A device and method for detecting the concentration of microbial communities in sewage sludge

Abstract

The application provides a sewage sludge microbial flora concentration detection device and method, and relates to the technical field of sewage detection. The detection device comprises a light-proof box body, a light-transmitting container, a detection assembly, a moving mechanism and a processor. The light-transmitting container is arranged in the light-proof box body and can be filled with sewage samples and added fluorescent agents. The detection assembly comprises a spectrophotometric detection module and a fluorescent detection module. The spectrophotometric detection module calculates total biomass by measuring transmitted light signals. The fluorescent detection module marks active flora by capturing specific wavelength fluorescent signals. The moving mechanism drives the detection assembly to move along the container axis to realize multi-point detection. The processor controls the moving mechanism, cooperatively analyzes the transmitted light and fluorescent data, establishes a quantitative relationship between the total biomass and the active bacteria signal, and finally calculates the active microbial concentration. The device solves the problem that traditional methods cannot distinguish between active and dead bacteria by fusing the data of spectrophotometric and fluorescent detection, and realizes accurate determination of the active flora concentration.

Description

Technical Field

[0001] This application relates to the field of wastewater testing technology, and in particular to a device and method for detecting the concentration of microbial communities in wastewater sludge. Background Technology

[0002] In wastewater treatment processes, biological treatment methods are widely used in various fields of industrial wastewater and domestic sewage treatment due to their mature and stable processes and lower operating costs compared to physical or chemical treatments. The main principle of biological treatment methods is to utilize the metabolic activity of microorganisms to remove pollutants such as organic matter, ammonia nitrogen, and total phosphorus from wastewater. Therefore, the health of the microorganisms directly determines the efficiency of pollutant removal.

[0003] During operation, the main indicators for routine monitoring of microorganisms in wastewater include sludge settling ratio (SV30), MLSS, or MLVSS. These indicators are primarily visual indicators and are easily affected by suspended solids and inorganic components in the water. The results can only be used as a reference and cannot accurately reflect the concentration of microbial communities in activated sludge. Currently, the main methods for determining microbial community concentration are plate counting, ATP method, and spectrophotometry. Among these, the plate counting method has the highest accuracy, but the testing cycle is relatively long, usually requiring 2-3 days to obtain results. The ATP method is currently only used for offline laboratory testing.

[0004] Spectrophotometry assesses microbial concentration by measuring the absorbance (OD value) of a sample at a specific wavelength. However, this method has a fundamental limitation: it cannot distinguish the optical signal differences between live and dead bacteria, leading to results that significantly deviate from the true live bacteria concentration. Because both live and dead bacteria produce similar absorbance signals at 590 nm, the OD value cannot accurately reflect the number of metabolically active microorganisms.

[0005] Experimental data show that although the dead bacterial suspension treated with high-temperature sterilization completely loses its metabolic activity, its OD value still deviates from the measurement results of the live bacterial culture medium by up to ±15%. This signal overlap phenomenon introduces systematic errors into the detection results relying solely on spectrophotometry. This limitation is particularly pronounced in wastewater treatment scenarios requiring precise activity monitoring.

[0006] Existing technologies attempt to distinguish between live and dead bacteria using fixed coefficient correction, but this method cannot adapt to the dynamic changes in the live / dead bacteria ratio in different samples. More seriously, as the sludge age increases, the proportion of dead bacteria increases, further amplifying the detection error of spectrophotometry. This inherent technical deficiency significantly limits the application of spectrophotometry in the detection of microbial activity. Summary of the Invention

[0007] In view of this, this application proposes a device and method for detecting the concentration of microbial communities in sewage sludge to solve the problem that existing spectrophotometric detection technologies cannot distinguish between live and dead bacteria signals, which leads to errors in the detection of the concentration of microbial communities in sewage sludge.

[0008] The technical solution of this application is implemented as follows:

[0009] On the one hand, this application provides a device for detecting the concentration of microbial communities in sewage sludge, comprising:

[0010] Light-proof enclosure;

[0011] The light-transmitting container is housed in a light-proof box. The light-transmitting container is equipped with an inlet and a filling port. The inlet is used to introduce wastewater samples, and the filling port is used to add fluorescent agents.

[0012] The detection assembly includes a spectroscopic detection module and a fluorescence detection module. The spectroscopic detection module has a spectroscopic light source and a spectroscopic receiver arranged opposite to each other along a first radial direction. The spectroscopic light source is used to emit a detection beam into a light-transmitting container, and the spectroscopic receiver is used to receive transmitted light signals. The fluorescence detection module has a fluorescence light source and a fluorescence receiver arranged opposite to each other along a second radial direction. The fluorescence light source is used to emit an excitation beam into the light-transmitting container, and a filter assembly is provided in front of the fluorescence receiver to receive and filter fluorescence signals of specific wavelengths. The first radial direction and the second radial direction do not coincide.

[0013] The moving mechanism, housed within the light-proof enclosure, is used to drive the detection component to move along the axial direction of the light-transmitting container;

[0014] The processor, electrically connected to the spectrophotometer, fluorescence receiver, and moving mechanism, is configured to control the moving mechanism to enable the detection component to perform detection at multiple axial positions, calculate the total biomass data of the sample based on the transmitted light signal, calculate the active microbial community labeling data based on the fluorescence signal, and calculate the concentration data of active microorganisms based on the total biomass data and the active microbial community fluorescence labeling data.

[0015] Based on the above technical solution, preferably, the spectral detection module further includes a first collimating lens disposed in the light-emitting direction of the spectral source and a first focusing lens disposed in the light-incident direction of the spectral receiver;

[0016] The fluorescence detection module further includes:

[0017] A second focusing lens is provided in the light-emitting direction of the fluorescent light source; and a collecting lens and a second collimating lens are provided in the light-incoming direction of the fluorescent receiver, the two being coaxially spaced apart;

[0018] The filter assembly includes a 520nm bandpass filter and a 620nm bandpass filter, used to detect the fluorescence signals of live and dead bacteria, respectively.

[0019] Based on the above technical solution, preferably, the spectral detection module further includes a first light source shield and a first receiving shield, the first light source shield covering the optical path between the spectral light source and the first collimating lens, and the first receiving shield covering the optical path between the first focusing lens and the spectral receiver.

[0020] The fluorescence detection module further includes a second light source shield and a second receiving shield. The second light source shield covers the optical path between the fluorescence light source and the second focusing lens, and the second receiving shield covers the optical path between the collecting lens and the fluorescence receiver.

[0021] Based on the above technical solution, preferably, the detection component further includes an annular mounting disk, which is coaxially sleeved on the outer wall of the light-transmitting container. The spectrophotometer and fluorescence detection modules are fixedly arranged along the circumference of the annular mounting disk, and the driving end of the moving mechanism is connected to the annular mounting disk.

[0022] Based on the above technical solution, preferably, it further includes a stirring mechanism, which includes:

[0023] The stirring shaft is coaxially installed inside the light-transmitting container, and its side wall has at least two symmetrically arranged storage slots.

[0024] The stirring rod is mounted in the storage tank at its upper end via a hinge shaft and can rotate around the hinge shaft.

[0025] A drive assembly, located on the outside of the light-transmitting container and connected to the stirring shaft, is used to drive the stirring shaft to rotate and move axially. When the stirring shaft rotates, the stirring rod unfolds from the receiving tank under the action of centrifugal force. When the stirring shaft stops rotating, the stirring rod retracts into the receiving tank under the action of gravity.

[0026] Based on the above technical solution, preferably, the driving component includes:

[0027] The mounting bracket is fixedly installed at the bottom of the light-transmitting container;

[0028] The limiting plate is fixedly installed on the mounting bracket, and has a through hole in the center;

[0029] A toothed disc is positioned between a limiting plate and the bottom surface of a light-transmitting container. The lower end of a stirring shaft moves through the toothed disc and a through hole. A guide groove is provided on the side wall of the stirring shaft along its axial direction. A guide protrusion that mates with the guide groove is provided on the inner wall of the toothed disc. A sealing element is fitted on the outer side of the stirring shaft. The sealing element is located between the toothed disc and the light-transmitting container.

[0030] The rotating motor is fixed on the mounting bracket, and the output end of the rotating motor meshes with the gear plate through the main gear;

[0031] The telescopic element has a fixed end and a telescopic end. The fixed end is fixed to the outside of the light-proof box, and the telescopic end is rotatably connected to the lower end of the stirring shaft.

[0032] Secondly, this application discloses a method for detecting the concentration of microbial communities in sewage sludge, which uses the sewage sludge microbial community concentration detection device described in the first aspect, and includes the following steps:

[0033] S1. The sludge sample to be tested is injected into the light-transmitting cavity after pretreatment.

[0034] S2: Start the spectrophotometer to measure the initial OD value of the sample at 590nm;

[0035] S3: Activate the fluorescence detection module and measure the background fluorescence intensity at 520nm. ;

[0036] S4: Inject fluorescent agent into the light-transmitting cavity and stir until homogeneous;

[0037] S5: Measure the fluorescence intensity at 520nm again. F 520 ;

[0038] S6: Calculate the viable bacteria concentration according to the formula: , where a is the calibration coefficient determined by the standard plate counting method.

[0039] Based on the above technical solution, preferably, in step S3... Calculated using the following formula: ,in, The fluorescence intensity at 520 nm before staining. The fluorescence intensity at 620 nm before staining. The background value of pure water at 620 nm is given, and β is the dead bacteria interference coefficient, calibrated to be 0.3-0.5. To extract specific signals from dead bacteria / organic matter in the 620nm channel, To convert the 620nm interference signal to an equivalent 520nm interference value proportionally, To subtract the contribution of dead bacteria / organic matter from the total signal at 520nm, the background signal of real live bacteria is obtained.

[0040] Based on the above technical solution, preferably, the... β The coefficients were determined by adding dead sludge in a gradient manner. The specific steps included: preparing sludge standard samples with dead bacteria percentages of 0%, 20%, 50%, and 80%; measuring the autofluorescence intensity of each standard sample at 520 nm / 620 nm; and fitting the coefficients. β This value minimizes the error in calculating the viable bacteria concentration.

[0041] Based on the above technical solution, preferably, an axial multi-point detection step is included after step S1:

[0042] S11. Drive the detection component to move along the axial direction of the light-transmitting container using the moving mechanism, and sequentially perform steps S2-S5 at at least three equally spaced positions, recording the OD at each position. 590 , F 520 and value;

[0043] S12, Measurements taken at each location F 520 and After taking the arithmetic mean separately, combine with the average OD 590 The value is then substituted into the formula in step S6 to calculate the final viable bacteria concentration.

[0044] This application has the following advantages over the prior art:

[0045] (1) The wastewater sludge microbial community concentration detection device disclosed in this application acquires the total biomass data of the sample through a spectrophotometric detection module, and simultaneously uses a fluorescence detection module to specifically identify the live bacteria labeling signal. The processor establishes a quantitative relationship between the total biomass data and the live bacteria labeling signal based on the synergistic analysis of the two, thereby achieving accurate calculation of the concentration of active microorganisms. The effectiveness of this technology stems from the inherent correlation between the total biomass benchmark provided by spectrophotometric detection and the live bacteria specific signal provided by fluorescence detection. Through data fusion processing by the processor, it effectively solves the technical problem that traditional methods cannot distinguish between live and dead bacteria.

[0046] (2) By optimizing the optical structure of the spectrophotometer and fluorescence detection modules, the detection accuracy and reliability are significantly improved. The spectrophotometer uses a first collimating lens to ensure parallel light beam incidence, and combines it with a first focusing lens to improve the signal-to-noise ratio of the transmitted light signal, making the OD value measurement more stable. The fluorescence detection module forms a uniform excitation spot through a second focusing lens, and at the same time adopts a combined optical path of a collecting lens + a second collimating lens, which greatly improves the collection efficiency of the fluorescence signal and optimizes the beam incident angle to match the optimal working conditions of the filter. The filter assembly (520nm / 620nm bandpass filter) accurately separates the fluorescence signals of live bacteria and dead bacteria, improves the specificity of live bacteria detection, and reduces interference from dead bacteria.

[0047] (3) By setting the stirring mechanism to a retractable structure, the stirring shaft is pushed into the light-transmitting container by the drive component during the stirring stage. The hinged stirring rod is unfolded by centrifugal force to achieve efficient mixing. After the stirring is completed, the stirring rod automatically retracts into the storage tank under the action of gravity, so that the stirring shaft can exit the detection area without obstruction. This not only ensures the uniform distribution of fluorescent dye, but also completely eliminates the obstruction of the light path by the traditional rigid stirring rod, realizing the non-interference collaborative operation of stirring and optical detection.

[0048] (4) The drive assembly achieves precise control of the rotation and axial movement of the stirring shaft through an innovative mechanical structure design. The gear disc and the stirring shaft are connected by a guide groove to ensure stable and reliable power transmission, while allowing the stirring shaft to rise and fall freely; the seals effectively prevent sewage leakage, and the telescopic element is rotatably connected to the stirring shaft to avoid motion interference. This design enables the stirring mechanism to smoothly complete the entire process of extending into the stirring and withdrawing to avoid obstacles, ensuring both the stirring effect and the uninterrupted light path during detection, resulting in stable and reliable overall operation.

[0049] (5) This application combines OD value (total biomass) with fluorescence signal (live bacteria specificity) and utilizes OD 590 As a baseline for total biomass, fluorescence difference ( Extracting the proportion of live bacteria while avoiding interference from dead bacteria. This is done using the unstained... (Including dead bacteria / organic matter signals) Corrected staining F 520 Significantly improves the specificity of live bacteria detection through experimental calibration. α It adapts to the characteristics of different sludge samples and reduces systematic errors. By fusing data from spectrophotometry (OD value) and fluorescence spectroscopy (520nm / 620nm dual channels), it overcomes the inherent limitation of traditional spectrophotometry in distinguishing between live and dead bacteria, greatly reducing the detection error of live bacteria concentration.

[0050] (6) Through 620nm signal and β Coordinated correction of coefficients quantizes the interference signal of dead bacteria / organic matter at 520nm. and from F auto520 Deduct from the middle to make the final It only reflects the true background value of viable bacteria. This method greatly reduces the detection error of viable bacteria concentration, and is especially suitable for complex sludge samples with a high proportion of dead bacteria or containing humic acid, significantly improving the accuracy and universality of the detection method in this embodiment.

[0051] (7) The moving mechanism drives the detection component to measure at least three equally spaced positions along the axis of the light-transmitting container. The average value of the data at each point is taken to calculate the final viable bacteria concentration. This method effectively eliminates the measurement deviation caused by uneven axial distribution of the sample, makes the detection results more representative, minimizes measurement errors, and enhances the system's adaptability to complex samples such as high concentrations or containing particulate matter. Attached Figure Description

[0052] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0053] Figure 1 This is a three-dimensional structural schematic diagram of the wastewater sludge microbial community concentration detection device disclosed in this application;

[0054] Figure 2 This is a schematic diagram of the internal structure of the wastewater sludge microbial community concentration detection device disclosed in this application;

[0055] Figure 3 This is a schematic diagram of the planar structure of the detection component disclosed in this application;

[0056] Figure 4 This is a first-view three-dimensional structural diagram of the wastewater and sludge microbial community concentration detection device disclosed in this application after removing the light-proof box.

[0057] Figure 5 This is a second-view three-dimensional structural diagram of the wastewater sludge microbial community concentration detection device disclosed in this application after removing the light-shielding box.

[0058] Figure 6 This is a three-dimensional structural diagram of the stirring mechanism disclosed in this application;

[0059] Figure 7 This is a top view of the wastewater sludge microbial community concentration detection device disclosed in this application;

[0060] Figure 8 for Figure 7 Planar sectional view at point AA;

[0061] Figure 9 for Figure 7 Plan view at point BB;

[0062] Figure 10 for Figure 9 Enlarged view of a section at point C;

[0063] Figure label:

[0064] 1. Light-shielding enclosure; 2. Light-transmitting container; 21. Liquid inlet; 22. Feed inlet; 3. Detection assembly; 30. Annular mounting plate; 31. Spectroscopic detection module; 311. Spectroscopic light source; 312. Spectroscopic receiver; 313. First collimating lens; 314. First focusing lens; 315. First light source shield; 316. First receiver shield; 32. Fluorescence detection module; 321. Fluorescence light source; 322. Fluorescence receiver; 323. Second focusing lens; 324. Collecting lens; 325. Second collimating lens; 326. Filter assembly; 327. Second light source shield; 328. Second receiver shield; 3261. 520nm bandpass filter; 3262. 620nm bandpass filter; 4. Moving mechanism; 5. Stirring mechanism; 51. Stirring shaft; 511. Storage groove; 512. Guide groove; 52. Stirring rod; 52. Drive assembly; 521. Mounting bracket; 5211. Limiting plate; 5212. Through hole; 522. Gear plate; 5221. Guide protrusion; 523. Rotating motor; 524. Main gear; 525. Telescopic element; 526. Seal; 527. Linkage element; 6. Mounting cylinder; 7. Fixing frame. Detailed Implementation

[0065] The technical solutions in the embodiments of this application will be clearly and completely described below with reference to the embodiments of this application. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0066] like Figure 1 As shown, combined with Figure 2-10 This application discloses a wastewater sludge microbial community concentration detection device, including a light-proof box 1, a light-transmitting container 2, a detection component 3, a moving mechanism 4, and a processor.

[0067] Among them, the light-shielding box 1 serves as the installation base for the entire detection device, providing a closed detection environment to eliminate ambient light interference, especially interference with fluorescence signals.

[0068] A light-transmitting container 2, housed within a light-shielding enclosure 1, is used to contain wastewater and sludge samples and to serve as an optical detection window. The container 2 is made of a light-transmitting material, requiring high transmittance of ultraviolet-visible light, such as quartz glass. In this embodiment, the container 2 is preferably cylindrical, allowing the wastewater and sludge samples to be uniformly dispersed within the container. The container 2 is equipped with an inlet 21 and a feed inlet 22. The inlet 21 is used to introduce the wastewater sample, and the feed inlet 22 is used to add a fluorescent dye, such as SYBR Green I.

[0069] The detection component 3 includes a spectrophotometer detection module 31 and a fluorescence detection module 32. The spectrophotometer detection module 31 has a spectrophotometer light source 311 and a spectrophotometer receiver 312 arranged opposite each other along a first radial direction. The spectrophotometer light source 311 emits a detection beam into the light-transmitting container 2, and the spectrophotometer receiver 312 receives the transmitted light signal. In this embodiment, the wavelength of the light emitted by the spectrophotometer light source 311 is 550nm-650nm, preferably a 590nm LED, which avoids common absorption peaks of sludge, such as the strong absorption of humic acid at 400-500nm. The spectrophotometer receiver 312 is selected as a silicon photodiode, with a corresponding range of 400nm-1100nm.

[0070] In this embodiment, the spectrophotometer 311 emits a detection beam, which then penetrates vertically through the wastewater sample in the light-transmitting container 2. The transmitted light is detected by the spectrophotometer receiver 312 (silicon photodiode), and the total biomass is calculated according to the Lambert-Beer light absorption law.

[0071] The fluorescence detection module 32 has a fluorescence light source 321 and a fluorescence receiver 322 arranged opposite to each other along a second radial direction. The fluorescence light source 321 is used to emit an excitation beam into the light-transmitting container 2, and the fluorescence receiver 322 is used to receive fluorescence signals. In this embodiment, the fluorescence light source 321 is selected as a 470nm blue light-emitting LED, matched with the optimal excitation wavelength of SYBR Green I. The fluorescence receiver 322 is a photomultiplier tube (PMT) or a high-sensitivity APD. A filter assembly 326 is provided in front of the fluorescence receiver 322. The filter assembly 326 includes a 520nm bandpass filter 3261 and a 620nm bandpass filter 3262, which are used to detect the fluorescence signals of live bacteria and dead bacteria, respectively.

[0072] A fluorescent dye is excited by 470nm excitation light. Live bacterial DNA binds to the dye and emits fluorescence at 520nm, while dead bacteria / impurities emit fluorescence at 620nm. By separating the signals using dual-wavelength filters, the marker data of the active bacterial community can be calculated.

[0073] In this embodiment, the first radial direction and the second radial direction do not coincide, so that the optical paths of the spectrophotometer detection module 31 and the fluorescence detection module 32 do not coincide. This allows the light beams to be emitted into the sample separately, thereby obtaining the corresponding data.

[0074] The moving mechanism 4, located inside the light-proof enclosure 1, drives the detection component 3 to move axially along the light-transmitting container 2. This configuration allows the spectrophotometer detection module 31 to move axially along the light-transmitting container 2, thereby detecting the total biomass data of wastewater samples at different positions within the container 2 along the axial direction. Correspondingly, the fluorescence detection module 32 can also move axially along the light-transmitting container 2 to detect the fluorescence signal of wastewater samples at different positions within the container 2 along the axial direction.

[0075] In this embodiment, dead bacteria, organic debris, or free dyes in the sludge sample spontaneously generate fluorescence (such as a 620nm signal), which is directly superimposed on the live bacteria-specific signal (520nm), resulting in a high false positive rate.

[0076] Therefore, before adding fluorescent dyes, it is necessary to measure the background signal. This involves measuring the autofluorescence of the sludge sample at wavelengths of 520 nm and 620 nm, specifically including the background fluorescence (main peak at 620 nm) produced by DNA / RNA fragments released from dead bacteria / cell debris and humic acid and other organic matter (the weak fluorescence of natural organic matter in the sludge at 520 nm). Measuring the background signal is used for subsequent data correction after staining.

[0077] Fluorescence measurements after dye addition include the total fluorescence response encompassing both live bacterial signal and residual background. Specifically, this includes live bacterial-specific signals: the dye binds to intact live bacterial DNA, emitting 520 nm fluorescence (intensity F). 520 ); Residual background signal: Non-specific binding of dead bacteria / organic matter to dye (still containing 620nm fluorescence).

[0078] The true live bacteria signal can be obtained by subtracting the background signal from the total fluorescence response data.

[0079] In actual processing, a spectral light source emits a 590nm beam, and a spectral receiver measures the transmitted light intensity (OD value) at that location; a fluorescent light source excites the sample, and a fluorescence receiver measures the 520nm fluorescence signal and background interference (such as the 620nm signal). The processor directly uses the OD value and fluorescence signal (520nm minus background) at the current point to calculate the viable bacterial concentration using a pre-calibrated calibration coefficient.

[0080] During data processing, this embodiment uses a processor electrically connected to a spectrophotometer receiver 312, a fluorescence receiver 322, and a moving mechanism 4. The processor is an embedded system or an industrial computer, configured to control the moving mechanism 4 to enable the detection component 3 to perform detection at multiple axial positions, and can measure data at different positions.

[0081] The total biomass data of the sample is calculated based on the transmitted light signal. The spectrophotometer 31 measures the intensity of transmitted light (OD value) of the sample at a wavelength of 590nm. This signal directly reflects the total biomass concentration of the sample (including live bacteria, dead bacteria and other light-absorbing substances).

[0082] Based on the fluorescence signal, the active bacterial community marker data are calculated. Specifically, the fluorescence detection module induces SYBR Green I dye to bind to the live bacterial DNA through 470nm excitation light, emitting a 520nm fluorescence signal. F 520 ), while detecting the 620nm background signal (F 背景 Viable microbial marker data were calculated using dual-wavelength difference: Viable microbial marker = Background subtraction eliminates interference from dead bacteria and impurities.

[0083] Based on total biomass data and fluorescent labeling data of viable bacterial communities, the concentration of viable microorganisms was calculated. The viable bacterial concentration can be calculated using the following formula: , where a is the calibration coefficient determined by the standard plate counting method.

[0084] In this embodiment, if F is used directly 520 Calculations show that samples with the same viable bacteria concentration can exhibit significant deviations in results due to differences in turbidity (such as varying sludge thickness). High-turbidity samples will attenuate fluorescence intensity (light scattering effect), requiring compensation using OD values, which can be achieved through differential calculations. Eliminate autofluorescence interference from the sample matrix to ensure that the detection signal reflects only the specific fluorescence of SYBR Green I dye binding to live bacterial DNA. OD 590 Includes dead bacteria absorbing light, via OD 590 (Total biomass) and fluorescence signal (F) 520 The inverse relationship between OD / F can be calculated to suppress interference; the higher the proportion of dead bacteria, the higher the OD / F ratio. 520 The larger the ratio, the more automatically the calculation result will be corrected.

[0085] The wastewater sludge microbial community concentration detection device disclosed in this application acquires total biomass data of the sample through a spectrophotometric detection module 31, and simultaneously uses a fluorescence detection module 32 to specifically identify live bacteria labeling signals. The processor establishes a quantitative relationship between the total biomass data and the live bacteria labeling signals through synergistic analysis, thereby achieving accurate calculation of the concentration of active microorganisms. The effectiveness of this technology stems from the inherent correlation between the total biomass benchmark provided by spectrophotometric detection and the live bacteria-specific signals provided by fluorescence detection. Through data fusion processing by the processor, it effectively solves the technical problem that traditional methods cannot distinguish between live and dead bacteria.

[0086] As some implementation methods, see the appendix. Figure 3 and 8 As shown, the spectral detection module 31 also includes a first collimating lens 313 and a first focusing lens 314.

[0087] The first collimating lens 313 is positioned in the light-emitting direction of the beam-splitting light source 311 to convert the divergent light source into a parallel beam, eliminating the measurement error of the OD value caused by the difference in beam angle and ensuring the applicability of Beer-Lambert's law (uniform optical path).

[0088] The first focusing lens 314 is located in the light-incident direction of the beam splitter 312, and its focal length matches the diameter of the light-transmitting container 2, converging the transmitted light onto the detection surface of the beam splitter 312 to improve the signal-to-noise ratio.

[0089] See attached document Figure 3 and 9 As shown, the fluorescence detection module 32 also includes a second focusing lens 323, a collecting lens 324, and a second collimating lens 325.

[0090] The second focusing lens 323 is positioned in the light-emitting direction of the fluorescence light source 321 and is configured as an aspherical lens to form an excitation spot of a certain diameter in the sample area. The collecting lens 324 and the second collimating lens 325 are positioned in the light-incident direction of the fluorescence receiver 322, with the second collimating lens 325 close to the fluorescence receiver 322. The collecting lens 324 is positioned as close as possible to the side wall of the light-transmitting container 2 to efficiently converge the scattered fluorescence emitted by the sample, thereby improving signal capture capability. The second collimating lens 325 re-collimates the scattered light into a parallel beam, ensuring that the light passes through the subsequent filter assembly 326 at the optimal incident angle, avoiding spectral shift caused by oblique incidence.

[0091] The filter assembly 326 includes a 520nm bandpass filter 3261 and a 620nm bandpass filter 3262, used to detect the fluorescence signals of live and dead bacteria, respectively. Specifically, the 520nm bandpass filter 3261 is used to extract the characteristic fluorescence (main peak at 520nm) of the fluorescent dye (SYBR Green I) binding to the DNA of live bacteria. The 620nm bandpass filter 3262 is used to monitor the interfering fluorescence of dead bacteria / organic matter (for background subtraction).

[0092] By optimizing the optical structures of the spectrophotometer detection module 31 and the fluorescence detection module 32, the detection accuracy and reliability are significantly improved. The spectrophotometer detection module 31 uses a first collimating lens 313 to ensure parallel light beam incidence, and combines this with a first focusing lens 314 to improve the signal-to-noise ratio of the transmitted light signal, making OD value measurement more stable. The fluorescence detection module 32 forms a uniform excitation spot through a second focusing lens 323, and simultaneously employs a combined optical path of a collecting lens 324 and a second collimating lens 325, significantly improving the collection efficiency of the fluorescence signal and optimizing the beam incident angle to match the optimal working conditions of the filter. The filter assembly 326 (520nm / 620nm bandpass filter 3262) accurately separates the fluorescence signals of live and dead bacteria, improving the specificity of live bacteria detection and reducing interference from dead bacteria. The overall solution, through precise optical design, achieves highly sensitive and highly repeatable detection of active microbial concentrations.

[0093] As some implementation methods, see the appendix. Figure 4 , 8As shown in Figure 9, the beam splitting detection module 31 also includes a first light source shield 315 and a first receiving shield 316. The first light source shield 315 covers the optical path between the beam splitting light source 311 and the first collimating lens 313, eliminating lateral leakage of LED divergent light and blocking ambient light from entering the optical path from the lens edge. The first receiving shield 316 covers the optical path between the first focusing lens 314 and the beam splitting receiver 312, preventing transmitted light from being contaminated by ambient light before reaching the detector and suppressing secondary interference from reflected light on the lens surface.

[0094] The fluorescence detection module 32 also includes a second light source shield 327 and a second receiving shield 328. The second light source shield 327 covers the optical path between the fluorescence light source 321 and the second focusing lens 323, ensuring that the excitation light enters the sample only through a preset optical path. The second receiving shield 328 covers the optical path between the collecting lens 324 and the fluorescence receiver 322, blocking background light interference from outside the sample cell and maintaining the dark-field detection environment of the filter assembly 326.

[0095] All of the above-mentioned sunshades are made of opaque material.

[0096] In order to enable the spectral detection module 31 and the fluorescence detection module 32 to move synchronously along the axial direction of the light-transmitting container 2, the detection component 3 in this embodiment also includes an annular mounting disk 30, which is coaxially sleeved on the outer wall of the light-transmitting container 2. The spectral detection module 31 and the fluorescence detection module 32 are fixedly arranged circumferentially along the annular mounting disk 30, and the driving end of the moving mechanism 4 is connected to the annular mounting disk 30.

[0097] In this embodiment, the optical paths of the spectrophotometer detection module 31 and the fluorescence detection module 32 are perpendicular to each other. This allows the components in the spectrophotometer detection module 31 and the fluorescence detection module 32 to be evenly distributed on the annular mounting plate 30. Simultaneously, the optical paths of the spectrophotometer detection module 31 and the fluorescence detection module 32 are coplanar. This allows the spectrophotometer detection module 31 and the fluorescence detection module 32 to simultaneously detect the sample at the same cross-section of the light-transmitting container 2. For example, before the addition of fluorescent dye, the spectrophotometer detection module 31 and the fluorescence detection module 32 can be alternately triggered for detection, with an alternation time difference of less than 1 ms, ensuring spatial-temporal synchronization of the spectrophotometer and fluorescence signals and reducing errors in the calculation of viable bacteria concentration. For instance, before the addition of fluorescent dye, during the synchronous movement of the spectrophotometer detection module 31 and the fluorescence detection module 32 along the axial direction of the light-transmitting container driven by the moving mechanism, the spectrophotometer detection module 31 and the fluorescence detection module 32 can be alternately triggered for detection, measuring the OD value at various axial positions of the wastewater sample. F 背景 .

[0098] In this embodiment, the moving mechanism 4 is a lead screw module, including a motor, a lead screw and a nut seat. The motor is fixed on the bottom surface inside the light-proof box 1. The output shaft of the motor is vertically upward and connected to the annular mounting plate 30 through the nut seat. This allows for efficient and precise control of the position of the detection component 3 in the axial direction of the light-transmitting container 2.

[0099] After adding fluorescent dye to the light-transmitting container 2, it needs to be stirred and mixed thoroughly. If stirring is not performed, the fluorescent dye (such as SYBR Green I) is prone to uneven distribution in the sludge, resulting in excessively high local concentrations and large deviations in the detection results. Traditional fixed stirrers will block the light path, causing distortion of OD measurement values ​​and scattering of fluorescence signals.

[0100] Therefore, this embodiment shows a stirring mechanism 5, specifically, see attached drawing. Figure 4-10 As shown, the stirring mechanism 5 includes a stirring shaft 51, a stirring rod 52, and a drive assembly 52.

[0101] The stirring shaft 51 is coaxially disposed inside the light-transmitting container 2, and its side wall is provided with at least two symmetrically arranged storage slots 511. The upper end of the stirring rod 52 is installed in the storage slot 511 through a hinge shaft and can rotate around the hinge shaft. Specifically, when the stirring shaft 51 rotates centrifugally, the upper end of the stirring rod 52 can rotate around the hinge shaft outside the stirring shaft 51. When there is no centrifugal force, the stirring rod 52 retracts into the storage slot 511 under the action of gravity.

[0102] In this embodiment, the length and width of the receiving groove are slightly larger than the length and width of the stirring rod 52. This ensures that the stirring rod 52 can easily retract into the receiving groove 511 under the action of gravity. By setting the depth of the receiving groove to be greater than the thickness of the stirring rod 52, the outer wall of the stirring rod 52 and the outer wall of the stirring shaft 51 are not coaxial after the stirring rod 52 is retracted into the receiving groove 511. This can prevent sludge from adhering to the receiving groove 511, so that the outer wall of the stirring rod 52 protrudes outside the receiving groove 511 after it is retracted into the receiving groove 511.

[0103] In this embodiment, the drive assembly 52 is disposed on the outside of the light-transmitting container 2 and connected to the stirring shaft 51, for driving the stirring shaft 51 to rotate and move axially. Specifically, the drive assembly 52 is disposed at the bottom of the light-transmitting container 2.

[0104] When stirring is required, the drive assembly 52 drives the stirring shaft 51 to extend into the light-transmitting container 2. After the stirring shaft 51 extends into the appropriate position in the light-transmitting container 2, the drive assembly 52 drives the stirring shaft 51 to rotate. The stirring rod 52 unfolds from the receiving tank 511 under the action of centrifugal force, thereby stirring and mixing the wastewater containing fluorescent dye.

[0105] After mixing is complete, if the stirring rod 52 and the stirring shaft 51 are rigidly connected, the stirring shaft 51 will be blocked by the stirring rod 52, and the stirring shaft 51 cannot be pulled out by the drive assembly 52. ​​Therefore, in this embodiment, the upper end of the stirring rod 52 and the stirring shaft 51 are hinged, and a receiving groove 511 is provided on the outside of the stirring shaft 51. After mixing is complete, the stirring rod 52 can automatically retract into the receiving groove 511 by gravity, preventing the stirring rod 52 from leaking out of the outside of the stirring shaft 51 and affecting the removal of the stirring shaft 51 from the light-transmitting container 2.

[0106] The stirring mechanism 5 in this embodiment adopts a retractable design. During the stirring stage, the driving component 52 pushes the stirring shaft 51 to extend into the light-transmitting container 2. Centrifugal force causes the hinged stirring rod 52 to unfold, achieving efficient mixing. After stirring is completed, the stirring rod 52 automatically retracts into the storage groove 511 under the action of gravity, allowing the stirring shaft 51 to exit the detection area without obstruction. This not only ensures the uniform distribution of fluorescent dye, but also completely eliminates the obstruction of the light path by the traditional rigid stirring rod 52, realizing interference-free collaborative operation of stirring and optical detection.

[0107] To enable the drive assembly 52 to both rotate and linearly move the stirring shaft 51, this embodiment presents a preferred structural configuration of the drive assembly 52. ​​Specifically, the drive assembly 52 includes a mounting bracket 521, a gear disc 522, a rotary motor 523, a main gear 524, and a telescopic element 525.

[0108] The mounting bracket 521 is fixedly installed at the bottom of the light-transmitting container 2. The mounting bracket 521 is used to position the gear disk 522 and the rotating motor 523. Specifically, a limiting plate 5211 is fixedly installed on the mounting bracket 521. The limiting plate 5211 is parallel to the bottom surface of the light-transmitting container 2, and there is a gap between the limiting plate 5211 and the light-transmitting container 2 to install the gear disk 522. The gap is adapted to the thickness of the gear disk 522, thereby ensuring that the gear disk 522 will not move axially after it is located between the limiting plate 5211 and the bottom surface of the light-transmitting container 2.

[0109] The limiting plate 5211 has a through hole 5212 for the stirring shaft 51 to pass through, and the stirring shaft 51 can move and rotate along the axial direction of the through hole 5212.

[0110] The toothed disc 522 is positioned between the limiting plate 5211 and the bottom surface of the light-transmitting container 2. The lower end of the stirring shaft 51 moves through the toothed disc 522 and the through hole 5212. The side wall of the stirring shaft 51 is provided with a guide groove 512 along its axial direction. The inner wall of the toothed disc 522 is provided with a guide protrusion 5221 that cooperates with the guide groove 512. Because the guide groove 512 and the guide protrusion cooperate, the toothed disc 522 and the stirring shaft 51 cannot rotate relative to each other, but can only rotate as a whole. At the same time, the stirring shaft 51 can move axially relative to the toothed disc 522.

[0111] Because of the presence of the guide groove 512 on the stirring shaft 51, the sewage inside the light-transmitting container 2 will flow out of the outside of the light-transmitting container 2 along the guide groove 512. Therefore, in this embodiment, a sealing member 526 is also sleeved on the outside of the stirring shaft 51. The sealing member 526 is located between the toothed disc 522 and the light-transmitting container 2. With this setting, the sealing member 526 can seal the bottom surface of the mounting hole of the stirring shaft 51 and the light-transmitting container 2, preventing sewage from flowing out of the outside of the light-transmitting container 2 along the guide groove 512.

[0112] The rotating motor 523 is fixedly mounted on the mounting bracket 521. The output end of the rotating motor 523 meshes with the gear disk 522 through the main gear 524. Thus, the rotating motor 523 drives the main gear 524 to rotate, and the main gear 524 drives the gear disk 522 to rotate, thereby causing the gear disk 522 to drive the stirring shaft 51 to rotate.

[0113] The telescopic element 525 has a fixed end and a telescopic end. The fixed end is fixed to the outside of the light-proof housing 1, and the telescopic end is rotatably connected to the lower end of the stirring shaft 51. Thus, when the stirring shaft 51 needs to move, the extension or retraction of the telescopic element 525 drives the stirring shaft 51 to move axially in the light-transmitting container 2. Specifically, when the stirring shaft 51 needs to be inserted into the light-transmitting container 2 for stirring, the telescopic end of the telescopic element 525 extends; when the stirring shaft 51 needs to be pulled out of the light-transmitting container 2, the telescopic end of the telescopic element 525 retracts.

[0114] During the movement of the stirring shaft 51, due to the cooperation between the guide groove 512 and the guide protrusion 5221, the stirring shaft 51 can only move axially relative to the toothed disc 522. At this time, the toothed disc 522 is limited between the limiting plate 5211 and the light-transmitting container 2. Therefore, the toothed disc 522 will not move axially. In this embodiment, the end of the stirring shaft 51 cannot be separated from the contact surface between the toothed disc 522 and the limiting plate 5211. This is because there is a sealing element 526 between the top surface of the toothed disc 522 and the bottom surface of the light-transmitting container 2, which can seal the mounting hole on the light-transmitting container 2 and prevent sewage from overflowing from the mounting hole to the outer periphery of the toothed disc 522.

[0115] Since the telescopic end of the telescopic element 525 is rotatably connected to the stirring shaft 51, the stirring shaft 51 can rotate relative to the telescopic end of the telescopic element 525 during rotation, which does not affect the rigid movement restriction of the telescopic element 525. In this embodiment, the telescopic element 525 is a cylinder, a hydraulic cylinder, or an electric push rod.

[0116] The drive assembly 52 in this embodiment achieves precise control of the rotation and axial movement of the stirring shaft 51 through an innovative mechanical structure design. The gear disc 522 engages with the stirring shaft 51 via a guide groove, ensuring stable and reliable power transmission while allowing the stirring shaft 51 to move freely up and down. The seal 526 effectively prevents sewage leakage, and the telescopic element 525 is rotatably connected to the stirring shaft 51, avoiding motion interference. This design enables the stirring mechanism 5 to smoothly complete the entire workflow of extending into the stirring area and retracting to avoid obstacles, ensuring both effective stirring and uninterrupted optical path during detection, resulting in stable and reliable overall operation.

[0117] In some implementations, the light-transmitting container 2 is fixed to the bottom surface of the light-shielding box 1 by a mounting cylinder 6. The mounting cylinder 6 has an opening at the bottom and a cavity of a certain depth. The bottom opening of the light-shielding box 1 corresponds to the opening of the mounting cylinder 6. The mounting bracket 521, the gear plate 522, the rotating motor 523 and the active gear are arranged in the cavity of the mounting cylinder 6, thereby protecting the drive assembly 52 and preventing the entire drive assembly 52 from being completely exposed to the outside of the light-shielding box 1.

[0118] In addition, in order to install the telescopic element 525, this embodiment also fixes a fixing bracket 7 at the bottom opening of the light-shielding box 1. On the one hand, it seals the bottom opening of the mounting cylinder 6. On the other hand, as the mounting base for the telescopic element 525, by setting the mounting cylinder 6 inside the light-shielding box 1, at least part of the drive component 52 can be hidden inside the light-shielding box 1, avoiding it being completely exposed at the bottom of the light-shielding box 1, which would cause the mechanical parts to occupy a large space at the bottom of the light-shielding box 1, and at the same time reduce the protection of the drive component 52.

[0119] This application discloses a method for detecting the concentration of microbial communities in sewage sludge, including the following steps:

[0120] S1. The sludge sample to be tested is injected into the light-transmitting cavity after pretreatment. The sludge sample is injected into the light-transmitting container after pretreatment (such as crushing, filtering, and dilution) to ensure sample homogeneity and avoid interference from particulate matter with optical detection.

[0121] S2: Start the spectrophotometer module and measure the initial OD of the sample at 590nm. 590 The initial OD value was measured using a wavelength of 590 nm, reflecting the total biomass of the sample (including viable bacteria, dead bacteria, and non-biological light-absorbing substances).

[0122] S3: Activate the fluorescence detection module and measure the background fluorescence intensity at 520nm. The autofluorescence intensity at 520 nm was measured without the addition of fluorescent dye. This is used to subsequently subtract interference signals from dead bacteria / organic matter.

[0123] S4: Inject the fluorescent agent into the light-transmitting cavity and stir evenly. Add fluorescent dyes such as SYBR Green I, and use a stirring mechanism to stir the fluorescent dye and wastewater to ensure that the fluorescent dye and live bacterial DNA are fully mixed.

[0124] S5: Measure the fluorescence intensity at 520nm again. F 520 Before proceeding to step S5, the stirring shaft needs to be removed from the transparent container to prevent it and the stirring rod from affecting the optical path. Measure the fluorescence intensity at 520 nm. F 520 At this point, the signal contains live bacteria-specific fluorescence and residual background.

[0125] S6: Calculate the viable bacteria concentration according to the formula: , where a is the calibration coefficient determined by the standard plate counting method.

[0126] Combining OD value (total biomass) with fluorescence signal (live bacteria specificity), utilizing OD 590 As a baseline for total biomass, fluorescence difference ( Extracting the proportion of live bacteria while avoiding interference from dead bacteria. This is done using the unstained... (Including dead bacteria / organic matter signals) Corrected staining F 520 Significantly improves the specificity of live bacteria detection through experimental calibration. α It adapts to the characteristics of different sludge samples and reduces systematic errors. By fusing data from spectrophotometry (OD value) and fluorescence spectroscopy (520nm / 620nm dual channels), it overcomes the inherent limitation of traditional spectrophotometry in distinguishing between live and dead bacteria, greatly reducing the detection error of live bacteria concentration.

[0127] The above method measures the fluorescence difference at 520 nm before and after staining. The viable bacterial concentration was calculated, but unstained sludge samples may exhibit autofluorescence at 520 nm due to dead bacteria, cell debris, or organic matter such as humic acid. F auto520 ), directly take it as This can lead to an overestimation of the live bacteria signal. This background subtraction method is particularly prone to significant errors when the proportion of dead bacteria in the sludge is high.

[0128] Therefore, this embodiment proposes a new... The calculation method, specifically, is in step S3. Calculated using the following formula: .

[0129] in, The fluorescence intensity at 520 nm before staining may include fluorescence interference from live bacteria, dead bacteria, and organic matter.

[0130] The fluorescence intensity at 620 nm before staining is mainly derived from dead bacteria / organic debris.

[0131] This is the background value of pure water at 620nm, used to subtract instrument noise.

[0132] β The dead bacteria interference coefficient, calibrated to 0.3-0.5, represents the proportion of interference of the 620nm signal to the 520nm channel.

[0133] in, To extract specific signals of dead bacteria / organic matter from the 620nm channel after subtracting the instrument background.

[0134] To convert the 620nm interference signal to an equivalent 520nm interference value proportionally, To subtract the contribution of dead bacteria / organic matter from the total signal at 520nm, the background signal of real live bacteria is obtained.

[0135] By introducing a dead bacteria-specific signal from a 620nm channel ( F auto620 ) and interference coefficient β Establish a dynamic correction model, from F auto520 The method precisely removes the contribution of dead bacteria, thus solving the problem of impure background signals in the above detection methods.

[0136] Through 620nm signal and β Coordinated correction of coefficients quantizes the interference signal of dead bacteria / organic matter at 520nm. and from F auto520 Deduct from the middle to make the final It only reflects the true background value of viable bacteria. This method greatly reduces the detection error of viable bacteria concentration, and is especially suitable for complex sludge samples with a high proportion of dead bacteria or containing humic acid, significantly improving the accuracy and universality of the detection method in this embodiment.

[0137] This embodiment also discloses the dead bacteria interference coefficient. β The specific calibration method and steps include:

[0138] Sludge standard samples with dead bacteria percentages of 0%, 20%, 50%, and 80% were prepared to simulate the changes in the proportion of dead bacteria that may be encountered in actual wastewater treatment.

[0139] The autofluorescence intensity of each standard sample at 520nm / 620nm can be measured. The unstained fluorescence intensity of each standard sample at 520nm and 620nm can be measured by the fluorescence detection module.

[0140] Fitting β The value minimizes the error in calculating the viable bacteria concentration. Based on experimental data, the value is adjusted. β The value (0.3-0.5) minimizes the error in calculating the viable bacteria concentration (e.g., by using least squares fitting).

[0141] The proportion of dead bacteria in sludge varies significantly between different wastewater treatment plants or at different stages of operation (e.g., younger sludge has fewer dead bacteria, while older sludge has more). This can be determined through experimental calibration. β Values ​​are used to ensure the algorithm can adapt to different samples and avoid fixed values. β Systematic errors caused by the value.

[0142] In the above embodiment, an axial multi-point detection step is further included after step S1:

[0143] S11. Drive the detection component to move along the axial direction of the light-transmitting container using the moving mechanism, and sequentially perform steps S2-S5 at at least three equally spaced positions, recording the OD at each position. 590 , F 520 and value;

[0144] S12, Measurements taken at each location F 520 and After taking the arithmetic mean separately, combine with the average OD 590 The value is then substituted into the formula in step S6 to calculate the final viable bacteria concentration.

[0145] Axial multi-point detection significantly improves the accuracy and reliability of viable bacteria concentration measurement. Its technical advantages are mainly reflected in the following: A moving mechanism drives the detection component to measure at least three equidistant positions along the axis of the transparent container. The final viable bacteria concentration is calculated by averaging the data from each point. This method effectively eliminates measurement deviations caused by uneven axial distribution of the sample, making the test results more representative and minimizing measurement errors. It also enhances the system's adaptability to complex samples such as those with high concentrations or containing particulate matter. Through coordinated operation with the stirring mechanism, a fully automated process from sample mixing to multi-point detection is achieved, greatly improving detection efficiency and result stability, and providing more reliable data support for the precise control of wastewater treatment processes.

[0146] It is worth noting that, in order to adapt to the structural characteristics of the detection device in this embodiment, before adding fluorescent dye, the detection component can be driven by a moving mechanism to perform multi-point testing along the axial direction of the light-transmitting container. During this process, the OD at different locations can be measured. 590 and This allows for the acquisition of data at multiple points along the axial direction of the sample. After the fluorescent dye is added, the spectrophotometer can be activated without activating the fluorescence detection module; in this case, the fluorescence detection module can be started, allowing for the measurement of values ​​at different locations. F 520 .

[0147] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A device for detecting the concentration of microbial communities in sewage sludge, characterized in that, The application relates to a device for detecting the concentration of active microorganisms in sewage, which comprises the following components: a light-proof box body; a light-transmitting container arranged in the light-proof box body, the light-transmitting container being provided with a liquid inlet and a feeding opening, the liquid inlet being used for feeding sewage samples, and the feeding opening being used for adding fluorescent dyes; a detection assembly, which comprises a light-splitting detection module and a fluorescence detection module, the light-splitting detection module being provided with a light-splitting light source and a light-splitting receiver which are oppositely arranged along a first radial direction, the light-splitting light source being used for emitting a detection light beam to the light-transmitting container, and the light-splitting receiver being used for receiving a transmitted light signal, the fluorescence detection module being provided with a fluorescence light source and a fluorescence receiver which are oppositely arranged along a second radial direction, the fluorescence light source being used for emitting an excitation light beam to the light-transmitting container, and the fluorescence receiver being provided with a filter assembly in front of the fluorescence receiver, the filter assembly being used for receiving and screening a fluorescence signal of a specific wavelength, wherein the first radial direction and the second radial direction are not coincident; a moving mechanism arranged in the light-proof box body and used for driving the detection assembly to move along the axis of the light-transmitting container; a processor which is electrically connected with the light-splitting receiver, the fluorescence receiver and the moving mechanism, and is configured to control the moving mechanism to drive the detection assembly to perform detection at multiple axial positions, calculate sample total biomass data according to the transmitted light signal, calculate active flora marker data according to the fluorescence signal, and calculate concentration data of active microorganisms based on the total biomass data and the active flora fluorescence marker data; Formula for calculating the concentration of viable microorganisms wherein, F 520 F520 is the fluorescence intensity measured at 520 nm after injection of the fluorescent agent, F520bg is the background fluorescence intensity measured at 520 nm, OD 590 F590 is the initial fluorescence intensity measured at 590 nm of the sample, OD F590bg is the background fluorescence intensity measured at 590 nm, α C is the calibration coefficient calibrated by the standard plate count method.​ 2. The apparatus for detecting concentration of sewage sludge microorganism group according to claim 1, wherein: the light-splitting detection module further comprises a first collimating lens arranged in the light-emitting direction of the light-splitting light source and a first focusing lens arranged in the light-receiving direction of the light-splitting receiver; the fluorescence detection module further comprises a second focusing lens arranged in the light-emitting direction of the fluorescence light source, a collecting lens and a second collimating lens which are coaxially and separately arranged in the light-receiving direction of the fluorescence receiver; the filter assembly comprises a 520 nm band-pass filter and a 620 nm band-pass filter, which are used for detecting the fluorescence signals of living bacteria and dead bacteria respectively.

3. The apparatus for detecting concentration of sewage sludge microorganism group according to claim 2, wherein: the light-splitting detection module further comprises a first light source light shield cover and a first receiving light shield cover, the first light source light shield cover covers the light path between the light-splitting light source and the first collimating lens, and the first receiving light shield cover covers the light path between the first focusing lens and the light-splitting receiver; the fluorescence detection module further comprises a second light source light shield cover and a second receiving light shield cover, the second light source light shield cover covers the light path between the fluorescence light source and the second focusing lens, and the second receiving light shield cover covers the light path between the collecting lens and the fluorescence receiver.

4. The apparatus for detecting concentration of sewage sludge microorganism group according to claim 1, wherein the detection assembly further comprises an annular mounting disc which is coaxially sleeved on the outer wall of the light-transmitting container, the light-splitting detection module and the fluorescence detection module are fixedly arranged along the circumference of the annular mounting disc, and the driving end of the moving mechanism is connected with the annular mounting disc.

5. The apparatus for detecting concentration of sewage sludge microorganism group according to claim 1, wherein the device further comprises a stirring mechanism, and the stirring mechanism comprises: a stirring shaft which is coaxially arranged in the light-transmitting container and is provided with at least two symmetrically arranged receiving grooves in the side wall; a stirring rod, the upper end of the stirring rod being mounted in the receiving groove through a hinge shaft and being rotatable about the hinge shaft; a driving assembly which is arranged outside the light-transmitting container and is connected with the stirring shaft, and is used for driving the stirring shaft to rotate and move axially; when the stirring shaft rotates, the stirring rod is unfolded from the receiving groove under the action of centrifugal force; when the stirring shaft stops rotating, the stirring rod is folded into the receiving groove under the action of gravity.

6. The apparatus for detecting concentration of sewage sludge microorganism group according to claim 5, wherein the driving assembly comprises: The installation support is fixedly arranged at the bottom of the light-transmitting container, and a limiting plate is fixedly arranged on the installation support. The gear disc is arranged between the limiting plate and the bottom surface of the light-transmitting container, the lower end of the stirring shaft is movably arranged through the gear disc and the through hole, a guide groove is arranged on the side wall of the stirring shaft along the axial direction, a guide protrusion matched with the guide groove is arranged on the inner wall of the gear disc, a sealing element is arranged on the outer side of the stirring shaft, and the sealing element is located between the gear disc and the light-transmitting container. The rotating motor is fixed on the installation support, and the output end of the rotating motor is engaged with the gear disc through a main gear. The telescopic element has a fixed end and a telescopic end, the fixed end is fixed to the outer side of the light-avoiding box body, and the telescopic end is rotatably connected with the lower end of the stirring shaft.

7. A method for detecting the concentration of a microbial population in sewage sludge, which uses the device for detecting the concentration of a microbial population in sewage sludge according to any one of claims 2 to 6, characterized by The method comprises the following steps: S1, injecting the pretreated sludge sample into the light-transmitting cavity; S2: Start the spectrophotometric detection module and measure the initial absorbance of the sample at 590 nm OD 590 value; S3: start the fluorescence detection module, measure the background fluorescence intensity at 520 nm ; S4, injecting the fluorescent agent into the light-transmitting cavity and stirring uniformly; S5: Measure 520 nm fluorescence intensity again F 520 ; S6: Calculate the viable cell concentration according to the formula: wherein α is the calibration factor for standard plate count method calibration.

8. The method for detecting concentration of sewage sludge microorganism group according to claim 7, wherein, In step S3 Calculated by the following formula: , wherein, F520 is the fluorescence intensity at 520 nm before staining, F620 is the fluorescence intensity at 620 nm before staining, F6200 is the background value of pure water at 620 nm, β F6200 is the background value of pure water at 620 nm, F6200 is the background value of pure water at 620 nm, F6200 is the background value of pure water at 620 nm, F6200 is the background value of pure water at 620 nm, 9. The method for detecting concentration of sewage sludge microorganism group according to claim 8, wherein, The dead bacteria interference coefficient β The specific steps include: preparing sludge standard samples with dead bacteria proportions of 0%, 20%, 50%, and 80%; measuring the autofluorescence intensities of the standard samples at 520 nm / 620 nm; fitting β The value minimizes the error of the live bacteria concentration calculation.

10. The method for detecting concentration of sewage sludge microorganism group according to claim 7, wherein After the step S1, an axial multi-point detection step is further included: S11, driving the detection assembly to move along the axis of the light-transmitting container by the moving mechanism, sequentially performing steps S2-S5 at at least three equidistant positions to record the OD values of the respective positions 590 、 F 520 and values; S12, measurements taken at each location F 520 and After taking the arithmetic mean separately, combine with the average OD 590 The value is then substituted into the formula in step S6 to calculate the final viable bacteria concentration.

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