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Lab-on-a-chip microfluidic device based on plasmonicdriven thermal sensing for rapid detection of Salmonella typhimurium in agro-food field.

Periodic Reporting for period 1 - HEATSENS_S (Lab-on-a-chip microfluidic device based on plasmonicdriven thermal sensing for rapid detection of Salmonella typhimurium in agro-food field.)

Reporting period: 2015-05-01 to 2015-10-31

HEATSENS_S is a high-sensitivity device that will detect Salmonella in chicken meat in less than one working day (specifically <5 hours). This has made it so that all the potential business customers that we contacted could learn first-hand about the market we are targeting, and they have shown great interest.
Salmonellosis is the second most frequent cause of gastrointestinal infections in the world, causing 93.8 million infections in humans and 155 thousand deaths every year. The cause of the disease is salmonella, a zoonotic bacterium, that is, a bacterium that is transmitted from animals to human beings, and which is found in all kinds of food, but is particularly common in the digestive system of birds (Salmonella Enteriditis infects 1 out of every 20,000 eggs), and thus has particular impact on the poultry industry.
The threshold for these controls is extremely low: meat is considered to be uncontaminated only if there is less than one colony-forming unit (CFU) in 25 mg of meat, and the total absence of bacteria must be guaranteed for the rest of samples, tools, and surfaces in contact with the animals.
If any of these tests gives a positive result for salmonella, the serotype (Enteriditis o Typhimurium) of the salmonella contaminating the sample must be determined. The serotype is the subspecies to which the bacteria belong. It is mandatory to establish the salmonella serotype found in chicken meat, mainly to have greater control over the origin of contamination.
To detect salmonella in chicken meat, the most common option nowadays, used by 80% of the market, is cell culture, the traditional detection system for all microbiological analyses. In addition, some alternative analysis techniques (such as PCR and VIDAS) are now appearing, and so far are being mainly used by large integrated firms. These two current techniques have two points in common:
1. Given that the threshold for salmonella is very low, they require an initial stage for the pre-enrichment of the sample, to allow potential salmonella strains to develop. Once the strains have grown, they can be measured (current methods are not sufficiently sensitive to smaller salmonella concentrations).
2. The analysis detects generic salmonella. If the result is positive, an additional test must be performed in the laboratory to establish the serotype of the contaminating bacteria..
On the basis of the time factor, and taking the entire process into account, the current results would take approximately 43 hours in the case of the quicker options (19 hours if the result is negative), and up to 90 hours (and 66 hours if the result is negative) in the case of slower but more widespread options. Thus there is currently no option that provides same day results.
Whereas, assessing them by the cost of analysis, the cell culture option is more attractive than the current options, whose cost for generic salmonella triples that of cell cultures. If positive, the cost of serotyping is further added.
Speeding up the analytical process means that chicken meat marketers have more days to sell a product, which increases their sales. Moreover, if a positive result is obtained on the farms (animal still alive), if the animals are not treated in time, the producer would have to run a logistical slaughter of chickens that may be contaminated, being able to market them only as frozen meat, incurring substantial losses on their investment. Or what is even worse: if the positive result arrives when the meat is already at the point-of-sale to the end customer (in the supermarket’s display case), this would mean serious harm to public health, high economic cost (product recall), and loss of reputation for the producer. This is why it is extremely important to speed up this process. So much so that when customers are asked directly as to what the ideal solution would be, they say that they will always choose the option that provides results in a shorter time, at a reasonable cost, whos
The work performed by the NIT team involved in the project resulted in successfully achieving the goals that were set, in addition to technically and economically assessing the viability of launching the new device, in which HEATSENS is integrated for the detection of Salmonella in chicken meat.
The contact with these companies has allowed us to define the exact user specifications of the HEATSENS_S device, and to know their views first-hand about our technology applied to the food industry. Once the needs of our end-users were identified, the technical team worked on the final specifications of the device.

The worked was performed on a technical, commercial and financial level and lasted for 6 months (May – October 2015):

 Technical level: After proving that the technology works for the identification of Salmonella in chicken meat, we identified the partners who would help us to integrate this technology into a device and the pre-treatment device best suited to the preparation of the sample to analyse it with our technology.

 Commercial level: The NIT marketing team carried out an in-depth study on the markets in which we planned to launch the device, by determining the size of the total market, specifying what sector will be the Company’s target objective.
In this regard, we have already contacted the relevant companies in the sector on a European level to get their views about the possibilities of the technology, and these companies have shown great interest in the benefits of the technology and provided us with different points of view that we took into consideration when designing the final device.

Financial level: The Finance Department has compiled the information necessary to identify the technical and financial resources that the Company will need to both carry out Phase 2 of the SME Tool, in which the manufacture of the first prototype of our device will be carried out, and during its commercial launch.

Moreover, after analysing the market, and with the estimates of units sold from the Marketing Department, a sales forecast for HEATSENS_S was developed considering the allowable profit margins. In turn, this has led to examining the profitability of HEATSENS_S as a line of business.

Furthermore, the different stages of the manufacturing process were considered to evaluate the suitability of manufacturing it ourselves or subcontracting, concluding that the contribution of the Company will focus on the part of the process, which lies in its know-how, and will collaborate with the more specialised partners in the rest of the phases.
After identifying the needs of the poultry sector and analysing each of the features of the HEATSENS technology applied to the detection of salmonella in chicken meat, the outcome is the following:
 Ultrasensitivity: The sensitivity of HEATSENS_S makes it possible to detect salmonella at a much lower concentration in the sample, so pre-enrichment is reduced to 4.5 h.
 High specificity: Recognition of the analyte is so specific that the same device makes it possible to carry out the serotyping, avoiding the need to send the samples to an external laboratory in the event of a positive result, so the positive result is provided 1 day sooner.
 Quick analysis: The analysis takes 30-40 minutes, so final results are provided on the same day in which the sample is taken. And makes the simultaneous analysis of 3 different samples of chicken meat possible.
 Low cost: The price will be below the actual price of analysis.
 Simple use: No complex procedures or laboratory techniques, requiring no qualified scientific staff, unlike current solutions.
Thus, the main benefit of HEATSENS_S is that it makes it possible to reduce the pre-treatment stage required by all the other current options, providing the first option in the market to achieve a final analysis of salmonella in chicken meat within one day, at a competitive price.
The reduction of the process to a single day is of crucial importance for chicken producers, as stated by all the professionals contacted by NANOIMMUNOTECH, as it allows them to speed up their production processes, so that they are able to deliver consignments of meat on the same day that the analysis is performed—proving that they are not contaminated—which is very important for a product with such a short lifespan. All the potential customers contacted have stated that this competitive advantage would be in itself suffice to make a purchase decision.
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