Mycotoxins - Risk management for reducing the problem

MICOTOXINS

RISK MANAGEMENT FOR REDUCING THE PROBLEM Dr. Carlos Augusto Mallmann, Dr. Paulo Dilkin, MSc Adriano Olnei Mallmann.

Universidade Federal de Santa Maria (UFSM), Department of Preventive Veterinary Medicine, Mycotoxicological Analysis Laboratory (LAMIC), Santa Maria, Rio Grande do Sul, Brasil. www.lamic.ufsm.br

In most cases, the presence of mycotoxins in raw materials is less than a 0,001%

Harvest

Reception of the grains

N

utrition, for most species, has currently reached high levels of improvement that have led to excellent productive results in livestock production.

Adequate nutrition and handling, as well as controlled health status and high genetic quality, have all led to the emergence of other limiting factors that impact production and that had not been noticed before.

Mycotoxins are one of these new challenges, and due to financial and health status reasons, they have become one of the factors that receive increasingly more attention.

The presence of mycotoxins in raw materials is not homogeneous, appearing in less than the 0,001% of grains. Such a minute quantity is quantified as a concentration of parts per billion (ppb) or million (ppm). This in corn, for example, represents the equivalent to the weight of 1 grain in a total mass of approximately 350 tons.

Unloading platform Grain storage

Conveyor belt

Feed sampling Consumed feed sampling

Sampling due to clinical suspicion

POSIBLE SAMPLING STAGES

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These circumstances alone represent a non-resolved problem in the precise diagnostics of mycotoxins.

Thus, decision making regarding the possible procedures to be applied when receiving the cereals and along the feed processing chain generates a certain level of uncertainty (Whitaker et al. 2011).

Due to the fact that the decision on the destination and the control measures regarding the raw material or feed depends on the results of the analysis, sampling represents the most critical step of the process, paying more attention to the samples collected for humidity evaluation, for example.

Hereafter, we present some of the steps between harvesting and the process of production in which sampling can be done.

Sample collection for mycotoxin control is the most critical step of the process and must be addressed with the utmost attention

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SAMPLING STAGES

STATIC SAMPLING ON THE TRUCK Sampling points

x

At harvest time Sampling at this moment offers the benefits of obtaining information in advance and allows for making decisions about the destination of the cereals with enough time.

However, the need for ultra-fast analytical detection methods impedes its use, with the exception of high-quality products that are usually not raw materials used in animal feed.

x

x A

x x

x x x x

x

x x

Loads of 20 to 32 Tm = 10 kg of global sample collected

x x

x x

B Loads of 13 to 20 Tm = 6 kg of global sample collected

Figure 1. Sampling points in trucks

At the Grain Reception Unit

Reception

Using sample collectors such as a manual shovel is usually the most used way of taking samples in most small units. t is an essential to collect samples from at least 6 or 10 points, depending on the size of the truck, and the recollection must always be even throughout all the load, following the indications described in Figure 1. (Mallmann et al. 2013) When using pneumatic sample collectors, the same rules must be followed.

Manual Sampling

Collected Samples STEPS IN THE SAMPLE COLLECTION PROCEDURE FOR MYCOTOXIN ANALYSIS

Milling Process Sample Reduction

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Samples for Analysis

SAMPLING STAGES On Unloading Platforms TYPES OF SAMPLE COLLECTORS Sample collecting when unloading the raw material has pros and cons.

Extraction speed Better uniformity as the recollection is done while the material is in movement

Manual Sample Collector

Difficulty separating different batches of cereals because they are unloaded in the same tank, which can have any kind of contamination The great amount unloaded makes it a costly process

Sampling in Cereal Storage Units Pneumatic Sample Collector It is possible to sample the silos and storage facilities with pneumatic probes that allow the recollection of all the samples profiles of the silo.

Currently, there are studies in this area because sampling at this point offers information in advance, allowing for better allocation of the raw materials adjustments in their nutritional matrix (Mallmann et al. 2014).

Figure 2. Images of the process of sample collection with manual and pneumatic sample collectors.

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SAMPLING STAGES During internal transportation Sampling whole and milled grains The movement of raw materials inside the processing systems (“Redler� type chain conveyors and belt conveyors) allow to homogenize the materials and, thanks to this, obtain samples that enable a more precise evaluation of the mycotoxins.

Collecting samples of whole grains is usually easier, but the representiveness of the sample collected is higher if it is milled beforehand, as it fragments the grain in several particles that are dispersed, making the results more reliable.

If extraction before milling is not possible, it is recommendable to do a global sample collection before reducing the volume in order to send a smaller sample for its nutritional analysis in the laboratory.

The most practical system is with a 45-degree hole in the screw conveyor passing through the spiral. This system allows small portions of the raw material to be collected while the material passes through the spiral.

Collecting samples of whole grains is usually easier, but the representiveness of the sample collected is higher if it is milled beforehand

Sample volume per batch in kg 6

It is advisable to use the system during the transport stage, when the mixture is homogenous, in other words, before the segregation stages.

When collecting samples in this dynamic process, we must respect the sample volume per batch in kg =√(20 x tonnes per batch). This way, with a later sample (in case of whole grains) we will obtain more representative samples (Mallmann et al. 2014) Figure 3.

=

20 x Tonnes/BATCH

SAMPLING STAGES DYNAMIC SAMPLING SYSTEM AT THE HOLE IN THE SCREW CONVEYOR The arrows (A and B) indicate the flow of the corn bran inside the spiral which is collected through tube C and stored in container D (global sample)

B

A

E (45ยบ) C

C D

D

Figure 3. Dynamic sampling system applied to raw materials in the movement for the analysis of mycotoxins .

Mycotoxin Risk Assesment -MRA- is a parameter to keep in mind when making decisions about using or not using an anti-mycotoxin additive (AMA)

Sampling finished feed This sampling allows the evaluation of the actual contamination of the food destined to consumption.

At this level, the sampling is closer to reality when evaluating the risk of mycotoxin contamination.

The samples collected will be more representative because the process of premixture allows for better dispersion of the contaminated particles.

The analytical methods based on liquid chromatography (HPLC) are the only ones that are precise enough for evaluating in advance and making decisions about monitoring the feed, but in reality, the result, in most cases, is only obtained when the animals have already consumed the ration.

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SAMPLING PHASES

Sampling at the point of feed consumption It is rare to collect samples at this point. The feed produced at a mill is distributed to different consumption points (livestock farms), which makes it impossible to control it at this stage.

It is only recommended to do it at this point in case of suspicion of poor conservation conditions or long storage periods, factors that increase the risk of mycotoxins being present.

Sampling based on clinical suspicion This type of sampling is only done in case of clinical signs that are compatible with the presence of mycotoxicosis and it is carried out as a compliment to a diagnostic analysis and in legal situations. In these legal cases the following aspects must be observed: The interested stakeholders and witnesses will be present Strict monitoring of the sampling After recollection, the samples must be stored and sent out in ideal conditions for their conservation and subsequent analysis through HPLC. Sampling and analysis in this stage are usually unnecessary, as it is directly related to the frequent monitorization of the raw materials at the production plant.

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ETAPAS DE ANĂ LISIS & CONTROL FREQUENCY OF THE ANALYSIS

The MRA -Mycotoxin Risk Assessment- must be based on the data history that is capable of showing the trends and critical points. Thus, the frequency should be of at least a daily sample. Weekly samples -at least five- will be used to establish the average contamination and percentage of positivity that will be used to calculate the MRA.

The volume of feed production

CRITERIA TO KEEP IN MIND WHEN PERFORMING PERIODIC ANALYSIS

In order for correct monitorization of the mycotoxins, the definition of the frequency of the analysis is equally as important as the sampling process previously described.

The heterogeneity of the material used as a sample The sensitivity of the species The age category The frequency of production of the feed

It is equally important to do a correct sampling as to know the necessary frequency of the analysis

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Mycotoxin analysis DIAGNOSTIC METHODS

The methods used for mycotoxin control are basically: ELISA kits (Enzyme Linked Immuno Sorbent Assay) HPLC (High Performance Liquid Chromatography) Although Thin-layer chromatography was widely used in the past, it is currently in disuse during routine monitoring.

ELISA kits are frequently used for routine monitoring due to its main advantages: The possibility of in situ analysis Low operating cost User-friendliness Thus, they are used as indicators of the presence of mycotoxins because their quantitative results are not exact and are more orientative.

This diagnostic system is to be used in raw materials (corn, wheat...) and is not recommended in the finished feed, which limits its reliability when making important decisions.

STAGES OF ANALYSIS & CONTROL ELISA kits offer the possibility of in situ analysis, at a low operational cost and easy usage

If later on a more thorough analysis is needed to determine the exact qualitative and quantitative contamination parameters, then chromatographic methods, such as modern liquid chromatography coupled to mass spectrometry (LC-MS/MS), offer reliable results that are more adequate for making decisions based on the results Image 1.

Obviously, for this, the methodology must preferentially be in compliance with the international regulations ISO/IEC 17.025.

The ELISA kits should only be used as indicators of the presence of mycotoxins

Image 1. Liquid Chromatography coupled with Sequential Mass Spectrometry (LC-MS/MS) equipment used for the quantification of mycotoxins.

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STAGES OF ANALYSIS & CONTROL

Sample Collection

+

Analysis

= MRA

The Mycotoxin Risk curve is the result of an algorithm for each mycotoxin that allows the evaluation of:

INFORMATION USAGE

The intensity of the contamination (average of weekly samples in ppb) The positivity (percentage of positivity of the week) The results obtained after collecting and analyzing the samples allow for the obtention of a curved known as “Mycotoxin Risk Assessment” (MRA).

The sampling factor (it is measured in situ and changes depending on the sampling plan used)

A follow-up period of no less than 6 months makes it possible to observe inflections in the MRA that change depending on the positivity and the contamination.

The Mycotoxin Risk Assessment -MRAcalculated is used as a parameter for making decisions about using or not using an anti-mycotoxin additive -AMA-

The information obtained will be used to:

MYCOTOXIN RISK 1000

Assign the correct destination to this material until obtaining the final product

HIGH Risk

750

Withdraw products

500

MEDIUM Risk

250

LOW Risk 1

7

13

19

25

29 2013

35

41

47

51 2

8

14 20 2014

26

30

Graphic 1. Weekly Mycotoxin Risk at a feed mill Use Anti-Mycotoxin Additives (AMA)

Discard the mycotoxin factor when diagnosing performance problems, etc.

Graphic 1 shows an example of a certain MR at a feed mill. The MR is used as a parameter when making decisions about using or not using an Anti-mycotoxin Additive (AMA) with tested effectiveness -in vivo evaluation following the recommendations from the Brasilian Agricultural and Livestock Ministry’s Task Force- against each mycotoxin in the different animal production species.

The decision about the acceptable MR limit for using or not an ingredient or an AMA varies depending on the situation of each feed mill, the target species, the production stage, nutrition, genetics, health, handling, and other associated factors.

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Mycotoxin Risk The mycotoxin risk determines the level of mycotoxicological risk that the entire feed mill production process is subjected to.

CONCLUSIONS The presence of mycotoxins in the animals’ diets can lead to important losses in the production chain.

Its control depends on the application of the correct policies regarding agricultural practices, and the production and storage systems, that are the root of the problem.

The investigation in these areas has improved, leading to better productive and financial results in livestock production, as well as improving feed security in the food destined to human consumption.

The considerable presence of mycotoxins in the main components of the diet makes it important to adopt continuous monitoring programs for raw materials and feed destined to feeding animals.

Monitoring allows for mycotoxin risk management, establishing the technical criteria for decision making.

Strategic use of Anti-Mycotoxin Additives in contaminated diets is important because once the toxin is formed, the process of purification is costly and impractical.

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