Enzymatic pretreatment mitigates the dissemination of antibiotic resistance genes via regulating microbial populations and gene expressions during food waste fermentation

Food waste(FW)has been recognized as essential reservoir for resource recovery via anaerobic fermentation,which could also bring the potential risk of antibiotic resistance genes(ARGs)dissemination.Although the structural deficiency of FW could be stimulated by enzymatic pretreatment to enhance fermentation efficiency,the influences of enzymatic pretreatment on ARGs fate and microbial metabolic pathways involved in ARGs dissemination have rarely been reported.This work proved that enzymatic pretreatment could effectively decrease the total abundance of ARGs(reduced by 13.8%-24.5%)during long-term FW fermentation.It was found that enzymatic pretreatment significantly reduced the ARGs belonging to the efflux pump,which might be ascribed to its ability to increase membrane permeability.Furthermore,enzymatic pretreatment was in favor of reducing microbial diversity and various potential ARGs host(e.g.,Methanosarcina,Clostridium,Prevotella,Parabacteroides).Also,this pretreatment remarkably up-regulated the genetic expressions involved in ABC transporter(e.g.,eryF and mntA)and down-regulated the genetic expressions that participated in DNA replication,two-component systems(e.g.,uphA and cckA),and quorum sensing(e.g.,rpfF and lsrG),thereby decreasing ARGs transmission.This study would expand the insight of the influences of pretreatment method on ARGs fate during FW fermentation,and offer practical guidance on the sustainable management of FW.

Effect of Soybean Peptides on in vitro Ruminal Fermentation and Microbial Population

This study was conducted to investigate the effect of Soybean Peptides(SPs) on ruminal fermentation and microbial diversity in vitro. Four levels of SPs supplements(0, 0.25%, 0.50% and 0.75% dry matter basis) were tested. p H, NH3-N, Neutral Detergent Fiber(NDF), Acid Detergent Fiber(ADF), Dry Matter Digestibility(DMD) and the Total Volatile Fatty Acid(TVFA) were measured at 6, 24, 48 h of in vitro mixed incubation the fluids. Microbial populations were determined at 24 h and Microbial Proteins(MCP) were determined at 24 and 48 h. The gas production after 48 h in vitro incubation presented linear growth(p<0.05) and the highest content was the level of 0.5% SPs(dry matter basis). NH3-N concentration reached the highest concentration with 0.75%(dry matter basis) at 48 h. p H linearly increased(linear, p<0.05) from 6 to 48 h. The digestion of DMD increased with increasing doses of SPs at 24 h. NDF and ADF linearly(p<0.01) decreased by adding SPs. The concentration of TVFA linearly increased(p<0.05) at 6, 24 and 48 h, and reached the highest concentration at the level of 0.75%(dry matter basis). SPs decreased(p<0.05) the molar proportion of acetate and propionate, respectively at 24 and 48 h. Acetat to Propionate ratio(A/P; linear, p<0.05) increased at 48 h, and reached the greatest value at the level of 0.75%(dry matter basis) at 48 h. The populations of rumen Ruminococcus flavefaciens(R. flavefaciens), Ruminococcusalbus(R. albus), Fibrobacter succinogenes(F. succinogenes), Butyrivibrio fibrisolvens(B. fibrisolvens), Streptococcus bovis(S.bovis), Ruminobacter amylophilus(R. amylophilus) and Succinimonas amylolytica(S. amylolytica) were analyzed based on the total rumen bacterial 16 S ribosomal deoxyribonucleic acid(r DNA). The relative abundance of R. flavefaciens and R. albus increased at 24 h, but the relative abundance of F. succinogenes decreased at this time. The incubation of SPs had no effect on the abundance of S. bovis and R. amylophilus. The relative abundance of B. fibrisolvens and S. amylolytica reached the greatest value(p<0.05) at middle doses of SPs inclusion at 24 h. The value of MCPs linearly increased(p<0.05) at 24 and 48 h. These results showed that SPs could improve in vitro fermentation and nutrient digestion of feed substrates, and had the ability to modulate the ruminal fermentation pattern by regulating the composition of functional rumen microbes. Hence, SPs might be a potential feed additive applied in the diets of ruminants.

Preliminary investigation on the role of microorganisms in the production of phosphine

The relationships between the phosphine content and various microbial populations,activities of different enzymes were investigated firstly.The results indicated that the phosphine content of samples from various environments was positively related to total anaerobic microorganisms,organic phosphate compound-dissolving bacteria,denitrifying bacteria,and the activities of alkaline phosphatase and dehydrogenase,with correlation coefficients (R^2) up to 0.93,0.90,0.69,0.79,and 0.82,respectively.Results also sh...

Antioxidant compounds and minerals in tomatoes by Trichodermaenriched biofertilizer and their relationship with the soil environments

The experiment was conducted to evaluate the efficacy of Trichoderma-enriched biofertilizer（BioF/compost）on antioxidants and minerals in ripe tomatoes and soil health improvements in terms of nutrient availability and microbial populations.The study was comprised of six treatments：control（zero input）;recommended doses of NPK（135.5,45.6 and 22.9 kg ha^-1,respectively）;100%BioF/compost;75%BioF/compost＋25%N;50%BioF/compost＋50%N;and 25%BioF/compost＋75%N.The recommended doses of P and K were used in the last three treatments.It was found that the application of 100%BioF/compost enhanced plant growth,leaf greenness,and produced 12.9%higher yield compared to the recommended doses of NPK and other treatments.Mineral contents（P,K,Ca,Mg,Cu,Fe,Mn and Zn）in tomato roots,shoots and fruits and antioxidant compounds,i.e.,ascorbic acid,β-carotine,and lycopene were increased significantly in fruits fertilized with100%BioF/compost.The high efficiency of Trichoderma compost might be the result of its potential of nutrient solubilization and harboring soil microorganisms.Collectively,BioF/compost increased soil fertility and favored growth of microbes in the rhizosphere which ultimately contributed to higher yield,antioxidant,and mineral concentrations in tomatoes.Thus,Trichoderma-enriched biofertilizer may reduce application of chemical fertilizers and therefore,can be considered as a noble practice in sustainable agriculture.

Causes of Deterioration and Changes of Aroma Components in Cigarette Raw Materials

The quality of the favoring agents and fragrances degenerated frequently during the storage and transportation process in a cigarette factory in Yunnan Province, causing huge economic losses. In order to ascertain the causes of deterioration, the microbial population, quantity and aroma components of the raw mate- rials were analyzed in the present study. The results showed that a total of 28 bacterial strains and one fungal strain were isolated and identified, belonging to 13 genera. The 29 strains were Bacillus sp. , Microbacterium sp. , Sphingomonas sp. , Leclercia sp. and Acinetobacter sp. isolated from normal ZY material; Bacil- lus sp. , Microbacterium sp. , Sphingomonas sp. , Brevundimonus sp. , Acinetobacter sp. , Agrobacterium sp. , Ralstonia sp. , Acetobacterium sp. , Saccharomyces sp. and Acidovorax sp. isolated from deteriorated ZY material; Agrobacterium sp. , Microbacterium sp. , Burkholderia sp. , Bacillus subtilis strain, Acetobacterium sp. and Leclercia sp. isolated from normal RZ material ; Agrobacterium sp. , Microbacterium sp. , Bacillus subtilis strain, Acetobacterium sp. , Lec/erc/a sp. , Enter- obacter sp. , Acinetobacter sp. and Saccharomyces sp. isolated from deteriorated RZ material. Microbacterium was the dominant genus by number in normal ZY ma- terial, and the population of this genus accounted for 55.6% of the total. In deteriorated ZY material, Acetobacterium, Bacillus and Saccharomyces were dominant genera, as the population of the three genera occupied 75.0% of the total. The total population of microorganisms in the deteriorated ZY material was 90 times of that in normal ZY material. In normal RZ material, the population of each of Microbacterium, Agrobacterium and Bacillus accounted for 25%, while that of each of Leclercia and Burkholderia accounted for 12.5%. In deteriorated RZ material, the population of Bacillus accounted for 65.0% of the total. The total population of the microorganisms in deteriorated RZ material was 29 times of that in normal RZ material, indicating that the deterioration of the materials was caused by microor- ganisms. The contents of aroma components were detected using two-dimensional gas chromatography-mass spectrometry （GC-MS）. The results showed that in nor- mal ZY material, the content of β-damascone was higher than that of other aroma components. And normal ZY material had higher contents of benzaldehyde, β-damascone, β-damascone, β-damascone, menthol and benzyl benzoate than deteriorated ZY material, and the differences in the contents of β-ddamascone and β-damascone were most significant between the two materials. In normal RZ material, the content of β-damascone was higher than that of other aroma components. And normal RZ material had higher contents of menthol, ethyl capratc, β-damascone, β-damascone and tabanane C than deteriorated RZ material, and the differ- ences in the contents of ethyl caprate and capric acid were most significant between the two materials. The results revealed that aroma components changed greatly in deteriorated tobacco materials.

Degradation of crude oil by indigenous microorganisms supplemented with nutrients

Different kinds of mineral nutrients(NO_3-N, NH_4-N and PO_4-P) were applied in the simulated oil-polluted seawater for enhancing oil biodegradation in the N/P ratio 10∶1 and 20:1 Although indigenous microorganisms have the ability to degrade oil, adding nutrients accelerated biodegradation rates significantly. For the group amended with NO_3-N and PO_4-P in the ratio 10∶1, the reaction rate coefficient was 4 times higher than the natural biodegradation. Chemical and microbiological analysis showed that the optimal N/P ratio in the system is 10∶1, and microorganisms tend to utilize nitrate rather than ammonium as N source.

Effects of Different Halophytes on Soil Microflora and Enzyme Acti-vities of Saline-alkali Soil

In the pot experiment,seven varieties of halophytes were grown in saline-alkali soil to investigate the responses of microflora and soil enzymes in the rhizosphere.The results showed that compared to the control,the population of bacterial colony(84.8%-95.6%),actinomycetes colony(12.0%-14.5%)and fungi colony(0.5%-1.1%)increased significantly(P<0.05).The population of ammonia bacteria,aerobic cellulose decomposition bacteria in the soil of Vicia sativa L.(201.99%and 395.49%),Medicago sativa(152.43%and 319.90%)and Sesbania cannabina(Retz.)Pori(193.14%and 396.08%)were higher significantly than that of Panicum virgatum L.(49%and 60%),Sorghum bicolor(L.)Moench(99%and 210%),Amaranthus hypochondriacus L.(75%and 36%)and Aneurotepidimu chinense(75%and 77%)(P<0.05).However,Sorghum bicolor(L.)Moench was evidently higher than Panicum virgatum L.,Amaranthus hypochondriacus L.and Aneurotepidimu chinense(P<0.05)in the soil.The population of ammonia bacteria and aerobic cellulose-decomposing bacteria was significantly correlated with the five enzymes(P<0.05),which could improve the microenvironment in saline-alkali soil to accelerate the element cycling and promote the sustainable development of agriculture through cultivating Medicago sativa,Vicia sativa L.,Sesbania cannabina(Retz.)Pori and Sorghum bicolor(L.)Moench.

Xylanase-producing microflora in Eastern Ghats of Andhra Pradesh,India

We screened soil samples collected from underneath shrubs and/or large trees at different locations in the Eastern Ghats of Andhra Pradesh for xylanase-producing microorganisms. Xylose-utilizing bacteria were numerically dominant in soils of most locations whereas xylose-and xylan-utilizing actinobacteria were minor components. Xylan-utilizing fungi constituted a major share of total microbial populations in soil samples collected at half of the sites, whereas xylan-utilizing bacteria were predominant at other sampling locations. Some of the isolates of fungi exhibited xylanase activity with a range of400–4000 U/ml, indicating great potential for their uses in paper, pulping and bioethanol industries for producing value-added products.

System of Rice Intensification Verses Conventional Rice System: Off-farm Field Studies

With inevitable growth of demand for human and industrial needs,water available for agriculture will become scarcer in the future.India is a highly water-stressed country.Hence,India needs to invest in improving its water productivity,and any capacity to produce more rice with less water.System of Rice Intensification(SRI)has attracted much attention in increasing rice yield per unit area.For this study,fifteen farmers were selected those were practicing SRI technology by themselves during the Boro-cultivation season(January-April).The study was continued for three consecutive years 2012 to 14 on the same fields.In addition to the SRI plots,a similar size of non-SRI plot was maintained in conventional cultivation for comparison purpose.On an average,the non-SR I ight increased by 12%,number of tillers per square meter by 85%,number of reproductive tillers per hill by 286%,weight of panicle per hill by 139%,number of seeds per panicle by 41%and test weight by 26% due to SRI practice over the non-SRI practice.Average increment in straw and grain yield due to SRI over the non-SRI is 70%and 59%respectively.The physico-chemical and biological properties of soil improved due to SRI practice.

Microbial responses to combined oxidation and catalysis treatment of 1,4-dioxane and co-contaminants in groundwater and soil

Post-treatment impacts of a novel combined hydrogen peroxide(H2O_(2))oxidation and WOx/ZrO_(2) catalysis used for the removal of 1,4-dioxane and chlorinated volatile organic compound(CVOC)contaminants were investigated in soil and groundwater microbial community.This treatment train removed~90%1,4-dioxane regardless of initial concentrations of 1,4-dioxane and CVOCs.The Illumina Miseq platform and bioinformatics were used to study the changes to microbial community structure.This approach determined that dynamic shifts of microbiomes were associated with conditions specific to treatments as well as 1,4-dioxane and CVOCs mixtures.The biodiversity was observed to decrease only after oxidation under conditions that included high levels of 1,4-dioxane and CVOCs,but increased when 1,4-dioxane was present without CVOCs.WO_(x)/ZrO_(2) catalysis reduced biodiversity across all conditions.Taxonomic classification demonstrated oxidative tolerance for members of the genera Massilia and Rhodococcus,while catalyst tolerance was observed for members of the genera Sphingomonas and Devosia.Linear discriminant analysis effect size was a useful statistical tool to highlight representative microbes,while the multidimensional analysis elucidated the separation of microbiomes under the low 1,4-dioxane-only condition from all other conditions containing CVOCs,as well as the differences of microbial population among original,post-oxidation,and post-catalysis states.The results of this study enhance our understanding of microbial community responses to a promising chemical treatment train,and the metagenomic analysis will help practitioners predict the microbial community status during the post-treatment period,which may have consequences for long-term management strategies that include additional biodegradation treatment or natural attenuation.

Effect of wastewater COD/N ratio on aerobic nitrifying sludge granulation and microbial population shift

The effect of COD/N ratio on the granulation process and microbial population succession was investigated.Four identical sequencing batch reactors,R1,R2,R3 and R4,were operated with various initial COD/N ratios ranging from 0/200 to 800/200（m/m）.Ethanol was fed as the source of COD.Aerobic granules were successfully cultivated in R2 and R3,operating with the COD/N ratio of 200/200 and 400/200,respectively.Scanning electron microscope observations indicated that short rod-shaped and spherical bacteria were dominant in R2,while granules produced in R3 were surrounded with a large amount of filamentous bacteria.The average specific nitritation rate in R2 and R3 were 0.019 and 0.008 mg N/（mg MLVSS.hr）,respectively.Fluorescence in situ hybridization results demonstrated that nitrifying bacteria population was enriched remarkably in R2.It indicated that nitrification ability and nitrifying bacteria population were enriched remarkably at low COD/N ratio.However,no granules were formed in R1 and R4 which might attribute to either limited or excessive extracellular polymeric substances production.This study contributed to a better understanding of the role of COD/N ratio in nitrifying sludge granulation.

Effects of feeding whole linseed on ruminal fatty acid composition and microbial population in goats

The objective of the present study was to evaluate the effect of feeding different levels of whole linseed,as a source of n-3 polyunsaturated fatty acids(PUFA), on ruminal fatty acid composition and microbial population in the goat. Twenty-four crossbred Boer goats were assigned to 3 dietary treatments: LO(control), L10 and L20 containing 0,10%, or 20% whole linseed, respectively. The ruminal pH and concentration of total volatile fatty acids(VFA) were not affected by dietary treatments. The feeding of L10 and L20 diets produced higher(P < 0.05) molar proportions of acetate and lower(P < 0.05) molar proportions of butyrate and valerate than the LO diet. Molar proportions of myristic acid(C14:0) and palmitic acid(C16:0) were lower(P < 0.05) in the rumen of goats offered L10 and L20 diets than the control diet. However, stearic acid(C18:0), vaccenic acid(C18:1 trans-11), conjugated linoleic acid(CLA,C18:2 trans-10, cis-12) and a-lenolenic acid(C18:3 n-3) were higher(P < 0.05) in the rumen of goats fed L10 and L20 than L0. Both inclusion levels of linseed in the diet(L10 and L20) reduced the ruminal total bacteria, methanogens, and protozoa compared with LO(P < 0.05). The effect of the dietary treatments on cellulolytic bacteria, varied between the individual species. Both inclusion levels of linseed resulted in a significant decrease(P < 0.05) in the population of Fibrobacter succinogenes, and Rumunococus flavefaciens compared with LO, with no significant difference between the groups fed linseed diets. The population of Rumunococus albus was not affected by the different dietary treatments. It was concluded that inclusion of whole linseed in the diet of goats could increase the concentration of PUFA in the rumen, and decrease the population of F. succinogenes, R. flavefaciens, methanogens and protozoa in rumen liquid of goats.

Ruminal fermentation and microbial community differently influenced by four typical subtropical forages in vitro

The present study evaluated the effects of 4 typical subtropical forages on ruminal microbial community composition to formulate a better diet for buffalo.Corn straw silage,elephant grass,cassava residues and sugarcane tail silage were used as substrates for in vitro fermentation.Eight replicates were set up for every substrate,and fermentation was carried out in a 100-m L glass syringe,using buffalo rumen inoculum.Every replicate was anaerobically dispensed with 10 m L of rumen inoculum,20 m L of Mc Dougall's buffer and 200 mg of dried substrate,and placed in a water bath at 39C.Gas production was recorded at 0,2,6,12,24,36,48 and 72 h of incubation.After 24 h,fermentation was ceased for 4 replicates and samples were collected.Volatile fatty acids(VFA) concentrations were measured using gas chromatography.Microbial populations were quantified using quantitative real-time PCR(q RT-PCR),and microbial community was analyzed using high throughput sequencing technology.The results showed,cassava residues as substrate had the highest gas production,acetate,propionate and total VFA concentrations(P < 0.05),and corn straw silage had the lowest acetate:propionate ratio(P < 0.05).The lowest numbers of fungi,Ruminococcus albus and Fibrobacter succinogenes,and the highest number of protozoa were observed with cassava residues(P < 0.05).The least abundances of bacterial phyla Firmicutes,Bacteroidetes and genus Prevotella,and substantially higher abundance of phylum proteobacteria(56%) and genus Succinivibrio(52%) were observed with cassava residues.The most abundances of Methanobrevibacter gottschalkii and Entodinium were observed with cassava residues.Spearman's correlations analysis showed,Succinivibrio had strong positive correlations with propionate,butyrate,Metadinium and M.gottschalkii,indicating fermentation products were related to microbial community.In conclusion,incubation with cassava residues resulted in lower number of fiber degrading microbes but higher protozoal population because of its low fiber contents.The microbial community was highly altered by in vitro incubation with cassava residues,whereas remained similar for the other 3 high fiber containing substrates.

Microbial population dynamics and changes in main nutrients during the acidification process of pig manures

This study evaluated the impact of pig manure acidification on anaerobic treatment and composition of the fecal microbial community. According to the different chemical oxygen demand （COD） in the anaerobic treatment processes, pig manure was diluted 4 times （×4）, 16 times （×16）, or 64 times （×64） and subjected to acidification. During the acidification process, pH, soluble chemical oxygen demand （SCOD）, volatile fatty acids （VFAs）, nitrogen （N）, phosphorus （P） and potassium （K） were determined along with microbial population dynamics. The pH of the three dilutions first declined, and then slowly increased. The total VFAs of x4 and xl6 dilutions peaked on day 15 and 20, respectively. The content of acetic acid, propanoic acid, butanoic acid and valeric acid of the × 4 dilution were 23.6, 11.4, 8.8 and 0.6 g/L respectively, and that of the xl6 dilution was 5.6, 2.3, 0.9 and 0.2 g/L respectively. Only acetic acid was detected in the ×64 dilution, and its level peaked on day 10. The results showed that the liquid pig manure was more suitable to enter the anaerobic methanogenic hioreactors after two weeks of acidification. During the acidification process, total P concentration increased during the first ten days, then dropped sharply, and rose again to a relatively high final concentration, while total N concentration rose initially and then declined. Based on the analysis of denaturing gradient gel electrophoresis （DGGE） and 16S rRNA gene clone library, we concluded that the acidification process reduced the number of pathogenic bacteria species in pig manure.

Temporal variation of microbial population in a thermophilic biofilter for SO_2 removal

The performance of a biofilter relies on the activity of microorganisms during the gas contaminant treatment process. In this study, SO2 was treated using a laboratory-scale biofilter packed with polyurethane foam cubes（PUFC）, on which thermophilic desulfurization bacteria were attached. The thermophilic biofilter effectively reduced SO2 within 10 months of operation time, with a maximum elimination capacity of 48.29 g/m^3/hr.Temporal shifts in the microbial population in the thermophilic biofilter were determined through polymerase chain reaction-denaturing gradient gel electrophoresis and deoxyribonucleic acid（DNA） sequence analysis. The substrate species and environmental conditions in the biofilter influenced the microbial population. Oxygen distribution in the PUFC was analyzed using a microelectrode. When the water-containing rate in PUFC was over 98%, the oxygen distribution presented aerobic–anoxic–aerobic states along the test route on the PUFC. The appearance of sulfate-reducing bacteria was caused by the anaerobic conditions and sulfate formation after 4 months of operation.

Effects of oxygen and water content on microbial distribution in the polyurethane foam cubes of a biofilter for SO_2 removal

The performance of a biofilter for off-gas treatment relies on the activity of microorganisms and adequate O_2 and H_2O. In present study, a microelectrode was applied to analyze O_2 in polyurethane foam cubes（PUFCs） packed in a biofilter for SO_2 removal. The O_2 distribution varied with the density and water-containing rate（WCR） of PUFCs. The O_2 concentration dropped sharply from 10.2 to 0.8 mg/L from the surface to the center of a PUFC with 97.20%of WCR. The PUFCs with high WCR presented aerobic–anoxic–aerobic areas.Three-dimensional simulated images demonstrated that the structure of PUFCs with high WCR consisted of an aerobic ＂shell＂ and an anoxic ＂core＂, with high-density PUFCs featuring a larger anoxic area than low-density PUFCs. Moreover, the H_2O distribution in the PUFC was uneven and affected the O_2 concentration. Whereas aerobic bacteria were observed in the PUFC surface, facultative anaerobic microorganisms were found at the PUFC core, where the O_2 concentration was relatively low. O_2 and H_2O distributions differed in the PUFCs, and the distribution of microorganisms varied accordingly.