during the nighttime and the most significant proportions of choices by nymphs and adults were for pollen and diet, with adults spending more time eating pollen. These findings indicate that the trophic choices of D. luteipes are relevant to understand its contribution as an agent to control pest insects and fungal diseases in corn.

Resumo Doru luteipes (Scudder, 1876) é um predador onívoro, que encontra na planta do milho diferentes recursos alimentares: ovos de Spodoptera frugiperda (J.E. Smith, 1797), uredósporos de Puccinia polysora (Underw, 1897) e pólen. Para definição da relevância desse predador como agente de controle biológico, conhecer a sobrevivência e preferência alimentar é essencial. Nós hipotetizamos que o comportamento de forrageamento e a capacidade de predação de D. luteipes podem ser afetados quando uma mesma planta oferece ovos, uredósporos e pólen, concomitantemente. A sobrevivência de D. luteipes na fase de ninfa e sua preferência entre os recursos alimentares, frequentemente disponíveis nas plantas de milho, foram determinados. Para verificar a sobrevivência de D. luteipes, ninfas recém eclodidas foram alimentadas exclusivamente com 1- uredósporos de P. polysora, 2- ovos de S. frugiperda, 3- pólen de milho, 4- combinação de uredósporos + ovos e 5- dieta artificial (controle). Em outro experimento, ninfas e adultos de D. luteipes com 24 e 48 horas de jejum foram liberadas individualmente, em recipientes contendo quatro dietas: 1- uredósporos de P. polysora, 2- ovos de S. frugiperda, 3- pólen de milho, 4- dieta artificial e mantidas durante 10 min, sendo avaliados a escolha pelo alimento e o tempo de alimentação. A alimentação exclusiva com ovos de S. frugiperda ocasionou baixa sobrevivência das ninfas (8%), porém a combinação de uredósporos de P. polysora + ovos de S. frugiperda possibilitou sobrevivência de 58,3%. D. luteipes preferiu se alimentar durante o período noturno e as maiores proporções de escolhas das ninfas e dos adultos ocorreram no pólen e na dieta, sendo que os adultos gastaram mais tempo se alimentando de pólen. Estas descobertas indicam que as escolhas tróficas de D. luteipes são relevantes para compreender sua contribuição como agente de controle de insetos-praga e doença fúngicas em milho.

How omnivory affects the survival and choices of earwig Doru luteipes (Scudder) (Dermaptera: Forficulidae)? / Como a onivoria afeta a sobrevivência e escolhas da tesourinha Doru luteipes (Scudder) (Dermaptera: Forficulidae)?

Doru luteipes (Scudder, 1876) is an omnivorous predator that finds different food resources in the corn plant: eggs of Spodoptera frugiperda (J.E. Smith, 1797), uredospores of Puccinia polysora (Underw, 1897), and pollen. Knowing the survival and food preferences of this predator is essential to define its relevance as a biological control agent. We hypothesize that the foraging behavior and predatory capacity of D. luteipes may be affected when several food resources, especially eggs of S. frugiperda, uredospores of P. polysora, and pollen are concurrently in the same plant. The survival of D. luteipes in the nymph stage and their preference among food resources, often available in corn plants, were determined. To verify the survival of D. luteipes, newly hatched nymphs were fed exclusively with 1- uredospores of P. polysora, 2- eggs of S. frugiperda, 3- corn pollen, 4- a combination of uredospores + eggs, and 5- artificial diet (control). In another experiment, nymphs and adults of D. luteipes with 24 and 48 hours of fasting were individually released in the center of a container with four diets: 1- uredospores of P. polysora, 2- eggs of S. frugiperda, 3- corn pollen, 4- artificial diet, and maintained for 10 minutes, to evaluate the food choice and feeding time. The exclusive feeding with S. frugiperda eggs caused low nymph survival (8%), but the combination of P. polysora uredospores + S. frugiperda eggs allowed 58.3% survival. D. luteipes preferred feeding during the nighttime and the most significant proportions of choices by nymphs and adults were for pollen and diet, with adults spending more time eating pollen. These findings indicate that the trophic choices of D. luteipes are relevant to understand its contribution as an agent to control pest insects and fungal diseases in corn.

Doru luteipes (Scudder, 1876) é um predador onívoro, que encontra na planta do milho diferentes recursos alimentares: ovos de Spodoptera frugiperda (J.E. Smith, 1797), uredósporos de Puccinia polysora (Underw, 1897) e pólen. Para definição da relevância desse predador como agente de controle biológico, conhecer a sobrevivência e preferência alimentar é essencial. Nós hipotetizamos que o comportamento de forrageamento e a capacidade de predação de D. luteipes podem ser afetados quando uma mesma planta oferece ovos, uredósporos e pólen, concomitantemente. A sobrevivência de D. luteipes na fase de ninfa e sua preferência entre os recursos alimentares, frequentemente disponíveis nas plantas de milho, foram determinados. Para verificar a sobrevivência de D. luteipes, ninfas recém eclodidas foram alimentadas exclusivamente com 1- uredósporos de P. polysora, 2- ovos de S. frugiperda, 3- pólen de milho, 4- combinação de uredósporos + ovos e 5- dieta artificial (controle). Em outro experimento, ninfas e adultos de D. luteipes com 24 e 48 horas de jejum foram liberadas individualmente, em recipientes contendo quatro dietas: 1- uredósporos de P. polysora, 2- ovos de S. frugiperda, 3- pólen de milho, 4- dieta artificial e mantidas durante 10 min, sendo avaliados a escolha pelo alimento e o tempo de alimentação. A alimentação exclusiva com ovos de S. frugiperda ocasionou baixa sobrevivência das ninfas (8%), porém a combinação de uredósporos de P. polysora + ovos de S. frugiperda possibilitou sobrevivência de 58,3%. D. luteipes preferiu se alimentar durante o período noturno e as maiores proporções de escolhas das ninfas e dos adultos ocorreram no pólen e na dieta, sendo que os adultos gastaram mais tempo se alimentando de pólen. Estas descobertas indicam que as escolhas tróficas de D. luteipes são relevantes para compreender sua contribuição como agente de controle de insetos-praga e doença fúngicas em milho.

Thermo-resistant enzyme-producing microorganisms isolated from composting / Microrganismos produtores de enzimas termo-resistentes isolados de compostagem

Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.

How omnivory affects the survival and choices of earwig Doru luteipes (Scudder) (Dermaptera: Forficulidae)?

Abstract Doru luteipes (Scudder, 1876) is an omnivorous predator that finds different food resources in the corn plant: eggs of Spodoptera frugiperda (J.E. Smith, 1797), uredospores of Puccinia polysora (Underw, 1897), and pollen. Knowing the survival and food preferences of this predator is essential to define its relevance as a biological control agent. We hypothesize that the foraging behavior and predatory capacity of D. luteipes may be affected when several food resources, especially eggs of S. frugiperda, uredospores of P. polysora, and pollen are concurrently in the same plant. The survival of D. luteipes in the nymph stage and their preference among food resources, often available in corn plants, were determined. To verify the survival of D. luteipes, newly hatched nymphs were fed exclusively with 1- uredospores of P. polysora, 2- eggs of S. frugiperda, 3- corn pollen, 4- a combination of uredospores + eggs, and 5- artificial diet (control). In another experiment, nymphs and adults of D. luteipes with 24 and 48 hours of fasting were individually released in the center of a container with four diets: 1- uredospores of P. polysora, 2- eggs of S. frugiperda, 3- corn pollen, 4- artificial diet, and maintained for 10 minutes, to evaluate the food choice and feeding time. The exclusive feeding with S. frugiperda eggs caused low nymph survival (8%), but the combination of P. polysora uredospores + S. frugiperda eggs allowed 58.3% survival. D. luteipes preferred feeding during the nighttime and the most significant proportions of choices by nymphs and adults were for pollen and diet, with adults spending more time eating pollen. These findings indicate that the trophic choices of D. luteipes are relevant to understand its contribution as an agent to control pest insects and fungal diseases in corn.

Resumo Doru luteipes (Scudder, 1876) é um predador onívoro, que encontra na planta do milho diferentes recursos alimentares: ovos de Spodoptera frugiperda (J.E. Smith, 1797), uredósporos de Puccinia polysora (Underw, 1897) e pólen. Para definição da relevância desse predador como agente de controle biológico, conhecer a sobrevivência e preferência alimentar é essencial. Nós hipotetizamos que o comportamento de forrageamento e a capacidade de predação de D. luteipes podem ser afetados quando uma mesma planta oferece ovos, uredósporos e pólen, concomitantemente. A sobrevivência de D. luteipes na fase de ninfa e sua preferência entre os recursos alimentares, frequentemente disponíveis nas plantas de milho, foram determinados. Para verificar a sobrevivência de D. luteipes, ninfas recém eclodidas foram alimentadas exclusivamente com 1- uredósporos de P. polysora, 2- ovos de S. frugiperda, 3- pólen de milho, 4- combinação de uredósporos + ovos e 5- dieta artificial (controle). Em outro experimento, ninfas e adultos de D. luteipes com 24 e 48 horas de jejum foram liberadas individualmente, em recipientes contendo quatro dietas: 1- uredósporos de P. polysora, 2- ovos de S. frugiperda, 3- pólen de milho, 4- dieta artificial e mantidas durante 10 min, sendo avaliados a escolha pelo alimento e o tempo de alimentação. A alimentação exclusiva com ovos de S. frugiperda ocasionou baixa sobrevivência das ninfas (8%), porém a combinação de uredósporos de P. polysora + ovos de S. frugiperda possibilitou sobrevivência de 58,3%. D. luteipes preferiu se alimentar durante o período noturno e as maiores proporções de escolhas das ninfas e dos adultos ocorreram no pólen e na dieta, sendo que os adultos gastaram mais tempo se alimentando de pólen. Estas descobertas indicam que as escolhas tróficas de D. luteipes são relevantes para compreender sua contribuição como agente de controle de insetos-praga e doença fúngicas em milho.

Thermo-resistant enzyme-producing microorganisms isolated from composting

Abstract Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Resumo Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.

Expression analysis of phosphate induced genes in contrasting maize genotypes for phosphorus use efficiency.

Phosphorus is an essential nutrient for plant growth and development. The ability of plants to acquire phosphate (Pi) from the rhizosphere soil is critical in the Brazilian Cerrado characterized by acidic soil. The induction of Pi transporters is one of the earliest molecular responses to Pi deficiency in plants. In this study, we characterize the transcriptional regulation of six (ZmPT1 to ZmPT6) high-affinity Pi transporters genes in four Pi-efficient and four Pi-inefficient maize (Zea mays) genotypes. The expression analysis indicated that Pi-starvation induced the transcription of all ZmPT genes tested. The abundance of transcripts was inversely related to Pi concentration in nutrient solution and was observed as early as five days following the Pi deprivation. The Pi-starved plants replenished with 250 µM Pi for four to five days resulted in ZmPT suppression, indicating the Pi role in gene expression. The tissue-specific expression analysis revealed the abundance of ZmPT transcripts in roots and shoots. The six maize Pi transporters were primarily detected in the upper and middle root portions and barely expressed in root tips. The expression profiles of the six ZmPTs phosphate transporters between and among Pi-efficient and Pi-inefficient genotypes showed an absence of significant differences in the expression pattern of the ZmPTs among Pi-efficient and Pi-inefficient genotypes. The results suggested that Pi acquisition efficiency is a complex trait determined by quantitative loci in maize.

In silico prediction of the enzymes involved in the degradation of the herbicide molinate by Gulosibacter molinativorax ON4T.

Gulosibacter molinativorax ON4T is the only known organism to produce molinate hydrolase (MolA), which catalyses the breakdown of the thiocarbamate herbicide into azepane-1-carboxylic acid (ACA) and ethanethiol. A combined genomic and transcriptomic strategy was used to fully characterize the strain ON4T genome, particularly the molA genetic environment, to identify the potential genes encoding ACA degradation enzymes. Genomic data revealed that molA is the only catabolic gene of a novel composite transposon (Tn6311), located in a novel low copy number plasmid (pARLON1) harbouring a putative T4SS of the class FATA. pARLON1 had an ANI value of 88.2% with contig 18 from Agrococcus casei LMG 22410T draft genome. Such results suggest that pARLON1 is related to genomic elements of other Actinobacteria, although Tn6311 was observed only in strain ON4T. Furthermore, genomic and transcriptomic data demonstrated that the genes involved in ACA degradation are chromosomal. Based on their overexpression when growing in the presence of molinate, the enzymes potentially involved in the heterocyclic ring breakdown were predicted. Among these, the activity of a protein related to caprolactone hydrolase was demonstrated using heterologous expression. However, further studies are needed to confirm the role of the other putative enzymes.

Oocyte in vitro maturation with eugenol improves the medium antioxidant capacity and total cell number per blastocyst.

This study investigates the impact of eugenol (EU) supplementation on bovine oocyte in vitro maturation (IVM) and antioxidant capacity, as well as in vitro embryo production and quality after conventional in vitro fertilization (IVF). A total of 1077 cumulus oocyte complexes were cultured in TCM-199+ without EU supplementation (control treatment) or supplemented with EU at the concentrations of 10 µM (EU-10), 20 µM (EU-20), or 40 µM (EU-40). After IVM, the oocytes were subjected to IVF and embryo culture. The addition of EU at 40 µM to the IVM medium improved (P < 0.05) the antioxidant capacity and cleavage rate when compared to the control treatment. Moreover, a positive correlation (r = 0.61, P < 0.03) was observed between cleavage rate and EU concentration. The addition of EU at concentrations of 10 and 20 µM decreased (P < 0.05) the calreticulin (CALR) levels in expanded blastocysts when compared to the control treatment and EU-40 treatment. However, the EU-10 and EU-20 treatments had a greater (P < 0.05) mean total cell number (TCN) per expanded blastocyst when compared to the control treatment and EU-40 treatment. In conclusion, the addition of EU to the enriched culture medium during IVM of bovine oocytes improved the antioxidant capacity of the spent medium, as well as the cleavage rate and embryonic quality (i.e., TCN/expanded blastocyst), and reduced the endoplasmic reticulum stress (i.e., CALR levels) in the embryos. Thus, we recommend enriching the IVM medium with 10 µM EU for in vitro bovine embryo production.

Microorganisms from corn stigma with biocontrol potential of Fusarium verticillioides.

The mycotoxigenic fungus Fusarium verticillioides is the primary maize pathogen and causes the maize stalk and ear rot diseases with significant economic losses. Furthermore, the excessive use of fungicides to control F. verticillioides constitutes threats to the environment and human health. Thus, sustainable alternatives such as biological control are needed to minimize the hazards associated with the current method. Although much is known about the vulnerability of the maize silks as a gateway for several fungal pathogens invading the developing grains, studies on the chemical properties of silk extracts and their resident microbiota are scarce. This study isolated and characterized bacteria and fungi that colonize the maize stigma to assess new potential biocontrol agents. The samples were collected from maize fields in the Brazilian localities of Sete Lagoas-MG, Sidrolândia-MS, Sertaneja-PR, and Goiânia-GO. One hundred sixty-seven microorganisms were isolated, 46% endophytic and 54% epiphytic. First, the antagonist activity was evaluated by the agar disc diffusion method performed in triplicate, and 83% of the isolates showed antagonist activity against F. verticillioides. Then, the 42 most efficient isolates were identified based on the partial sequencing of the bacterial 16S rRNA gene and fungi ITS region. The bacteria belong to the genera Bacillus (57.1%), Burkholderia (23.8%), Achromobacter (7.1%), Pseudomonas (2.4%), and Serratia (2.4%), while the fungi are Penicillium (2.4%), Candida (2.4), and Aspergillus (2.4%). The results showed that microorganisms from maize stigma might represent new promising agents for F. verticillioides control.

In vitro- and in vivo-derived early antral follicles have comparable in vitro follicular growth and oocyte maturation rates in goats.

Recent in vitro follicle culture (IVFC) studies in caprine have yielded lower maturation rates using late preantral follicles compared to early antral follicles. Thus, research focusing on developing stage-specific customized culture systems able to improve the efficiency of IVFC for late preantral follicles are warranted. This study aimed to compare the morphometric features, estradiol production, and gene expression between early antral caprine follicles produced in vitro and in vivo. In vitro-derived early antral follicles were produced after a 6-day in vitro culture of late preantral follicles, while in vivo-derived early antral follicles were yielded immediately after isolation from the ovaries; antral follicles were, thereafter, cultured for 18 days. In vitro-derived antral follicles were cultured either in a medium developed for preantral follicles (PF medium) or in a medium developed for antral follicles (AF medium). In vivo-derived early antral follicles, on the other hand, were cultured in AF medium (Control treatment). Results demonstrated that in vitro-derived antral follicles cultured in PF medium produced higher estradiol concentration, and m-RNA expression for matrix metalloproteinase-9 (MMP-9), and insulin receptor when compared to both in vitro- and in vivo-derived antral follicles cultured in AF medium. Remarkably, in vitro-derived antral follicles cultured in PF medium had similar MII and oocytes ≥110 µm rates compared with in vivo-derived antral follicles (Control treatment). In conclusion, when cultured in a single and appropriate medium (i.e., PF medium), in vitro-derived early antral follicles had comparable oocyte maturation rates to the in vivo-derived early antral follicles.

Eugenol Improves Follicular Survival and Development During in vitro Culture of Goat Ovarian Tissue.

This study evaluated the effects of different concentrations (10, 20, or 40 µM) of eugenol (EUG 10, EUG 20, or EUG 40), ascorbic acid (50 µg/mL; AA) or anethole (300 µg/mL; ANE 300) on the in-vitro survival and development of goat preantral follicles and oxidative stress in the cultured ovarian tissue. Ovarian fragments from five goats were cultured for 1 or 7 days in Alpha Minimum Essential Medium (α-MEM+) supplemented or not with AA, ANE 300, EUG 10, EUG 20 or EUG 40. On day 7 of culture, when compared to MEM, the addition of EUG 40 had increased the rate of follicular development, as observed by a decrease in the proportion of primordial follicles alongside with an increase in the rate of normally developing follicles. Furthermore, EUG 40 significantly increased both follicular and oocyte diameters. Subsequently, ovarian fragments from three goats were cultured for 1 or 7 days in α-MEM+ supplemented or not with AA, ANE 300 or EUG 40. All tested antioxidants, except ANE 300, were able to significantly decrease the levels of reactive oxygen species in the ovarian tissue, but EUG 40 could most efficiently neutralize free radicals. All ovarian tissues cultured in the presence of antioxidants, especially EUG 40, presented a significant decrease in H3K4me3 labeling, indicating a silencing of genes that play a role in the inhibition of follicular activation and apoptosis induction. When compared to cultured control tissues, both EUG 40 and ANE 300 significantly increased the intensity of calreticulin labeling in growing follicles. The mRNA relative expression of ERP29 and KDM3A was significantly increased when the culture medium was supplemented with EUG 40, indicating a response to ER stress experienced during culture. In conclusion, EUG 40 improved in-vitro follicle survival, activation and development and decreased ROS production, ER stress and histone lysine methylation in goat ovarian tissue.

Interference of fixatives and fixation period on the morphologic analysis of ovarian preantral follicles.

Ovine ovarian fragments (3 × 3 × 1 mm) were fixed in neutral buffered formalin (NBF), Carnoy's solution (CAR), Davidson's solution (DAV), or paraformaldehyde (PFA) for 12 h or 24 h. After this fixation time, each fragment was prepared for histological analysis. Although fixative and fixation period did not affect follicular and stromal cells density, the percentages of morphologically normal primordial and primary follicles was affected by the fixative type and period of fixation. Paraformaldehyde was not indicated as a fixative for ovarian fragments. Formalin was a suitable fixative only when the period of fixation was 12 h, while Carnoy was efficient after a fixation period of 24 h. In conclusion, the most indicated fixative for the morphological evaluation of ovarian preantral follicles was DAV, regardless of the fixation period, that is 12 or 24 h.