In Vitro Food Diagnostics

Company
Overview
ProGnosis Biotech at a glance

Established in 2011 in Larissa, Greece, 100% privately owned company

Specialized in producing in vitro diagnostics for the food quality sector

Global presence (more than 40 countries) & continuously expanding distribution network

ISO 9001:2015 - Certified company with external recognition (ILVO)

Rep. Offices in Beijing, China (est. 2015) and Madrid, Spain (est. 2019)

Subsidiary in Cyprus - Lateral Logic LTD - (est.2016)

Current position
4 continents | 40 countries
 Overview
Mycotoxins

Toxins produced by Aspergillus species of fungi, i.e A. Flavus. A.Fusarium, A.

Penicilium.
 25% of world agricultural commodities are contaminated by Mycotoxins
 More than 400 mycotoxins exist
 6 major types of mycotoxins according to international organizations and legislators

Aflatoxins

T-2/HT-2 DON

Mycotoxins

OTA
Fumonisin

ZON/ZEA
Overview

Influential factors:
Mold growth and environmental factors in the
consequently mycotoxin field and during storage,
production, is promoted by water activity
stress conditions (high
humidity and extreme
temperatures)

Any temperature above 7 Mycotoxins are more

degrees Celsius and below prevalent in tropical and
40, is enough to promote subtropical climates
fungi growth and
subsequently excretion of
toxins

Areas receiving high

amounts of precipitations
followed by drought, or vice
versa, are in the red zone
 The characteristics

 Mycotoxins are colorless and odorless

 Mycotoxicosis is called any condition or disease caused by fungal toxins
 75% of the samples worldwide are contaminated by at least 2
mycotoxins

Contamination routes of mycotoxins

Pre-Harvest Harvest Storage Transport

 Mycotoxin Occurrence

 Matrices demonstrate different behavior under different conditions, i.e. corn/maize are
almost by default susceptible due to high humidity in the field and in the kernels
themselves
 Mycotoxins can be found in maize; barley; wheat; milk; nuts; spices; fruits; cocoa;
soybeans; cottonseeds; wine and more

High Risk Moderate Risk Low Risk

Maize Figs Soybeans
Peanuts/groundnuts Almonds Barley
Cottonseed Rice
Pecans
Brazil nuts Oats
Pistachio nuts Walnuts Wheat
Copra Sultana Pulses
 Aflatoxins-subgroup of Mycotoxins

By-products (secondary metabolites)

of the fungi of the genus Aspergillus

Aflatoxin B1 is the most natural

carcinogenic which has mutagenic and
immunosuppressive effects

It is produced in susceptible grains and

crops (corn, groundnuts, cottonseed)

AFB1 is converted in animals’ liver to

AFM1 and excreted to milk
The relationship between the amount
of AFB1 and AFM1

 4 out of 10.000 corn seeds can result in

corn having more than 10μg/kg (10 ppb)

 The relationship between the amount of

AFB1 and AFM1 in milk is quite variable

 The transfer of aflatoxins to milk is

between 1 and 3%

 However, transfer factors up to 5-6%

have been reported
 The Impacts of Mycotoxyns

Mycotoxins affect

Animals Humans Rural Economy

Impacts of Mycotoxins on Animals

 Ruminants are more tolerant to mycotoxins than monogastrics

(poultry, swine)
 Very high levels of Aflatoxins can cause acute toxicosis and
death
 Chronic consumption of lower levels can cause significant
organ damage
 Main long-term symptoms are:
I. Gastrointestinal dysfunction resulting in reduced feed intake
II. Oestrogenic anomalies (reproduction)
III. Mastitis
IV. Laminitis
V. Decreased growth and significant reduced milk and meat
production
Impacts of Mycotoxins on Humans

 It is asymptomatic in the short-term

 Aflatoxicosis provokes a variety of symptoms in the long term
(chronic exposure)
 AFB1 is linked to primary liver cancer and is officially considered to
be a human carcinogen by the World Health Organization (WHO)
 The largest risk in humans is associated with hepatocellular
carcinomas, which might be compounded by Hepatitis B
 Chronic exposure of Aflatoxins can also cause:
I. Organ damage
II. Immune suppression
III. Higher rates of illness
IV. Stunting for infants
V. Problems in the reproductive system and gene suppression
(teratogenic effect)
Impacts of Mycotoxins on Rural Economy

 FAO estimates global losses of foodstuffs due to aflatoxins, are in the

range of 1-1.5 billion tones per year

 Aflatoxins lead to direct spoilage and the decrease in nutritional value

of the cereals with subsequent yield losses

 Significant reduced milk production (14-47% loss per day)

 Increased somatic cell counts, reported up to 300%

 Financial losses due to the cost of regulatory programs designed to

reduce risks to animal and human health
SAMPLING PROCEDURES AND PROTOCOLS

A STUDY FOR THE Grain INDUSTRY

Sampling sites

“One sample is never the same with another“

Grain silos in At factory gate, Grain silos in Production line At factory exit,
collection reception per factories dispatch per
centers incoming truck outgoing truck
EU Procedures & Protocols

• EU Approach - EC (European Commission) Regulation 401/2006

 Minimum aggregate sample depending on Lot weight (1-10 kg)
 Minimum incremental sample for cereals (100 gr)
 Different treatment of smaller and bigger kernel matrices, such as figs and nuts (300 gr)
 Different Sampling protocols for different types of matrix groups
 Acknowledgment of heterogeneity

Takes into account Recovery method ranges per Mycotoxin

 Acceptable recovery ranges between 60-120% (varies per toxin)
 In some cases 70-110% (also varies per toxin)
 Recoveries more than 60% are considered as acceptable
 Only in milk, ISO 14675:2003 states that acceptable recoveries are 80-120%
 There is no ISO for recoveries in grain matrices
Sampling Process

 Per 30,000 kg an aggregate sample of at least 10 kg must be taken

 These 10 kg must be collected from at least 12 different spots
 Minimum 100 gr from each spot (incremental sample)

Grind

Mix

Sub-Sampling
Sub-sampling Process

 Conducted always in the lab

 Grind all the sample before sub-sampling
 Mix the bag containing the ground cereals
 Take small spoon samples from many different spots
 Final sample must be at least 5 gr and maximum 100 gr (due to solvent cost)
 Bigger sample is better, but more expensive
Aspects to consider: extraction solvent, size of falcon/tubes, time needed to transfer and liquid quantities

 pH of extracted sample should always be tested before analysis

 Methods used for Mycotoxins Analysis

HPLC-FLD

Instrumental Analysis
(Chromatographic Methods)
LC MS-MS

ELISA

Immunoassays
Lateral Flow Sticks
(Rapid Tests)
Technologies we use

Immunoassays

Enzyme-linked Lateral flow

immunosorbent assay immunochromatographic assay
(ELISA)
PRODUCTS
PORTFOLIO
 Our Products

Bio-Shield Series 1-Standard Series Rapid Test Series Symmetric Series

ELISA kits for: ELISA kits for: Lateral Flow tests for: Lateral Flow tests for:
 Mycotoxin Analysis  Mycotoxin Analysis  Detection of Milk  Mycotoxin Analysis
in Milk in Grains Adulteration in Milk
 Mycotoxin Analysis (Quantitatively &  Mycotoxin
in Grains Qualitatively) Analysis in Grains
 Histamine Analysis  Histamine Analysis
in Fish & Seafood in Fish & Seafood
 Quantification of
Milk Adulteration
Product
5 minutes ELISA Series
Innovation

Innovative
Common Accuracy and Τime-saving and
ELISA series
extraction for all reliability cost-effective
with 5 min
mycotoxins unaltered method
protocol
Product
1-Standard ELISA Method
Innovation

The method uses

Innovative Cost-effective Τime-saving
a pre-calibrated
ELISA series method method: Low
curve to provide
using only the producing highly procedure time
quantification
first standard accurate results of 5 mins
results
 Our
Software
Software for ELISA Kits
ProGnosis Data Reader
 Software platform for the ELISA kits
 Free of charge and easy-to-use statistics analysis program
 Write directly to the program or import xls or Open Office files
 Provides full analytic report (pdf)
 Product
Innovation
Symmetric Green Series

 One extraction for all grain

mycotoxins (B1, Total
Aflatoxin, Deoxynivalenol,
Zearalenone, Ochratoxin Α,
Fumonisin, T-2/HT-2)

 No use of organic solvents

 The same easy 5-minute

protocol for all mycotoxins

 Up to 6 different product
sticks per scan
 Symmetric Quantitative Rapid Test

Key Features

▶ High accuracy and repeatability

▶ Very sensitive, Limit of Quantification (LOQ) at 70 ppt.
▶ Special Software for Full Traceability (S-Flow)
▶ No sample preparation is required
▶ Can be used on raw, UHT and pasteurized milk
▶ Can be used on milk of different species i.e. cow, buffalo, ewe, goat etc.
▶ No influence by milk temperature (4 to 20 ºC)
 Our
Software
Software for Lateral Flow Kits

S-Flow Software

 Flexible software platform

 Quantitative and semi-quantitative interpretation for L.F strips
 Can read one or more L.F strips
 Use of advanced algorithms
 Provides full analytic report (pdf)
Thank you for your
attention!

Company
Overview