WO2021242911A1

WO2021242911A1 - Wastewater system to monitor pathogens and methods of use

Info

Publication number: WO2021242911A1
Application number: PCT/US2021/034331
Authority: WO
Inventors: Bryan Walser
Original assignee: Pangolin Llc
Priority date: 2020-05-26
Filing date: 2021-05-26
Publication date: 2021-12-02

Abstract

Provided herein are systems and methods to monitor waste water, detect a pathogen, and control the outbreak and spread of diseases associated with the pathogen.

Description

TITLE

WASTEWATER SYSTEM TO MONITOR PATHOGENS AND METHODS OF USE

BACKGROUND

[001] The invention generally relates to wastewater sampling and pathogen surveillance.

[002] COVID-19 is a highly pathogenic respiratory disease, which exhibited an outbreak after its first appearance in Wuhan, China in December 2019. COVID-19 is caused by a novel coronavirus namely SARS-CoV-2, which causes respiratory illness with elevated fatality rate in patients, including patients with one or more of comorbidities such as obesity, hypertension and diabetes. Cases of COVID-19 in which the patient shows no symptoms of infection appear asymptomatic but may still infect or transmit the virus to the community, state, or country.

[003] The Centers for Disease Control and Prevention confirms that the virus has been found in the feces of patients diagnosed with COVID-19, and is thus gathering in city sewers. The surveillance of sewage or waste water for COVID-19 may provide details on the epidemiology and accelerate governments’ efforts to contain the virus, outbreak, and save human lives.

[004] The present invention attempts to solve these problems, as well as others.

SUMMARY OF THE INVENTION

[005] Provided herein are methods and systems to monitor waste water and detect a pathogen. [006] The systems and methods are set forth in part in the description which follows, and in part will be obvious from the description, or can be learned by practice of the methods, apparatuses, and systems. The advantages of the systems and methods will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the systems and methods as claimed.

[007] Accordingly, it is an object of the invention not to encompass within the invention any previously known product, process of making the product, or method of using the product such that Applicants reserve the right and hereby disclose a disclaimer of any previously known product, process, or method. It is further noted that the invention does not intend to encompass within the scope of the invention any product, process, or making of the product or method of using the product, which does not meet the written description and enablement requirements of the USPTO (35 U.S.C. § 112, first paragraph) or the EPO (Article 83 of the EPC), such that Applicants reserve the right and hereby disclose a disclaimer of any previously described product, process of making the product, or method of using the product. It may be advantageous in the practice of the invention to be in compliance with Art. 53(c) EPC and Rule 28(b) and (c) EPC. All rights to explicitly disclaim any embodiments that are the subject of any granted patent(s) of applicant in the lineage of this application or in any other lineage or in any prior filed application of any third party is explicitly reserved. Nothing herein is to be construed as a promise.

BRIEF DESCRIPTION OF THE DRAWINGS

[008] In the accompanying figures, like elements are identified by like reference numerals among the several preferred embodiments of the present invention.

[009] FIG. 1A is a perspective view of a wastewater system.

[010] FIG. IB is a schematic flow chart of the wastewater pathogen detection system.

[Oil] FIG. 2A is a schematic flow chart a continuous policy feedback loop.

[012] FIG. 2B is a schematic flow chart for an afferent clinical feedback loop.

DETATEED DESCRIPTION OF THE INVENTION

[013] The foregoing and other features and advantages of the invention are apparent from the following detailed description of exemplary embodiments, read in conjunction with the accompanying drawings. The detailed description and drawings are merely illustrative of the invention rather than limiting, the scope of the invention being defined by the appended claims and equivalents thereof.

[014] Embodiments of the invention will now be described with reference to the Figures, wherein like numerals reflect like elements throughout. The terminology used in the description presented herein is not intended to be interpreted in any limited or restrictive way, simply because it is being utilized in conjunction with detailed description of certain specific embodiments of the invention. Furthermore, embodiments of the invention may include several novel features, no single one of which is solely responsible for its desirable attributes or which is essential to practicing the invention described herein.

[015] The use of the terms “a” and “an” and “the” and similar referents in the context of describing the invention are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. It will be further understood that the terms “comprises,” “comprising,” “includes,” and/or “including,” when used herein, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.

[016] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. The word “about,” when accompanying a numerical value, is to be construed as indicating a deviation of up to and inclusive of 10% from the stated numerical value. The use of any and all examples, or exemplary language (“e.g ” or “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any nonclaimed element as essential to the practice of the invention.

[017] References to “one embodiment,” “an embodiment,” “example embodiment,” “various embodiments,” etc., may indicate that the embodiment(s) of the invention so described may include a particular feature, structure, or characteristic, but not every embodiment necessarily includes the particular feature, structure, or characteristic. Further, repeated use of the phrase “in one embodiment,” or “in an exemplary embodiment,” do not necessarily refer to the same embodiment, although they may.

[018] As used herein the term “method” refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, biological, industrial, electrical, software, and mechanical arts, as well as public health, public policy, and healthcare. Unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification. [019] System

[020] The present invention is a system that applies to separate sanitary sewer systems, combined sewer systems, or standalone sewer systems. Separate sanitary sewer systems are designed to transport sewage alone in underground pipe or tunnel system from houses and commercial buildings to treatment facilities or disposal. In municipalities served by sanitary sewers, separate storm drains may convey surface runoff directly to surface waters. Sanitary sewers are part of an overall system called a sewage system or sewerage. Sanitary sewers are distinguished from combined sewers, which combine sewage with storm water runoff in one pipe. A combined sewer system is a sewage collection system of pipes and tunnels designed to simultaneously collect surface runoff and sewage water in a shared system. A standalone sewer system may be an open system or septic tank where sewage is collected for home or small building where there is no separate or combined sewer system.

[021] “Wastewater system” herein means a separate sewer system, a combined sewer system, or a standalone sewer system that is defined by a geographical region, generally shown with the system 100 in FIG. 1A. “Wastewater” herein means the type of wastewater that is produced by a community of people. Wastewater is characterized by volume or rate of flow, pressure within the enclosed pipe, physical condition, chemical and toxic constituents, and its bacteriologic, virological, or pathogenic status (which organisms it contains and in what quantities). Wastewater consists mostly of greywater (from sinks, bathtubs, showers, dishwashers, and clothes washers), blackwater (the water used to flush toilets, combined with the human waste that it flushes away); soaps and detergents; and toilet paper. The wastewater system is a pressurized system with variable wastewater flow rates due to levels of diluent in the system, blockages, etc. In alternative embodiments, the system may incorporated for testing finished drinking water, treated water, disinfected water, irrigation water, and water obtained from wells, rivers, lakes and recreational waters such as swimming pools. Other embodiments, the system samples and analyzes food (such as fruits, vegetables, meat and prepared food items), swabs taken from slaughter lines, and meat surfaces, as well as swabs taken from environmental surfaces from slaughter houses, and meat preparation facilities, soil and clinical and veterinary samples including stool and biopsy samples.

[022] Generally speaking, the system 100 to monitor sewage or untreated wastewater for pathogen states in a city, state, or country and comprises detecting a pathogen in the wastewater system and providing feedback for actionable containment measures or feedback to adjust to varying parameters to optimize detection of the pathogen in wastewater. A pathogen is a bacterium, virus, or other microorganism that can cause disease. Pathogens detectable by the system, include, but are not limited to: prions, bacterium, viruses, fungi, algae, or parasites. In one embodiment, the pathogen is the virus COVID-19.

[023] As shown in FIG. IB, the system 100 comprises accessing the wastewater system 110, retrieving wastewater samples 120, detecting the pathogen in wastewater 130, data generation 140, data retrieval 150, data interpretation 160, data presentation 170, implementation 180, implementation evaluation 190, and updating the detector 132. The methods and steps for the system 100 may be combined in any particular order or combination to provide for wastewater pathogen detection and appropriate response for the population or adaptation of the pathogen detection. In one embodiment, accessing the wastewater 110 comprises using a mapping algorithm to provide geolocation in the wastewater system to sample for the pathogen or where to place the detectors in the wastewater system, including separate standalone locations and topographical locations factoring volumetric wastewater flow rates and velocities of the wastewater. In another embodiment, accessing the wastewater system 110 comprises using a timing algorithm to process and compute the flow of wastewater, pressure of wastewater, the season of detection, and contaminants in the wastewater. The algorithms may estimate wastewater loading or fecal loading using historic wastewater meter data and population density, according to one embodiment.

[024] The system 100 is operably coupled with a server and a network communicating information from the retrieving wastewater samples 120, detecting the pathogen in wastewater 130, data generation 140, data retrieval 150, data interpretation 160, data presentation 170, implementation 180, implementation evaluation 190, and updating the detector 132. Each part of the system may operate through a computer or program module to execute a set of instructions, protocols, or parameters and be updated through an Artificial Intelligence (AI) system or distributed Internet of Things (IoT) system. The server may be operably coupled with a database to store information.

[025] The system 100 further comprises retrieving the samples from the wastewater 120 including retrieving samples to test the pathogen at a water treatment plant, a manhole, or along upstream stations within the wastewater system, according to one embodiment. The retrieved wastewater samples may be stored at stable conditions for further testing or analysis of the pathogen, analytes, or wastewater conditions. In another embodiment, the retrieving the samples from the wastewater 120 comprises a modular detector to detect the pathogen in the wastewater remotely in the wastewater system.

[026] The system 100 comprises detecting the pathogen in the wastewater system 130. In one embodiment, the detector to detect the pathogen may be placed at any location along the wastewater system with access to the wastewater. The locations along the wastewater system include, junction, unusual blockages in the pipes, upstream facilities, nursing homes, hospitals, industrial complexes, residential areas, including the most distal wastewater sites in the city, state, or country. The detector may employ molecular methods to detect the pathogen, including Polymerase Chain Reaction (PCR). PCR detects bacteria, viral, and protozoan pathogens with high sensitivity in wastewater, regardless if the pathogen is alive or dead. PCR includes primers specific for the pathogen. In one embodiment, the primer is specific for the (a) the region spanning NSP1, NSP2, NSP3 of the SARS-CoV-2 gene (Accession No. YP_009725297.1, Accession No. P89070, or Accession No. B5APV2); (b) the region spanning Nl, N2, and N3 loci of the SARS-CoV-2 nucleocapsid gene; (c) region spanning E gene (SARS-Cov2 Wuhan-Hu isolate sequence 124 NC_045512.); the region spanning the SARS-CoV-2 S gene (GenBank under the accession numbers MT072688 and NC_045512); or primer having any arbitrary 8-mer 10-mer 12-mer 64-mer 128-mer specific along the coding and non-coding regions for the SARS- CoV-2 gene.. PCR conditions to remove interfering substances in the wastewater include, but are not limited to: inhibition avoidance, chemical avoidance, and physical avoidance, as described in commonly assigned U.S. provisional application serial no 63/032,565, filed May 30, 2020. The detection may be performed by nucleic acid microarrays or antibody/receptor technologies to detect multiple pathogens simultaneously. Nucleic acid capture optimization and electrochemical detection of the pathogen may also be deployed. Combining these multiplexed methods with fiberoptic sensors and lab-on-a-chip technology allows the system to rapidly screen, identify, and quantify multiple pathogens in real time.

[027] The system 100 comprises a data generator 140 to indicate the level of the pathogen in a city, state, or country, accounting for the population and levels of the wastewater. The data generator 140 may indicate rising levels of the pathogen above an infection level, threshold or rate of detection indicates that the city, state, or country may require containment measures to delay or stop the spread of the pathogen. The threshold level of pathogen considers the population above N people, N% of relevant population, and size of population. In one embodiment, the infection level or threshold of the pathogen may be above about 0.1 million, about 0.2 million, about 0.3 million, about 0.4 million, about 0.5 million, or about 0.6 million genome units fL of wastewater, according to one embodiment. In another embodiment, the infection level or threshold of the pathogen may be above about 10, about 20, about 30, about 40, or about 50 genome units per mL of wastewater. In one embodiment, for the COVID-19 pathogen, as many as about 600,000 to about 30,000,000 viral genomes of SARS-CoV-2 per mL of fecal material, assuming a fecal load of about 100-400 g feces/day/person with a density of about 1.06 g/mL. Normalization of population of the pathogen’s infection level in the wastewater ensure that a significant increase in pathogen concentration in a wastewater sample does not correspond to an increase in population in the serviced wastewater area or a decrease in the amount of diluent as might occur during a dry season or drought. The normalization of the pathogen’s infection level includes factoring the cultural practices, concentration of the pathogen in the wastewater, considering seasonal and rain effects on the levels of diluent, which may be higher due to hurricanes, monsoons, or rainy seasons, or lower due to drought, cultural practices, or dry season.

[028] The data generator 140 may correlate with clinical data and the containment measures. Containment measures may include stay at home orders from the government, social distancing in public places, allowing only essential businesses to remain open, a requirement to wear masks in public, creation of non-acute and acute care sites for hospital patient overflow, production and demand for pharmaceuticals with pathogen treatment, activation of vaccine development and production, or a requirement for all health care workers to wear Personal Protective Equipment (PPE) whenever interacting with an admitted or sick patient. Falling levels of the pathogen below an infection level or particular threshold indicates that the community, state, or country no longer has the risk of pathogen transmission and may remove the containment measures for the city, state, or country. The infection levels of the pathogen may correlate with clinical data from the number of positive cases or deaths in the community, state, or country. The containment measures are to ensure the epidemiological curve of the pathogen remains low and the number of critically ill patients does not overburden the health system of the city, state, or country. Containment measures and pathogen detection may ensure second wave of epidemics or pandemics are prevented.

[029] In one embodiment, the system 100 comprises providing a continuous policy feedback by the detection of the pathogen in the wastewater system and establishing a clinical afferent loop, according to one embodiment. In one embodiment, the continuous policy feedback 200 is shown in FIG. 2A, where the detection of the pathogen in the wastewater 210 is continuous and in real time and allows for the detection of the levels of the pathogen indicating a disease state 220 in the city, state, or country. If the detection of the pathogen is above an infection level or threshold 230, then containment measures are deployed in real time 240, as indicated above. In the case of asymptomatic conditions, clinical testing of the pathogen of admitted or deceased patients 250 proceeds to confirm the presence of the pathogen in admitted or deceased patients. If the detection of the pathogen is not below the infection level or threshold 340, then continuous pathogen detection 210 proceeds or adapts to detection levels through sampling, attributes and detector feedback, as indicated herein. If the detection of the pathogen level or threshold in the wastewater is not below an infection level or disease state, then additional containment measures are deployed or extended in the in the city, state, or country 240. Once the levels of the pathogen are below an infection level or level 260, then the containment measures may be removed 270. [030] A clinical afferent loop coincides with the levels or thresholds of the pathogen in the wastewater and provides a city, state, or country actionable or containment measures to curb the spread of the pathogen and prevent outbreaks and transmission of the pathogen. In one embodiment, the clinical afferent loop 300 is shown in FIG. 2B, and comprises detection of the pathogen in the wastewater 310 and allows for the detection of the infection levels of the pathogen indicating a disease state 320 in the city, state, or country. If the detection of the pathogen is above a level or threshold 320, then clinical testing of the pathogen of admitted patients 330 proceeds to confirm the presence of the pathogen in admitted patients. Testing of admitted patients or deceased patients for the pathogen confirms presence of the pathogen correlated with the pathogen detection in the wastewater. Comparison and correlation with clinical data in the surrounding cities, states, or countries confirms the spread or containment of the pathogen, where the comparison or correlation is processed by Artificial Intelligence systems operably coupled with the server or database. AI systems may provide prospective information or retrospective information regarding the levels of the pathogen, considering factors of the wastewater conditions, population, type of pathogen, clinical data, hospital capacity, population, and the like.

[031] If the detection of the pathogen is not above of level or threshold 320, then continuous pathogen detection 310 proceeds or adapts to the detection levels through sampling, attributes, AI systems, and detector feedback, as indicated herein. If the detection of the pathogen a level or threshold is not below an infection level or disease state 340, then additional clinical testing of admitted patients initializes 330, as well as other proactive measures such as creating non-acute and acute care sites. Once the levels of the pathogen are below are threshold or level 340, then the clinical testing of admitted patients requirement may be removed.

[032] In another embodiment, the system comprises establishing an afferent virological loop and an afferent biological loop. The afferent virological loop establishes levels or thresholds of the pathogen in the wastewater, and analyses and interprets the impact of virus molecular evolution, degradation, virus load in urine, feces, or saliva, site of entry in the host, and epidemiology in a city, state, or country. The virological loop may also consider the pathogen’s degradation in wastewater, the shedding rate of the pathogen, where the shedding rate is the number of pathogens or viruses excreted by infected humans. In one embodiment, COVID-19 includes a persistence at 4°C in wastewater for more than 20 days, or a persistence of at least 1 or 2 days at summer temperatures. In another embodiment, COVID-19 includes a half-life at ambient conditions (20 °C) to range between approximately 4.8 h and 7.2 h. The afferent virological loop considers the maximum time a virus load can spend in the sewer line on route to the detector or sampling location before the virus load falls below an estimated limit of detection. In another embodiment, where wastewater flow is at a temperature of 20 °C, at least 25% of the virus load should remain even in situations where the average in-sewer travel time is long at least 10 h and the virus stability is relatively low (/ ₀.5 = 4.8 h), for when; e.g. where

ti/2, 1 is the initial half-life, 71 is the temperature at which initial half-life

was derived, ti/2,2 is the half-life at seasonally- and spatially-adjusted wastewater temperature, 72 is the calculated temperature to which initial half-life is adjusted to, and Q 10 is a factor of temperature-dependent of rate change, ranging between 2 and 3 for most biologic systems.

[033] The afferent biological loop establishes levels or thresholds of the pathogen in the wastewater, and analyses the biological feedback in a city, state, or country considering the season, wastewater temperature, population density, and the like. The afferent biological loop normalizes the pathogen detection with other biomarkers in the wastewater, where the biomarkers substances that are natural excreted by humans in constant quantities. In one embodiment, at least 0.88% of the population is detected for the pathogen in the wastewater system and if infected the pathogen (1 in 114 individuals) and in another embodiment, at least 0.00005% of the population is detected for the pathogen in the wastewater system (1 infected case in about 2 million non-infected individuals). In one embodiment, population size may be normalized to certain hydrochemical parameters, such as chemical oxygen demand (COD), biological oxygen demand (BOD) or ammonium (NH4+), which will aid in estimating populations at a particular time period. The sample collection and analytical variability may be normalized to spatial and temporal comparisons in the wastewater. Biological factors that affect concentration of the pathogen in sewage including the following: site of replication in the host GI, upper respiratory, nose, skin, internal organs; duration of release from the host; site of entry in the host; site of infection in the host; duration of infection in the host; concentration in the source; incidence of infection or circulation in the population; cultural practices of toilet use/toilet paper use/flushing wipes or soap, water use per capita; season; and survival in the wastewater system. Other factors for the wastewater field consider environmental and cultural factors where pathogens may be concentrated in the wastewater or more diluted in the wastewater. The biological factors in are input variables for the AI system in the biological afferent loop where

[034] In one embodiment, the system 100 comprises data retrieval 150 including a plurality of collection sites in the wastewater system for the detection of the pathogen or modular detectors that are collected and replaced. In another embodiment, the data retrieval 150 is through a plurality of detectors with a communication protocol or wireless transmitter to connect to the internet, wireless hub, satellite device, or Internet of Things (IoT) devices. The Internet of things (IoT) is a system of interrelated computing devices, mechanical and digital machines provided with unique identifiers (UIDs) and the ability to transfer data over a network without requiring human-to-human or human-to-computer interaction. In one embodiment, the system comprises a distributional IoT system including a plurality of attributes and solutions in a communication protocol. The IoT system uses of a variety of processing software and data storage, where data can be stored at the edge of the network or stored centrally; and the software processing can take place centrally (in cloud services), or processing can take place at the edge of the network, such as IoT gateways. The distributed IoT system is a collection of mostly autonomous processors (nodes) communicating over a communication network and having the following features: 1) Common goal: the nodes are working to achieve a common goal that cannot be achieved using a single processor; 2) Loosely coupled: the nodes on the network should be loosely coupled and should be able to communicate via a messaging system using a standard protocol; 3) No direct synchronization with other nodes on the network: the nodes on the network should not be synchronized via a common clock; 4) Distributed memory: the nodes on the network should not have a shared memory space if possible, but shared distributed memory space for the entire system can be provided as an abstraction; 5) Geographical distribution: the nodes on the network should be geographically distributed and should avoid any centralization; 6) Autonomy and heterogeneity: the nodes on the network should be autonomous and heterogeneous in nature.

[035] In one embodiment, the plurality of attributes and solutions of the distributional IoT includes: embedded connectivity for the detector to make the potential to make every detector a smart detector to capture and integrate the appropriate data, decipher and analyze the pathogen correctly, and take downstream or upstream actions, including automation processes, corrective adjustments, remote monitoring of wastewater conditions to see how the wastewater system is performing or to track pathogen through wastewater system. Alternative plurality of attributes and solutions of the distributional IoT includes: tracking and tracing of pathogens so all times the exact whereabouts of the pathogen in the wastewater system, the condition of the pathogen and if the pathogen has degraded; monitor detectors to ensure they are working properly or will soon need maintenance; ensure temperature pathogen detection; track performance and productivity of detectors; and establish and measure detector parameters for continuous improvement of the system. Alternative plurality of attributes and solutions of the distributional IoT includes: monitor detectors in real-time to determine if a detector is being over-utilized or when the detector is idle indicating possible pipe blockage; or monitor detectors for i) other pathogens, ii) other biological entities, iii) other biological compounds, or iv) other substances of relevance to public health, epidemiology, or other population-based studies.

[036] In one embodiment, the data retrieval 150 through IoT uses detectors to aggregate foreground parameters in the wastewater including, but not limited to, timing of detection, location of detection, flow or pressure of wastewater, season of detection, sampling type, detector type, or other conditions and variability of the wastewater. In another embodiment, the data retrieval 150 by IoT aggregates background parameters, including, but not limited to, population status, population extent, population background, health or public policy, state of containment measures, and other conditions in the city, state, or country.

[037] In one embodiment, the data retrieval 150 detects at the most distal location of the city, state, or country and includes non-wastewater system collection (septic tanks), non-wastewater system customers, for the most distal location of the city, state, or country. In one embodiment, the timing of the pathogen detection is less than about 12 hours, less than about 24 hours, less than about 48 hours, since pathogens have a rapid generation time. In one embodiment, the generation time N is a 8 generations, alternatively about 10 generations, alternatively about 12, alternatively N < X, where X is arbitrary or distinct for the pathogen. For COVID-19, generation time is about 3 days = 24; depends on Ro and surface type dwell life,

[038] In one embodiment, the data interpretation 160 comprises interpreting data through an algorithm or Artificial Intelligence (AI) platform. The AI platform may interpret data over time and assess foreground variables and background variables for the detection of the pathogen in wastewater. In another embodiment, the data interpretation 160 comprises digital health options for the population or at upstream facilities.

[039] The system 100 comprises data presentation 170 and implementation 180. Implementation 180 comprises In Vitro Diagnostic Multivariate Index Assay (IVDMIA) confirmation or correlation of the pathogen in wastewater through separate testing. IVDMIA is a device that 1) combines the values of multiple variables using an interpretation function to yield a single, patient-specific result (e.g., a “classification,” “score,” “index,” etc.), for the diagnosis of disease or other conditions, or in the cure, mitigation, treatment or prevention of disease, and 2) provides a result whose derivation can be considered non-transparent and cannot be independently derived or verified by the end user. IVDMIA components may include a static assessment or a dynamic assessment, according to one embodiment. In one embodiment, implementation 180 comprises inclusion of background elements in the city, state, or country. [040] The system 100 comprises implementation evaluation 190 including input data of the timing of pathogen detection, analyte of the pathogen, and wastewater conditions, according to one embodiment. In another embodiment the implementation evaluation 190 includes delivering output data for the service levels of the wastewater and timing of pathogen detection. The timing of the pathogen detection in the water is critical for considering the generation time of the pathogen.

[041] The system 100 further comprises updating the detector 132. The detector may be updated or adjusted with data including, but not limited, service levels of the wastewater, the number and location of interlocks or interceptors in the wastewater system, a plurality of input parameters, a plurality of Output parameters, and In Vitro Diagnostic Multivariate Index Assay (IVDMIA) confirmation of the pathogen in wastewater through separate testing. The detectors may be updated and ensured data identity, strength, quality, and signal of the pathogen detection by current Good Manufacturing Practices (cGMP). The system comprises defined input parameters and defined operations to achieve an implied defined output. Data, AI implementation, cGMP, and algorithmic interpretation methods may be used as described in commonly assigned U.S. provisional application serial no. 63/033,627, filed June 2, 2020.

[042] Detecting the pathogen or sampling the wastewater at each interlock or interceptor and mapping each interceptor to the specific neighborhoods it serves, the system provides pathogen occurrence data collection representative of each serviced area of the city, state, or country. If pathogen concentrations detected are higher in one interceptor than the rest in the wastewater system, the corresponding serviced area could be quarantined and subject to pharmaceutical or non-pharmaceutical treatments, and nearby areas subject to the containment measures for risk of a potential viral outbreak. Detecting the pathogen or sampling wastewater in rural areas, the system determines where in the environment to sample based upon watershed modeling and microbial source tracking.

[043] Pathogens

[044] A wide variety of pathogenic organisms pass through municipal waste-water treatment systems. Any type of infection within a community is likely to lead to pathogen excretion in bodily fluids/substances and therefore, transported into the community sewage system. Infections may be classified into symptomatic infections, and asymptomatic infections. Symptomatic infections may result in death, severe illness, moderate severity, and mild illness-all of which are clinical diseases. Asymptomatic infections may be infection without clinical illness and exposure including colonization showing no illness.

[045] Table 1 classifies pathogen in categories Category A pathogens require the most intensive public preparedness efforts due to the potential for mass causalities, public fear, and civil disruption. Category B pathogens are also moderately easy to spread, but have lower mortality rates. Category C pathogens do not present a high public health threat, but could emerge as future threats.

[046] Table 1: The center for disease control select agents

[047] Agents causing enteric and respiratory infections are released in large numbers in feces and respiratory secretions. Many of the enteric viruses such as the enteroviruses and adenoviruses may replicate both in the intestinal and respiratory tract. The number of enteric viruses detected can approach peak concentrations of 10¹² organisms per gram of stool while protozoa can approach 10⁶-10⁷ per gram. Cultivatable enteric bacterial pathogens such as Salmonella may also occur in concentrations as large as 10¹¹ per gram. The concentration of respiratory viruses ranges from 10⁵ to 10⁷ per ml of respiratory secretion. Blood-borne viruses such as HIV will be found in the feces of infected persons and many viruses will occur in the urine during infection of the host, although these excreted viruses may not be infectious. The total amount of virus released by a person is, of course, also related to the amount of feces, urine, respiratory secretion, and skin that is released by the person. On average, a person excretes between 100 g to 400 g of feces and 700-2000 ml of urine per day.

[048] Non-pathogens

[049] Non-biological entities detected by the system include, but are not limited to drug compounds such as pharmaceuticals, opioids, tracer chemicals, inorganic molecules, and the like. Biological entities detected by the system include, but are not limited to biological or chemical compounds such as simple or complex organic molecules, indicator molecules, peptides, proteins (e.g. antibodies) or a polynucleotides (e.g. anti-sense), microbes, and other organisms. Biological compounds detectable by the system may include or substances of relevance to public health, epidemiology, or other population-based studies.

[050] Examples

[051] The previous examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and/or methods claimed herein are made and evaluated, and are intended to be purely exemplary of the invention and are not intended to limit the scope of what the inventors regard as their invention. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

[052] Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in ° C. or is at ambient temperature, and pressure is at or near atmospheric. [053] System hardware and software

[054] As used in this application, the terms “system” may refer to a computer-related entity, either hardware, a combination of hardware and software, software, or software in execution. For example, a system can be, but is not limited to being, a process running on a processor, a processor, an object, an executable, a thread of execution, a program, and/or a computer. By way of illustration, both an application running on a server and the server can be a component. One or more components can reside within a process and/or thread of execution, and a component can be localized on one computer and/or distributed between two or more computers.

[055] Generally, program modules include routines, programs, components, data structures, etc., that perform particular tasks or implement particular abstract data types. Moreover, those skilled in the art will appreciate that the inventive methods can be practiced with other computer system configurations, including single-processor or multiprocessor computer systems, minicomputers, mainframe computers, as well as personal computers, hand-held computing devices, microprocessor-based or programmable consumer electronics, and the like, each of which can be operatively coupled to one or more associated devices.

[056] The illustrated aspects of the innovation may also be practiced in distributed computing environments where certain tasks are performed by remote processing devices that are linked through a communications network. In a distributed computing environment, program modules can be located in both local and remote memory storage devices.

[057] A computer typically includes a variety of computer-readable media. Computer-readable media can be any available media that can be accessed by the computer and includes both volatile and nonvolatile media, removable and non-removable media. By way of example, and not limitation, computer-readable media can comprise computer storage media and communication media. Computer storage media includes volatile and nonvolatile, removable and non-removable media implemented in any method or technology for storage of information such as computer-readable instructions, data structures, program modules or other data. Computer storage media includes, but is not limited to, RAM, ROM, EEPROM, flash memory or other memory technology, CD-ROM, digital versatile disk (DVD) or other optical disk storage, magnetic cassettes, magnetic tape, magnetic disk storage or other magnetic storage devices, or any other medium which can be used to store the desired information and which can be accessed by the computer. [058] Communication media typically embodies computer-readable instructions, data structures, program modules or other data in a modulated data signal such as a carrier wave or other transport mechanism, and includes any information delivery media. The term “modulated data signal” means a signal that has one or more of its characteristics set or changed in such a manner as to encode information in the signal. By way of example, and not limitation, communication media includes wired media such as a wired network or direct-wired connection, and wireless media such as acoustic, RF, infrared and other wireless media. Combinations of the any of the above should also be included within the scope of computer-readable media.

[059] Software includes applications and algorithms. Software may be implemented in a smart phone, tablet, or personal computer, in the cloud, on a wearable device, or other computing or processing device. Software may include logs, journals, tables, games, recordings, communications, SMS messages, Web sites, charts, interactive tools, social networks, VOIP (Voice Over Internet Protocol), e-mails, and videos.

[060] In some embodiments, some or all of the functions or process(es) described herein and performed by a computer program that is formed from computer readable program code and that is embodied in a computer readable medium. The phrase “computer readable program code” includes any type of computer code, including source code, object code, executable code, firmware, software, etc. The phrase “computer readable medium” includes any type of medium capable of being accessed by a computer, such as read only memory (ROM), random access memory (RAM), a hard disk drive, a compact disc (CD), a digital video disc (DVD), or any other type of memory.

[061] All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[062] While the invention has been described in connection with various embodiments, it will be understood that the invention is capable of further modifications. This application is intended to cover any variations, uses or adaptations of the invention following, in general, the principles of the invention, and including such departures from the present disclosure as, within the known and customary practice within the art to which the invention pertains.

Claims

CLAIMS What is claimed is:

1. A method for detecting a biological entity in wastewater comprises: a. detecting the biological entity in a wastewater system; b. generating data to indicate the level of the biological entity in a city, a state, or a country, accounting for the population and the levels of biological entity in the wastewater, wherein the city, the state, or the country deploys a plurality of containment measures to delay or stop the spread of the biological entity when the levels of the biological entity rise above an infection level; and c. clinical testing of the biological entity of admitted or deceased patients in hospitals to confirm the presence of the biological entity in admitted or deceased patients, wherein the biological entity is selected from the group consisting of: pathogens, biological compounds, or substances of relevance to public health, epidemiology, or other population-based studies.

2. The method of Claim 1, wherein the threshold level of biological entity comprises the population above N people, N% of relevant population, and size of population.

3. The method of Claim 2, where the infection level of the biological entity is above about 0.1 million, about 0.2 million, about 0.3 million, about 0.4 million, about 0.5 million, or about 0.6 million genome units fL of wastewater,

4. The method of Claim 2, where the infection level of the biological entity may be above about 10, about 20, about 30, about 40, or about 50 genome units per mL of wastewater.

5. The method of Claim 3 or 4, wherein the biological entity is SARS-CoV-2.

6. The method of Claim 2, wherein detecting the biological entity is at a plurality of locations in the wastewater system selected from the group consisting of: junctions, unusual blockages in the pipes, upstream facilities, nursing homes, hospitals, industrial complexes, residential areas, including the most distal wastewater sites in the city, state, or country.

7. The method of Claim 2, wherein detecting the biological entity is by including Polymerase Chain Reaction (PCR) and non-PCR methods.

8. The method of Claim 2, wherein the containment measures are selected from the group consisting of: stay at home orders from the government, social distancing in public places, allowing only essential businesses to remain open, a requirement to wear masks in public, creation of non-acute and acute care sites for hospital patient overflow, and a requirement for all health care workers to wear Personal Protective Equipment (PPE) whenever interacting with an admitted or sick patient.

9. The method of Claim 2, further comprising comparing the infection levels of the biological entity in the surrounding cities, states, or countries confirms the spread or containment of the biological entity, where the comparing the infection levels is processed by Artificial Intelligence systems operably coupled with a server.

10. The method of Claim 9, further comprising comparing the infection levels of the biological entity with clinical data of surrounding cities, states, or countries confirms the spread or containment of the biological entity, where the comparing the infections levels is processed by Artificial Intelligence systems operably coupled with a server.

11. The method of Claim 2, further comprising interpreting the impact of virus molecular evolution, a biological entity degradation in the wastewater, a biological entity load in the wastewater, a site of entry of the biological entity in the host, shedding rate of the biological entity, and an epidemiology of the biological entity in the city, the state, or the country.

12. The method of Claim 2, further comprising analyzing the biological feedback in the city, the state, or the country, wherein the biological feedback includes the season, wastewater temperature, population density, spatial and temporal comparisons in the wastewater; and normalizing the biological entity detection with other biomarkers in the wastewater, and detecting the biological entity in at least N% of the population in the wastewater system.