INTRODUCTION:

Many products that have made people's lives easier and more convenient have been made possible by technological breakthroughs, while at the same time, causing environmental damage. These objectives have led to the development of numerous novel tools and methods for the effective detection and management of environmental pollutants. At the moment, nano biotechnology is being used to create sensors that effectively identify different types of pollutants, and methods are also being created to effectively remove the pollutants from the environment.¹

In addition to its broad applications in the environmental field, nano biotechnology is also widely used in the biomedical, agricultural, robotics, sensors, electronics, and other fields. Nano biotechnology is a special combination of nanotechnology and biotechnology that uses biotechnological procedures and nano particles to create new techniques and ways that can improve human lives. Over time, the broad range of applications for nanobiosensors has become visible, since they provide a large platform for the detection of different analytes through the use of biomolecules and nanoparticles. Furthermore, it has been shown that biosensors made with nanoparticles regularly show improved capabilities.²

Due to the rapid outbreak of severe acute respiratory syndrome coronavirus-19 (SARS-CoV-2), there is a greater need for nanobiosensors. As a result, more nanobiosensors are being developed to detect SARS-CoV-2 in both the environment and biological medium. Since SARS-CoV-2 has not only harmed people but also the environment, it is necessary to create quick and precise nanobiosensors that can identify SARS-CoV-2 particle.

All living things have a tendency to detect changes in their environment and adapt to them in order to live. These beings have the capacity to sense beyond everyone's conception scientists. The senses of the eels and the animals' capacity to smell ability of a tiny variation in water tons, the butterflies' ability to detect the secretions of their partners, the Toxin sensing in algae is one instance found in nature. The fundamentals of the biosensor idea were first explored through in vitro research that imitated the sensory abilities of living things.³ Biosensor technology came up with this concept and advanced quickly. The definition of a biosensor is later provided by the International Union of Pure and Applied Chemistry (IUPAC), which formed a commission in 1996 to categorize and name the devices. The biosensor is described as “a device that uses specific biochemical reactions mediated by immune systems, tissues, organelles, or whole cells to detect chemical compounds, usually by electrical, thermal, or optical signals.” As the backbone of biosensor systems, which have the sense processes of sight, hearing, smell, taste, and touch, smaller, less expensive, and more precise devices than microelectronics could be made. The revolution in sensor technologies comes about by nano-biosensors, which allow for the quick analysis of several samples at the appropriate location and time. Many novel signal transduction methods have lately been applied, demonstrating excellent performance in selectivity, biocompatibility, non-toxicity, reversibility, quick reaction, and the sensitivity of determination by using nanomaterials. From a financial point of view, a more efficient chemistry lowers reagent use and total costs. The performance of various bioassays can be improved by nano materials.⁴

Advancements in micro- and nanofabrication techniques may make it easier to create smaller devices that can be applied in the field. Low sample volume, less reagent use, less invasive sample collection techniques, multiple analyte detection, and quick analysis periods are just a few of the benefits that biosensors offer. They also offer real-time decision-making for targeted therapy, in addition to these characteristics. As a result, biosensor monitoring has several benefits like in conjunction with current technology, and their elevated degree of compatibility might lead to notable advancements. The current analysis sought to highlight advancements in the methods of sample preparation and important detection in nanobiosensors.

In microbial biological sensors, a microbe or its enzymes and organelles are attached to electrodes and hence to a transducer that converts this biological reaction into electrical signals. The different enzymes available inside the microbial cell is the bio-element. The enzymes inside the cell bring about a very specific and selective reaction to the substance. Gene biosensors are most widely used and also since pure enzymes are being used.

The specificity and the response are far better than the other biosensors; however, it is a very long process and also very expensive as multiple enzymes and cofactors are needed to be purified. While microbial sensors, on the other hand, are little or less time-consuming compared to others with no need for purification and additionally a quick response was obtained.

The microorganisms should be immobilized in the transducer so that the sensor is to give reliable results. Microbial cells are immobilized by employing various chemical and physical strategies including adsorption, encapsulation, covalent binding, cross linking etc.

Table 1: The Properties of the Biosensors.⁵

Analyte	Bio- Component	Transducer Based on
Hormones	Enzymes	Electrochemical
Enzyme	Antibody	Semiconductors
Coenzymes	Cell	Optics
Substrate	Tissue	Photometric
Activator	Receptors	Flourometric
Inhibitors	Microorganisms	Flourometric
Antibody- Antigen	Nucleic Acid	Peizoelectric
Nucleic Acid	Lipids	Quartz Crystal Microbalance (QCM)
Microorganisms	Organelles	Microcantilever
Vitamin B₁₂	Aptamer	Aptasensors

The History of Biosensors:

The first biosensor was designed by Clark and Lyons in 1962 using an enzyme electrode. The target material in this system is glucose and its oxidation reaction was mediated by glucose oxidase.

The voltage between electrodes was sufficient for the reduction of O₂, and electric current was measured by the rate of concentration of O₂. The reduction of electric current directly depended on glucose concentration.

Another significant innovation is the developed biosensors with the designed potentiometric urea electrode. The classification of such types of electrodes was classified as first-generation biosensors.

According to this second generation of biosensors, auxiliary enzymes and/or co-reactants are immobilized with the analyte-converting enzyme to improve the analytical quality and simplify the performance.^5,6

Fig. 1: Schematic illustration of biosensors.⁷

A surface of the transducer is chemically modified with annexations next to the cladding material ELISA (enzyme-linked immunosorbent assay) electrodes belong to this class.

The biomolecules get involved in them toward the biosensing material. For example, this might occur in SPR (Surface Plasmon Resonance) biosensors.

Lastly, huge amounts of features are expected in the 4th generation with the developments in the MEMS /NEMS /BioNEMS, nanotechnology and biotechnology. With the contributions of engineers and scientists from a variety of fields, there are many recent, creative, and versatile opportunities to develop production techniques for biosensors. These biosensors are also used for the determination of biological and chemical effective substances in agricultural production, food analyzes and environmental monitoring in addition to mining, bioprocess, bio war and homeland security.

Microbial Nanobiosensors:

The term "microbial NBSs" refers to a class of biologically-based sensors (NBSs) that measure the electrical activity of a microorganism's metabolism when it consumes a "biofuel," such as glucose. These sensors use the same principle of measuring metabolism activity in the presence of analyte using immobilized microorganisms from which the metabolism resulting products are measured. Their potential to be genuine NBSs has been overlooked since they are being developed as a distinct field of electrochemical cells that produce power through the use of biofuels. We shall address these two courses independently, emphasizing their contributions competences in relation to the prerequisites for NBS classification. Microbiological Immobilized microorganisms and a considerably more varied transduction chain are seen in NBSs.

Microbial NBSs follow the same pattern as NBSs from enzymes. The plan is to immobilize whole cells in close contact to a particular transducer that translates metabolic products or effects into a biochemical signal. Microbial NBSs are used in biochemical and microbiological industrial processes where fermentation and electron transfer reactions are key components, such as in the food, pharmaceutical, and wastewater treatment industries, as well as in the generation of energy. Microbial NBS implementation has significant promise in the clinical laboratory fields of microbiology and parasitology, where culture media are utilized widely.⁸

Principle of Nanobiosensors:

Combining traditional biosensors with the fast-expanding field of nanotechnology led to the development of nanobiosensors. Nanobiosensors are devices that detect biological substances at the nanoscale through a transduction unit and a biological recognition element. Physicochemical transducers and receptors make up nanobiosensors. Biosensors are based on the recognition of molecules. Only when a particular molecular recognition is shared by the receptor and the bacteria can biological receptors identify them. The greatest illustrations of the interaction between an antibody and an antigen in molecular recognition are lock and key models. The components of the sensor that communicate with the target are called bioreceptors. In order to bind the target entity (enzymes, antibodies, deoxyribonucleic acid (DNA), cells, and aptamers) steadily under varied storage circumstances, bio-receptors are immobilely fixed on the transducer's surface.⁹

Measurable signals are converted into energy from the receptor, acting as an interface. Transducers modified with nanoparticles are the highlights of nanobiosensors, allowing rapid detection in a short period. Compared to simple biosensors, nanobiosensors can detect the quantity and presence of analytes.

Mechanism:

Analytical instruments containing a biological sensor and a physicochemical converter are called nano biosensors, or Nano BioSS. One of Nano BioSS's primary functions, it creates a digital electrical signal that is directly correlated with the total of one or a number of compounds are being examined. These nanobiosystems are supporting some important analytical developments that are receiving support also bolstered by developments in nanotechnology, contributing to the proof that they are increasing their respective applications and supporting equipment. This NanoBioSS/BioSS can accurately and quickly identify nanomaterials (NMs), which makes It is helpful in many industrial, environmental, and agricultural clinical, biological, medical. The Design/Fabrication Process of NanoBioSS is equally varied as its uses, with every NanoBioSS category that includes its benefits and drawbacks as a due to restrictions based on the uses.¹⁰

Fig. 2: Schematic mechanism of Nanobiosensors.¹¹

Nanobiosensors Types:

Fig. 3: Various types of Nanobiosensors.¹²

a. Nanoparticles based sensor:

1. Acoustic waves: Surface acoustic wave (SAW) devices, acoustic plate mode (APM) devices, flexural plate wave (FPW) devices, and thickness shear mode (TSM) resonators are examples of acoustic wave devices that have been employed for materials characterization and sensing applications. One port is utilized for both the input and the output terminals in one-port acoustic devices like TSM, while two-port devices like SAW, APM, and FPW use one port for input and the other for output. An acoustic wave produced by the input signal travels to a receiving transducer, where it is converted back into an electrical signal at the output port. The relative signal levels and phase delay between the input and output ports are used to calculate the sensor response.¹³

2. Magnetic Biosensors:

Using the magnetoresistance effect, magnetic biosensors utilize microparticles of 5–300nm and 300–500nm, respectively, in microfluidic channels. These particle surfaces are functionalized and altered to identify particular compounds with enticing sensitivity. Researchers are interested in magnetic biosensors because they provide many advantages over fluorescent-based techniques. Magnetic probes can be utilized for long-term labeling tests without producing background noise effects since they are more stable over time in culture. External magnetic fields allow for remote biological environment evaluation and control.¹⁴

3. Electrochemical Biosensor: Analytical tools known as electrochemical biosensors convert biological processes like the interaction of an enzyme with a substrate or an antibody with an antigen into electrical signals like current, voltage, impedance, etc. Since Clark created the first electrochemical biosensor for measuring blood glucose, several biosensor kinds have been gradually released and brought to market for a range of uses.¹⁵ An electrode is a crucial part of this electrochemical biosensor; it serves as a firm foundation for the movement of electrons and the immobilization of biomolecules like enzymes, antibodies, and nucleic acids. A variety of chemical modification techniques are used for this, including thiol (maleimide), aldehyde (hydrazide), and carboxyl (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, depending on the chemical groups on the electrode and whether or not supporting materials are present. Given that ineffective immobilization may result in inactivity.

b. Nanotube: A common nanomaterial in material science and optoelectronics is carbon nanotubes. After their discovery in the late 1990s, their outstanding characteristics have attracted attention from all over the world. Their electrical conductivity, flexible geometries, and dynamic physicomechanical qualities-such as high aspect ratios, superior functionalization abilities, and high mechanical folding and strength properties—are their most crucial characteristics. These properties have led to the utilization of one- and multi-wall nanotubes in the development of improved biosensors.² Among the most widely used sensing developments in recent years is the development of glucose biosensors that use nanotubes as the glucose oxidase enzyme's inhibiting surface. Using this enzyme, the concentration of glucose in various bodily fluids can be determined.

c. Nanowires: Nanowires are cylinder-shaped structures with diameters and lengths ranging from a few micrometers to centimeters. One-dimensional nanostructures with superior electron transport capabilities are called nanowires.² The migration of charge carriers is a key link between solid substances and nanowires. Although there aren't many nanowire sensors, there are a few significant instances in the scientific record of how nanowires have enhanced biological function and detection. Cui and Lieber reported on the performance of biosensors for detecting biological and chemical species utilizing silicon nanowires doped with boron. In addition to being thoroughly studied, semiconductor nanowires have been used to couple different proteins into particular substrates in order to facilitate identification. Silicon nanowires coated with streptavidin molecules have been used to identify and separate them from a mixture.

d. Optical: The phenomena of optical nanostructures interacting with light is the base of optical nanobiosensors. Certain biomolecules cause modifications to the optical characteristics of light, including absorbance or fluorescence detection, when they attach to the analyte in the sample. The presence and quantity of the analyte are ascertained by detecting and quantifying these changes.¹⁶ By utilizing this phenomena of light-matter interaction, optical nanobiosensor guarantee exact detection in applications such as food and water handling, all while offering great sensitivity and selectivity. There are important uses for the Surface Plasmon Resonance (SPR) phenomenon in the identification of pathogens in food and water.

Advantages and Limitations of Nanobiosensors:

Advantages:	Limitations
High Sensitivity	Complex Fabrication
High Specificity	Cost & Optimization
Rapid Detection	Sample Complexity
Multiplexing	Regulatory Approval Hurdles

Detection Method for Nano biosensor:

1. Detection of E. coli: As an important and well-known Gram-negative bacterium belonging to the Enterobacteriaceae family, E. Coli poses a health risk and causes significant financial losses to the animal industries because it is linked to a number of illnesses and syndromes in both humans and a range of farm animals, including cattle, pigs, sheep, goats, and poultry. This bacteria has been discovered with the aid of different bioreceptors, including antibodies, aptamers, bacteriophages, and DNA probes, using^[17] optical, electrochemical, and mass-sensitive biosensors. A important and well-known Gram-negative bacterium belonging to the Enterobacteriaceae family, E. coli causes a number of diseases and syndromes in both people and a range of farm animals, including cattle, pigs, sheep, goats, chickens, and others.

Le and colleagues developed a quick, easy, and economical colorimetric test using chitosan-coated iron oxide magnetic nanoparticles (CS-MNPs) that allowed for the 10-minute detection of S. aureus and E. coli bacterial cells. The technique they developed was predicated on the peroxidase-like activity of iron oxide magnetic nanoparticles, which is comparatively hindered once bacterial cells connect to the CS-MNPs complexes, leading to a decreased colorimeteric reply. Using spectrophotometry and the human eye to track the response, the colorimetric assay's detection limits were found to be 102 and 104 CFU/ml, respectively. Joung developed a highly sensitive SPR biosensor device that uses peptide nucleic acid probes and gold nanoparticles to detect E. Coli 16s rRNA.

Fig. 4: Utilizing an electrochemical immunoassay based on Cu-Au NPs for identification and counting the quantity of Escherichia coli cells.¹⁷

Zhang and colleagues described an anodic stripping voltammetric (ASV) technique that used core-shell Cu-AuNPs as anti-E. coli antibody labels to quickly identify E. coli cells (in 2 hours). They utilized Cu-AuNPs coupled with anti-E. coli antibodies to describe a series of standard dilutions of E. coli cells linked to a PS-modified-ITO (Polystyrene-modified Indium-doped tin oxide chip). The immobilized Cu-Au NPs labels were indirectly detected by measuring the liberated Cu2+ ions following oxidative treatment in an acidic solution, following the washing away of the unbound Cu-AuNPs anti-E. coli antibody labels. Their findings demonstrated a linear link between the logarithmic value of E. coli concentrations and the elimination current response of the released Cu2+ ions from Cu‐AuNPs. In two hours, this method could identify E Coli. with a limit detection of 3 × 101CFU/ml.

2. Detection of B. anthracis: The severe zoonotic illness known as "Anthrax" in people and animals is caused by the Gram-positive spore-forming bacteria B. anthracis. Serological tests, molecular diagnostics, and blood cultures are among the conventional techniques used to diagnose anthrax. But it might take a day or^[17] more for these tests to provide a conclusive diagnosis. Electrochemical-based sensors are innovative diagnostic tools that can be used to create workable, low-cost methods of disease identification. Damrow and associates using DNA displacement to diagnose B. anthracis using an amperometric biosensor. In this method, a ferrocene‐marked signal probe was displaced by sample DNA following hybridization, resulting in a higher redox current which was measured in ampere.

3. Detection of Campylobacter spp: Another genus of Gram-negative bacteria is Campylobacter, which has a number of species that may essentially infecting gastrointestinal system, since campylobacteriosis, a common foodborne infection caused by Campylobacter, affects millions of individuals annually in various regions of the world. Furthermore, genital and reproductive issues can be brought on by animal strains of these bacteria, especially in ruminants. As of right now, sample enrichment, sample preparation, or specific laboratory conditions are required for all traditional microbiological procedures including ELISA, PCR, and real-time PCR that are used to identify Campylobacter species or their infections. gastrointestinal system, since campylobacteriosis, a common foodborne¹⁷ infection caused by Campylobacter, affects millions of individuals annually in various regions of the world. Animal strains of these bacteria have the additional potential to induce genital and reproductive problems, especially in ruminants. At the moment, every single one of Conventional microbiological techniques like ELISA, PCR, and real-time PCR require sample enrichment or sample preparation and can only be used in certain laboratory circumstances to identify Campylobacter species or their illnesses. However, in addition to optical, electrochemical, and mass-sensitive transducers, other identification components such as aptamers, antibodies and bacteriophages are also employed to identify these bacteria.

Application of Microbial Nanobiosensors:¹⁸

Biochemical, biomedical, pharmacological, agricultural, environmental, electronics, energy harvesting, and food technology are just a few of the disciplines of research that can profit from nanomaterials. It is used in practically every branch of research and technology.¹⁹ Furthermore, the popularity of nanomaterials has skyrocketed, particularly for POC applications, with the advent of biosensors. From then on, biosensors and nanomaterials are frequently referred to as nanobiosensors. These nanobiosensors are incredibly adaptable and have several uses for tracking and identifying analytes. Biochemical, biomedical, pharmacological, agricultural, environmental, electronics, energy harvesting, and food technology are just a few of the disciplines of research that can profit from nanomaterials.¹⁸ It is used in practically every branch of research and technology. Furthermore, the popularity of nanomaterials has skyrocketed, particularly for POC applications, with the advent of biosensors. From then on, biosensors and nanomaterials are frequently referred to as nanobiosensors.²⁰

1. Food Industry: The food sector prioritizes developing new, strong, and sensitive technologies. Nanobiosensor based approaches are capable of detecting, processing, and producing correct signals. The food sector prioritizes developing new, strong, and sensitive technologies.^21,22 Nanobiosensor based approaches are capable of sensing, processing, and producing precise signals.²¹

The primary analytical methods include the use of functionalized plasmonic nanoparticles, gold and silver nanoparticles with DNA to detect changes in optical characteristics, and cantilever detection of changes in mass resonance frequency.²³ Loop mediated isothermal amplification (LAMP), microfluidics, and clever packaging are a few modern methods that have increased detection. The majority of nanobiosensors that have been created thus far have improved the detection limit for the sensitive solution.²⁰ To protect the quality and safety of food products, the food industry has created a range of aptamers and microfluidic approaches-based nanobiosensors to satisfy its various criteria for detecting heavy metal leftovers toxins, pesticides, and infections. Nanobiosensors utilizing silver nanoparticles and the laser treatment detection systems have been created. Due to the safety factor, silver metal nanoparticles have been chosen above other metals. Moreover, high intensity lasers may detect very little change in concentration.^21,24

2. Agriculture:

Because they have greater sensitivity and specificity than conventional approaches, nanobiosensors are one of the new tools needed to maintain agricultural practices sustainability.²⁵ Nanobiosensors can alleviate a few issues with traditional approaches, include the inability to handle a large number of samples at once, interference with different matrices, and lengthy sample intake times for analysis.^26,27 As a result, nanobiosensors offer effective, more precise detection as well as ongoing monitoring at a lower cost and time commitment. Ukhurebor and Adetunji suggest that nanobiosensors can be used to use eco-friendly fertilizers that are less expensive.^28,29 Using nanobiosensors properly can improve agricultural output and mitigate environmental impacts, promoting sustainability. Nanobiosensors can help manage money and natural resources in agriculture. Nanobiosensors, according to Ukhurebor and Adetunji, are also used to use low-cost, environmentally friendly fertilizers.^30,31 Proper usage of nanobiosensors can improve agricultural output and mitigate environmental impacts, promoting stability in the sector. Nanobiosensors could enhance agri-management and control.¹⁹

3. Disease Diagnosis:

Numerous recurring plant diseases that result from deficiencies or infections have a substantial negative influence on agricultural productivity and lower crop yields. It's common knowledge that prevention is always preferable to treatment. Therefore, estimating the disease's appearance in its early stages can aid in removing the main causes of its progression.³² Currently used techniques for pathogen identification and diagnosis include polymerase chain reaction (PCR), enzyme linked immunosorbent assay (ELISA), and microbiological procedures. These methods can take a lot of time, even if they are frequently precise and complex.³³ Farmers can receive real-time data on crop health via nanobiosensors, which can provide an alternate means of detecting infections, nutrient deficiencies, and other issues pertaining to soil and water resources.³⁴ For instance, diseases like Xylella fastidiosa subspecies pauca strain CoDiRO can infect other plant species and cause olive rapid decline syndrome (OQDS).Numerous recurring plant diseases that result from deficiencies or infections have a substantial negative influence on agricultural productivity and lower crop yields. It's common knowledge that prevention is always preferable to treatment.³⁵ Therefore, estimating the disease's appearance in its early stages can aid in removing the main causes of its progression.³⁶ Currently used techniques for pathogen identification and diagnosis include polymerase chain reaction (PCR), enzyme linked immunosorbent assay (ELISA), and microbiological procedures. These methods can take a lot of time, even if they are frequently precise and complex.^37,38

FUTURE PROSPECTIVES:

The world' s population is forecast to expand gradually to reach 9.8 billion people by 2050, from 8.5 billion in 2030, according to the data.¹⁸ This could have an effect on the healthcare industry by creating additional difficulties, such as the need for more diagnostic equipment for every patient, a shortage of testing facilities, and longer turnaround times due to high demand. The world' The goal of nanobiosensor research is to create novel technologies for detection and monitoring applications that can significantly benefit the biomedical, biochemical, environmental, agricultural, and food sectors. Furthermore, the goal of the nanobiosensor is to advance society and humanity. The advancement of nanotechnology has opened up new possibilities for the development of nanobiosensors with submicron-sized dimensions appropriate for intracellular use, depending on the application. Investigating several special effects that are unique to the production of nanostructured materials and are fundamentally their most attractive feature, such as dimension, quantum size, and surface effect, should be a priority. Furthermore, in order to demonstrate even greater qualities for biosensing applications, new nanomaterials must be developed. Biosensors based on nanotechnology should ideally be completely incorporated into microscopic microfluidic devices with on-chip sample.³⁹

CONCLUSION:

In the fields of food safety, healthcare, and environmental monitoring, microbial nanobiosensors offer a bright future. Their capacity to identify certain biomarkers with high sensitivity and specificity has the potential to completely transform how we diagnose and treat diseases by allowing for real-time monitoring and quick reactions to microbiological threats. Furthermore, the use of nanotechnology improves these biosensors' effectiveness and usability, enabling them to be used in a variety of settings and circumstances. For their widespread implementation, it will be crucial to address issues including stability, scalability, and regulatory frameworks as research advances. In the end, microbial nanobiosensors have the potential to greatly influence public health and safety in addition to their creative design, opening the door for a more proactive and responsive approach to managing microbial threats.

ACKNOWLEDGEMENT:

Authors are highly thankful to Minerva College of Pharmacy, Indora, Kangra, HP India for providing the necessary facility

CONFLICT OF INTEREST:

None.

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Received on 08.10.2025 Revised on 21.11.2025

Accepted on 29.12.2025 Published on 30.01.2026

Available online from February 05, 2026

Res. J. Pharma. Dosage Forms and Tech.2026; 18(1):57-64.

DOI: 10.52711/0975-4377.2026.00010

This work is licensed under a Creative Commons Attribution-Non Commercial-Share Alike 4.0 International License. Creative Commons License.