Silver-nanoparticle-modified nanocellulose synthesized by pyroligneous acid: cytotoxicity towards HaCat cells.

In the present study, pyroligneous acid, also known as wood vinegar, has been employed as reducing and stabilizing agent in the synthesis of silver nanoparticles (AgNPs) anchored on nanocellulose (NC). The idea is to confer the latter bactericidal properties for its typical uses such as in cosmetics and food-packing. It has been demonstrated that AgNPs can be directly produced onto NC in one-pot fashion while dramatically enhancing the kinetics of AgNPs synthesis (2 h for reaction completion) in comparison to the NC-less counterpart (10 days for reaction completion). Furthermore, NC allowed for a narrower size distribution of AgNPs. NC-supported and non-supported AgNPs had sizes of 5.1 ± 1.6 nm and 16.7 ± 4.62 nm, respectively. Immortalized human keratinocytes (HaCat) cells were then employed as model to evaluate the cytotoxicity of the AgNPs-NC compound. The latter was found not to impact cell proliferation at any formulation, while decreasing the viability by only 6.8% after 72 h. This study contributes to the development of more environmentally benign routes to produce nanomaterials and to the understanding of their impact on cells.

The Potential of Wood Vinegar to Replace Antimicrobials Used in Animal Husbandry-A Review.

The indiscriminate use of antimicrobials in animal husbandry can result in various types of environmental contamination. Part of the dose of these products is excreted, still active, in the animals' feces and urine. These excreta are widely used as organic fertilizers, which results in contamination with antimicrobial molecules. The impacts can occur in several compartments, such as soil, groundwater, and surface watercourses. Also, contamination by antimicrobials fed or administrated to pigs, chickens, and cattle can reach the meat, milk, and other animal products, which calls into question the sustainability of using these products as part of eco-friendly practices. Therefore, a search for alternative natural products is required to replace the conventional antimicrobials currently used in animal husbandry, aiming to mitigate environmental contamination. We thus carried out a review addressing this issue, highlighting wood vinegar (WV), also known as pyroligneous acid, as an alternative antimicrobial with good potential to replace conventional products. In this regard, many studies have demonstrated that WV is a promising product. WV is a nontoxic additive widely employed in the food industry to impart a smoked flavor to foods. Studies have shown that, depending on the WV concentration, good results can be achieved using it as an antimicrobial against pathogenic bacteria and fungi and a valuable growth promoter for poultry and pigs.

Antimicrobial activity and chemical profile of wood vinegar from eucalyptus (Eucalyptus urophylla x Eucalyptus grandis - clone I144) and bamboo (Bambusa vulgaris).

Microbial resistance to drugs is a public health problem; therefore, there is a search for alternatives to replace conventional products with natural agents. One of the potential antimicrobial agents is wood vinegar derived from the carbonization of lignocellulosic raw materials. The objectives of the present work were to evaluate the antibacterial and antifungal action of two kinds of wood vinegar (WV), one of Eucalyptus urograndis wood and another of Bambusa vulgaris biomass, and determine their chemical profile. The antimicrobial effect was assessed against Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enteritidis, Escherichia coli, Streptococcus agalactiae, and Candida albicans. The minimum inhibitory concentration and the minimum bactericidal and fungicidal concentrations were determined. Micrographs of the microorganisms before and after exposure to both kinds of wood vinegar were obtained by scanning electron microscopy. The chemical profile of the eucalyptus and bamboo vinegar was carried out by gas chromatography and mass spectrometry (GC/MS). Both types of WV presented significant antimicrobial activity, with the bamboo one having a higher efficiency. Both studied pyroligneous extracts seem promising for developing natural antimicrobials due to their efficiency against pathogens. GC/MS analyses demonstrated that the chemical profiles of both kinds of WV were similar but with some significant differences. The major component of the eucalyptus vinegar was furfural (17.2%), while the bamboo WV was phenol (15.3%). Several compounds in both WVs have proven antimicrobial activity, such as acetic acid, furfural, phenol, cresols, guaiacol, and xylenols. Together, they are the major in the chemical composition of the organic fraction of both WVs. Bamboo vinegar had a more expressive content of organic acids. Micrographs of microorganisms taken after exposure to both kinds of wood vinegar displayed several cell modifications. The potential of both types of wood vinegar as a basis for natural antimicrobial products seems feasible due to their proven effect on inhibiting the microorganisms' growth assessed in this experiment.

Understanding the multifunctionality of pyroligneous acid from waste biomass and the potential applications in agriculture.

Efforts have been directed to the development of environmentally friendly processes and manufacturing of green products, use of renewable energy and more sustainable agricultural practices. Pyroligneous acid (PA) is a byproduct of biomass pyrolysis that consists of a complex mixture of bioactive substances. The complexity and richness of PA composition have opened a window for PA application in agriculture and mitigation of environmental pollution. This review brings a brief historical on the use of PA and regulatory policies adopted in Brazil, China, Japan and Thailand for PA application in agriculture. The composition and stability of PAs of several origins are presented, together with a discussion of the use of PA to boost plant growth and crop productivity, remove toxic metals from soil, inhibit soil ureases, mitigate the emission of greenhouse gases, control phytopathogen proliferation and weed dissemination. A great variety of biomass types are reported as feedstock to produce PA with distinct chemically diverse and active substances at wide-ranging concentrations. PA has been shown to successfully improve farming practices in a more sustainable fashion. The disclosure of the mechanisms of action that drive the PA's effects, together with the pursue of safety and efficacy data in a case-by-case way to address toxicity and shelf stability, will be valuable to expand the use of PA worldwide for food production.

Dual action of pyroligneous acid in the eco-friendly synthesis of bactericidal silver nanoparticles.

In the present study, we demonstrate that pyroligneous acid (PA), also known as wood vinegar, functions efficiently as both reducing and stabilizing agent in the synthesis of silver nanoparticles (AgNPs). The synthesis and stabilization of AgNPs take place in the following fashion: 1) in alkaline environment, oxygenated species (phenols in the present case) contained in PA reduce silver ions to metallic silver; 2) acetic acid, abundantly present in PA, adsorb onto the AgNPs conferring electrostatic stabilization. This mechanism is supported by GC-MS and RAMAN analysis, with the former revealing the compounds lacking in PA after nanoparticle synthesis and the latter demonstrating acetic acid adsorbed on the nanoparticles. The AgNPs produced via this method were quite stable up to 150 days (zeta potential = -56 mV). The AgNPs were then found to inhibit the growth of Escherichia coli and Staphylococcus aureus. Concerning PA, we showed that it displays bactericidal properties only under acidic conditions. This study contributes to the development of more environmentally benign routes to produce nanomaterials.

Pyroligneous acid and antibacterial activity: criticism of a paper by Araújo et al. (2018).

AIMS: A paper by Araújo et al. (2018) claims that a variety of compounds present in pyroligneous acid (guaiacol, phenols and furfural) are responsible for the observed antimicrobial activity. We show, on the other hand, that the low pH due to acetic acid present in pyroligneous acid is the real cause of the activity. METHODS AND RESULTS: Pyroligneous acid (PA) was synthesized according to a previous method (Medeiros et al. 2019) with its inhibition activity tested on Escherichia coli and Staphylococcus aureus via the agar diffusion method. The activity of acetic acid at different concentrations was also evaluated for comparison. As expected, crude PA (pH 3.0) and acetic acid produced inhibition halos whose diameters varied according to their employed concentration. However, any PA inhibitory activity completely vanished upon neutralization (pH 7.0), a behaviour also observed for neutralized acetic acid. CONCLUSIONS: The claim that guaiacol, phenols and furfural are responsible for any inhibitory activity is unsubstantiated. The authors should have neutralized the pyroligneous acid if any activity was to be discovered. SIGNIFICANCE AND IMPACT OF STUDY: To increase awareness that interfering species may play a detrimental role on the interpretation of results. In this case, the action of acetic acid is vastly more important for the inhibitory activity than any other compound present in PA.

Pyroligneous acid from Mimosa tenuiflora and Eucalyptus urograndis as an antimicrobial in dairy goats.

AIMS: To evaluate the applicability of the Mimosa tenuiflora and Eucalyptus urograndis pyroligneous acids (PAs) as alternative antiseptics in dairy goats. METHODS AND RESULTS: Cytotoxicity was evaluated in vitro using bacteria, as well as in vivo using goats, and the influence of PAs on the physicochemical parameters of fresh milk were examined. The cytotoxicity of PAs was evaluated in terms of morphology, cell viability and metabolic activity of goat tegumentary cells. The PA of M. tenuiflora had results similar to those of 2% iodine. For the in vitro tests, strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were used with the well technique, demonstrating inhibition halos greater than 9 mm. In the in vivo test, 15 animals were used per phase of the experiment, and the plate counting technique showed that there was antiseptic action of both extracts, with emphasis on the M. tenuiflora PA. Physicochemical analysis of the milk showed that neither PAs interfered with its physical-chemical parameters. CONCLUSIONS: The PA of M. tenuiflora presented potential as an alternative antiseptic in dairy goats. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the use of PA as an antimicrobial agent in animals.

Chemical Characterization and Antiradical Properties of Pyroligneous Acid from a Preserved Bamboo, Guadua angustifolia Kunth

Abstract Pyroligneous acid (PA) was obtained by condensation of the vapors produced in the thermal decomposition of culms residues from Guadua angustifolia Kunth (G. angustifolia) cultivated in Colombia, with and without previous preservation treatment with borax salts. Chemical characterization by GC-MS showed that PA extracts has high content of phenolic compounds. Mequinol, isocreosol, 4-ethylphenol, 4-ethyl-2-methoxyphenol, 3,5-dimethoxy-4-hydroxytoluene and 2,6-dimethoxyphenol were the most abundant substances, identified. The total phenolic content (TPC) and DPPH free radical scavenging activity, were investigated. TPC showed a concentration of 1.959 mg GA g-1±0.010 and 3.844 mg GA g-1±0.027 to PAC and PAS samples. These samples also exhibited high DPPH activity of 70.975%±0.921 and, 16.667%±0.298, respectively. The chemical composition, TPC and DPPH results indicate that the PA extracts obtained from G. angustifolia may be used as a raw material in the food industry as natural preservative, in medicine as alternative to antibiotics and in agriculture as insect repellent and foliar fertilizer.

Evaluation of pyroligneous acid as a therapeutic agent against Salmonella in a simulated gastrointestinal tract of poultry.

Pyroligneous acid (PA) was evaluated as a potential alternative to therapeutic antibiotics in poultry. Antimicrobial activity of PA was studied at acidic pH (2.0) and neutral pH (7.0) of the liquid against Salmonella enterica and Lactobacillus acidophilus. Acidic PA gave a MIC value of 0.8% (v/v) and 1.6% (v/v), and neutralized PA gave a MIC value of 1.6% (v/v) and 3.2% (v/v) against S. enterica and L. acidophilus respectively. Acidic PA was evaluated at different concentrations in a simulated poultry digestive tract and cecal fermentation to study its effect on the cecal microflora and fermentation profile. PA at a concentration of 1.6% (v/v) completely inhibited S. enterica and was also found to have a similar effect on lactobacilli count as compared with the control (p = 0.17). Additionally, PA at this concentration was found not to have a significant effect on acetic acid production after 24 h of cecal fermentation (p = 0.20). Graphical abstract.

Chemical Composition of Pyroligneous Acid Obtained from Eucalyptus GG100 Clone.

The present study aimed to characterize the chemical composition of pyroligneous acid (PA) obtained from slow pyrolysis of the clone GG100 of Eucalyptus urophylla × Eucalyptus grandis. The efficiency of extraction of organic compounds by using different solvents-dichloromethane (DCM), diethyl ether (DE) and ethyl acetate (EA)-was evaluated. Wood discs were collected and carbonized at a heating rate of 1.25 °C/min until 450 °C. Pyrolysis gases were trapped and condensed, yielding a crude liquid product (CLP), which was refined to obtain pure PA. Then liquid-liquid extraction was carried out. Each extracted fraction was analyzed by GC-MS and the chemical compounds were identified. Experimental results showed that a larger number of chemical compounds could be extracted by using DCM and EA in comparison to diethyl ether DE. A total number of 93 compounds were identified, with phenolic compounds being the major group, followed by aldehydes and ketones, furans, pyrans and esters. Higher contents of guaiacol, phenol, cresols and furfural seem to explain the antibacterial and antifungal activity shown by PA, as reported previously in the literature. Experimental data indicated that the organic phase extracted from GG100 PA consists of a mixture of compounds similar to liquid smokes regularly used in the food industry.

Antibacterial and antifungal activities of pyroligneous acid from wood of Eucalyptus urograndis and Mimosa tenuiflora.

AIMS: This work aimed to evaluate the antibacterial and antifungal activities of two types of pyroligneous acid (PA) obtained from slow pyrolysis of wood of Mimosa tenuiflora and of a hybrid of Eucalyptus urophylla × Eucalyptus grandis. METHODS AND RESULTS: Wood wedges were carbonized on a heating rate of 1·25°C min-1 until 450°C. Pyrolysis smoke was trapped and condensed to yield liquid products. Crude pyrolysis liquids were bidistilled under 5 mmHg vacuum yielding purified PA. Multi-antibiotic-resistant strains of Escherichia coli, Pseudomonas aeruginosa (ATCC 27853) and Staphylococcus aureus (ATCC 25923) had their sensitivity to PA evaluated using agar diffusion test. Two yeasts were evaluated as well, Candida albicans (ATCC 10231) and Cryptococcus neoformans. GC-MS analysis of both PAs was carried out to obtain their chemical composition. Regression analysis was performed, and models were adjusted, with diameter of inhibition halos and PA concentration (100, 50 and 20%) as parameters. Identity of regression models and equality of parameters in polynomial orthogonal equations were verified. Inhibition halos were observed in the range 15-25 mm of diameter. CONCLUSIONS: All micro-organisms were inhibited by both types of PA even in the lowest concentration of 20%. SIGNIFICANCE AND IMPACT OF THE STUDY: The feasibility of the usage of PAs produced with wood species planted in large scale in Brazil was evident and the real potential as a basis to produce natural antibacterial and antifungal agents, with real possibility to be used in veterinary and zootechnical applications.

Collard greens yield with mineral and organic fertilization / Produção de couve brassica oleracea L. var. acephala com adubação mineral e orgânica

The development of the collard greens crop was studied in the seedling phase and after planting in vessels, which received different treatments of fertilization which included mineral fertilizer (as standard), Bokashi compost (Bo) and EM4, earthworms humus (Hu), and piroligneous acid (AP), combined in ten treatments for each crop phase. The standard treatment included commercial substrate for seedlings (SC) and after planting with the formula 08-28-16. The length of the leaf was assessed 25 days after emergency. The leaf length and width was assessed 85 days after planting. The fresh and dry weight of leaf and petiole were assessed 144 days after seedling. Inferior development was observed in plants with humus. In general, in the treatments which included Bokashi, the results were similar or close to the standard treatment.    

Foi estudado o desenvolvimento da cultura da couve-de-folha na fase de muda e após o transplantio em vasos, submetida a diferentes tratamentos de adubação, os quais incluíam fertilizante mineral (como padrão), composto Bokashi (Bo) e EM-4, húmus de minhoca (Hu) e ácido pirolenhoso (AP), combinados em dez tratamentos para cada fase. O tratamento padrão incluiu substrato comercial para mudas (SC) e adubação após o transplantio com a fórmula 08-28-16. O comprimento do limbo foliar foi avaliado 25 dias após a emergência. O comprimento e largura do limbo foliar foram avaliados 85 dias após o transplantio. O peso fresco e seco do limbo e pecíolo foi avaliado aos 144 dias após a semeadura. Desenvolvimento inferior foi observado em plantas com Hu. De maneira geral, nos tratamentos com Bo e EM-4, os resultados foram similares ou apresentaram desenvolvimento próximo ao tratamento padrão.   

Collard greens yield with mineral and organic fertilization / Produção de couve brassica oleracea L. var. acephala com adubação mineral e orgânica

The development of the collard greens crop was studied in the seedling phase and after planting in vessels, which received different treatments of fertilization which included mineral fertilizer (as standard), Bokashi compost (Bo) and EM4, earthworms humus (Hu), and piroligneous acid (AP), combined in ten treatments for each crop phase. The standard treatment included commercial substrate for seedlings (SC) and after planting with the formula 08-28-16. The length of the leaf was assessed 25 days after emergency. The leaf length and width was assessed 85 days after planting. The fresh and dry weight of leaf and petiole were assessed 144 days after seedling. Inferior development was observed in plants with humus. In general, in the treatments which included Bokashi, the results were similar or close to the standard treatment.

Foi estudado o desenvolvimento da cultura da couve-de-folha na fase de muda e após o transplantio em vasos, submetida a diferentes tratamentos de adubação, os quais incluíam fertilizante mineral (como padrão), composto Bokashi (Bo) e EM-4, húmus de minhoca (Hu) e ácido pirolenhoso (AP), combinados em dez tratamentos para cada fase. O tratamento padrão incluiu substrato comercial para mudas (SC) e adubação após o transplantio com a fórmula 08-28-16. O comprimento do limbo foliar foi avaliado 25 dias após a emergência. O comprimento e largura do limbo foliar foram avaliados 85 dias após o transplantio. O peso fresco e seco do limbo e pecíolo foi avaliado aos 144 dias após a semeadura. Desenvolvimento inferior foi observado em plantas com Hu. De maneira geral, nos tratamentos com Bo e EM-4, os resultados foram similares ou apresentaram desenvolvimento próximo ao tratamento padrão.