Selectivity of herbicides applied to Brazilian tree species at post-emergence

The presence of weeds in silvicultural systems has been considered one of the main obstacles to the success of projects designed to recover degraded areas. The aim of this study was to evaluate the selectivity of herbicides applied at post-emergence in the initial growth of seedlings of capixingui (Croton floribundus), açoita-cavalo (Luehea divaricata), and guaritá (Astronium graveolens), in the municipalities of Jaboticabal and Junqueirópolis, state of São Paulo. The experimental design was completely randomized, with four replications, and the treatments consisted of herbicides (g a.i.·ha-1) clethodim + phenoxaprop-p-ethyl (50 + 50), sethoxydim (184), quizalofop-p-ethyl (75), nicosulfuron (50), fluazifop-p-butyl (125), fomesafen (225), haloxyfop-methyl (48), bentazon (720), chlorimuron-ethyl (15), in addition to control without herbicide. The characteristics analyzed were: plant height increase and visual phytointoxication at 7, 14, 21, 28, 35, and 42 days after herbicide application. At the end of the experiment, the shoots of the plants were removed to assess shoot dry matter. The herbicides clethodim + fenoxaprop-p-ethyl, fluazifop-p-butyl, and quizalofop-p-ethyl showed selective potential for the species capixingui, açoita-cavalo, and guaritá. The herbicide chlorimuron-ethyl caused mild intoxication symptoms when applied to seedlings of capixingui and açoita-cavalo; also, it was not selective for the guaritá species grown in Jaboticabal. All species showed selectivity to the herbicides sethoxydim, fomesafen, haloxyfop-methyl, and nicosulfuron, as their growth and initial development were not influenced. The herbicide bentazon caused high percentages of injury to açoita-cavalo plants in both cultivation places, but it did not influence the growth and development of the species.

Pasture weed biology and management in Brazil / Biologia e manejo de plantas daninhas em pastagens no Brasil

The livestock production model historically practiced in Brazil has a strong extractive bias, wherein the premise is to produce livestock with absolutely no concern for the preservation or renewal of environmental resources. The absence of technical criteria for the use of pastures has generated low productivity rates, making the activity unsustainable from both economic and environmental points of view. This scenario led the several sectors linked to the production chain to develop a package of strategies to solve the problems faced by livestock farmers. This package of strategies is conventionally called postmodern or corporate farming, in which the extractive process gives way to the business logic of avoiding waste and recovering profit margins mainly through pasture perpetuation. However, there is still a technical gap in corporate cattle farming related to problems caused by pasture weeds because all the concepts applied are derived or copied from concepts generated in agriculture. Furthermore, few researchers have studied or scientific articles written on elucidating the real problem of weeds in livestock production. Thus, the goal of the present review was to present some aspects related to weed ecology, their interference, and management alternatives in pasture areas, thereby collaborating with corporate livestock farming in Brazil because solutions to weed problems are crucial to increase commitment in all sectors of the production chain.(AU)

O modelo de atividade pecuária historicamente praticado no Brasil está alicerçado, em sua grande maioria, em forte viés extrativista, onde a premissa é produzir sem absolutamente nenhuma preocupação quanto à preservação ou renovação dos recursos ambientais. A ausência de critérios técnicos na utilização das pastagens gerou e ainda gera baixos índices de produtividade e torna a atividade insustentável do ponto de vista econômico e ambiental. Tal cenário levou os diversos setores ligados à cadeia produtiva a desenvolver um pacote de estratégias visando solucionar os problemas enfrentados pelos pecuaristas. A esse pacote de estratégias convencionou-se chamar de pecuária pós-moderna ou empresarial, onde o processo extrativista cede lugar à lógica empresarial de evitar desperdícios e recuperar as margens de lucros através, principalmente, da perenização das pastagens. Entretanto, ainda existe uma lacuna técnica na pecuária empresarial referente aos problemas ocasionados pelas plantas daninhas em áreas de pastagem, uma vez que todos os conceitos aplicados são provenientes ou copiados daqueles conceitos gerados na agricultura. Além disso, são raros os pesquisadores e artigos científicos dedicados à compreensão da real problemática das plantas daninhas na atividade de produção animal. Assim, a presente revisão tem como objetivo apresentar alguns aspectos ligados à ecologia de plantas daninhas, sua interferência e alternativas de manejo em áreas de pastagens, visando, dessa forma, colaborar com a pecuária empresarial no Brasil, uma vez que soluções para o problema da ocorrência de plantas daninhas é um ponto crucial para proporcionar aumento do empenho em todos os setores da cadeia produtiva.(AU)

Weed coexistence in carioca bean inoculated with Rhizobium tropici or nitrogen-fertilized / Convivência das plantas daninhas no feijão carioca inoculado com Rhizobium tropici ou com adubação nitrogenada

The supply of nitrogen (N) to the carioca bean plant via inoculation with Rhizobium tropici can prevent competition with the weed community by allowing the crop to absorb the nutrient available in the soil. On this basis, this study proposes to examine the period before weed interference (PBI) in the carioca bean plant following inoculation with R. tropici or N topdressing. The experiments were carried out under field conditions during the summer seasons of 2014 and 2015. A randomized-block experimental design with four replicates was adopted, in a 2 × 11 factorial arrangement (common bean plant inoculated or topdressed with N × 11 periods of coexistence with weeds, namely, 0, 7, 14, 21, 28, 35, 42, 49, 56, 63, or 90 days after emergence [DAE]). Nitrogen topdressing increased the crop's tolerance to coexist with weeds from 6 to 14 DAE, compared with inoculation with R. tropici The PBI for the inoculated common bean plant was 24 and 16 DAE in the years 2014 and 2015, respectively. For the N-topdressed plant, the PBI was 30 DAE in both years.(AU)

O fornecimento do nitrogênio ao feijoeiro carioca via inoculação com Rhizobium tropici pode evitar a competição com a comunidade de plantas daninhas pela absorção do nutriente disponível no solo. Portanto, o objeto deste estudo foi avaliar o período anterior à interferência (PAI) no feijoeiro carioca com inoculação com R. tropici e com adubação nitrogenada em cobertura. Os experimentos foram conduzidos em condições de campo durante as safras de verão de 2014 e de 2015. O delineamento experimental foi de blocos casualizados com quatro repetições em esquema fatorial 2x11 (feijoeiro com inoculação ou com adução nitrogenada em cobertura x 11 períodos de convívio com as plantas daninhas: 0, 7, 14, 21, 28, 35, 42, 49, 56, 63 e 90 dias após a emergência (DAE). A adubação nitrogenada em cobertura aumentou a capacidade da cultura em conviver com as plantas daninhas de 6 a 14 DAE quando comparada a inoculação com R. tropici. O PAI para o feijoeiro inoculado foi de 24 e 16 DAE em 2014 e 2015, respectivamente. Para o feijoeiro com adubação nitrogenada em cobertura o PAI foi de 30 DAE em ambos os anos.(AU)

Evaluation of the Viral Interference between Lentogenic Newcastle Disease Virus (Lasota) and Avian Influenza Virus (H9N2) using Real-Time Reverse Transcription Polymerase Chain Reaction in SPF Chicken

ABSTRACT Lentogenic Newcastle disease virus (lNDV) such as Lasota strain and low pathogenicity avian influenza such as H9N2 virus are two of the most economically important viruses affecting poultry worldwide, and little attention in recent years has been paid to simultaneous infections in chickens with these two viruses for the reason that co-infection do occur but are not easily detected. In the present study, chickens were inoculated with lNDV (Lasota) and LPAIV (A/chicken/Tehran/ZMT-173/99(H9N2)) simultaneously or sequentially three days apart. Oropharyngeal and cloacal swabs were collected from chickens from 1 to 14 days after inoculation. RRT-PCR for AIV and NDV detection was performed. The rate of viral shedding was measured within 14 days. No clinical symptoms were observed during the experiment however the pattern of virus shed was different with co-infection, thus comparing the results obtained from viral shedding showed that AIV is a much stronger agent than NDV in the occurrence of viral interference. This is due to the fact that in simultaneous inoculation, AIV replication delayed and reduced NDV replication, while replication of Lasota in simultaneous or pre-inoculated inoculation could not significantly disrupt H9N2 virus replication. These findings indicate that the infection with one virus can interfere with the replication of another, modifying the pathogenesis of the viruses. So, infection of the host with both viral agents simultaneously causes higher shedding of LPAIV than lNDV in OP and CL areas. In conclusion, co-infection with LPAVI in chickens did not impact clinical signs but affected the replication dynamics of these viruses.

Evaluation of the Viral Interference between Lentogenic Newcastle Disease Virus (Lasota) and Avian Influenza Virus (H9N2) using Real-Time Reverse Transcription Polymerase Chain Reaction in SPF Chicken

Lentogenic Newcastle disease virus (lNDV) such as Lasota strain and low pathogenicity avian influenza such as H9N2 virus are two of the most economically important viruses affecting poultry worldwide, and little attention in recent years has been paid to simultaneous infections in chickens with these two viruses for the reason that co-infection do occur but are not easily detected. In the present study, chickens were inoculated with lNDV (Lasota) and LPAIV (A/chicken/Tehran/ZMT-173/99(H9N2)) simultaneously or sequentially three days apart. Oropharyngeal and cloacal swabs were collected from chickens from 1 to 14 days after inoculation. RRT-PCR for AIV and NDV detection was performed. The rate of viral shedding was measured within 14 days. No clinical symptoms were observed during the experiment however the pattern of virus shed was different with co-infection, thus comparing the results obtained from viral shedding showed that AIV is a much stronger agent than NDV in the occurrence of viral interference. This is due to the fact that in simultaneous inoculation, AIV replication delayed and reduced NDV replication, while replication of Lasota in simultaneous or pre-inoculated inoculation could not significantly disrupt H9N2 virus replication. These findings indicate that the infection with one virus can interfere with the replication of another, modifying the pathogenesis of the viruses. So, infection of the host with both viral agents simultaneously causes higher shedding of LPAIV than lNDV in OP and CL areas. In conclusion, co-infection with LPAVI in chickens did not impact clinical signs but affected the replication dynamics of these viruses.(AU)

Evaluation of the Viral Interference between Lentogenic Newcastle Disease Virus (Lasota) and Avian Influenza Virus (H9N2) using Real-Time Reverse Transcription Polymerase Chain Reaction in SPF Chicken

Lentogenic Newcastle disease virus (lNDV) such as Lasota strain and low pathogenicity avian influenza such as H9N2 virus are two of the most economically important viruses affecting poultry worldwide, and little attention in recent years has been paid to simultaneous infections in chickens with these two viruses for the reason that co-infection do occur but are not easily detected. In the present study, chickens were inoculated with lNDV (Lasota) and LPAIV (A/chicken/Tehran/ZMT-173/99(H9N2)) simultaneously or sequentially three days apart. Oropharyngeal and cloacal swabs were collected from chickens from 1 to 14 days after inoculation. RRT-PCR for AIV and NDV detection was performed. The rate of viral shedding was measured within 14 days. No clinical symptoms were observed during the experiment however the pattern of virus shed was different with co-infection, thus comparing the results obtained from viral shedding showed that AIV is a much stronger agent than NDV in the occurrence of viral interference. This is due to the fact that in simultaneous inoculation, AIV replication delayed and reduced NDV replication, while replication of Lasota in simultaneous or pre-inoculated inoculation could not significantly disrupt H9N2 virus replication. These findings indicate that the infection with one virus can interfere with the replication of another, modifying the pathogenesis of the viruses. So, infection of the host with both viral agents simultaneously causes higher shedding of LPAIV than lNDV in OP and CL areas. In conclusion, co-infection with LPAVI in chickens did not impact clinical signs but affected the replication dynamics of these viruses.

Short interfering RNAs targeting a vampire-bat related rabies virus phosphoprotein mRNA

The aim of this study was to assess the in vitro and in vivo effects of short-interfering RNAs (siRNAs) against rabies virus phosphoprotein (P) mRNA in a post-infection treatment for rabies as an extension of a previous report (Braz J Microbiol. 2013 Nov 15;44(3):879-82). To this end, rabies virus strain RABV-4005 (related to the Desmodus rotundus vampire bat) were used to inoculate BHK-21 cells and mice, and the transfection with each of the siRNAs was made with Lipofectamine-2000™. In vitro results showed that siRNA 360 was able to inhibit the replication of strain RABV-4005 with a 1 log decrease in virus titter and 5.16-fold reduction in P mRNA, 24 h post-inoculation when compared to non-treated cells. In vivo, siRNA 360 was able to induce partial protection, but with no significant difference when compared to non-treated mice. These results indicate that, despite the need for improvement for in vivo applications, P mRNA might be a target for an RNAi-based treatment for rabies.(AU)

Periods of weed interference in maize crops cultivated in the first and second cycles / Períodos de interferência de plantas daninhas no milho de primeira e segunda safra

The interference of weeds in maize production may be reflected in grain yield losses that vary as a function of the density, stage and degree of aggressiveness of the species present. In the agricultural ecosystem, crops and weeds demand light, water, nutrients and space, which are frequently not available in sufficient quantities, leading to competition. The aim of this work was to determine the period of interference of weed plants, in particular of naked crabgrass (Digitaria nuda) on maize crop in the first and second harvest. The treatments were defined as increasing periods of coexistence and increasing control of weed community (7, 14, 21, 28, 35, 42, 49, and 56 days), two more controls, a control including one with weed control until the end of the culture cycle and another with coexistence until the harvest. For each period, were evaluated the stand of maize plants, length of ear, number of grains per row, number of rows per ear, cob, 100-grain weight, and grain productivity. The data obtained were subjected to analysis of variance using the F test, with average treatments compared using Tukeys test at 5% probability. Crop productivity was evaluated by means of regressions, the critical periods of interference were estimated. The critical timing of weed removal was 25 days for both harvests. The critical weed free period was 54 and 27 days for the first and second harvest respectively. For the conditions of the first and second harvest, the critical period of weed control was of 29 and 2 days respectively.

A interferência de plantas daninhas no milho pode refletir em perdas na produtividade de grãos que variam em função da densidade, estágio e grau de agressividade das espécies presentes. No ecossistema agrícola, a cultura e planta daninha possuem demandas por luz, água, nutrientes e espaço, que na maioria das vezes, não estão disponíveis em quantidades suficientes, estabelecendo-se a competição. O objetivo desse trabalho foi determinar o período de interferência de plantas de plantas daninhas em especial do capim-colchão (Digitaria nuda) na cultura do milho na primeira e segunda safra. Os tratamentos foram divididos em períodos crescentes de convivência e de controle da comunidade infestante (7; 14; 21; 28; 35; 42; 49 e 56 dias), mais duas testemunhas, uma com controle até o fim do ciclo da cultura e outra com convivência até a colheita. Para cada período foram avaliadas o estande de plantas de milho, comprimento das espigas, número de grãos por fileiras, número de fileiras por espiga, massa de 100 grãos e produtividade de grãos. Os dados obtidos foram submetidos à análise de variância pelo teste F, sendo as médias dos tratamentos comparadas pelo teste Tukey a 5% de probabilidade. A produtividade da cultura foi avaliada por meio de regressões, onde foram estimados os períodos críticos de interferência. O período anterior à interferência foi de 25 dias para ambas as safras. O período total de prevenção da interferência foi de 54 e 27 dias para primeira e segunda safra respectivamente. Para as condições de primeira e segunda safra, o período crítico de prevenção a interferência foi de 29 e 2 dias respectivamente.

Periods of weed interference in maize crops cultivated in the first and second cycles / Períodos de interferência de plantas daninhas no milho de primeira e segunda safra

The interference of weeds in maize production may be reflected in grain yield losses that vary as a function of the density, stage and degree of aggressiveness of the species present. In the agricultural ecosystem, crops and weeds demand light, water, nutrients and space, which are frequently not available in sufficient quantities, leading to competition. The aim of this work was to determine the period of interference of weed plants, in particular of naked crabgrass (Digitaria nuda) on maize crop in the first and second harvest. The treatments were defined as increasing periods of coexistence and increasing control of weed community (7, 14, 21, 28, 35, 42, 49, and 56 days), two more controls, a control including one with weed control until the end of the culture cycle and another with coexistence until the harvest. For each period, were evaluated the stand of maize plants, length of ear, number of grains per row, number of rows per ear, cob, 100-grain weight, and grain productivity. The data obtained were subjected to analysis of variance using the F test, with average treatments compared using Tukeys test at 5% probability. Crop productivity was evaluated by means of regressions, the critical periods of interference were estimated. The critical timing of weed removal was 25 days for both harvests. The critical weed free period was 54 and 27 days for the first and second harvest respectively. For the conditions of the first and second harvest, the critical period of weed control was of 29 and 2 days respectively.(AU)

A interferência de plantas daninhas no milho pode refletir em perdas na produtividade de grãos que variam em função da densidade, estágio e grau de agressividade das espécies presentes. No ecossistema agrícola, a cultura e planta daninha possuem demandas por luz, água, nutrientes e espaço, que na maioria das vezes, não estão disponíveis em quantidades suficientes, estabelecendo-se a competição. O objetivo desse trabalho foi determinar o período de interferência de plantas de plantas daninhas em especial do capim-colchão (Digitaria nuda) na cultura do milho na primeira e segunda safra. Os tratamentos foram divididos em períodos crescentes de convivência e de controle da comunidade infestante (7; 14; 21; 28; 35; 42; 49 e 56 dias), mais duas testemunhas, uma com controle até o fim do ciclo da cultura e outra com convivência até a colheita. Para cada período foram avaliadas o estande de plantas de milho, comprimento das espigas, número de grãos por fileiras, número de fileiras por espiga, massa de 100 grãos e produtividade de grãos. Os dados obtidos foram submetidos à análise de variância pelo teste F, sendo as médias dos tratamentos comparadas pelo teste Tukey a 5% de probabilidade. A produtividade da cultura foi avaliada por meio de regressões, onde foram estimados os períodos críticos de interferência. O período anterior à interferência foi de 25 dias para ambas as safras. O período total de prevenção da interferência foi de 54 e 27 dias para primeira e segunda safra respectivamente. Para as condições de primeira e segunda safra, o período crítico de prevenção a interferência foi de 29 e 2 dias respectivamente.(AU)

Delayed mortality of juvenile shrimp Penaeus vannamei challenged to White spot syndrome virus (WSSV) previously exposed to Infectious hypodermal and haematopoietic necrosis virus (IHHNV) or inactivated WSSV

In Ecuador, the development and sustainability of the cultured white shrimp, Penaeus vannamei, has been threatened by the occurrence of several viral pathogens, Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and White spot syndrome virus (WSSV) mainly. The aim of the present study was to evaluate the exposition of P. vannamei juveniles to IHHNV and formalin-inactivated viruses (inactWSSV or inactIHHNV) to induce a protective response in P. vannamei juveniles against WSSV infection. P. vannamei were challenged to WSSV by intramuscular injection. Shrimp mortalities appeared at day 1 post-injection (p.i.) in positive control and inactIHHNV treatment, while IHHNV and inactWSSV treatments presented onset of mortalities at day 2 p.i. Positive control and inactIHHNV treatment presented 100% mortality at day 4 p.i., while IHHNV and inactWSSV treatments reached similar mortality at day 6 p.i. Statistical analysis revealed that WSSV-induced mortalities in juvenile P. vannamei of IHHNV and inactWSSV treatments had a significant delay (P 0.05) compared to both the inactIHHNV-treatment and positive control. Our results showed thatpreliminary exposure to IHHNV or to formalin-inactivated WSSV can induce delayed mortality in Penaeus vannameifollowing challenge with WSSV via intramuscular injection. In case of IHHNV infection, viral interference could be thebiological phenomenon involved, mediated by competition between IHHNV and WSSV. Regarding to WSSV inactivatedby formalin, a “vaccination” response would be responsible for the delay, evidencing a possible specific antiviral immuneresponse from the host.

Delayed mortality of juvenile shrimp Penaeus vannamei challenged to White spot syndrome virus (WSSV) previously exposed to Infectious hypodermal and haematopoietic necrosis virus (IHHNV) or inactivated WSSV

In Ecuador, the development and sustainability of the cultured white shrimp, Penaeus vannamei, has been threatened by the occurrence of several viral pathogens, Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and White spot syndrome virus (WSSV) mainly. The aim of the present study was to evaluate the exposition of P. vannamei juveniles to IHHNV and formalin-inactivated viruses (inactWSSV or inactIHHNV) to induce a protective response in P. vannamei juveniles against WSSV infection. P. vannamei were challenged to WSSV by intramuscular injection. Shrimp mortalities appeared at day 1 post-injection (p.i.) in positive control and inactIHHNV treatment, while IHHNV and inactWSSV treatments presented onset of mortalities at day 2 p.i. Positive control and inactIHHNV treatment presented 100% mortality at day 4 p.i., while IHHNV and inactWSSV treatments reached similar mortality at day 6 p.i. Statistical analysis revealed that WSSV-induced mortalities in juvenile P. vannamei of IHHNV and inactWSSV treatments had a significant delay (P 0.05) compared to both the inactIHHNV-treatment and positive control. Our results showed thatpreliminary exposure to IHHNV or to formalin-inactivated WSSV can induce delayed mortality in Penaeus vannameifollowing challenge with WSSV via intramuscular injection. In case of IHHNV infection, viral interference could be thebiological phenomenon involved, mediated by competition between IHHNV and WSSV. Regarding to WSSV inactivatedby formalin, a “vaccination” response would be responsible for the delay, evidencing a possible specific antiviral immuneresponse from the host.(AU)

Balloniscus glaber (Crustacea, Isopoda, Balloniscidae), a habitat specialist species in a disturbed area of Brazil / Balloniscus glaber (Crustacea, Isopoda, Balloniscidae), uma espécie especialista de hábitat em uma área perturbada do Brasil

Balloniscus glaber Araujo & Zardo, 1995 (Balloniscidae), a Neotropical Oniscidea, has been recorded historically in environments with low or no human interference. In one of these areas, it was determined as aK-strategist. Recently, however, this species was documented in a disturbed forest within urban limits. The present work revealed that the population in the urban area has high density, high number of ovigerous females and mancae in the population, a long reproductive period, and early sexual maturity. These results suggest that modified environments may provide favorable conditions and that the species is not negatively affected by human influence.(AU)

Balloniscus glaber Araujo & Zardo, 1995 (Balloniscidae), um oniscídeo Neotropical, tem sido registrado historicamente em ambientes com baixa ou sem interferência humana. Em uma dessas áreas, a espécie foi classificada como K-estrategista. No entanto, recentemente foi registrada em uma floresta perturbada dentro dos limites urbanos. O presente trabalho revela que a população na área urbana tem densidade alta, elevado número de fêmeas ovígeras e mancas, período reprodutivo longo e maturidade sexual precoce. Estes resultados sugerem que um ambiente modificado pode proporcionar condições favoráveis e que a espécie não é afetada negativamente pela influência humana.(AU)

Acúmulo de macronutrientes por plantas daninhas e de milho cultivadas em convivência em solo com diferentes manejos de fertilidade / Accumulation of macronutrients by weed and corn in coexistence in soil with different fertility managements

Nos sistemas agrícolas o entendimento da interferência de plantas daninhas sobre as culturas é de grande importância. No entanto, pouco se sabe sobre a influência do manejo de fertilidade do solo nas relações de competição entre plantas. Assim, objetivou-se avaliar os efeitos da interferência mútua entre plantas daninhas e de milho no acúmulo de macronutrientes em solo com diferentes manejos de fertilidade. O ensaio foi realizado em casa de vegetação, incluindo quatro manejos de fertilidade do solo (com silicato de cálcio e magnésio e adubação; com calcário e adubação; sem correção de acidez, mas com adubação; sem correção de acidez e sem adubação) e onze cultivos (cinco arranjos de competição entre Zea mays e as plantas daninhas Urochloa brizantha, Ipomoea grandifolia, Conyza canadensis, Hyptis suaveolens e Bidens pilosa, acrescido das seis plantas em monocultivo). Sob interferência de U. brizantha, o milho apresentou reduções superiores a 50% no conteúdo de todos os macronutrientes. Adicionalmente, U. brizantha e B. pilosa em competição com o milho apresentaram elevado acúmulo de nutrientes nas quatro condições de solo. Silicato de cálcio e magnésio e calcário exerceram influência variável sobre o acúmulo de nutrientes pelas plantas daninhas em monocultivo ou sob interferência. O milho cultivado no solo corrigido com a fonte silicatada sofreu maior concorrência com as plantas...(AU)

The understanding of weed interference on crops is very important in agricultural systems. However, there is little known about the influence of soil fertility management in competitive relations between plants. The objective of this study was to evaluate the effects of mutual interference between weeds and corn on the content of macronutrients under different soil fertility managements. The experimental test was conducted in a greenhouse, considering four soil fertility managements (with calcium and magnesium silicate and fertilization; with limestone and fertilization; without correction of acidity, but with fertilization; without correction of acidity and without fertilization) and eleven crops (five arrangements of competition between Zea mays and the weeds Urochloa brizantha, Ipomoea grandifolia, Conyza canadensis, Hyptis suaveolens and Bidens pilosa, plus the six species in monoculture). Corn had reductions exceeding 50% in the content of all macronutrients under interference of U. brizantha. Additionally, U. brizantha and B. pilosa in competition with corn has high ability to extract macronutrients in the four soil conditions. Calcium and magnesium silicate and limestone influenced variable on nutrient accumulation by weeds in monoculture or under interference. Corn grown in soil amended with silicate suffered greater competition with weeds compared to grown in soil...(AU)

Balloniscus glaber (Crustacea, Isopoda, Balloniscidae), a habitat specialist species in a disturbed area of Brazil / Balloniscus glaber (Crustacea, Isopoda, Balloniscidae), uma espécie especialista de hábitat em uma área perturbada do Brasil

Balloniscus glaber Araujo & Zardo, 1995 (Balloniscidae), a Neotropical Oniscidea, has been recorded historically in environments with low or no human interference. In one of these areas, it was determined as aK-strategist. Recently, however, this species was documented in a disturbed forest within urban limits. The present work revealed that the population in the urban area has high density, high number of ovigerous females and mancae in the population, a long reproductive period, and early sexual maturity. These results suggest that modified environments may provide favorable conditions and that the species is not negatively affected by human influence.

Balloniscus glaber Araujo & Zardo, 1995 (Balloniscidae), um oniscídeo Neotropical, tem sido registrado historicamente em ambientes com baixa ou sem interferência humana. Em uma dessas áreas, a espécie foi classificada como K-estrategista. No entanto, recentemente foi registrada em uma floresta perturbada dentro dos limites urbanos. O presente trabalho revela que a população na área urbana tem densidade alta, elevado número de fêmeas ovígeras e mancas, período reprodutivo longo e maturidade sexual precoce. Estes resultados sugerem que um ambiente modificado pode proporcionar condições favoráveis e que a espécie não é afetada negativamente pela influência humana.

Detection of specific antibodies in cows after injection of PPD / Detecção de anticorpos específicos em vacas após a injeção de PPD

The diagnosis of bovine tuberculosis aims to identify the immune response against mycobacterial antigens. Although Single Intradermal Comparative Cervical Tuberculin test (SICCT) is broadly used for first identification of the disease, the performance of ELISAs has been investigated for diagnosis improvement. The present study expected to find out the influence of intradermal skin tests on the results of ELISAs using the recombinant proteins MPB70 and MPB83 as antigens on cows from a naturally infected herd. Results were analyzed by the F-test, Mann-Whitney and Friedman tests Although comparable to both proteins, results showed that positive animals presented a tendency of augment reactivity to MPB70, representing a tendency for a booster effect, but not to MPB83.(AU)

O diagnóstico da tuberculose bovina baseia-se na identificação da resposta imune do animal frente aos antígenos de Mycobacterium bovis. Embora o teste intradérmico comparativo cervical seja o mais empregado como primeiro teste diagnóstico, outras técnicas, como ELISA, tem sido investigadas para aumentar a detecção de animais infectados. O presente estudo analisou se os testes intradérmicos influenciariam os resultados de ELISAs que utilizaram as proteínas MPB70 e MPB83 como antígenos de captura em um rebanho naturalmente infectado. Os resultados foram analisados por meio de testes estatísticos de comparação e correlação de resultados: teste-F, Mann-Whitney e Friedman. O desempenho de ambos os ELISAs foi comparável; no entanto, os resultados demonstraram que entre os animais positivos, houve um aumento de reatividade ao MPB70, dado este que não foi observado junto ao ELISA-MPB83.(AU)

In vitro and in vivo inhibition of rabies virus replication by RNA interference

Rabies is a zoonotic disease that affects all mammals and leads to more than 55,000 human deaths every year, caused by rabies virus (RABV) (Mononegavirales: Rhabdoviridae: Lyssavirus). Currently, human rabies treatment is based on the Milwaukee Protocol which consists on the induction of coma and massive antiviral therapy. The aim of this study was to assess the decrease in the titer of rabies virus both in vitro and in vivo using short-interfering RNAs. To this end, three siRNAs were used with antisense strands complementary to rabies virus nucleoprotein (N) mRNA. BHK-21 cells monolayers were infected with 1000 to 0.1 TCID50 of PV and after 2 hours the cells were transfected with each of tree RNAs in separate using Lipofectamine-2000. All three siRNAs reduced the titer of PV strain in a least 0.72 logTCID50/mL and no cytotoxic effect was observed in the monolayers treated with Lipofectamine-2000. Swiss albino mice infected with 10.000 to 1 LD of PV strain by the intracerebral route were also transfected after two hours of infection with a pool 3 siRNAs with Lipofectamine-2000 by the intracerebral route, resulting in a survival rate of 30% in mice inoculated with 100 LD50, while the same dose led to 100% mortality in untreated animals. Lipofectamine-2000 showed no toxic effect in control mice. These results suggest that intracerebral administration of siRNAs might be an effective antiviral strategy for rabies.(AU)

Tecnologia de Phage Display no mapeamento de Mimetopos de Haemonchus contortus e uso de RNA de interferência no silenciamento do Gene codificante da protéina Gliceraldeído-3-fosfato desidrogenase (GAPDH) do parasita

Haemonchus contortus é um nematoda hematófago de pequenos ruminantes, extremamente importante em escala mundial, sendo responsável por muitos danos à saúde dos animais, além de perdas produtivas e econômicas aos criadores. O panorama atual é de resistência do parasita à maioria das classes de anti-helmínticos disponíveis no mercado, o que demonstra necessidade urgente de métodos alternativos de controle. Portanto, os objetivos deste estudo foram selecionar mimetopos de H. contortus e avaliar o uso de RNA de interferência no silenciamento do gene codificante da proteína gliceraldeído-3-fosfato desidrogenase (GAPDH) do parasita. A presente tese está dividida em 3 capítulos apresentados na forma de introdução geral e 2 manuscritos. O manuscrito 1 consistiu no uso de anticorpo policlonal de ovinos para mapear mimetopos de H. contortus usando a biblioteca de phage display. O clone 6 mimetopo de GAPDH e clone 17 - mimetopo da família do músculo desorganizado (Dim 1) foram selecionados para ensaio de imunização de ovinos. Peptídeos mimetopos (sintéticos) foram avaliados como moléculas antigênicas por meio de ensaio imunoenzimático (ELISA). No ensaio de imunização constatou-se aumento dos títulos de IgG anti-fago M13, mas não ocorreu aumento de IgG anti-peptídeos mimetopos. Os peptídeos mimetopos sintéticos foram reconhecidos por IgG de ovinos naturalmente infectados por H. contortus. Este é o primeiro relato de uso bem sucedido da biblioteca de phage display para a identificação de mimetopos de H. contortus. O manuscrito 2 teve o objetivo de avaliar o uso de RNA de interferência (RNAi) para silenciamento do gene codificante da proteína GAPDH de H. contortus. Larvas infectantes foram incubadas com RNAi por 3; 6; 24 e 48h. Os resultados revelaram ausência de transcrição gênica em todas os períodos avaliados. Este estudo relata pela primeira vez o silenciamento de GAPDH em H. contortus, confirmando o gene como passível ao silenciamento por RNAi. Os dados expostos nesta tese apontam resultados promissores para uso em novas pesquisas voltadas ao desenvolvimento de terapias para o controle de H. contortus.

Haemonchus contortus is a hematophagous nematoda of small ruminants, extremely important on a worldwide scale, being responsible for severe impact to the animals health, besides productive and economic losses to the breeders. The current panorama is parasite resistance to the most anthelminthics classes commercially available for its control. This demonstrates the urgent need of alternative control methods. Therefore, the goals of this study were to select mimotopes from H. contortus and to evaluate the use of RNA interference (RNAi) to silence the gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of the parasite. This thesis is divided into 3 chapters presented as 1 general introduction and 2 manuscripts. The goal of the first manuscript was to evaluate polyclonal antibody to map H. contortus mimetopos using the phage display library. Clone 6 - GAPDH mimetope and clone 17 - mimetope from the disorganized muscle family (Dim 1) were selected for sheep an immunization assay. Peptide mimotopes (synthetic) were evaluated as antigenic molecules by enzyme-linked immunosorbent assay (ELISA). In the immunization assay, IgG titers anti-phage M13 showed increase, but IgG anti-peptide mimotopes did not increased. Peptide mimotopes (synthetic) were recognized by IgG from sheep naturally infected by H. contortus. This is the first report of successful use of the phage display library for identification of H. contortus mimotopes. The aim of the manuscript 2 was to evaluate the RNAi to silence the GAPDH gene of H. contortus. Infective larvae were incubated with RNAi for 3; 6; 24 and 48h. The results showed absence of gene transcription in all evaluated periods. This study reports for the first time the silencing of GAPDH gene in H. contortus, confirming the gene as susceptible to silencing by RNAi. The data presented in this thesis point out promising results for use in new researches to development H. contortus control therapies.

Efficient RNAi-induced protein knockdown in somatic cells using diced or chemically produced small interfering RNAs (siRNA)

Background: RNA interference (RNAi) is a post-transcriptional gene silencing process in which double-stranded RNA (dsRNA) directs the degradation of a specific corresponding target mRNA. The mediators of this process are small dsRNAs of approximately 21 to 23 bp in length, called small interfering RNAs (siRNAs), which can be prepared in vitro and used to direct the degradation of specific mRNAs inside cells. Hence, siRNAs represent a powerful tool to study and control gene and cell function. Rapid progress has been made in the use of siRNA as a means to attenuate the expression of any protein for which the cDNA sequence is known. Individual siRNAs can be chemically synthesized, in vitro-transcribed, or expressed in cells from siRNA expression vectors. However, screening for the most efficient siRNAs for post-transcriptional gene silencing in cells in culture is a laborious and expensive process. In this study, the effectiveness of two siRNA production strategies for the attenuation of abundant proteins for DNA repair were compared in human cells: (a) the in vitro production of siRNA mixtures by the Dicer enzyme (Diced siRNAs); and (b) the chemical synthesis of very specific and unique siRNA sequences (Stealth RNaiTM). Materials, Methods & Results: For in vitro-produced siRNAs, two segments of the human Ku70 (167 bp in exon 5; and 249 bp in exon 13; NM001469) and Xrcc4 (172 bp in exon 2; and 108 bp in exon 6; NM003401) genes were chosen to generate dsRNA for subsequent "Dicing" to create mixtures of siRNAs. The Diced fragments of siRNA for each gene sequence were pooled and stored at -80ºC. Alternatively, chemically synthesized Stealth siRNAs were designed and generated to match two very specific gene sequence regions for each target gene of interest (Ku70 and Xrcc4). HCT116 cells were plated at 30% confluence in 24- or 6-well culture plates. The next day, cells were transfected by lipofection with either Diced or Stealth siRNAs for Ku70 or Xrcc4, in duplicate, at various doses, with blank and sham transfections used as controls. Cells were harvested at 0, 24, 48, 72 and 96 h post-transfection for protein determination. The knockdown of specific targeted gene products was quantified by Western blot using GAPDH as control. Transfection of gene-specific siRNA to either Ku70 or Xrcc4 with both Diced and Stealth siRNAs resulted in a down regulation of the targeted proteins to approximately 10 to 20% of control levels 48 h after transfection, with recovery to pre-treatment levels by 96 h. Discussion: By transfecting cells with Diced or chemically synthesized Stealth siRNAs, Ku70 and Xrcc4, two highly expressed proteins in cells, were effectively attenuated, demonstrating the great potential for the use of both siRNA production strategies as tools to perform loss of function experiments in mammalian cells. In fact, down-regulation of Ku70 and Xrcc4 has been shown to reduce the activity of the non-homologous end joining DNA pathway, a very desirable approach for the use of homologous recombination technology for gene targeting or knockout studies. Stealth RNAiTM was developed to achieve high specificity and greater stability when compared with mixtures of enzymatically-produced (Diced) siRNA fragments. In this study, both siRNA approaches inhibited the expression of Ku70 and Xrcc4 gene products, with no detectable toxic effects to the cells in culture. However, similar knockdown effects using Diced siRNAs were only attained at concentrations 10-fold higher than with Stealth siRNAs. The application of RNAi technology will expand and continue to provide new insights into gene regulation and as potential applications for new therapies, transgenic animal production and basic research.

Efficient RNAi-induced protein knockdown in somatic cells using diced or chemically produced small interfering RNAs (siRNA) / Efficient RNAi-induced protein knockdown in somatic cells using diced or chemically produced small interfering RNAs (siRNA)

Background: RNA interference (RNAi) is a post-transcriptional gene silencing process in which double-stranded RNA (dsRNA) directs the degradation of a specifi c corresponding target mRNA. The mediators of this process are small dsRNAs of approximately 21 to 23 bp in length, called small interfering RNAs (siRNAs), which can be prepared in vitro and used to direct the degradation of specifi c mRNAs inside cells. Hence, siRNAs represent a powerful tool to study and control gene and cell function. Rapid progress has been made in the use of siRNA as a means to attenuate the expression of any protein for which the cDNA sequence is known. Individual siRNAs can be chemically synthesized, in vitro-transcribed, or expressed in cells from siRNA expression vectors. However, screening for the most effi cient siRNAs for post-transcriptional gene silencing in cells in culture is a laborious and expensive process. In this study, the effectiveness of two siRNA production strategies for the attenuation of abundant proteins for DNA repair were compared in human cells: (a) the in vitro production of siRNA mixtures by the Dicer enzyme (Diced siRNAs); and (b) the chemical synthesis of very specifi c and unique siRNA sequences (Stealth RNaiTM).Dicing to create mixtures of siRNAs. The Diced fragments of siRNA for each gene sequence were pooled and stored at -80o C. Alternatively, chemically synthesiz

Background: RNA interference (RNAi) is a post-transcriptional gene silencing process in which double-stranded RNA (dsRNA) directs the degradation of a specifi c corresponding target mRNA. The mediators of this process are small dsRNAs of approximately 21 to 23 bp in length, called small interfering RNAs (siRNAs), which can be prepared in vitro and used to direct the degradation of specifi c mRNAs inside cells. Hence, siRNAs represent a powerful tool to study and control gene and cell function. Rapid progress has been made in the use of siRNA as a means to attenuate the expression of any protein for which the cDNA sequence is known. Individual siRNAs can be chemically synthesized, in vitro-transcribed, or expressed in cells from siRNA expression vectors. However, screening for the most effi cient siRNAs for post-transcriptional gene silencing in cells in culture is a laborious and expensive process. In this study, the effectiveness of two siRNA production strategies for the attenuation of abundant proteins for DNA repair were compared in human cells: (a) the in vitro production of siRNA mixtures by the Dicer enzyme (Diced siRNAs); and (b) the chemical synthesis of very specifi c and unique siRNA sequences (Stealth RNaiTM).Materials, Methods & Results: For in vitro-produced siRNAs, two segments of the human Ku70 (167 bp in exon 5; and 249 bp in exon 13; NM001469) and Xrcc4 (1

Detecção de Proteus mirabilis nas fezes de camundongos spf / /Detection of Proteus mirabilis in specifc pathogen free mice feces

O presente estudo teve como objetivo apresentar os resultados de pesquisa de positividade do microorganismo Proteus mirabilis em camundongos das linhagens C57BL/6, BALB/c e BALB/c Nude, originários do setor de criação de camundongos livres de patógenos específicos (SPF) do Centro de Bioterismo da Faculdade de Medicina da USP e relatar a importância do isolamento deste microorganismo em colônias de animais. Para tanto, amostras de fezes de 71 camundongos da linhagem C57BL/6, 48 camundongos da linhagem BALB/c e 26 camundongos da linhagem BALB/c Nude, foram semeadas em meios específicos para bactérias gram negativas e diagnosticado no Enterokit B da Probac do Brasil. Nossos resultados mostraram que, as linhagens de camundongos C57BL/6, BALB/c e BALB/c Nude apresentaram P.mirabilis nas fezes, nas porcentagens de 80,3%, 62,5% e 73,1% respectivamente, sendo inadequadas para utilização em protocolos experimentais das áreas de radiobiologia, imunologia, oncologia e cirurgias experimentais invasivas, por interferirem potencialmente nos resultados destas áreas. CEUA nº 251/11.

This study aimed to present the positive results of Proteus mirabilis research from strains inbred mice C57BL/6, BALB/c and BALB/c Nude, originating from the sector of Specific Pathogen Free (SPF) mice - Animal Facilities of Faculdade de Medicina (USP – University of São Paulo) and report the importance of isolation of this microorganism in colonies of animals. For this, stool samples from: 71 C57BL/6, 48 BALB/c and 26 BALB /c Nude inbred mice, were inoculated in cultural bacteria specific media and diagnosed in the B Enterokit Probac-Brasil. Our results showed that the strains of C57BL/6, BALB/c and BALB/c Nude presented P.mirabilis in stool, percentages of 80.3%, 62.5% and 73.1% respectively, being unsuitable for use in experimental protocols in the fields of radiobiology, immunology, oncology, and experimental invasive surgeries by interfering mainly on the results of these areas. CEUA No. 251/11.