Lab News

Three posters were presented by the BioNano Lab members of the University of Guelph at the Nano Ontario 2015 Conference held at the University of Ottawa on November 5th and 6th, 2015.

Redox Active Graphene Oxide Nanosheets: Dual-Functional Biosensor for On-Farm Monitoring of Dairy Cow Metabolic Diseases by Murugan Veerapandian and Robert Hunter

Methylene Blue Sensitized Screen Printed Immunoelectrode: Rapid Detection of Avian Influenza Virus by Rob and Murugan

SERS based Point-of-care Biosensing System for Food-Borne Pathogen Detection by Mungroo and Gustavo

Big Smile by Robert at the Ottawa Nano Ontario Annual Event

GryphSens - Equipping Farmers to Test For Severe Cattle Diseases

 IP Licensing OpportunityPDF

Up until recently, dairy farmers have been hindered in quickly detecting possible reasons for a reduction in milk production among their herd. One such reason is sub-clinical ketosis (SCK) when an affected cow appears to be well but only becomes observably under stress when SCK becomes full ketosis affecting major organs, or it could be other metabolic diseases. The only true method of detecting these abnormalities was by taking vials of the affected cow’s blood and sending samples to a lab for diagnosis. However, Bionanolab of the School of Engineering has advanced this procedure by putting the ability to diagnose a cow’s health in the hands of farmers themselves, thereby saving crucial time in detecting certain bovine metabolic irregularities, and thereby providing earlier treatment.

By detecting certain enzymes in blood, the newly developed device can pick out these biomarkers present in miniscule amounts and help identify diseases. A combination of the device’s unique composition of electrodes, a plant enzyme, and the correct amount of electric current were discovered by the bionano research team to be the winning formula used collectively in the device known as a “Gryphsens”.

A New Tool on the Farm

Through the implementation of a hand held sensor, a dairy farmer can rapidly detect whether a cow has sub-clinical ketosis or other metabolic diseases through a small amount of blood being taken and having the sample analyzed in real-time through interfacing with the Internet by a smart phone. Such technology not only allows a dairy farmer to rapidly determine a cause for a reduction in milk production, but it also allows early detection of metabolic diseases that can then be treated, facilitating a cow to return to its normal milk volume levels in a shorter period of time. The cost saving of such early detection is substantial for small and larger dairy herds.

In the case of larger dairy herds, this Canadian invention that uses a unique electrochemical measurement of samples, can be engineered for use with in-line robotic milking machines to monitor a herd individually, yet collectively, to avoid the repetitive and time-consuming method of testing each cow separately. The sensor, developed at Guelph’s Bionano laboratory of the School of Engineering by a team headed by Dr. Suresh Neethirajan is able to detect minute electrochemical activity in biological fluids that indicate biomarkers for certain irregularities and diseases. These markers flag slightly elevated levels of non-esterified fatty acids (NEFA), and a ketone prevalent in cows, β-hydroxybutyrate (BHBA), that at higher levels can both signal the early onset of ketosis and other metabolic diseases.

A Critical Time Line

Historically, the calving period is a stressful time for cows, described as a time of negative energy balance (NEB), when the onset of ketosis or other metabolic diseases are most prominent. Although the levels of NEFA and BHBA are miniscule at the beginning, early detection can reduce complications and a faster recuperation period, otherwise a later detection could lead to fatty liver, ketosis, displaced abomasum (twisted stomach), inflammation of the uterus or a retained placenta. Dairy farmers are highly cognizant that charting a cow’s NEFA and BHBA levels is the litmus test for the animal’s overall health. One of the great advantages of testing for on-farm dairy cow diseases using our developed biosensor is that it not only significantly reduces the stress on the animals due to relying on a drop of blood instead of vials sent, but also provides instant test results.

The Lab Comes to the Barn

Traditionally, these levels are determined through expensive and lengthy tests performed in laboratories. Through electrochemistry and nanotechnology the University of Guelph’s bionano team has made it possible for dairy farmers from all scales of operation to ascertain for themselves their herd’s health. Although humans have similar devices for measuring glucose levels for diabetes, the cow’s organism presents a further challenge by having 11 major blood groups versus four. The challenge was developing the sensor’s electrode that could simultaneously detect both NEFA and BHBA in variable metabolic conditions that can include a number of interfering components, which could alter the test results. Ability to detect multiple disease biomarkers from just a droplet of blood sample that could be used by untrained farmers is unique.

Disposable screen-printed carbon electrodes (SPE) were chosen for their affordability and for their compatibility with hand held and in-line robotic milking devices. From previous discoveries, the compounds used on the electrodes favoured redox active hybrid graphene oxide (GO) materials that have proven very effective in biochemical applications. In particular for this application to identify NEFA and BHBA, electrodes incorporating soybean based enzyme was layered on the GO material. This proved to be the defining chemical for detecting both NEFA and BHBA by having superior and durable redox properties of the critical biomarkers compared to untreated GO electrodes.

A Plant with the Answer

An issue the University of Guelph team had to surmount was the insulating property of GO that hindered the electrochemical function crucial for the biosensor. A particular enzyme from the soybean plant was integral in solving this problem and was incorporated into the dual electrodes. Although lipoxygenase is found in both animal and plant species, using soybean lipoxygenase-1 (SLO) was itself a first for catalyzing direct electrochemical oxidation of NEFA in conjunction with [Ru(bpy)3]2+-GO. In this situation, SLO breaks down the fatty acids found in metabolic lipids to produce fatty acid hydroperoxide. When used in conjunction with electrochemistry, NEFA became oxidized and detectable by the electrode.

On the sensor’s other electrode for detecting the ketone BHBA, it uses another enzyme (HBDH) that is produced naturally by an organism under stress and is detected through electrochemical oxidation. The isolation to enable it to be detected was engineered through the electrochemical oxidation of coenzyme NAD+ found in organisms to produce NADH. As HBDH is dependent on the coenzyme, the electrode can readily identify the enzyme through covalent bonding of atoms at the molecular level.

Using the correct micro voltage supplied by the sensor to the electrodes, it further enhances the provision of efficient and reliable readings. The result is a biosensor that has a dual function of detecting NEFA and BHBA in less than a minute on-site by the dairy farmer using a small sample of whole blood. Dairy cattle is an investment that must be kept in prime condition to maintain optimal production levels, and the Canadian biosensor is the latest tool for dairy farmers and large scale operations to monitor the health of their herds. The dual sensor can be an important part of routine screening used by farmers in the dairy barn. The ease of use and convenience will significantly enhance the management of dairy herd health.

SERS based Point-of-care Biosensing System for Foodborne Pathogen Detection

Monday, November 2, 2015
1:00 PM
Room 1002, Thornbrough Building
by
Nawfal Mungroo MASc Candidate

 

Chair: Dr. R. Dony, P.Eng
Advisor: Dr. S. Neethirajan, P.Eng
Committee Member: Dr. S. Yang

ABSTRACT   

Foodborne illnesses are ubiquitous and life-threatening. The number and severity of foodborne outbreaks could be significantly reduced by the early detection of pathogenic species during food processing and handling. However, most techniques that detect pathogenic species, such as Escherichia coli, Salmonella and Listeria species, are time-consuming, costly, and impractical for use at agricultural sites, warranting material transport to an offsite laboratory; this adds to time and cost, while increasing the likelihood of false positives. Therefore, it is imperative that onsite technology capable of rapidly and cost-effectively detecting and differentiating pathogenic species be developed. Biosensors are one approach that might meet these goals. No current biosensor can faithfully differentiate pathogenic species within the food industry, although Surface Enhanced Raman Spectroscopy (SERS) shows potential for bacterial detection. To overcome current limitations in detection, a microfluidic platform and a protocol has been developed that enhances pathogenic bacterial species detection using silver nanoparticles and chemometric data analysis platforms utilizing a combination of principle component analysis and linear discriminant analysis. Our procedure can successfully distinguish eight key foodborne pathogens (E. coli, S. typhimirium, S. enteritis, Pseudomonas aeruginosa, L. monocytogenes, L. innocua, MRSA 35 and MRSA 86), which holds great promise for food safety detection applications.

2015 Bio-Instrumentation Trade SHOW 

Venue:  School of Engineering - Engineering Atrium - Thornbrough Building

Date:  November 18, 2015 ,  Wednesday 

Time: 11:00 am to 13:00 pm 

 

All are Welcome!  

Connect with industries and learn what prototypes and products the 4th year Biological and Biomedical Engineering students have designed and developed to solve practical problems in biomedical, health care, food and agricultural industry. 

 

Graphene oxide chemically decorated with Ag-Ru/chitosan nanoparticles: Fabrication, Electrode Processing and Immunosensing Property

Murugan Veerapandian and Suresh Neethirajan

RSC Advances Journal DOI: 10.1039/C5RA15329H  (Link)

Abstract

Nanosheets of graphene oxide is chemically decorated with hybrid nanoparticles of silver-ruthenium bipyridine complex (Ag@[Ru(bpy)3]2+) core and chitosan shell. Oxygenated groups of graphene oxide and abundant amine groups of chitosan layer on the surface of hybrid nanoparticles allowed the functionalization reaction. Changes in the optical, chemical and structural properties of graphene oxide due to hybrid nanoparticles were characterized using spectroscopy and microscopic techniques. Electrodes modified with hybrid nanoparticles-graphene oxide (HNPs-GO) displayed an amplified steady-state anodic (IpA) and cathodic (IpC) peak currents, with a correlation coefficients of 0.9987 (IpA1), 0.9952 (IpA2) and 0.9964 (IpC1). Using monoclonal antibody, the HNPs-GO immunosensor could specifically detect Listeria monocytogenes from buffer and milk, with a concentration range from 102 to 107 cells/mL and detection limit of 2 cells/mL. Our results suggest that selective optimization of bio-recognition elements on the HNPs-GO electrode may find prospective use in food safety applications.

 

 

 

 

Contact Us

Bionanotechnology Laboratory
Suresh Neethirajan

School of Engineering
University of Guelph
Guelph, Ontario
Canada N1G 2W1

Office:
Room 3513 - Richards Building
50 Stone Road East

Lab: THRN 2133 BioNano Lab

Phone: (519) 824-4120 Ext 53922
Fax: (519) 836-0227

E-mail: sneethir@uoguelph.ca

 
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