Effect of Enhanced Hydrolytic Acidification Process on the Treatment of Azo Dye Wastewater.

The hydrolysis acidification process is an economical and effective method, but its efficiency is still low in treating azo dye wastewater. It is therefore crucial to find more suitable and efficient means or techniques to further strengthen the process of treating azo dye wastewater by a hydrolytic acidification process. In this study, a hydrolytic acidification aerobic reactor was used to simulate the azo dye wastewater process. The change of wastewater quality during the reaction process was monitored, and the deep enhancement effect of single or composite biological intensification technology on the treatment of azo dye wastewater by the hydrolytic acidification process was also explored. Co-substrate strengthening and the addition of fructose co-substrate can significantly improve the efficiency of hydrolytic acidification. Compared with the experimental group without the addition of fructose, the decolorization ratio of wastewater was higher (93%) after adding fructose co-substrate. The immobilization technology was strengthened, and the immobilized functional bacteria DDMZ1 pellet was used to treat the simulated azo dye wastewater. The results showed that the composite technology experimental group with the additional fructose co-matrix had a better decolorization efficiency than the single immobilized bio-enhancement technology, with the highest decolorization ratio of 97%. As a composite biological intensification method, the fructose co-matrix composite with immobilized functional bacteria DDMZ1 technology can be applied to the treatment of azo dye wastewater.

Co-metabolic biodegradation of structurally discrepant dyestuffs by Klebsiella sp. KL-1: A molecular mechanism with regards to the differential responsiveness.

In this study, an attempt was made to decipher the underlying differential response mechanism of Klebsiella sp. KL-1 induced by exposure to disparate categories of dyestuffs in xylose (Xyl) co-metabolic system. Here, representative reactive black 5 (RB5), remazol brilliant blue R (RBBR) and malachite green (MG) belonging to the azo, anthraquinone and triphenylmethane categories were employed as three model dyestuffs. Klebsiella sp. KL-1 enabled nearly 98%, 80% or 97% removal of contaminants in assays Xyl + RB5, Xyl + RBBR or Xyl + MG after 48 h, which was respectively 16%, 11% or 22% higher than those in the assays devoid of xylose. LC-QTOF-MS revealed an increased formation of smaller molecular weight intermediates in assay Xyl + RB5, whereas more metabolic pathways were deduced in assay Xyl + RBBR. Metaproteomics analysis displayed remarkable proteome alteration with regards to the structural difference effect of dyestuffs by Klebsiella sp. KL-1. Significant (p-value<0.05) activation of pivotal candidate NADH-quinone oxidoreductase occurred after 48 h of disparate dyestuff exposure but with varying abundance. Dominant FMN-dependent NADH-azoreductase, Cytochrome d terminal oxidase or Thiol peroxidase were likewise deemed to be responsible for the catalytic cleavage of RB5, RBBR or MG, respectively. Further, the differential response mechanism towards the structurally discrepant dyestuffs was put forward. Elevated reducing force associated with the corresponding functional proteins/enzymes was transferred to the exterior of the cell to differentially decompose the target contaminants. Overall, this study was dedicated to provide in-depth insights into the molecular response mechanism of co-metabolic degradation of refractory and structurally discrepant dyestuffs by an indigenous isolated Klebsiella strain.

Biochemical characterization of a novel azo reductase named BVU5 from the bacterial flora DDMZ1: application for decolorization of azo dyes.

One of the main mechanisms of bacterial decolorization and degradation of azo dyes is the use of biological enzymes to catalyze the breaking of azo bonds. This paper shows the expression and properties of a novel azo reductase (hybrid-cluster NAD(P)-dependent oxidoreductase, accession no. A0A1S1BVU5, named BVU5) from the bacterial flora DDMZ1 for degradation of azo dyes. The molecular weight of BVU5 is about 40.1 kDa, and it contains the prosthetic group flavin mononucleotide (FMN). It has the decolorization ability of 80.1 ± 2.5% within 3 min for a dye concentration of 20 mg L-1, and 53.5 ± 1.8% even for a dye concentration of 200 mg L-1 after 30 min. The optimum temperature of enzyme BVU5 is 30 °C and the optimum pH is 6. It is insensitive to salt concentration up to a salinity level of 10%. Furthermore, enzyme BVU5 has good tolerance toward some metal ions (2 mM) such as Mn2+, Ca2+, Mg2+ and Cu2+ and some organic solvents (20%) such as DMSO, methanol, isopentyl, ethylene glycol and N-hexane. However, the enzyme BVU5 has a low tolerance to high concentrations of denaturants. In particular, it is sensitive to the denaturants guanidine hydrochloride (GdmCl) (2 M) and urea (2 M). Analysis of the dye substrate specificity shows that enzyme BVU5 decolorizes most azo dyes, which is indicating that the enzyme is not strictly substrate specific, it is a functional enzyme for breaking the azo structure. Liquid chromatography/time-of-flight/mass spectrometry (LC-TOF-MS) revealed after the action of enzyme BVU5 that some intermediate products with relatively large molecular weights were produced; this illustrates a symmetric or an asymmetric rapid cleavage of the azo bonds by this enzyme. The potential degradation pathways and the enzyme-catalyzed degradation mechanism are deduced in the end of this paper. The results give insight into the potential of a rapid bio-pretreatment by enzyme BVU5 for processing azo dye wastewater.

Application of a Spiral Symmetric Stream Anaerobic Bioreactor for treating saline heparin sodium pharmaceutical wastewater: Reactor operating characteristics, organics degradation pathway and salt tolerance mechanism.

In this study, a Spiral Symmetry Stream Anaerobic Bioreactor (SSSAB) was adopted for treating actual saline heparin sodium pharmaceutical wastewater (HSPW). After adaptation, under the influent COD of 8731 mg/L, OLR of 6.98 kg COD/(m³â€¢d) and salinity of 3.57 wt%, the COD removal reached up to 82%. This value is much higher than the reported for the other reactors at similar salinity. Benzenes are the major organic compounds in HSPW. The main rate-limiting steps are the degradations of phenol and p-cresol. In addition, the degradation pathways of typical benzenes in HSPW were analyzed. After adaptation, the soluble salt content in the granular sludge increased, and the bacterial extracellular polymers (EPS), especially tightly-bound EPS also significantly increased. 16S rRNA analysis revealed that the microbial community in the anaerobic granular sludge (AGS) had become adapted to the HSPW treatment since Mesotoga (12.4%), Anaerophaga (9.0%), Oceanotoga (6.1%) and Aminobacterium (4.1%) increased from previously below 1.0% values. The relative abundance of Methanosarcina in the upper layer of the reactor (68.7%) is significantly higher than that at the bottom (3.8%). This proves the superiority of the SSSAB structure. Finally, a model for salt-tolerant microorganisms is given, which proposes a mechanism for this study and provides reference for other anaerobic biological treatments of high-salt containing wastewater.

Variation and comparison of biotoxicity during typical biological treatment of dyeing wastewater.

In present study, dyeing wastewater samples were collected from three typical dyeing wastewater treatment plants in Wujiang, Shengze and Shanghai, China. Physicochemical properties and biotoxicity indicators (luminescent bacteria acute toxicity and umu genotoxicity) were tested and the relationships among them were analyzed. The results revealed that two biotoxicity indicators varied significantly among different treatment units of three plants. After treatment by plant A, luminescent bacteria acute toxicity of dyeing wastewater reduced effectively, while umu genotoxicity increased significantly. Two biotoxicity indicators exhibited decrease and increase trends during the treatment processes of plant B and plant C, respectively. Correlation analysis indicated that there was little correlation among biotoxicity indicators and physicochemical properties, meanwhile two kinds of biotoxicity indicators were relatively independent. Therefore, it was recommended that comprehensive evaluation of dyeing wastewater toxicity needs the combination of various biotoxicity indicators, and the relationship among biotoxicity indicators and physicochemical properties of dyeing wastewater should be established individually. The results of this study would offer a general understanding and evaluation of biotoxicity during actual dyeing wastewater treatment processes and provide database for toxicity reduction and management of dyeing wastewater.

Deciphering the mechanism of carbon sources inhibiting recolorization in the removal of refractory dye: Based on an untargeted LC-MS metabolomics approach.

In this study, the biological decolorization of reactive black 5 (RB5) by Klebsiella sp. KL-1 in yeast extract (YE) medium was captured the recolorization after exposure to O2, which induced a 15.82% reduction in decolorization efficiency. Similar result was also observed in YE + lactose medium, but not in YE + glucose/xylose media (groups YE + Glu/Xyl). Through biodegradation studies, several degradation intermediates without quinoid structure were produced in groups YE + Glu/Xyl and differential degradation pathways were deduced in diverse groups. Metabolomics analysis revealed significant variations in up-/down-regulated metabolites using RB5 and different carbon sources. Moreover, the underlying mechanism of recolorization inhibition was proposed. Elevated reducing power associated with variable metabolites (2-hydroxyhexadecanoic acid, 9(R)-HODE cholesteryl ester, linoleamide, oleamide) rendered additional reductive cleavage of C-N bond on naphthalene ring. This study provided a new orientation to inhibit recolorization and deepened the understanding of the molecular mechanism of carbon sources inhibiting recolorization in the removal of refractory dyes.

Co-metabolic degradation of refractory dye: A metagenomic and metaproteomic study.

Fructose was utilized as an additional co-substrate to systematically investigate the molecular mechanism of its boosting effect for the degradation of refractory dye reactive black 5 (RB5) by a natural bacterial flora DDMZ1. A decolorizing rate of 98% was measured for sample YE + FRU(200) (with 3 g/L fructose additionally to yeast extract medium, 10% (v/v) inoculation size of flora DDMZ1, 200 mg/L RB5) after 48 h. This result was 21% and 77%, respectively, higher than those of samples with only yeast extract or only fructose. Fructose was found to significantly stimulated both intracellular and extracellular azoreductase secretion causing enhanced activity. Metagenomic sequencing technology was used to analyze the functional potential of genes. A label-free quantitative proteomic approach further confirmed the encoding of functional proteins by the candidate genes. Subsequently, the molecular mechanism of RB5 degradation by candidate genes and functional proteins of the dominant species were proposed. This study provides important perspectives to the molecular mechanism of co-metabolic degradation of refractory pollutants by a natural bacterial flora.

Sugar sources as Co-substrates promoting the degradation of refractory dye: A comparative study.

Four sugar sources were used as co-substrates to promote the degradation of a selected refractory dye reactive black 5 (RB5) by the natural bacterial flora DDMZ1. The boosting performance of the four sugar sources on RB5 decolorization ranked as: fructose > sucrose > glucose > glucose + fructose. Kinetic results of these four co-metabolism systems agreed well with a first-order kinetic model. Four sugar sources stimulated the extracellular azoreductase secretion causing enhanced enzyme activity. An increased formation of low molecular weight intermediates was caused by the addition of sugar sources. The toxicity of RB5 degradation products was significantly reduced in the presence of sugar sources. The bacterial community structure differed remarkably as a result of sugar sources addition. For a fructose addition, a considerably enriched population of the functional species Burkholderia-Paraburkholderia and Klebsiella was noted. The results enlarge our knowledge of the microkinetic and microbiological mechanisms of co-metabolic degradation of refractory pollutants.

Unveiling the activating mechanism of tea residue for boosting the biological decolorization performance of refractory dye.

Conventional microbial treatments are challenged by new synthetic refractory dyes. In this work, tea residue was found serving as an effective activator to boost the decolorization performance of anthraquinone dye (reactive blue 19, RB19) by a new bacterial flora DDMY2. The unfermented West Lake Longjing tea residue showed the best enhancement performance. Seventeen main kinds of components in tea residue had been selected to take separate and orthogonal experiments on decolorization of RB19 by DDMY2. Results suggested epigallocatechin gallate (EGCG) in tea residue played important roles in boosting the treatment performance. Illumina MiSeq sequencing results confirmed that EGCG and tea residue pose similar impact on the change of DDMY2 community structure. Some functional bacterial genera unclassified_o_Pseudomonadales, Stenotrophomonas and Bordetella were enriched during the treatment of RB19 by EGCG and tea residue. These evidences suggested EGCG might be the key active component in tea residue that responsible for the enhancement effect on decolorization performance. These results revealed the activating mechanism of tea residue from the perspective of composition.

Fructose as an additional co-metabolite promotes refractory dye degradation: Performance and mechanism.

In this work, the performance and mechanism for the boosting effects of fructose as an additional co-metabolite towards the biological treatment of reactive black 5 were systematically investigated. A decolorization efficiency of 98% was obtained in sample FRU200 (with 3 g/L fructose added based on 3 g/L yeast extract), which was 21% higher than that without fructose. Several intermediates with low molecular weight generated in sample FRU200 and different metabolic pathways were deduced. The bacterial community structure significantly changed due to fructose addition. Label-free quantitative proteomic approach suggested that several up-regulated proteins in sample FRU200 might play essential roles during the degradation. Furthermore, the mechanisms of RB5 degradation by proteins/enzymes of the dominant species in flora DDMZ1 were proposed. This work deepens our understanding of the molecular and ecological mechanism of fructose as co-metabolite enhancing the biodegradation of refractory organic pollutants by a natural bacterial flora.

Highly efficient biodegradation of reactive blue 19 under the activation of tea residue by a newly screened mixed bacterial flora DDMY2.

In this study, a newly screened mixed bacterial flora DDMY2 had high decolorization capacity for anthraquinone dye reactive blue 19 (RB19) and the decolorization efficiency of 300 mg L-1 RB19 could reach up to 98% within 48 h in the presence of tea residue. Results indicated that RB19 could be efficiently decolorized by flora DDMY2 in wide ranges of pH values (5.0-9.0), temperatures (30-40 °C) and initial dye concentrations (50-500 mg L-1) under the activation of tea residue. Concentration of tea residue had been proved to significantly impact the decolorization performance. UV-vis spectrophotometry, Fourier transform infrared spectrometry and liquid chromatography/time-of-flight/mass spectrometry analysis showed three identified degradation products and the possible degradation pathway of RB19 was speculated. High-throughput sequencing analysis revealed the community structures of bacterial flora before and after domestication by tea residue. Based on the result, it was inferred that unclassified_o_Pseudomonadales, Brevibacillus, Stenotrophomonas and Bordetella activated by tea residue were responsible for the excellent decolorization performance. Results of this research deepen our understanding of the biodegradation process of anthraquinone dyes by bacterial flora and broaden the knowledge of utilizing tea residue as a bioactivator in biological treatment.

Comparative study of antiestrogenic activity of two dyes after Fenton oxidation and biological degradation.

In present study, two methods (Fenton oxidation and biological degradation) were used to degrade azo dye (Reactive Black 5, RB5) and anthraquinone dye (Remazol Brilliant Blue R, RBBR). The changes of antiestrogenic activities of these two dyes through two degradation methods were detected using the yeast two-hybrid assay method. Fluorescence spectroscopy together with gas chromatography-mass spectrometry (GC-MS) method was performed to analyze the metabolites of RB5 and RBBR after Fenton oxidation and biological degradation. Results indicated that by Fenton oxidation, the decolorization of RB5 and RBBR were 99.31% and 96.62%, respectively, which were much higher than that by biological degradation. Dissolved organic carbon (DOC) reduction rates of RB5 and RBBR after Fenton oxidation were also much higher than that after biological degradation. By Fenton oxidation, the antiestrogenic activities of RB5 and RBBR all decreased below detection limit after degradation, while by biological degradation all of them increased significantly after degradation. Fluorescence spectroscopy analysis and GC-MS analysis confirmed the degradation effects of RB5 and RBBR by these two degradation methods. In addition, fluorescence spectroscopy analysis revealed that the metabolites humic acid-like substances might contribute to the increasing of antiestrogenic activity of RB5 and RBBR after biological degradation.

Illumina MiSeq sequencing reveals microbial community in HA process for dyeing wastewater treatment fed with different co-substrates.

In present study, a hydrolysis acidification (HA) reactor was used for simulated dyeing wastewater treatment. Co-substrates included starch, glucose, sucrose, yeast extract (YE) and peptone were fed sequentially into the HA reactor to enhance the HA process effects. The performance of the HA reactor and the microbial community structure in HA process were investigated under different co-substrates conditions. Results showed that different co-substrates had different influences on the performance of HA reactor. The highest decolorization (50.64%) and COD removal rate (60.73%) of the HA reactor were obtained when sucrose was as the co-substrate. And it found that carbon co-substrates starch, glucose and sucrose exhibited better decolorization and higher COD removal efficiency of the HA reactor than the nitrogen co-substrates YE and peptone. Microbial community structure in the HA process was analyzed by Illumina MiSeq sequencing. Results revealed different co-substrates had different influences on the community structure and microbial diversity in HA process. It was considered that sucrose could enrich the species such as Raoultella, Desulfovibrio, Tolumonas, Clostridium, which might be capable of degrading the dyes. Sucrose was considered to be the best co-substrate of enhancing the HA reactor's performance in this study. This work would provide deep insight into the influence of many different co-substrates on HA reactor performance and microbial communities in HA process.

Smartphone-based colorimetric chiral recognition of ibuprofen using aptamers-capped gold nanoparticles.

Sensitive and fast detection of ibuprofen enantiomers is very critical for required routine monitoring and risk assessment of trace pollutants in water samples. Here a simple, rapid and highly sensitive android smartphone application for chiral recognition was developed. Aptamer-capped gold nanoparticles (AuNPs) was demonstrated as an efficient detection platform for (S)-(+)-ibuprofen (S-Ibu) and (R)-(-)-ibuprofen (R-Ibu). Detachment of an enantioselective aptamer from the AuNPs surface and binding with an enantiomer of Ibu lead to AuNPs aggregation, which allows a rapid enantiodiscrimination of Ibu by monitoring the absorbance changes of AuNPs solution in the UV-vis spectrum. Under optimal conditions, the limit of detection for S-Ibu and R-Ibu was 1.24 and 3.91 pg/mL, respectively. These probes showed good chiral recognition ability in mixed samples (i.e. S-Ibu + R-Ibu) and environmental samples. These advantages can be further developed by quantitative measurement with smartphone, which opens new opportunities for on-site detection of trace chiral pollutants in a simple and practical manner.

Phosphorus (P) recovery coupled with increasing influent ammonium facilitated intracellular carbon source storage and simultaneous aerobic phosphorus & nitrogen removal.

Under decreasing C/N (from 8.8 to 3.5) conditions, an alternating anaerobic/aerobic biofilter (AABF) was used to remove nitrogen and accumulate/recover phosphorus (P) from synthetic wastewater. The AABF was periodically (every 10 days) fed with an additional carbon source (10 L, chemical oxygen demand (COD) = 900 mg L-1 sodium acetate (NaAC) solution) in the anaerobic phase to induce the release of P sequestered in the biofilm. An increase in PHA storage in the biofilm was observed and characterized with TEM and a GC-MS method. The accumulation of P and removal of total nitrogen occurred primarily in the aerobic phase. As the NH4+-N loading rate increased from 0.095 to 0.238 kg m-3 d-1 at a total empty bed retention time (EBRT) of 4.6 h, the TN removal in AABF was reduced from 91.2% to 43.4%, while the P removal or recovery rate remained unaffected. The high-throughput community sequencing analysis indicated that the relative abundance of Candidatus Competibacter, Nitrospira and Arcobacter increased while the Accumulibacter phosphatis decreased with an increase of ammonium loading rate within a short operational period (30 days). A putative N and P removal pattern via simultaneous nitrification and PHA-based denitrification, as well as P accumulation in the biofilm was proposed. The research demonstrated that an efficient N removal and P recovery process, i.e., simultaneous nitrification and denitrification, P accumulation and carbon source-regulated P recovery can be achieved by the symbiotic functional groups in a single biofilm reactor.

Performance and microbial community structures of hydrolysis acidification process treating azo and anthraquinone dyes in different stages.

In this study, performance of hydrolysis acidification process treating simulated dyeing wastewater containing azo and anthraquinone dyes in different stages was investigated. The decolorization ratio, CODCr removal ratio, BOD5/CODCr value, and volatile fatty acids (VFAs) production were almost better in stage 1 than that in stage 2. Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS) confirmed the biodegradation of Reactive Black 5 (RB5) and Remazol Brilliant Blue R (RBBR) in hydrolysis acidification process. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analyses revealed that significant difference of microbial community structures existed in stage 1 and 2. The dominant species in stage 1 was related to Bacteroidetes group, while the dominant species in stage 2 was related to Bacteroidetes and Firmicutes groups. From the results, it could be speculated that different dyes' structures might have significant influence on the existence and function of different bacterial species, which might supply information for bacteria screening and acclimation in the treatment of actual dyeing wastewater.

Enhanced phosphorus recovery and biofilm microbial community changes in an alternating anaerobic/aerobic biofilter.

The operation of an alternating anaerobic/aerobic biofilter (AABF), treating synthetic wastewater, was modified to enhance recovery of phosphorus (P). The AABF was periodically fed with an additional carbon source during the anaerobic phase to force the release of biofilm-sequestered P which was then harvested and recovered. A maximum of 48% of the total influent P was found to be released in the solution for recovery. Upon implementation of periodic P bio-sequestering and P harvesting, the predominant bacterial communities changed from ß-Proteobacteria to γ-Proteobacteria groups. The genus Pseudomonas of γ-Proteobacteria was found to enrich greatly with 98% dominance. Dense intracellular poly-P granules were found within the cells of the biofilm, confirming the presence of P accumulating organisms (PAOs). Periodic addition of a carbon source to the AABF coupled with intracellular P reduction during the anaerobic phase most probably exerted environmental stress in the selection of Pseudomonas PAOs over PAOs of other phylogenic types. Results of the study provided operational information on the selection of certain microbial communities for P removal and recovery. This information can be used to further advance P recovery in biofilm systems such as the AABFs.

Characteristics, process parameters, and inner components of anaerobic bioreactors.

The anaerobic bioreactor applies the principles of biotechnology and microbiology, and nowadays it has been used widely in the wastewater treatment plants due to their high efficiency, low energy use, and green energy generation. Advantages and disadvantages of anaerobic process were shown, and three main characteristics of anaerobic bioreactor (AB), namely, inhomogeneous system, time instability, and space instability were also discussed in this work. For high efficiency of wastewater treatment, the process parameters of anaerobic digestion, such as temperature, pH, Hydraulic retention time (HRT), Organic Loading Rate (OLR), and sludge retention time (SRT) were introduced to take into account the optimum conditions for living, growth, and multiplication of bacteria. The inner components, which can improve SRT, and even enhance mass transfer, were also explained and have been divided into transverse inner components, longitudinal inner components, and biofilm-packing material. At last, the newly developed special inner components were discussed and found more efficient and productive.

Bioleaching of arsenic-rich gold concentrates by bacterial flora before and after mutation.

In order to improve the bioleaching efficiency of arsenic-rich gold concentrates, a mixed bacterial flora had been developed, and the mutation breeding method was adopted to conduct the research. The original mixed bacterial flora had been enrichedin acid mine drainage of Dexing copper mine, Jiangxi Province, China. It was induced by UV (ultraviolet), ultrasonic, and microwave, and their combination mutation. The most efficient bacterial flora after mutation was collected for further bioleaching of arsenic-rich gold concentrates. Results indicated that the bacterial flora after mutation by UV 60 s combined with ultrasonic 10 min had the best oxidation rate of ferrous, the biggest density of cells, and the most activity of total protein. During bioleaching of arsenic-rich gold concentrates, the density of the mutant bacterial cells reached to 1.13 × 108 cells/mL at 15 days, more than 10 times compared with that of the original culture. The extraction of iron reached to 95.7% after 15 days, increased by 9.9% compared with that of the original culture. The extraction of arsenic reached to 92.6% after 12 days, which was increased by 46.1%. These results suggested that optimum combined mutation could improve leaching ability of the bacterial flora more significantly.

Re-feeding evokes reproductive overcompensation of food-restricted Brandt's voles.

In natural conditions, animals have to cope with fluctuations of food resources. Animals having experienced prolonged decrease in feeding opportunities may increase their reproductive success when meeting abundant food. Though food restriction is well known to reduce reproductive success of animals, it is not clear whether re-feeding can restore or even overcompensate the reproductive success. In this study, we investigated the differences in reproductive parameters between food-restricted and refed (FR-RF) group and control group of Brandt's vole (Lasiopodomys brandtii). For 4 weeks, FR-RF voles were provided with 70% of their normal daily food intake and then they were fed ad libitum for the next 4 weeks. Voles of control group were fed ad libitum for 8 weeks. Females (FR-RF or control) were mated to non-littermate males of the same group (FR-RF or control), and we found that the mean litter size and survival rate of F1 pups of FR-RF group were significantly higher than those of control group. We also provided a field example showing that the litter size of Brandt's voles tended to be higher if they experienced two consecutive dry and wet months than that of voles didn't have this experience. Our results suggest that re-feeding may have evoked an overcompensatory mechanism of food-restricted voles in reproductive success. This may be an adaptive strategy for Brandt's voles (with oscillating populations) to cope with the fluctuating food resources in natural conditions by adjusting their reproductive success.