Method for harmlessly pretreating organic solid waste based on combination of calcium ion and hydrothermal treatment

Abstract

A method for harmlessly pretreating an organic solid waste based on combination of calcium ion and hydrothermal treatment, including: preliminarily screening the organic solid waste followed by addition of calcium ions and hydrothermal treatment to make the organic solid waste harmless.

Claims

1. A method for harmlessly pretreating an organic solid waste, comprising; (1) screening preliminarily the organic solid waste to remove impurities; and feeding a calcium ion-containing reagent to the organic solid waste followed by adjustment to a water content of 85±5% to form a reaction system; (2) subjecting the reaction system obtained in step (1) to thermal hydrolysis in a closed environment; wherein the thermal hydrolysis is performed at 140-170° C. and 0.8±0.2 MPa for 60±30 min; (3) after the thermal hydrolysis is completed, cooling the reaction system to 60° C. or less, followed by pressure releasing to obtain a treated product; wherein a gas released during the pressure releasing is centralizedly absorbed or collected; and (4) detecting a content of antibiotics, antibiotic resistance genes and viruses in the treated product obtained in step (3); wherein the calcium ion-containing reagent is calcium chloride; a dry weight ratio of the calcium ions to the organic solid waste is 0.5:1; the organic solid waste is biological sewage sludge, biological wet waste, agricultural straw, manure and dead body of livestock and poultry, industrial bacterial residue, industrial oil sludge, or a combination thereof; and the organic solid waste contains fluoroquinolone antibiotics.

2. The method of claim 1, wherein the water content of the reaction system is adjusted to 85%.

3. The method of claim 1, wherein the thermal hydrolysis is performed at 170° C. and 0.8 MPa for 90 min.

4. The method of claim 1, wherein the gas for pressure releasing is absorbed by using an alkali liquor, or is collected by using a gas film or gas bag.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) To render the technical solutions of the embodiments of this disclosure or the prior art clearer, the drawings used in the description of the embodiments of this disclosure or the prior art will be briefly described below. Obviously, presented in the following drawings are merely some embodiments of the disclosure. Other drawings can be obtained by those skilled in the art based on the drawings provided herein without paying any creative effort.

(2) This FIGURE is a flow chart of a method for harmlessly pretreating an organic solid waste based on combination of calcium ion and hydrothermal treatment according to an embodiment of the present disclosure.

DETAILED DESCRIPTION OF EMBODIMENTS

(3) Various exemplary embodiments of the present disclosure will be described in detail below. The detailed description should not be considered to limit the present disclosure, but should be understood as a more detailed description of certain aspects, characteristics, and embodiments of the present disclosure.

(4) It should be understood that the terms described in the present disclosure are only used to describe specific embodiments and are not intended to limit the present disclosure. In addition, with respect to the numerical range used herein, it should be understood that each intermediate value between the upper limit and the lower limit of the range has also been disclosed specifically. Intermediate values within any stated value or range and each smaller range between any other stated value or intermediate value within the range are also included in the present disclosure. Upper and lower limits of these smaller ranges of can be independently included or excluded from the range.

(5) Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although described herein are only preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present disclosure. All documents mentioned herein are incorporated by reference to disclose and describe methods and/or materials related to the documents.

(6) Without departing from the scope or spirit of the present disclosure, various improvements and changes can be made to the specific embodiments of the present disclosure, which are obvious to those skilled in the art. Other embodiments derived from the description of the present disclosure are obvious to those skilled in the art. The description and embodiments of this application are only exemplary.

(7) As used herein, the terms “comprising”, “including”, “having”, “containing” all have an open meaning, which means including but not limited to.

(8) The organic solid waste used herein is a mixture of sludge, livestock and poultry manure, and industrial bacteria residue, where a ratio of the organic substance content in these components is 1:1:1. Antibiotic standards of ofloxacin, norfloxacin, ciprofloxacin, lomefloxacin are purchased from CNW Technologies Gmbh Company (Germany). Solid-phase extraction cartridges are purchased from Waters Corporation (USA). Chromatographic reagents such as methanol and acetonitrile are purchased from Shanghai Anpu Company and Shanghai Sinopharm Company. The experimental water is deionized water and Millipore water.

(9) The primer sequences used herein are listed as follows:

(10) TABLE-US-00001 qnrS: (SEQ ID NO: 1) GTGAGTAATCGTATGTACTTTTGCAAACACCTCGACTTAAGTCT; qnrA: (SEQ ID NO: 2) TTCTCACGCCAGGATTTGCCATCCAGATCGGCAAA; intl1: (SEQ ID NO: 3) CGAACGAGTGGCGGAGGGTGTACCCGAGAGCTTGGCACCCA; human adenovirus: (SEQ ID NO: 4) GGACGCCTCGGAGTACCTGAGACRGTGGGGGTTTCTGAACTTGTT; human polyomavirus: (SEQ ID NO: 5) ATGTTTGCCAGTGATGATGAAAAGGAAAGTCTTTAGGGTCTTCTACCT; human herpes virus: (SEQ ID NO: 6) CGGCCGTGTGACACTATCGCTCGTAAAATGGCCCCTCC; pox virus: (SEQ ID NO: 7) TAATACTTCGATTGCTCATCCAGGACTTCTCACAAATGGATTTGAAAAC; enterovirus: (SEQ ID NO: 8) GATTGTCACCATAAGCAGCCCCCTGAATGCGGCTAATC; human Coronavirus HKU1: (SEQ ID NO: 9) GTTGGTTGTATGATGCGTTTGTTCTTCTACAAATAAACTAGCATCAACAT CATCGT;

(11) The detection of antibiotic content is described below. 0.2 g of a sample is added with 20 mL of a basic extraction solution followed by shaking for 5 min to obtain a mixture, where the basic extraction solution is a mixture of triethylamine, methanol, and water in a ratio of 5:25:75. The mixture is subjected to cell disruption at a gradient disruption power, extraction for 5 min, and centrifugation at 5000 r/min for 10 min to collect a supernatant. The above steps are repeated once to obtain another supernatant, and then the supernatants are combined. The combined supernatant is adjusted to pH 3.0 with formic acid, and filtered with a 0.45 μm filter to obtain a filtrate. The filtrate is subjected to dilution, and extraction using a hydrophile-lipophile balance (HLB) solid-phase extraction column. The HLB solid-phase extraction column is activated with 10 mL of methanol and 10 mL of ultrapure water. The liquid sample is passed through the HLB solid-phase extraction column at a flow rate of 5 mL/min, eluted with 5 mL of 5% methanol solution, vacuumed for 10 min under negative pressure, and eluted with 6 ml of 6% ammonia/methanol eluent to obtain an elution solution. The elution solution is dried with nitrogen in a water bath at 35° C., diluted to a volume of 1 mL using the initial mobile phase, filtered with the 0.45 μm pinhole-filtration membrane, and stored in chromatographic vials to be tested. The test is performed on an Agilent technologies 1260 high-performance liquid chromatography (HPLC) equipped with a fluorescence detector, where the mobile phase consists of phosphoric acid-triethylamine solution (A) and methanol (B) in a ratio of 80%:20%; the pH of the phosphoric acid-triethylamine solution is 3.0; an Agilent XDB-c18 column (150 mm*4.6 mm, 5.0 μm) is employed; the injection volume is 20 μm; the flow rate is 1 mL/min; the column temperature is 30° C.; and the analysis time for each sample is 18 min; the scanning wavelength is programmed as follows: 0-9.5 min: the excitation wavelength and emission wavelength are respectively 295 nm and 500 nm; 9.5-14 min: the excitation wavelength and emission wavelength are respectively 278 nm and 445 nm; and 14-18 min: the excitation wavelength and emission wavelength are respectively 285 nm and 470 nm.

(12) The detection of antibiotic resistance genes is performed as the following steps. 10 g of each homogeneous sample (original sample or hydrothermal product) is freeze-dried in a −80° C. ultra-low temperature refrigerator for 48 h. 0.25 g of the freeze-dried sample is subjected to extraction using a Fast DNA∘R Spin kit (MP Bimedicals). The DNA concentration is monitored using the QuantiFluor ∘R dsDNA system (Promega). The ViiATM 7 real-time PCR system is configured to perform quantitative PCR reaction. FQs resistance genes (qnrS and qnrA), an integrase gene intL1 and a 16S rRNA-encoding gene are selected for detection.

(13) The extraction and detection of viruses are performed as follows. 10 g of the organic solid waste is mixed with 50 mL of an eluent (10% beef extract, pH=7.2), stirred at 5000 r/min at room temperature for 30 min, and centrifuged at 4° C. and 10000 g for 30 min to collect a supernatant liquid. The supernatant is adjusted to pH 7.2. The supernatant liquid is filtered by a 0.22 μm filter membrane to remove residual bacterial fragments. The filtration fluid is added with 8% polyethylene glycol followed by mixing, standing at 4° C. for 12 h, centrifugation at 4° C. and 12000 g for 30 min. After that, the precipitation is collected to perform the extraction and detection of DNA and RNA directly.

EXAMPLE 1

(14) Provided herein was a method for harmlessly pretreating an organic solid waste, which was specifically described as follows (as shown in the FIGURE).

(15) (S1) Blending with Calcium Ions

(16) An organic solid waste was screened preliminarily to remove impurities. 70 g of the screened organic solid waste was added with 1 g of calcium oxide to form a reaction system. A water content of the reaction system was adjusted to 80%.

(17) (S2) Harmless Disinfection by Hydrothermal Treatment

(18) The reaction system obtained in step (1) was subjected to thermal hydrolysis in a closed container, where the thermal hydrolysis was performed at 140° C. and 0.6 MPa for 30 min.

(19) (S3) Cooling and Pressure Releasing

(20) After the thermal hydrolysis was completed, the reaction system was cooled to 60° C. or less, followed by opening a pressure-releasing valve to perform pressure releasing, where a gas released during the pressure releasing was centralizedly absorbed by using an alkali liquor, or was collected by using a gas film or gas bag.

(21) (S4) Detection of a Content of Antibiotics

(22) Antibiotics, resistance genes and viruses in the treated product obtained in step (3) were detected according to the above-mentioned detection methods.

EXAMPLE 2

(23) Provided herein was a method for harmlessly pretreating an organic solid waste, which was specifically described as follows.

(24) (S1) Blending with Calcium Ions

(25) An organic solid waste was screened preliminarily to remove impurities. 70 g of the screened organic solid waste was added with 97 g of calcium chloride to form a reaction system. A water content of the reaction system was adjusted to 85%.

(26) (S2) Harmless Disinfection by Hydrothermal Treatment

(27) The reaction system obtained in step (1) was subjected to thermal hydrolysis in a closed container, where the thermal hydrolysis was performed at 170° C. and 0.8 MPa for 90 min.

(28) (S3) Cooling and Pressure Releasing

(29) After the thermal hydrolysis was completed, the reaction system was cooled to 60° C. or less, followed by opening a pressure-releasing valve to perform pressure releasing, where a gas released during the pressure releasing was centralizedly absorbed or collected.

(30) (S4) Detection of a Content of Antibiotics

(31) Antibiotics, resistance genes and viruses in the treated product obtained in step (3) were detected according to the above-mentioned detection methods.

EXAMPLE 3

(32) Provided herein was a method for harmlessly pretreating an organic solid waste, which was specifically described as follows.

(33) (S1) Blending with Calcium Ions

(34) An organic solid waste was screened preliminarily to remove impurities. 70 g of the screened organic solid waste was added with 90 g of calcium oxide to form a reaction system. A water content of the reaction system was adjusted to 90%.

(35) (S2) Harmless Disinfection by Hydrothermal Treatment

(36) The reaction system obtained in step (1) was subjected to thermal hydrolysis in a closed container, where the thermal hydrolysis was performed at 160° C. and 1.0 MPa for 60 min.

(37) (S3) Cooling and Pressure Releasing

(38) After the thermal hydrolysis was completed, the reaction system was cooled to 60° C. or less, followed by opening a pressure-releasing valve to perform pressure releasing, where a gas released during the pressure releasing was centralizedly absorbed or collected.

(39) (S4) Detection of a Content of Antibiotics Antibiotics, resistance genes and viruses in the treated product obtained in step (3) were detected according to the above-mentioned detection methods.

EXPERIMENTAL RESULTS

(40) The experimental results were described below.

(41) TABLE-US-00002 TABLE 1 Effects of combination of calcium ion and hydrothermal treatment on degradation of antibiotics in organic solid wastes Antibiotic content (mg/kg) Samples Lomefloxacin Ciprofloxacin Norfloxacin Ofloxacin Original organic 0.32 0.61 1.56 8.04 solid waste Example 1 0.24 0.45 1.48 6.43 Example 2 0 0 1.31 3.61 Example 3 0.12 0.13 1.33 4.34

(42) It can be seen from Table 1 that the residual amount of antibiotics in the organic solid waste had been decreased significantly after combination of calcium ion and hydrothermal treatment, proving that combination of calcium ion and hydrothermal treatment could accelerate the degradation rate of antibiotics in the organic solid waste.

(43) TABLE-US-00003 TABLE 2 Effects of combination of calcium ion and hydrothermal treatment on absolute abundance of resistance genes of quinolone antibiotics in organic solid wastes Absolute abundance of resistance genes (copies/g) 16S Samples qnrS qnrA Intl1 rDNA gene Original organic 1.09 × 10.sup.11 1.01 × 10.sup.10 0.98 × 10.sup.8 10.sup.12 solid waste Example 1 1.03 × 10.sup.7  1.07 × 10.sup.7  2.11 × 10.sup.5 10.sup.9  Example 2 1.03 × 10.sup.5  — — 10.sup.7  Example 3 8.92 × 10.sup.5  2.08 × 10.sup.5  3.81 × 10.sup.3 10.sup.8 

(44) It can be seen from Table 2 that the absolute abundance of resistance genes of quinolone antibiotics in organic solid wastes had been greatly reduced after combination of calcium ion and hydrothermal treatment, proving that the combination of calcium ion and hydrothermal treatment could significantly reduce the resistance genes of quinolone antibiotics of organic solid wastes.

(45) TABLE-US-00004 TABLE 3 Effect of combination of calcium ion and hydrothermal treatment on absolute abundance of resistance genes of quinolone antibiotics in organic solid waste DNA virus Adenovirus RNA virus Samples huminis Herpesvirus Polyomavirus Poxvirus Enterovirus Coronavirus Original + + + + + + organic solid waste Example 1 − − − − − − Example 2 − − − − − − Example 3 − − − − − − Notes: “+” represents positive; “−” represents negative.

(46) Common viruses in organic solid wastes, including four kinds of DNA viruses (human adenovirus, herpes virus, polyoma virus, and pox virus) and 2 kinds of RNA viruses (enterovirus and human coronavirus) were detected herein to evaluate the effects of combination of calcium ion and hydrothermal treatment on virus killing.

(47) The experimental results showed that after treated through the combination of calcium ion and hydrothermal treatment, the organic solid waste was tested negative for the 6 kinds of viruses, indicating that the method disclosed herein could effectively eliminate viruses in the organic solid waste.

(48) Described above are merely preferred embodiments of this application, which are not intended to limit the application. It should be understood that modifications and replacements made by those skilled in the art without departing from the spirit of the application should fall within the scope of the application defined by the appended claims.