The use of biodegradable plastics can reduce the accumulation of environmentally persistent plastic wastes. The rate of degradation of biodegradable plastics depends on environmental conditions and is highly variable. Techniques for achieving more consistent degradation are needed. However, only a few microorganisms involved in the degradation process have been isolated so far from the environment. Here, we show that Pseudozyma spp. yeasts, which are common in the phyllosphere and are easily isolated from plant surfaces, displayed strong degradation activity on films made from poly-butylene succinate or poly-butylene succinate-co-adipate. Strains of P. antarctica isolated from leaves and husks of paddy rice displayed strong degradation activity on these films at 30°C. The type strain, P. antarctica JCM 10317, and Pseudozyma spp. strains from phyllosphere secreted a biodegradable plastic-degrading enzyme with a molecular mass of about 22 kDa. Reliable source of biodegradable plastic-degrading microorganisms are now in our hands.
Pseudozyma; Biodegradable plastic; Phyllosphere; Yeast
Freshwater shortage is the main problem in Heilonggang lower-lying plain, while a considerable amount of underground saline water is available. We wanted to find an effective way to use the brackish water in winter wheat production. Surface mulch has significant effect in reducing evaporation and decreasing soil salinity level. This research was aimed at comparing the effect of different mulch materials on winter wheat production. The experiment was conducted during 2002~2003 and 2003~2004. Four treatments were setup: (1) no mulch, (2) mulch with plastic film, (3) mulch with corn straw, (4) mulch with concrete slab between the rows. The result indicated that concrete mulch and straw mulch was effective in conserving soil water compared to plastic film mulch which increased soil temperature. Concrete mulch decreases surface soil salinity better in comparison to other mulches used. Straw mulch conserved more soil water but decreased wheat grain yield probably due to low temperature. Concrete mulch had similar effect with plastic film mulch on promoting winter wheat development and growth.
Mulch; Plastic film; Corn straw; Concrete; Saline water; Wheat growth
The gene encoding a poly(dl-lactic acid) (PLA) depolymerase from Paenibacillus amylolyticus strain TB-13 was cloned and overexpressed in Escherichia coli. The purified recombinant PLA depolymerase, PlaA, exhibited degradation activities toward various biodegradable polyesters, such as poly(butylene succinate), poly(butylene succinate-co-adipate), poly(ethylene succinate), and poly(ε-caprolactone), as well as PLA. The monomeric lactic acid was detected as the degradation product of PLA. The substrate specificity toward triglycerides and p-nitrophenyl esters indicated that PlaA is a type of lipase. The gene encoded 201 amino acid residues, including the conserved pentapeptide Ala-His-Ser-Met-Gly, present in the lipases of mesophilic Bacillus species. The identity of the amino acid sequence of PlaA with Bacillus lipases was no more than 45 to 50%, and some of its properties were different from those of these lipases.
Three crops of cucumber were grown in succession in beds by use of trickle irrigation, plastic film mulch, and soil chemical treatments over a 17-month period, including a fallow winter season. Total yield for the three crops was highest (1208 quintals/ha) in film-mulched plots treated with MBR-CP, and next-highest in film-mulched plots treated with DD-MENCS (1094 quintals/ha); total yield was only 456 quintals/ha in film-mulched control (untreated) plots. Yield in untreated film-mulched plots was 256% of that in untreated unmulched plots (178 quintals/ha). Plant growth and yields were greatest when populations of nematodes and soil-borne fungi were suppressed to very low levels. The residual control by soil treatments lasted longest on Meloidogyne incognita and Fusarium solani.
multiple cropping; multiple pest control
Okra was grown in field plots of Tifton loamy sand naturally infested with the nematodes Meloidogyne incognita and Criconemoides ornalus and the pathogenic fungi Fusarium oxysporum, F. solani, F. roseum, and Pythium spp. Plots were treated with various soil pesticides and left exposed or covered with biodegradable paper film mulch under trickle irrigation. Soil was assayed for nematodes and fungi, and plant roots were examined for root-rot and insect damage. Fewer nematodes and fungi generally were recovered from soil treated with DD-MENCS (with and without film mulch) or methyl bromide-chloropicrin (2:1) (MBC) and film mulch than from nontreated soil. Funfigation with DD-MENCS or MBC suppressed populations of M. incognita, C. ornatus, F. oxysporum, F. solani, F. roseum, and Pythium spp. Ethoprop (alone or combined with other pesticides), sodium azide, and chloroneb were less effective than DD-MENCS and MBC. Plant growth anti yield were greatest when nematodes and pathogenic fungi were controlled. Yield was increased 3-fold by DD-MENCS + film mulch or MBC + film mulch in comparison with the average yield of okra produced in Georgia. The root-knot nematode-Fusarium wilt complex was most severe in nonfuntigated soil.
Abelmoschus esculentus; multiple-pest control
A new class of biocomposites based on oil palm empty fruit bunch fiber and poly(butylene adipate-co-terephthalate) (PBAT), which is a biodegradable aliphatic aromatic co-polyester, were prepared using melt blending technique. The composites were prepared at various fiber contents of 10, 20, 30, 40 and 50 wt% and characterized. Chemical treatment of oil palm empty fruit bunch (EFB) fiber was successfully done by grafting succinic anhydride (SAH) onto the EFB fiber surface, and the modified fibers were obtained in two levels of grafting (low and high weight percentage gain, WPG) after 5 and 6 h of grafting. The FTIR characterization showed evidence of successful fiber esterification. The results showed that 40 wt% of fiber loading improved the tensile properties of the biocomposite. The effects of EFB fiber chemical treatments and various organic initiators content on mechanical and thermal properties and water absorption of PBAT/EFB 60/40 wt% biocomposites were also examined. The SAH-g-EFB fiber at low WPG in presence of 1 wt% of dicumyl peroxide (DCP) initiator was found to significantly enhance the tensile and flexural properties as well as water resistance of biocomposite (up to 24%) compared with those of untreated fiber reinforced composites. The thermal behavior of the composites was evaluated from thermogravimetric analysis (TGA)/differential thermogravimetric (DTG) thermograms. It was observed that, the chemical treatment has marginally improved the biocomposites’ thermal stability in presence of 1 wt% of dicumyl peroxide at the low WPG level of grafting. The improved fiber-matrix surface enhancement in the chemically treated biocomposite was confirmed by SEM analysis of the tensile fractured specimens.
oil palm EFB fiber; poly(butylene adipate-co-terephthalate); biocomposite; fiber esterification; thermal and mechanical properties; succinic anhydride
The biodegradabilities of poly(butylene succinate) (PBS) powders in a controlled compost at 58 °C have been studied using a Microbial Oxidative Degradation Analyzer (MODA) based on the ISO 14855-2 method, entitled “Determination of the ultimate aerobic biodegradability of plastic materials under controlled composting conditions—Method by analysis of evolved carbon dioxide—Part 2: Gravimetric measurement of carbon dioxide evolved in a laboratory-scale test”. The evolved CO2 was trapped by an additional aqueous Ba(OH)2 solution. The trapped BaCO3 was transformed into graphite via a serial vaporization and reduction reaction using a gas-tight tube and vacuum manifold system. This graphite was analyzed by accelerated mass spectrometry (AMS) to determine the percent modern carbon [pMC (sample)] based on the 14C radiocarbon concentration. By using the theory that pMC (sample) was the sum of the pMC (compost) (109.87%) and pMC (PBS) (0%) as the respective ratio in the determined period, the CO2 (respiration) was calculated from only one reaction vessel. It was found that the biodegradabilities determined by the CO2 amount from PBS in the sample vessel were about 30% lower than those based on the ISO method. These differences between the ISO and AMS methods are caused by the fact that part of the carbons from PBS are changed into metabolites by the microorganisms in the compost, and not changed into CO2.
biodegradation; poly(butylenes succinate); ISO 14855-2; accelerator mass spectrometry; radiocarbon
Field plots in Tifton loamy sand were treated with various soil pesticides in 1973 and 1974 and either left exposed or covered with biodegradable flint mulch. Test crops were cantaloup, slicing and pickling cucumber, squash, and sweet corn. Overhead sprinkler irrigation was used in 1973, and trickle irrigation under the film mulch was used on sweet corn in 1974. Soil was assayed for nematodes, and roots of plants were evaluated for damage by root-knot nematodes. Nematode populations were reduced by soil treatment with an organic phosphate or carbamate nematicide-herbicide-fungicide combination (NHF), DD-MENCS, methyl bromide-chloropicrin (MBR-CP), ethoprop, carbofuran, and sodium azide + ethoprop or carbofuran. Sodium azide, sodium azide + ethoprop or carbofuran, ethoprop, and carbofuran were less effective than DD-MENCS, MBR-CP, attd the NHF combination. The NHF combination controlled grasses and broadteaf weeds as effectively as the herbicide alone. Growth and yield were greatest when nematodes and weeds were controlled. Yields of marketable vegetables were highest from plants in plots treated with DD-MENCS with a film mulch.
Meloidogyne incognita; Macroposthonia ornata; multiple pest control
The effects of different-colored polyethylene mulches on the quantity and spectra of reflected light, earliness of fruit set, fruit yield and quality, and root-knot disease were studied in field-grown, staked tomato (Lycopersicon esculentum). White mulch reflected more photosynthetic light and a lower far-red-to-red ratio than red mulch, whereas black mulch reflected less than 5 percent of any color. Soil temperatures and fruit yields were recorded for tomato plants inoculated with Meloidogyne incognita race 3 at initial populations of 0, 1,000, 10,000, 50,000, or 100,000 eggs/plant and grown over black, white, or red plastic mulch in both spring and fall. Soil temperatures were lower under white mulch than under red or black mulch. Tomato yields declined as inoculum level increased. Plants grown over red mulch in the spring and inoculated with 50,000 eggs of M. incognita had greater early marketable yields than similarly inoculated plants grown over black or white mulch. Tomato plants inoculated with 100,000 eggs and grown over white mulch or red mulch in the spring had greater total yields per plot than similar plants grown over black mulch (7.39 kg and 7.71 kg vs. 3.65 kg, respectively).
colored mulch; light quality; Lycopersicon esculentum; Meloidogyne incognita; nematode; photomorphogenesis; photospectrum; physiology; root-knot nematode; tomato
We have developed a mulch sheet made by inflation molding of PLA, Ecoflex® and modified starch, which all have different biodegradabilities. A field test of use as an agricultural mulch sheet for mandarin oranges was carried out over two years. The mechanical properties of the mulch sheet were weakened with time during the field test, but the quality of the mandarin oranges increased, a result of the controlled degradation of the sheet. The most degradable modified starch degraded first, allowing control of the moisture on the soil. Accelerator mass spectroscopy was used for evaluation of the biomass carbon ratio. The biomass carbon ratio decreased by degradation of the biobased materials, PLA and modified starch in the mulch sheet.
poly(lactic acid); modified starch; mulch sheet; mandarin orange; biodegradable materials; biomass carbon ratio
Placement of a 3-m-wide, black, polyethylene film mulch down rows of peach (Prunus persica 'Red Haven' on 'Lovell' rootstock) and almond (Prunus dulcis 'Nonpareil' on 'Lovell') trees in the San Joaquin Valley of California resulted in irrigation water conservation of 75%, higher soil temperature in the surface 30 cm, a tendency toward greater root mass, elimination of weeds, and a greater abundance of Meloidogyne incognita second-stage juveniles in soil but reduced root galling when compared to the nonmulched control. Population levels of Pratylenchus hexincisus, a nematode found within tree roots, were reduced by mulching, as were those of Tylenchulus semipenetrans, which survived on old grape roots remaining from a previously planted vineyard, and Paratrichodorus minor, which probably fed on roots of various weed species growing in the nonmulched soil. Populations of Pythium ultimum were not significantly changed, probably also due to the biological refuge of the old grape roots and moderate soil heating level. Trunk diameters of peach trees were increased by mulching, but those of almond trees were reduced by the treatment. Leaf petiole analysis indicated that concentrations of mineral nutrients were inconsistent, except for a significant increase in Ca in both tree species.
almond; irrigation management; Meloidogyne incognita; mulching; nematode; peach; Paratrichodorus minor; Paratylenchus hamatus; Pratylenchus hexincisus; Prunus dulcis; Prunus persica; Pythium ultimum; soil heating; solarization; Tylenchulus semipenetrans
The effect of different-colored polyethylene mulches on quantity and spectra of reflected light, plant morphology, and root-knot disease was studied in tomato (Lycopersicon esculentum) grown in simulated planting beds. Tomato plants were inoculated with Meloidogyne incognita at initial populations (Pi) of 0, 1,000, 10,000, or 50,000 eggs/plant, and grown in a greenhouse for 50 days over white, red, or black mulch. Soil temperature was kept constant among the mulch treatments by placing an insulation barrier between the colored mulch and the soil surface. Soil temperature varied less than 0.5 °C between soil chambers at solar noon. Tomatoes grown over white mulch received more reflected photosynthetic light and had greater shoot weights (27%), root weights (32%), and leaf area (20%) than plants grown over black mulch. Plants grown over red mulch received a higher far-red-to-red ratio in the reflected light. Mulch color altered the plant's response to root-knot nematode infection by changing the distribution of mass in axillary shoots. At high Pi, axillary leaf area and leaf weight were greater in tomato grown over white mulch than when grown over red mulch. The root-gall index was lower for plants grown over white mulch than similar plants grown over red mulch.
host-parasite relationship; light quality; Meloidogyne incognita; nematode; photomorphogenesis; plastic mulch; polyethylene; root-knot; tomato
Traditional and novel techniques were tested and compared for their usefulness in evaluating biodegrad-ability claims made for newly formulated “degradable” plastic film products. Photosensitized polyethylene (PE), starch-PE, extensively plasticized polyvinyl chloride (PVC), and polypropylene (PP) films were incorporated into aerobic soil. Biodegradation was measured for 3 months under generally favorable conditions. Carbon dioxide evolution, residual weight recovery, and loss of tensile strength measurements were supplemented, for some films, by gas chromatographic measurements of plasticizer loss and gel permeation chromatographic (GPC) measurement of polymer molecular size distribution. Six- and 12-week sunlight exposures of photosensitized PE films resulted in extensive photochemical damage that failed to promote subsequent mineralization in soil. An 8% starch-PE film and the plasticized PVC film evolved significant amounts of CO2 in biodegradation tests and lost residual weight and tensile strength, but GPC measurements demonstrated that all these changes were confined to the additives and the PE and PVC polymers were not degraded. Carbon dioxide evolution was found to be a useful screening tool for plastic film biodegradation, but for films with additives, polymer biodegradation needs to be confirmed by GPC. Photochemical cross-linking of polymer strands reduces solubility and may interfere with GPC measurements of polymer degradation.
In this study, two strains of Aspergillus sp. and Lysinibacillus sp. with remarkable abilities to degrade low-density polyethylene (LDPE) were isolated from landfill soils in Tehran using enrichment culture and screening procedures. The biodegradation process was performed for 126 days in soil using UV- and non-UV-irradiated pure LDPE films without pro-oxidant additives in the presence and absence of mixed cultures of selected microorganisms. The process was monitored by measuring the microbial population, the biomass carbon, pH and respiration in the soil, and the mechanical properties of the films. The carbon dioxide measurements in the soil showed that the biodegradation in the un-inoculated treatments were slow and were about 7.6% and 8.6% of the mineralisation measured for the non-UV-irradiated and UV-irradiated LDPE, respectively, after 126 days. In contrast, in the presence of the selected microorganisms, biodegradation was much more efficient and the percentages of biodegradation were 29.5% and 15.8% for the UV-irradiated and non-UV-irradiated films, respectively. The percentage decrease in the carbonyl index was higher for the UV-irradiated LDPE when the biodegradation was performed in soil inoculated with the selected microorganisms. The percentage elongation of the films decreased during the biodegradation process. The Fourier transform infra-red (FT-IR), x-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to determine structural, morphological and surface changes on polyethylene. These analyses showed that the selected microorganisms could modify and colonise both types of polyethylene. This study also confirmed the ability of these isolates to utilise virgin polyethylene without pro-oxidant additives and oxidation pretreatment, as the carbon source.
In this study, we show that degradable particles of a hydrophobic polymer can effectively deliver drugs to tumors after i.v. administration. Free-standing nanoparticles with diameters of 100–300 nm were successfully fabricated from highly hydrophobic, biodegradable poly(ω-pentadecalactone- co-butylene-co-succinate) (PPBS) copolyesters. PPBS copolymers with various compositions (20–80 mol% PDL unit contents) were synthesized via copolymerization of ω-pentadecalactone (PDL), diethyl succinate (DES), and 1,4-butanediol (BD) using Candida antarctica lipase B (CALB) as the catalyst. Camptothecin (CPT, 12–22%) was loaded into PPBS nanoparticles with high encapsulation efficiency (up to 96%) using a modified oil-in-water single emulsion technique. The CPT-loaded nanoparticles had a zeta potential of about −10 mV. PPBS particles were non toxic in cell culture. Upon encapsulation, the active lactone form of CPT was remarkably stabilized and no lactone-to-carboxylate structural conversion was observed for CPT-loaded PPBS nanoparticles incubated in both phosphate-buffered saline (PBS, pH = 7.4) and DMEM medium for at least 24 hr. In PBS at 37 °C, CPT-loaded PPBS nanoparticles showed a low burst CPT release (20–30%) within the first 24 hrs followed by a sustained, essentially complete, release of the remaining drug over the subsequent 40 days. Compared to free CPT, CPT-loaded PPBS nanoparticles showed a significant enhancement of cellular uptake, higher cytotoxicity against Lewis lung carcinoma and 9L cell lines in vitro, a longer circulation time, and substantially better antitumor efficacy in vivo. These results demonstrate the potential of PPBS nanoparticles as long-term stable and effective drug delivery systems in cancer therapy.
The biodegradability of poly(tetramethylene succinate) (PTMS), a synthetic aliphatic polyester with a high melting point, was evaluated. The ecological study showed that the distribution of PTMS-degrading microorganisms in soil environments was quite restricted compared with the distribution of microorganisms that degrade poly((epsilon)-caprolactone) (PCL), a polyester with a low melting point. However, in soil samples in which the formation of a clear zone was observed, PTMS-degrading microorganisms constituted 0.2 to 6.0% of the total number of microorganisms, which is very close to the percentage (0.8 to 8.0%) observed for PCL-degrading microorganisms. Five strains were isolated from colonies which formed distinct clear zones on agar plates with emulsified PTMS. In liquid cultures of the isolates with ground PTMS powder, strain HT-6, an actinomycete, showed the highest PTMS degrading activity. It assimilated about 60% of the ground PTMS powder after 8 days of cultivation. When a PTMS emulsion was used, a higher degradation rate was observed and more than 90% of the PTMS was assimilated in 6 days. PTMS degradation products were analyzed by gas chromatography, and it was found that 1,4-butanediol, 4-hydroxy n-butyrate, and succinic acid accumulated during cultivation. Degradation of PTMS film by the strain occurred in two steps: fragmentation and then the formation of hemispherical holes on the surface of the film. Strain HT-6 was also able to assimilate PCL and poly((beta)-hydroxybutyrate) (PHB). The crude enzyme showed a wide range of substrate specificity, being able to degrade low-molecular-weight PTMS, PCL, PHB, and even high-molecular-weight PTMS.
There is much interest in the role that agricultural practices might play in sequestering carbon to help offset rising atmospheric CO2 concentrations. However, limited information exists regarding the potential for increased carbon sequestration of different management strategies. The objective of this study was to quantify and contrast carbon dioxide exchange in traditional non-mulching with flooding irrigation (TF) and plastic film mulching with drip irrigation (PM) cotton (Gossypium hirsutum L.) fields in northwest China. Net primary productivity (NPP), soil heterotrophic respiration (Rh) and net ecosystem productivity (NEP) were measured during the growing seasons in 2009 and 2010. As compared with TF, PM significantly increased the aboveground and belowground biomass and the NPP (340 g C m−2 season−1) of cotton, and decreased the Rh (89 g C m−2 season−1) (p<0.05). In a growing season, PM had a higher carbon sequestration in terms of NEP of ∼ 429 g C m−2 season−1 than the TF. These results demonstrate that conversion of this type of land use to mulching practices is an effective way to increase carbon sequestration in the short term in cotton systems of arid areas.
The specific aim of the present study was to investigate the biodegradation and biocompatibility characteristics of rosin, a natural film-forming polymer. Both in vitro as well as in vivo methods were used for assessment of the same. The in vitro degradation of rosin films was followed in pH 7.4 phosphate buffered saline at 37°C and in vivo by subdermal implantation in rats for up to 90 days. Initial biocompatibility was followed on postoperative days 7, 14, 21, and 28 by histological observations of the surrounding tissues around the implanted films. Poly (DL-lactic-co-glycolic acid) (PLGA) (50∶50) was used as reference material for biocompatibility. Rate and extent of degradation were followed in terms of dry film weight loss, molecular weight (MW) decline, and surface morphological changes. Although the rate of in vitro degradation was slow, rosin-free films showed complete degradation between 60 and 90 days following subdermal implantation in rats. The films degraded following different rates, in vitro and in vivo, but the mechanism followed was primarily bulk degradation. Rosin films demonstrated inflammatory reactions similar to PLGA, indicative of good biocompatibility. Good biocompatibility comparable to PLGA is demonstrated by the absence of necrosis or abscess formation in the surrounding tissues. The study provides valuable insight, which may lead to new applications of rosin in the field of drug delivery.
biodegradation; biocompatibility; rosin
Biodegradation is increasingly being considered as a less expensive alternative to physical and chemical means of decomposing organic pollutants. Pathways of biodegradation have been characterized for a number of heterotrophic microorganisms, mostly soil isolates, some of which have been used for remediation of water. Because cyanobacteria are photoautotrophic and some can fix atmospheric nitrogen, their use for bioremediation of surface waters would circumvent the need to supply biodegradative heterotrophs with organic nutrients. This paper demonstrates that two filamentous cyanobacteria have a natural ability to degrade a highly chlorinated aliphatic pesticide, lindane (gamma-hexachlorocyclohexane); presents quantitative evidence that this ability can be enhanced by genetic engineering; and provides qualitative evidence that those two strains can be genetically engineered to degrade another chlorinated pollutant, 4-chlorobenzoate.
Complicated nano-patterns with linewidth less than 18 nm can be automatically hammered by using atomic force microscopy (AFM) tip in tapping mode with high speed. In this study, the special sample was thin poly(styrene-ethylene/butylenes-styrene) (SEBS) block copolymer film with hexagonal spherical microstructures. An ordinary silicon tip was used as a nano-hammer, and the entire hammering process is controlled by a computer program. Experimental results demonstrate that such structure-tailored thin films enable AFM tip hammering to be performed on their surfaces. Both imprinted and embossed nano-patterns can be generated by using a vibrating tip with a larger tapping load and by using a predefined program to control the route of tip movement as it passes over the sample’s surface. Specific details for the fabrication of structure-tailored SEBS film and the theory for auto-hammering patterns were presented in detail.
Automatic nanolithography; Atomic force microscopy; Poly(styrene-ethylene/butylenes-styrene)
Hydrophobic surface binding protein A (HsbA) is a secreted protein (14.5 kDa) isolated from the culture broth of Aspergillus oryzae RIB40 grown in a medium containing polybutylene succinate-co-adipate (PBSA) as a sole carbon source. We purified HsbA from the culture broth and determined its N-terminal amino acid sequence. We found a DNA sequence encoding a protein whose N terminus matched that of purified HsbA in the A. ozyzae genomic sequence. We cloned the hsbA genomic DNA and cDNA from A. oryzae and constructed a recombinant A. oryzae strain highly expressing hsbA. Orthologues of HsbA were present in animal pathogenic and entomopathogenic fungi. Heterologously synthesized HsbA was purified and biochemically characterized. Although the HsbA amino acid sequence suggests that HsbA may be hydrophilic, HsbA adsorbed to hydrophobic PBSA surfaces in the presence of NaCl or CaCl2. When HsbA was adsorbed on the hydrophobic PBSA surfaces, it promoted PBSA degradation via the CutL1 polyesterase. CutL1 interacts directly with HsbA attached to the hydrophobic QCM electrode surface. These results suggest that when HsbA is adsorbed onto the PBSA surface, it recruits CutL1, and that when CutL1 is accumulated on the PBSA surface, it stimulates PBSA degradation. We previously reported that when the A. oryzae hydrophobin RolA is bound to PBSA surfaces, it too specifically recruits CutL1. Since HsbA is not a hydrophobin, A. oryzae may use several types of proteins to recruit lytic enzymes to the surface of hydrophobic solid materials and promote their degradation.
Micro- and nanospheres composed of biodegradable polymers show promise as versatile devices for the controlled delivery of biopharmaceuticals. Whereas important properties such as drug release profiles, biocompatibility, and (bio)degradability have been determined for many types of biodegradable particles, information about particle degradation inside phagocytic cells is usually lacking. Here, we report the use of confocal Raman microscopy to obtain chemical information about cross-linked dextran hydrogel microspheres and amphiphilic poly(ethylene glycol)-terephthalate/poly(butylene terephthalate) (PEGT/PBT) microspheres inside RAW 264.7 macrophage phagosomes. Using quantitative Raman microspectroscopy, we show that the dextran concentration inside phagocytosed dextran microspheres decreases with cell incubation time. In contrast to dextran microspheres, we did not observe PEGT/PBT microsphere degradation after 1 week of internalization by macrophages, confirming previous studies showing that dextran microsphere degradation proceeds faster than PEGT/PBT degradation. Raman microscopy further showed the conversion of macrophages to lipid-laden foam cells upon prolonged incubation with both types of microspheres, suggesting that a cellular inflammatory response is induced by these biomaterials in cell culture. Our results exemplify the power of Raman microscopy to characterize microsphere degradation in cells and offer exciting prospects for this technique as a noninvasive, label-free optical tool in biomaterials histology and tissue engineering.
biodegradation; microspheres; Raman microscopy; phagocytosis; drug delivery
The cyclic nitramine explosive CL-20 (2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane) was examined in soil microcosms to determine whether it is biodegradable. CL-20 was incubated with a variety of soils. The explosive disappeared in all microcosms except the controls in which microbial activity had been inhibited. CL-20 was degraded most rapidly in garden soil. After 2 days of incubation, about 80% of the initial CL-20 had disappeared. A CL-20-degrading bacterial strain, Agrobacterium sp. strain JS71, was isolated from enrichment cultures containing garden soil as an inoculum, succinate as a carbon source, and CL-20 as a nitrogen source. Growth experiments revealed that strain JS71 used 3 mol of nitrogen per mol of CL-20.
In a field release experiment, an isolate of Pseudomonas fluorescens, which was chromosomally modified with two reporter gene cassettes (lacZY and Kan(supr)-xylE), was applied to spring wheat as a seed coating and subsequently as a foliar spray. The wild-type strain was isolated from the phylloplane of sugar beet but was found to be a common colonizer of both the rizosphere and phylloplane of wheat as well. The impact on the indigenous microbial populations resulting from release of this genetically modified microorganism (GMM) was compared with the impact of the unmodified, wild-type strain and a nontreated control until 1 month after harvest of the crop. The release of the P. fluorescens GMM and the unmodified, wild-type strain resulted in significant but transient perturbations of some of the culturable components of the indigenous microbial communities that inhabited the rhizosphere and phylloplane of wheat, but no significant perturbations of the indigenous culturable microbial populations in nonrhizosphere soil were found. Fast-growing organisms that did not produce resting structures (for example, fluorescent pseudomonads and yeasts) seemed to be most sensitive to perturbation. In terms of hazard and risk to the environment, the observed microbial perturbations that resulted from this GMM release may be considered minor for several reasons. First, the recombinant P. fluorescens strain caused changes that were, in general, not significantly different from those caused by the unmodified wild-type strain; second, perturbations resulting from bacterial inoculations were mainly small; and third, the release of bacteria had no obvious effects on plant growth and plant health.
The biodegradability of three aliphatic adipic acid diesters and a 1,3-butylene glycol adipic acid polyester was determined in acclimated, activated sludge systems. Rapid primary biodegradation from 67 to 99+% was observed at 3- and 13-mg/liter feed levels for di-n-hexyl adipate, di(2-ethylhexyl) adipate, and di(heptyl, nonyl) adipate in 24 h. When acclimated, activated sludge microorganisms were employed as the seed for two carbon dioxide evolution procedures, greater than 75% of the theoretical carbon dioxide was evolved for the three diesters and the polyester in a 35-day test period. The essentially complete biodegradation observed in these studies suggests that these esters would not persist when exposed to similar mixed microbial populations in the environment.