New opportunities in plant microbiome engineering for increasing agricultural sustainability under stressful conditions.

Plant microbiome (or phytomicrobiome) engineering (PME) is an anticipated untapped alternative strategy that could be exploited for plant growth, health and productivity under different environmental conditions. It has been proven that the phytomicrobiome has crucial contributions to plant health, pathogen control and tolerance under drastic environmental (a)biotic constraints. Consistent with plant health and safety, in this article we address the fundamental role of plant microbiome and its insights in plant health and productivity. We also explore the potential of plant microbiome under environmental restrictions and the proposition of improving microbial functions that can be supportive for better plant growth and production. Understanding the crucial role of plant associated microbial communities, we propose how the associated microbial actions could be enhanced to improve plant growth-promoting mechanisms, with a particular emphasis on plant beneficial fungi. Additionally, we suggest the possible plant strategies to adapt to a harsh environment by manipulating plant microbiomes. However, our current understanding of the microbiome is still in its infancy, and the major perturbations, such as anthropocentric actions, are not fully understood. Therefore, this work highlights the importance of manipulating the beneficial plant microbiome to create more sustainable agriculture, particularly under different environmental stressors.

Effects of different acetylsalicylic acid doses on body organs, histopathology, and serum biochemical parameters in broiler birds / Efeito de diferentes doses de ácido acetil salicílico nos orgãos corpóreos, histopalogia e parâmetros bioquímicos sorológicos em frangos de corte

The poultry industry suffers from various diseases or stresses. In poultry, apart from being antipyretic, acetylsalicylic acid (ASA) is widely used to cope with different issues including heat stress, and respiratory and digestive orders. This study evaluated the beneficial and toxic effects of ASA at different dose levels in broiler birds. To evaluate these toxic and beneficial effects it is necessary to examine the physical and serum biochemical parameters as well as the histopathological changes with tissue sections taken from broiler birds under experimental trial. This study was conducted on 60 one-day-old broiler chicks purchased from a local market in Faisalabad. Chicks were reared for the first 14 days under similar conditions. On the 15th day, birds were randomly divided into 4 groups (1-4) with 15 birds in each group. Group 4 was kept as control, while groups 1, 2, and 3 were treated with ASA at the dose of 300, 600, and 1200 mg/L of drinking water for 21 days. There were 3 samplings performed at 21, 28, and 35 days post-treatment. The live body weight and carcass weight were noted on each sampling. All the visceral organs were recorded for gross pathological changes. The serum samples were collected for biochemical evaluation. Histopathology of all the visceral organs was performed to observe the microscopic changes. A significant (P<0.05) increase in live body weight at a 300mg/L dose was noted after the first 2 samplings. A significant (P<0.05) increase in the relative organ weight was recorded at 1200 mg/L. The groups treated with ASA 600 and 1200 mg/L showed increased (P<0.05) AST, ALT, and creatinine levels from that of the control group. The group treated with 1200 mg/L of ASA showed increased (P<0.05) urea, serum total protein, and albumin level in all the samplings. Histopathological changes revealed swollen hepatocytes, increased sinusoidal spaces in the liver, congestion and abnormal glomerular spaces in the kidney, congestion and alveolar disruption in the lungs, and generation of villi and cellular degeneration in the intestine in a high-dose group. The study concluded that ASA at a low dose can be used for a long time in broilers and has a growth promontory role, while high-level doses cause hepatorenal toxicity.(AU)

A indústria avícola é afetada por diversas doenças ou estresses. Particularmente devido às aves serem antipiréticas. O ácido acetil salicílico (AAS) é largamente utilizado com diferentes objetivos que incluem o controle do estresse calórico, bem como a atividade respiratória e digestiva. O propósito deste estudo foi a avaliação dos efeitos benéficos e tóxicos do emprego de diferentes dosagens do AAS em frangos de corte. As variáveis analisadas foram: exame físico, parâmetros bioquímicos, bem como as alterações histopatológica em seções de tecidos colhidas das aves em um ensaio experimental. O estudo foi conduzido em 60 frangos de corte com um dia de idade adquiridos em um mercado local de Faisalabad que foram recriados nos primeiros 14 dias em idênticas condições. Então no 15º dia as aves foram distribuídas randomicamente em quatro grupos identificados pelos números 1 a 4, com 15 aves em cada grupo. O grupo 4 foi mantido como grupo controle e os grupos 1, 2 e 3 foram tratados com AAS, respectivamente, nas doses de 300, 600 e 1200 mg/L de água de bebida, durante 21 dias. Foram realizadas três amostragens nos dias 21, 28 e 35 pós-tratamento. O peso vivo corpóreo e da carcaça foi registrado em cada amostragem. Em todos os órgãos viscerais foi analisada a presença de alterações patológicas. As amostras de soro sanguíneo foram colhidas para a avaliação bioquímica. O exame histopatológico de todos os órgãos viscerais foi realizado para a observação de alterações microscópicas. A partir da segunda amostragem foi observado um aumento significante (p<0,05) no peso corpóreo na dosagem de 300mg/L. Um significante aumento no peso relativo dos órgãos foi registrado na dosagem de 1200 mg/L. Os grupos tratados com dosagens de 600 e 1200 mg/L de AAS, apresentaram aumento significante dos níveis de AST, ALT e de creatinina quando comparados ao grupo controle. O grupo tratado com 1200 mg/L de AAS apresentou um aumento significante (P,0,05) de uréia, proteína sérica total e de albumina em todas as amostragens. No grupo de alta dosagem foram observados alterações histopatológicas constituídas por aumento dos hepatócitos, aumento dos espaços sinusoidais no fígado, congestão e anormalidades nos espaços glomerulares nos rins, congestão e ruptura alveolar nos pulmões, degeneração das vilosidades e celular nos intestinos A conclusão obtida foi que em frangos de corte uma baixa dosagem do AAS pode ser utilizada por um período de longa duração, apresentando um efeito promotor do crescimento, contudo as doses elevadas determinam toxicidade hepática e renal.(AU)

Biosynthesis of Graphene and Investigation of Antibacterial Activity of Graphene-parthenium hysterophorous Nanocomposite

Abstract There is a great interest to use carbon-based material like graphene and graphene oxide in biomedical applications due to its flexibility to be functionalized with bio-active molecules. Herein, graphene and graphene-based nanocomposites were biosynthesized by liquid-phase exfoliation of graphite using aqueous extract of Parthenium hysterophorous (P-H) as a surfactant. A set of five thin film samples of graphene was prepared from graphene suspension by vacuum filtration method. Samples were characterized by UV-vis spectroscopy, Raman spectroscopy, SEM, and XRD, which revealed successful synthesis of graphene. Graphene/P-H(G/P-H) nanocomposites comprising varied ratios of graphene and P-H were prepared and their antibacterial activity was investigated by agar well diffusion method. The experimental results indicated that G/P-H nanocomposite have higher antibacterial activity than graphene alone, and bioactivity of G/P-H nanocomposite was found to be controlled by the fraction of graphene in the composite.

Tillage systems and nitrogen timings effect on growth, stay green and grain quality in maize (Zea mays L) / Sistemas de preparo do solo e efeitos do tempo de nitrogênio no crescimento, manutenção da qualidade de grãos e verde no milho (Zea mays L)

Nitrogen (N), being mobile in soil is exposed to various losses owing to unwise use of nitrogen fertilizer, and conventional soil and crop management practices which can be minimized by temporal nitrogen application and different tillage practices. This study was conducted to elucidate the effect of different tillage systems and nitrogen timings on growth, stay green and grain quality in maize. Three tillage systems viz. T1: tillage with cultivator, T2: mouldboard plough + 2-cultivations, T3: chisel plough + 2-cultivations; and five nitrogen timings viz. N1: whole at sowing, N2: ½ at sowing+½ at V5 (5-leaf stage), N3: ½ at sowing+½ at tasseling, N4: ½ at V5+½ at tasseling, N5: 1/3 at sowing+1/3 at V5+1/3 at tasseling). Tillage systems and nitrogen application had significant effect on leaf area per plant, specific leaf area and leaf area ratio. Tillage systems had non-significant effect on stay green and grain quality parameters except for oil contents. However, nitrogen timings had significant effect on chlorophyll a, b and total contents as well as grain quality parameters. The higher a, b and total chlorophyll contents were noted with three splits i.e. 1/3 at sowing+1/3 at V5+1/3 at tasseling compared with other treatments. The results suggest to grow maize by preparing the field through chisel plough and applying N in three splits to improve its growth, chlorophyll contents and grain quality.

Nitrogênio (N), sendo móvel no solo, está exposto a várias perdas devido ao uso imprudente de fertilizantes nitrogenados e práticas convencionais de manejo do solo e das culturas, que podem ser minimizadas pela aplicação de nitrogênio temporal e diferentes práticas de preparo do solo. Este estudo foi realizado para elucidar o efeito de diferentes sistemas de preparo do solo e tempos de nitrogênio no crescimento, permanência e qualidade do grão verde no milho. Três sistemas de lavoura, viz. T1: preparo do solo com cultivador, T2: arado de aiveca + 2-cultivos, T3: escarificador + 2-cultivos; e cinco temporizações de azoto, viz. N1: todo na sementeira, N2: ½ na sementeira + ½ na V5 (fase de 5 folhas), N3: ½ na sementeira + ½ no pendoamento, N4: ½ na V5 + ½ na pompa, N5: 1/3 na sementeira + 1/3 em V5 + 1/3 no pendoamento). Sistemas de preparo do solo e aplicação de nitrogênio tiveram efeito significativo na área foliar por planta, área foliar específica e área foliar. Os sistemas de preparo do solo não tiveram efeito significativo na manutenção dos parâmetros de qualidade verde e de grãos, exceto para os teores de óleo. Entretanto, os tempos de nitrogênio tiveram efeito significativo sobre os teores de clorofila a, b e total, bem como parâmetros de qualidade de grãos. Os maiores teores de clorofila a, b e total foram observados com três fendas, isto é, 1/3 na semeadura + 1/3 em V5 + 1/3 no pendoamento comparado com outros tratamentos. Os resultados sugerem o cultivo do milho preparando o campo através do arado de cinzel e aplicando o N em três fendas para melhorar o seu crescimento, o teor de clorofila e a qualidade do grão.

Tillage systems and nitrogen timings effect on growth, stay green and grain quality in maize (Zea mays L) / Sistemas de preparo do solo e efeitos do tempo de nitrogênio no crescimento, manutenção da qualidade de grãos e verde no milho (Zea mays L)

Nitrogen (N), being mobile in soil is exposed to various losses owing to unwise use of nitrogen fertilizer, and conventional soil and crop management practices which can be minimized by temporal nitrogen application and different tillage practices. This study was conducted to elucidate the effect of different tillage systems and nitrogen timings on growth, stay green and grain quality in maize. Three tillage systems viz. T1: tillage with cultivator, T2: mouldboard plough + 2-cultivations, T3: chisel plough + 2-cultivations; and five nitrogen timings viz. N1: whole at sowing, N2: ½ at sowing+½ at V5 (5-leaf stage), N3: ½ at sowing+½ at tasseling, N4: ½ at V5+½ at tasseling, N5: 1/3 at sowing+1/3 at V5+1/3 at tasseling). Tillage systems and nitrogen application had significant effect on leaf area per plant, specific leaf area and leaf area ratio. Tillage systems had non-significant effect on stay green and grain quality parameters except for oil contents. However, nitrogen timings had significant effect on chlorophyll a, b and total contents as well as grain quality parameters. The higher a, b and total chlorophyll contents were noted with three splits i.e. 1/3 at sowing+1/3 at V5+1/3 at tasseling compared with other treatments. The results suggest to grow maize by preparing the field through chisel plough and applying N in three splits to improve its growth, chlorophyll contents and grain quality. (AU)

Nitrogênio (N), sendo móvel no solo, está exposto a várias perdas devido ao uso imprudente de fertilizantes nitrogenados e práticas convencionais de manejo do solo e das culturas, que podem ser minimizadas pela aplicação de nitrogênio temporal e diferentes práticas de preparo do solo. Este estudo foi realizado para elucidar o efeito de diferentes sistemas de preparo do solo e tempos de nitrogênio no crescimento, permanência e qualidade do grão verde no milho. Três sistemas de lavoura, viz. T1: preparo do solo com cultivador, T2: arado de aiveca + 2-cultivos, T3: escarificador + 2-cultivos; e cinco temporizações de azoto, viz. N1: todo na sementeira, N2: ½ na sementeira + ½ na V5 (fase de 5 folhas), N3: ½ na sementeira + ½ no pendoamento, N4: ½ na V5 + ½ na pompa, N5: 1/3 na sementeira + 1/3 em V5 + 1/3 no pendoamento). Sistemas de preparo do solo e aplicação de nitrogênio tiveram efeito significativo na área foliar por planta, área foliar específica e área foliar. Os sistemas de preparo do solo não tiveram efeito significativo na manutenção dos parâmetros de qualidade verde e de grãos, exceto para os teores de óleo. Entretanto, os tempos de nitrogênio tiveram efeito significativo sobre os teores de clorofila a, b e total, bem como parâmetros de qualidade de grãos. Os maiores teores de clorofila a, b e total foram observados com três fendas, isto é, 1/3 na semeadura + 1/3 em V5 + 1/3 no pendoamento comparado com outros tratamentos. Os resultados sugerem o cultivo do milho preparando o campo através do arado de cinzel e aplicando o N em três fendas para melhorar o seu crescimento, o teor de clorofila e a qualidade do grão.(AU)

Assessments on the catalytic and kinetic properties of beta-glucosidase isolated from a highly efficient antagonistic fungus Trichoderma harzianum / Avaliações das propriedades catalíticas e cinéticas da beta-glucosidase isoladas de um fungo antagonísta altamente eficiente Trichoderma harzianum

Due to the toxicity and inefficiency of chemical fungicides to control infestation of Macrophomina phaseolina (Tassi) Goid which causes charcoal rot in plants, a biotechnological approach using - glucosidase (EC.3.2.1) as the alternative bioactive ingredient in fungicide is hereby, proposed. The extracellular enzyme was isolated from a highly efficient fungal antagonist, Trichoderma harzianum T12. The highly similar molecular masses obtained using SDS-PAGE (96 kDa) and MALDI-TOF mass spectrometry (98.3 kDa) affirmed that the -glucosidase was purified to homogeneity. Consequently, optimum catalytic parameters that rendered the highest enzyme activity were found to be: 45°C, pH 7, inoculum size of 10 % (w/v), supplementation with metal ions Zn2+ and Mn2+ ions, and Tween 80. Addition of wheat bran and (NH4)2SO4 as carbon and nitrogen sources also improved enzyme activity. BLASTn showed the sequence of -glucosidase T12 was highly identical to other -glucosidases viz. T. harzianum strain IOC-3844 (99%), T. gamsii and T. virens bgl1 (86 %) as well as T. reesei strain SJVTR and T. viride strain AS 3.3711 (84 %). Kinetic assessment showed that -glucosidase T12 catalyzes hydrolytic activity is characterized by a Km of 0.79 mM and Vmax of 8.45 mM min-1 mg-1 protein, with a corresponding kcat of 10.69 s-1.

Devido à toxicidade e ineficiência dos fungicidas químicos para controlar a infestação de Macrophomina phaseolina (Tassi) Goid que causa o apodrecimento das plantas, uma abordagem biotecnológica usando - glicosidase (EC.3.2.1) como o ingrediente bioativo alternativo do fungicida é por este meio, proposto. A enzima extracelular foi isolada de um antagonista fúngico altamente eficiente, o Trichoderma harzianum T12. As massas moleculares altamente similares obtidas usando SDS-PAGE (96 kDa) e espectrometria de massa MALDI-TOF (98,3 kDa) afirmaram que a -glicosidase foi purificada até a homogeneidade. Consequentemente, os parâmetros catalíticos ótimos que apresentaram a maior atividade enzimática foram: 45°C, pH 7, tamanho do inóculo de 10% (p / v), suplementação com íons de metais Zn2+ e Mn2+, e Tween 80. Adição de farelo de trigo e (NH4) 2SO4 como fontes de carbono e nitrogênio também melhoraram a atividade enzimática. O BLASTn mostrou que a sequência da -glicosidase T12 era altamente idêntica a outras -glicosidase viz. A estirpe T. harzianum IOC-3844 (99%), T. gamsii e T. virens bgl1 (86%) assim como a estirpe T. reesei SJVTR e a estirpe T. viride AS 3.3711 (84%). A avaliação cinética mostrou que -glicosidase T12 catalisa a actividade hidrolítica caracterizada por um Km de 0,79 mM e Vmax de 8,45 mM min-1 mg-1 de proteína, com um correspondente kcat de 10,69 s-1.

Characterization Of A Novel Hydrolytic Enzyme Producing Thermophilic Bacterium Isolated From The Hot Spring Of Azad Kashmir-Pakistan

ABSTRACT A thermophilic bacterium (TP-2) was isolated from the Tatta Pani hot spring in Azad Kashmir and was characterized using phenotypic and genotypic characters. The strain developed cream colored, round, smooth, flat and slimy colonies while the cells were Gram positive rods that ranged in size from about 2.1-3.6 μm to 0.2-0.3 μm in width. Sequence analysis of its 16S rRNA gene showed that isolate TP-2 had 89% homology with Geobacillus debilis. It grew within pH range of 5.5 to 8.5 with optimum growth at pH 7.0. The isolate showed optimum growth at 65ºC and gave positive results for gelatin hydrolysis (GEL), ortho nitrophenyl-β-D-galactopyranosidase (ONPG), and nitrate production and produced acid from sucrose, glucose and maltose. It utilized glucose, fructose, maltose, lactose, sucrose, xylan, starch, filter paper and carboxymethylcellulose as sole carbon source. Isolate TP-2 produced significant amount of industrially important enzymes i.e. extracellular α-amylase, CMCase, FPase, Xylanase, Protease and Lipase and intracellular CMCase and FPase.

Inhibitors of ORAI1 Prevent Cytosolic Calcium-Associated Injury of Human Pancreatic Acinar Cells and Acute Pancreatitis in 3 Mouse Models.

BACKGROUND & AIMS: Sustained activation of the cytosolic calcium concentration induces injury to pancreatic acinar cells and necrosis. The calcium release-activated calcium modulator ORAI1 is the most abundant Ca(2+) entry channel in pancreatic acinar cells; it sustains calcium overload in mice exposed to toxins that induce pancreatitis. We investigated the roles of ORAI1 in pancreatic acinar cell injury and the development of acute pancreatitis in mice. METHODS: Mouse and human acinar cells, as well as HEK 293 cells transfected to express human ORAI1 with human stromal interaction molecule 1, were hyperstimulated or incubated with human bile acid, thapsigargin, or cyclopiazonic acid to induce calcium entry. GSK-7975A or CM_128 were added to some cells, which were analyzed by confocal and video microscopy and patch clamp recordings. Acute pancreatitis was induced in C57BL/6J mice by ductal injection of taurolithocholic acid 3-sulfate or intravenous' administration of cerulein or ethanol and palmitoleic acid. Some mice then were given GSK-7975A or CM_128, which inhibit ORAI1, at different time points to assess local and systemic effects. RESULTS: GSK-7975A and CM_128 each separately inhibited toxin-induced activation of ORAI1 and/or activation of Ca(2+) currents after Ca(2+) release, in a concentration-dependent manner, in mouse and human pancreatic acinar cells (inhibition >90% of the levels observed in control cells). The ORAI1 inhibitors also prevented activation of the necrotic cell death pathway in mouse and human pancreatic acinar cells. GSK-7975A and CM_128 each inhibited all local and systemic features of acute pancreatitis in all 3 models, in dose- and time-dependent manners. The agents were significantly more effective, in a range of parameters, when given at 1 vs 6 hours after induction of pancreatitis. CONCLUSIONS: Cytosolic calcium overload, mediated via ORAI1, contributes to the pathogenesis of acute pancreatitis. ORAI1 inhibitors might be developed for the treatment of patients with pancreatitis.