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  • 刊频： Quarterly
  • NLM标题： Braz J Microbiol
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  • 1.

    【2hr】Illumina sequencing and assessment of new cost-efficient protocol for metagenomic-DNA extraction from environmental water samples

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    机译 Illumina测序和评估从环境水样中提取宏基因组DNA的新成本有效方案
    摘要：In this study, the development and assessment of a modified, efficient, and cost-efficient protocol for mDNA (metagenomic DNA) extraction from contaminated water samples was attempted. The efficiency of the developed protocol was investigated in comparison to a well-established commercial kit (Epicentre, Metagenomic DNA Isolation Kit for Water). The comparison was in terms of degree of shearing, yield, purity, duration, suitability for polymerase chain reaction and next-generation sequencing in addition to the quality of next-generation sequencing data. The DNA yield obtained from the developed protocol was 2.6 folds higher than that of the commercial kit. No significant difference in the alpha (Observed species, Chao1, Simpson and PD whole tree) and beta diversity was found between the DNA samples extracted by the commercial kit and the developed protocol. The number of high-quality sequences of the samples extracted by the developed method was 20% higher than those obtained by the samples processed by the kit. The developed economic protocol successfully yielded high-quality pure mDNA compatible with complex molecular applications. Thus we propose the developed protocol as a gold standard for future metagenomic studies investigating a large number of samples.
  • 2.

    【2hr】Plant growth promoting rhizobacteria reduce aphid population and enhance the productivity of bread wheat

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    机译 促进根瘤菌生长的植物减少了蚜虫种群并提高了面包小麦的生产力
    摘要：Plant growth promoting rhizobacteria increase plant growth and give protection against insect pests and pathogens. Due to the negative impact of chemical pesticides on environment, alternatives to these chemicals are needed. In this scenario, the biological methods of pest control offer an eco-friendly and an attractive option. In this study, the effect of two plant growth promoting rhizobacterial strains (Bacillus sp. strain 6 and Pseudomonas sp. strain 6K) on aphid population and wheat productivity was evaluated in an aphid susceptible (Pasban-90) and resistant (Inqlab-91) wheat cultivar. The seeds were inoculated with each PGPR strain, separately or the combination of both. The lowest aphid population (2.1 tiller−1), and highest plant height (85.8 cm), number of spikelets per spike (18), grains per spike (44), productive tillers (320 m−2), straw yield (8.6 Mg ha−1), and grain yield (4.8 Mg ha−1) were achieved when seeds were inoculated with Bacillus sp. strain 6 + Pseudomonas sp. strain 6K. The grain yield of both varieties was enhanced by 35.5–38.9% with seed inoculation with both bacterial strains. Thus, the combine use of both PGPR strains viz. Bacillus sp. strain 6 + Pseudomonas sp. strain 6K offers an attractive option to reduce aphid population tied with better wheat productivity.
  • 3.

    【2hr】Combined application of bio-organic phosphate and phosphorus solubilizing bacteria (Bacillus strain MWT 14) improve the performance of bread wheat with low fertilizer input under an arid climate

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    机译 生物有机磷酸盐和磷增溶细菌（芽孢杆菌MWT 14）的组合施用可改善干旱气候下低施肥量的面包小麦的性能
    摘要：This study was aimed to investigate the effect of bio-organic phosphate either alone or in combination with phosphorus solubilizing bacteria strain (Bacillus MWT-14) on the growth and productivity of two wheat cultivars (Galaxy-2013 and Punjab-2011) along with recommended (150–100 NP kg ha−1) and half dose (75–50 NP kg ha−1) of fertilizers. The combined application of bio-organic phosphate and the phosphorous solubilizing bacteria strain at either fertilizer level significantly improved the growth, yield parameters and productivity of both wheat cultivars compared to non-inoculated control treatments. The cultivar Punjab-2011 produced the higher chlorophyll contents, crop growth rate, and the straw yield at half dose of NP fertilizer; while Galaxy-2013, with the combined application of bio-organic phosphate and phosphorous solubilizing bacteria under recommended NP fertilizer dose. Combined over both NP fertilizer levels, the combined use of bio-organic phosphate and phosphorous solubilizing bacteria enhanced the grain yield of cultivar Galaxy-2013 by 54.3% and that of cultivar Punjab-2011 by 83.3%. The combined application of bio-organic phosphate and phosphorous solubilizing bacteria also increased the population of phosphorous solubilizing bacteria, the soil organic matter and phosphorous contents in the soil. In conclusion, the combined application of bio-organic phosphate and phosphorous solubilizing bacteria offers an eco-friendly option to harvest the better wheat yield with low fertilizer input under arid climate.
  • 4.

    【2hr】Dynamics and diversity of the bacterial community during the spontaneous decay of açai (Euterpe oleracea) fruits

    包量

    机译 阿萨伊（Euterpe oleracea）果实自发腐烂过程中细菌群落的动态和多样性
    摘要：The biodiversity and evolution of the microbial community in açai fruits (AF) between three geographical origins and two spontaneous decay conditions were examined by applying culture-independent methods. Culture-independent methods based on 16S rRNA from fifteen samples revealed that Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes and Acidobacteria were the most abundant phyla. At the genus level, Massilia (taxon with more than 50% of the sequences remaining constant during the 30 h of decay), Pantoea, Naxibacter, Enterobacter, Raoultella and Klebsiella were identified, forming the carposphere bacterial microbiota of AF. AF is fibre-rich and Massilia bacteria could find a large quantity of substrate for its growth through cellulase production. Beta diversity showed that the quality parameters of AF (pH, soluble solids, titratable acidity and lipids) and elemental analysis (C, N, H and C/N ratio) were unable to drive microbial patterns in AF. This research offers new insight into the indigenous bacterial community composition on AF as a function of spontaneous postharvest decay.
  • 5.

    【2hr】Evaluation of skimmed milk flocculation method for virus recovery from tomatoes

    包量

    机译 脱脂牛奶絮凝方法从番茄中回收病毒的评价
    摘要：This study aimed to evaluate the elution-concentration methodology based on skimmed milk flocculation from three varieties of tomatoes (Solanum lycopersicum L. [globe], Solanum lycopersicum var. cerasiforme [cherry] and hybrid cocktail [grape tomato]) for further monitoring of field samples. Spiking experiments were performed to determine the success rate and efficiency recovery of human norovirus (NoV) genogroup II, norovirus murine-1 (MNV-1) used as sample process control virus and human adenovirus (HAdV). Mean values of 18.8%, 2.8% and 44.0% were observed for NoV GII, MNV-1 and HAdV, respectively with differences according to the types of tomatoes, with lower efficiency for cherry tomatoes. Analysis of 90 samples, obtained at commercial establishments in the metropolitan region of Rio de Janeiro State, revealed 4.5% positivity for HAdV. Bacterial analysis was also performed with no detection of Salmonella spp., L. monocytogenes and fecal coliforms. Data demonstrated that the skimmed milk flocculation method is suitable for recovering HAdV from tomatoes and highlights the need for considering investigation in order to improve food safety.
  • 6.

    【2hr】Endophytic Bacillus strains enhance pearl millet growth and nutrient uptake under low-P

    包量

    机译 低磷条件下内生芽孢杆菌菌株可促进珍珠粟的生长和养分吸收
    摘要：Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.
  • 7.

    【2hr】Diversity and communities of culturable endophytic fungi from different tree peonies (geoherbs and non-geoherbs), and their biosynthetic potential analysis

    包量

    机译 不同树牡丹（geoherbs和非geoherbs）可培养内生真菌的多样性和群落组成及其生物合成潜力分析
    摘要：To know more about the potential roles of endophytic fungi in the formation mechanism of Daodi medicinal material, diversity and communities of culturable endophytic fungi in three types of tree peonies were investigated. Endophytic fungi of three types of tree peonies were isolated and identified. The diversity was analyzed. Bayesian trees constructed by MrBayes 3.2.6 after phylogenetic analysis of the ITS sequences. The endophytic fungi potential for synthesis of natural products was assessed by means of detecting NRPS and PKS gene sequences. In total, 364 endophytic fungi isolates representing 26 genera were recovered from Paeonia ostii ‘Feng Dan’, Paeonia ostii ‘Luoyang Feng Dan’, and Paeonia suffruticosa ‘Luoyang Hong’. More culturable endophytic fungi appeared in P. suffruticosa ‘Luoyang Hong’ (206) compared with P. ostii ‘Feng Dan’ (60) and P. ostii ‘Luoyang Feng Dan’ (98). The fungal community of P. ostii ‘Feng Dan’ had the highest richness and diversity. PKSs and NRPS detection rates of endophytic fungi from P. ostii ‘Feng Dan’ are both the highest among the three types of tree peonies. Results indicate that endophytic fungus is an important factor of Daodi Cortex Moutan forming, and endophytic fungi in peony are related to genuineness of Cortex Moutan.
  • 8.

    【2hr】Screening of endophytic fungi stored in a culture collection for taxol production

    包量

    机译 筛选培养物中收集的内生真菌以生产紫杉醇
    摘要：In this work, four isolates of endophytic fungi (Alternaria alternata, Colletotrichum gloesporioides, Glomerella cingulata and Nigrospora sphaerica), deposited in the culture collection ‘University Recife Mycologia’ (URM) at the Universidade Federal de Pernambuco, were characterized for the genes ITS 1 and 4 (region 5.8 S) and evaluated for taxol production.
  • 9.

    【2hr】Pleurotus spp. cultivation on Brachiaria sp. straw treatment with alkaline water

    包量

    机译 杏鲍菇腕足菌的栽培。碱性水秸秆处理
    摘要：The aim of this research was to evaluate the efficiency of aqueous alkali-treated Brachiaria straw for the cultivation of appropriate species of oyster mushroom. The substrate used in the cultivation of various Pleurotus spp. was soaked for 20 min by using two different procedures: (i) 0.5–2.0% Ca(OH)2 in 100 L water, and (ii) 50–250 L water. As a result, 1% Ca(OH)2 dissolved in 100 L water and 3.5 kg of Brachiaria straw presented the best production. The most suitable species for the application of the present method were P. pulmonarius and P. sapidus. The success of this technique is directly related to the concentration of Ca(OH)2 and water, the species, and the origin and quality of raw material used as the substrate in the production of oyster mushroom.
  • 10.

    【2hr】Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus

    包量

    机译 基于重组衣壳蛋白的间接ELISA的开发以检测牛白血病病毒的抗体
    摘要：Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp® plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD450 nm) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a κ value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.
  • 11.

    【2hr】Survey on pathogenic Escherichia coli and Salmonella spp. in captive cockatiels (Nymphicus hollandicus)

    包量

    机译 致病性大肠杆菌和沙门氏菌调查。在圈养小形鹦鹉（Nymphicus hollandicus）中
    摘要：We surveyed healthy captive cockatiels (Nymphicus hollandicus) for Escherichia coli and Salmonella spp. Cloacal swabs were collected from 94 cockatiels kept in commercial breeders, private residencies and pet shops in the cities of São Paulo/SP and Niterói/RJ (Brazil). Three strains of E. coli from each individual were tested for the presence of ExPEC-, APEC- and DEC-related genes. We evaluated the blaTEM, blaSHV, blaOXA, blaCMY, blaCTX-M, tetA, tetB, aadA, aphA, strAB, sul1, sul2, sul3, qnrA, qnrD, qnrB, qnrS, oqxAB, aac (6)′-Ib-cr, qepA resistance genes and markers for plasmid incompatibility groups. Salmonella spp. was not detected. E. coli was isolated in 10% of the animals (9/94). Four APEC genes (ironN, ompT, iss and hlyF) were detected in two strains (2/27–7%), and iss (1/27–4%) in one isolate. The highest resistance rates were observed with amoxicillin (22/27–82%), ampicillin (21/27–79%), streptomycin (18/27–67%), tetracycline (11/27–41%). Multiresistance was verified in 59% (16/27) of the isolates. We detected strAB, blaTEM, tetA, tetB, aadA, aphaA, sul1, sul2, sul3 resistance genes and plasmid Inc groups in 20 (74%) of the strains. E. coli isolated from these cockatiels are of epidemiological importance, since these pets could transmit pathogenic and multiresistant microorganisms to humans and other animals.
  • 12.

    【2hr】Duplex nested-PCR for detection of small ruminant lentiviruses

    包量

    机译 双重巢式PCR检测小反刍动物慢病毒
    摘要：Small ruminant lentiviruses (SRLV) have high genetic variability which results in different viral strains around the world. This create a challenge to design sensible primers for molecular diagnosis in different regions. This work proposes a protocol of duplex nested-PCR for the precise diagnosis of SRLV. The technique was designed and tested with the control strains CAEV Co and MVV 1514. Then, field strains were submitted to the same protocol of duplex nested-PCR. Blood samples of sheep and goats were tested with AGID and nested PCR with specific primers for pol, gag and LTR. The AGID results showed low detection capacity of positive animals, while the nested PCR demonstrated a greater capacity of virus detection. Results demonstrated that LTR-PCR was more efficient in detecting positive sheep samples, whereas gag-PCR allowed a good detection of samples of positive goats and positive sheep. In addition, pol-PCR was more efficient with goat samples than for sheep. Duplex nested PCR performed with standard virus samples and field strains demonstrated that the technique is more efficient for the detection of multiple pro-viral DNA sequences. This study demonstrated a successful duplex nested PCR assay allowing a more accurate diagnosis of SRLV.
  • 13.

    【2hr】Characterization of Klebsiella pneumoniae associated with cattle infections in southwest China using multi-locus sequence typing (MLST), antibiotic resistance and virulence-associated gene profile analysis

    包量

    机译 利用多位点序列分型（MLST），抗生素抗性和毒力相关基因谱分析对中国西南地区与牛感染相关的肺炎克雷伯菌的鉴定
    摘要：Klebsiella pneumoniae is important human and animal pathogen that causes a wide spectrum of infections. In this study, isolates from cattle nasal swabs samples were identified by 16S rRNA, and to evaluate the antimicrobial susceptibility, virulence gene carrying levels, and multilocus sequence typing of K. pneumoniae isolates. 33 isolates of K. pneumoniae were isolated and identified in 213 nasal swabs samples, of which 12 were hypervirulent K. pneumoniae strains. Extended Spectrum Beta-Lactamases genes were found in 93.4% of the strains. Of which, TEM was the most prevalent (93.4%), followed by CTX-M and SHV were 57.6% and 39.4%, respectively. A main mutation pattern of quinoloneresistance-determining region, Thr83-Ieu and Asp87-Asn in gyrA and Ser87-Ile in parC, was detected in 33 K. pneumoniae isolates. All the isolates harbored at least two virulence factor genes, with ureA (97.0%) and wabG (91.0%) exhibiting high carriage rates in 33 K. pneumoniae isolates. MLST revealed 7 sequence types, of which 3 STs (2541, 2581 and 2844) were newly assigned. Using eBURST, ST2844 and ST2541 were assigned to new clonal complex 2844. Our study provides evidence and biological characteristics of K. pneumoniae isolates from cattle upper respiratory tract in Southwest China.
  • 14.

    【2hr】The use of Prevotella bryantii 3C5 for modulation of the ruminal environment in an ovine model

    包量

    机译 布鲁氏丙酸杆菌3C5在绵羊模型中调节瘤胃环境的用途
    摘要：In the Southern Hemisphere, ruminants are mostly raised in grazing systems where animals consume forage and are supplemented with low amounts of concentrates. Concentrates are usually given separately and are rapidly ingested. This practice leads to changing rumen environment conditions during the day, may alter the rumen microbial metabolism and could affect host performance. The native ruminal Prevotella bryantii strain 3C5 was administered every 48 h to wethers under experimental conditions simulating Southern-Hemisphere feeding to evaluate its potential as a rumen fermentation modulator. The inoculum potential was assessed on day 17. The ammonia nitrogen (NH3-N), volatile fatty acids and ruminal pH were monitored on a 24-h basis 19 days after the beginning of the experiment, and the microbial community structure was assessed by pyrosequencing. The administration of P. bryantii modified the fermentation products and daily pH values compared to the control. The NH3-N concentration in the rumen of treated animals was significantly higher than that of the untreated animals. Modification of the ruminal environment and fermentation pathways was achieved without altering the general structure of the microbial community or the potential methane production. P. bryantii 3C5 could be considered in potential probiotic formulations for ruminants in semi-intensive systems.
  • 15.

    【2hr】Characterization of avian pathogenic Escherichia coli isolated from free-range helmeted guineafowl

    包量

    机译 从散地头盔盔鸡分离的禽致病性大肠杆菌的特征
    摘要：Avian pathogenic Escherichia coli (APEC) isolates from apparently healthy free range helmeted guineafowl were characterized. Most of them had a high frequency of virulence associated genes, multi drug resistance and high pathogenicity. We demonstrated that helmeted guineafowl have potential to transmit antibiotic resistant APEC to other species including humans.
  • 16.

    【2hr】Antimicrobial effect of copper surfaces on bacteria isolated from poultry meat

    包量

    机译 铜表面对禽肉中细菌的抗菌作用
    摘要：Poultry meat is a food product that usually carries high rates of microbial contamination, including foodborne pathogens. The poultry industry has established different systems to minimize these hazards. In recent years, extensive literature has demonstrated the antimicrobial activity of different contact surfaces made of copper to effectively reduce microbial loads. The aim of the present study was to evaluate the antibacterial effect of copper surfaces on the transmission of two foodborne pathogens – Salmonella enterica and Listeria monocytogenes – and a poultry native microbiota bacterial species – Enterobacter cloacae. We also evaluated the impact of the poultry meat matrix on the antimicrobial activity of a copper surface. Our results indicated that copper surfaces reduced the bacterial load quickly (<than 4 min) when the microorganisms were exposed to polished copper surfaces. Even when bacteria were inoculated on copper surfaces soiled with the organic matrix (washing water from poultry carcasses) and survival rates were significantly higher, an antimicrobial effect was still observed. Survival rates of two microorganisms simultaneously exposed to copper did not show significant differences. We found an antimicrobial effect over pathogenic and non-pathogenic microorganisms. Results suggest a potential role for copper surfaces in the control of microbiological hazards in the poultry industry.
  • 17.

    【2hr】Carbon metabolism influenced for promoters and temperature used in the heterologous protein production using Pichia pastoris yeast

    包量

    机译 碳代谢影响使用毕赤酵母生产异源蛋白质的启动子和温度
    摘要：Nowadays, it is necessary to search for different high-scale production strategies to produce recombinant proteins of economic interest. Only a few microorganisms are industrially relevant for recombinant protein production: methylotrophic yeasts are known to use methanol efficiently as the sole carbon and energy source. Pichia pastoris is a methylotrophic yeast characterized as being an economical, fast and effective system for heterologous protein expression. Many factors can affect both the product and the production, including the promoter, carbon source, pH, production volume, temperature, and many others; but to control all of them most of the time is difficult and this depends on the initial selection of each variable. Therefore, this review focuses on the selection of the best promoter in the recombination process, considering different inductors, and the temperature as a culture medium variable in methylotrophic Pichia pastoris yeast. The goal is to understand the effects associated with different factors that influence its cell metabolism and to reach the construction of an expression system that fulfills the requirements of the yeast, presenting an optimal growth and development in batch, fed-batch or continuous cultures, and at the same time improve its yield in heterologous protein production.
  • 18.

    【2hr】Antimicrobial potential of pyroligneous extracts – a systematic review and technological prospecting

    包量

    机译 焦木提取物的抗菌潜力–系统评价和技术前景
    摘要：Pyroligneous extract is applied in diverse areas as an antioxidant, an antimicrobial, and an anti-inflammatory agent. The discovery of new cost-effective antimicrobial agents of natural origin remains a challenge for the scientific community. This study aimed to conduct a systematic review and a technological forecasting of the existent evidence regarding the use of pyroligneous extract as a potential antimicrobial agent. Studies were identified through an investigation of various electronic databases: PubMed, SciFinder, Web of Science, Scopus, Scielo, Google scholar, and ProQuest. Patents were searched through INPI, Google patents, Espacenet, Patents online, USPTO, and WIPO. The literature on antimicrobial activity of pyroligneous extract are limited given the short duration of studies and variability in study design, use of pyroligneous preparations, and reports on results. However, evidence suggests the potential of pyroligneous extract as a natural antimicrobial agent. The most studied activity was the role of PE as a food preservative. However, pyroligneous extracts are also effective against pathogenic bacteria in the oral microflora and treatment of candidal infections. Further research is needed using standardized preparations of pyroligneous extracts to determine their long-term effectiveness and ability as antimicrobial agents.
  • 19.

    【2hr】Ethanol production from sweet sorghum by Saccharomyces cerevisiae DBKKUY-53 immobilized on alginate-loofah matrices

    包量

    机译 固定在藻酸盐-丝瓜络基质上的酿酒酵母DBKKUY-53从甜高粱中生产乙醇
    摘要：Ethanol production from sweet sorghum juice (SSJ) using the thermotolerant Saccharomyces cerevisiae strain DBKKUY-53 immobilized in an alginate-loofah matrix (ALM) was successfully developed. As found in this study, an ALM with dimensions of 20 × 20 × 5 mm3 is effective for cell immobilization due to its compact structure and long-term stability. The ALM-immobilized cell system exhibited greater ethanol production efficiency than the freely suspended cell system. By using a central composite design (CCD), the optimum conditions for ethanol production from SSJ by ALM-immobilized cells were determined. The maximum ethanol concentration and volumetric ethanol productivity obtained using ALM-immobilized cells under the optimal conditions were 97.54 g/L and 1.36 g/L h, respectively. The use of the ALM-immobilized cells was successful for at least six consecutive batches (360 h) without any loss of ethanol production efficiency, suggesting their potential application in industrial ethanol production.
  • 20.

    【2hr】Cost-effective production of bacterial cellulose using acidic food industry by-products

    包量

    机译 利用酸性食品工业副产品经济高效地生产细菌纤维素
    摘要：To reduce the cost of obtaining bacterial cellulose, acidic by-products of the alcohol and dairy industries were used without any pretreatment or addition of other nitrogen sources. Studies have shown that the greatest accumulation of bacterial cellulose (6.19 g/L) occurs on wheat thin stillage for 3 days of cultivation under dynamic conditions, which is almost 3 times higher than on standard Hestrin and Schramm medium (2.14 g/L). The use of whey as a nutrient medium makes it possible to obtain 5.45 g/L bacterial cellulose under similar conditions of cultivation. It is established that the pH of the medium during the growth of Gluconacetobacter sucrofermentans B-11267 depends on the feedstock used and its initial value. By culturing the bacterium on thin stillage and whey, there is a decrease in the acidity of the waste. It is shown that the infrared spectra of bacterial cellulose obtained in a variety of environments have a similar character, but we found differences in the micromorphology and crystallinity of the resulting biopolymer.
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