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									                                  GH. SADEGHI
                                 University of Kurdistan




Use of Antimicrobial and Detoxifier

 Agents For Poultry Feed Safety

               By:
         GH. SADEGHI
      PhD of Poultry Nutrition
      University of Kurdistan
         Kurdistan, IRAN
                                              GH. SADEGHI
                                             University of Kurdistan




               Basic Fact
• In addition to the essential nutrients, feeds may
  contain organic and inorganic materials that
  have various effects on poultry:
  – beneficial, detrimental or negligible
  – they can affect growth, health or the processed
    product
  – may be naturally occurring, intentionally or
    unintentionally added
  – can be produced via microbial growth
                                        GH. SADEGHI
                                       University of Kurdistan




        Feed Ingredients Contamination
• Contamination of feeds may be take place in
  different stages:
• Pre-harvest (pesticides and other agrichemicals)
• Post-harvest
  • During Feed Processing
  • During Feed Storage
                                              GH. SADEGHI
                                             University of Kurdistan




         Contaminants of Feeds
• Chemicals:
  • Dioxins, pesticides and other agrichemicals
  • Drug Residues
• Heavy metals:
  • lead, mercury, and cadmium.
• Pathogen Microbs:
  •Salmonella
• Microbial Toxins:
  •Mycotoxins: Aflatoxin, Ochratoxin, etc.
                                               GH. SADEGHI
                                              University of Kurdistan




             Pathogens in Feed
• Most important pathogen in poultry feed is Salmonella.
• The major sources of salmonella for poultry are the bird
  themselves, feed and environment.
• Establish HACCP programs across the animal feed
  industry to minimize Salmonella.
• Implement a Salmonella-negative standard for animal
  feed.
                Drug Residues
• Medicated feeds left over in the equipment from
 one batch may contaminate the next batch of feed
 that is not medicated.
                                               GH. SADEGHI
                                              University of Kurdistan




                     Gizzerosin
•This is a toxic compound found in fish meal, a
 typical feed ingredient
• It is formed by the reaction of amino group of
 lysine with the imidazolyl ethyl group of
 histamine during the dry heat processing
•Comes from improper storage of raw fish prior to
 production of fish meal
•Causes a reduction in growth rate
•Usually comes from fish with“dark” or “red” meat and
free histidine in the soluble fraction of protein
                                             GH. SADEGHI
                                            University of Kurdistan




                    Gizzerosin
• Gizzerosin stimulate H2-receptors in proventriculus ,
 causes an excessive HCl secretion and sever
 decrease in pH at the proventriculus and gizzard,
 causes damage to koilin layer and submucosal of
 gizzard
• several dietary treatments for gizzrosine toxicosis
 have been tested including supplementation of:
   • H2-histamin receptor antagonist cimetidin
   • Antacid compounds sodium bicarbonate and
     magnesium trisilicate
                                          GH. SADEGHI
                                         University of Kurdistan




                 Bioterrorism?
Bioterrorism: What is it?
The intentional or threatened use of microorganisms
or biological toxins to kill or incapacitate people,
animals, or crops.
Toxins as Bioweapons:
Toxin: Any toxic substance produced by an
 animal,plant, or microbe. Some toxins are
produced by molecular biology techniques or
Chemical synthesis.
                                            GH. SADEGHI
                                           University of Kurdistan




 Microbial Toxins in Commercial Feeds
•Usually produce by microbes
• Microbes need enough CHO, moisture and O2 to
 produce these toxins
•Commercially-processed feeds are less likely to have
 these toxins
•Not destroyed by steam pelleting or extrusion
•Most important microbial toxins in poultry feed
 industry are mycotoxins
                                        GH. SADEGHI
                                       University of Kurdistan




                Mycotoxins
•The more important toxins affecting animal
   feeding are those associated with molds
•These are called “mycotoxins”
•Three important genera are:
    • Aspergillus
    • Penicillium
    •Fusarium
•They exist and grow anywhere as long as there is
  enough COH substrate, no less than 14%
  moisture, adequate temperature, oxygen
                                         GH. SADEGHI
                                        University of Kurdistan




                 Mycotoxins
•Usually produced in feedstuffs prior to harvest,
      but also result from poor storage
•Most important naturally occurring mycotoxins
are :
            Aflatoxin
            Ochratoxin
             Trichothecenes (Deoxynivalenol)
            Zearalenone
            Fumonisin
                                       GH. SADEGHI
                                      University of Kurdistan




               Mycotoxins
Mycotoxins may be:
Carsinogenic: Aflatoxin B1, Ochratoxin
Nephrotoxic : Ochratoxin, Citrinin
Nerotoxic : Fumonsin B1
Estrogenic : Zearalenone
Dermonecrotic : Thricothecens
                                                    GH. SADEGHI
                                                   University of Kurdistan




                     Mycotoxins
•Tow type of fungi can be recognized in contaminated
grains:
   •Those that preferably infect stored grains (saprophytic)
   • Those which attack the plant in the field (Phtopathogenic)
•The environmental factors that determine fungal
growth and mycotoxin production in feeds are related
to:
   • substrate itself (intrinsic factors)
   • storage condition of substrate ( extrinsic factors)
                                                                      GH. SADEGHI
                                                                     University of Kurdistan



       Factors affecting mold growth and mycotoxin
                 production in creral grains
            Intrinsic factors                          Extrinsic factors

Ingredient composition(lipids, CHO, trace   Relative humidity: RH greater than 70%
metals,etc.                                 favors fungal growth

pH: Fungal growth decrease with             Environmental temperature: the optimal
decreasing pH                               temprature for fungal growth and
                                            mycotoxin production varies with the
                                            spices.
Grain moisture: optimal grain moisture is   O2/CO2: Fungi are aerobic organisms.
>13% for saprophytic and >20% for           Studies with mycotoxigenic Fusaria
phytopathogenic fungi                       showed that in storage atmosphere
                                            containing 5% O2 and 40% CO2, fungi
                                            growth affected and no mycotoxin are
                                            produced
Water activity (aw): this parameter is      Surface area of grain: smaller particles
related to the water that is actually       have increased surface area which
available for fungi growth                  predisposes mold growth
                                                  Adapted from Lesson et al. 2001
                                        GH. SADEGHI
                                       University of Kurdistan




    The affects of mycotoxins on poultry
•Mycotoxins can result in vomiting, diarrhea,
 internal bleeding
•Mycotoxins can impair absorption of A, D, E,
 and K

• Spiking syndrome causes mortality in broilers,
  suggested mycotoxins levels are high, thiamin
  should be increased
                                                    GH. SADEGHI
                                                   University of Kurdistan




     The affects of mycotoxins on poultry
• Fungi can also adversely affect poultry performance
 by altering the nutritional composition of the substrate
 during their primary metabolism, without producing
 secondary metabolites (Mycotoxins)
• Fungal growth can:
   •Decreases the density and energy content of the grain(
   mold use TG and produce CO2)
   • Alter grain vitamin content (thiamin by producing
   thiaminase)
   • Alter grain amino acid content
                GH. SADEGHI
               University of Kurdistan




 AFLATOXIN
AFLATOXIN B1
                                         GH. SADEGHI
                                        University of Kurdistan




                  Aflatoxin
• Aflatoxins are a family of mycotoxins that are
       produced mainly by toxigenic strains of
       Aspergillus flavus and A. parasiticus.
• The name aflatoxin is derived from the bacteria
       that produce it:
       Aspergillus (a-), flavus (-fla-), and toxin.
• In 1960, aflatoxins made a name for
       themselves by killing 100,000 turkeys in
       England that were found to be eating
       moldy ground peanuts.
                                           GH. SADEGHI
                                          University of Kurdistan




                    Aflatoxin
•Aflatoxin is the mycotoxin of greatest concern in
feeding of poultry
•both outright toxic and carcinogenic
•liver (hepatoma) and blood clotting problems
•traditionally, sources include corn, cottonseed and
peanuts
•aflatoxin contamination varies year to year
•Aflatoxins (moldy feed) inhibit microbial intestinal
synthesis
             GH. SADEGHI
            University of Kurdistan




Aflatoxin
                                                   GH. SADEGHI
                                                  University of Kurdistan




       The Affects of Aflatoxins on Poultry
• A dose of 0.25ppm in turkey poults and ducklings
     impairs growth,
• A dose of 1.5ppm in broilers and 4ppm in Japanese quail
    also has a negative affect on growth.
• An increase in blood clotting time increases the
    susceptibility of the carcass to bruising even at doses
    below that to have an affect on growth.
• In poultry, aflatoxins impair the availability of bile salts,
   which decreases Vitamin D3 production.
    This causes a decrease in the absorption of
        fat-soluble vitamins.
                                               GH. SADEGHI
                                              University of Kurdistan




      The Affects of Aflatoxins on Poultry
• Aflatoxins also decrease the production of Vitamin A
   in the liver, and it has secondary effects such as
 decreased blood calcium levels, decreased bone
 strength, and a decreased tissue and serum tocopherol
 level.
   This decrease in tocopherol levels can lead to
     Vitamin A and E deficiencies.
                                       GH. SADEGHI
                                      University of Kurdistan




             Trichothecenes
• T-2 toxin and other trichothecenes (HT-2) can
  cause mouth and intestinal lesions as well as
  impair the birds immune response, causing egg
  production declines, decreased feed
  consumption, weight loss, and altered feather
  patterns.
• While much is yet to be learned, T-2 toxin and
  related compounds are currently thought to be
  most potent from Fusarium mycotoxins for
  poultry
                                          GH. SADEGHI
                                         University of Kurdistan




                 Ochratoxin
•These are compounds produced by Aspergillus
 and Penicillium molds
•Widely found in nature
•Typically associated with kidney toxicity
•Most mold toxins also destroy nutrients in feeds
   •example: Pseudomonas can separate
   glutamic acid from folic acid, making it
   ineffective
                                         GH. SADEGHI
                                        University of Kurdistan




               FUMONISINS
• Fumonisins are the most recently discovered
group.
• They are produced by the fungus Fusarium
moniliforme
• Fumonisin B1 is the most predominant molecule
• Fumonisin inhibit of sphingolipid biosynthesis or
sphingosin turnover because of the similarity of
polyhydric alcohol miotey of FB1 to the complex
amino alcohol sphingosine
                              GH. SADEGHI
                             University of Kurdistan




STEREOCHEMISTRY OF B SERIES FUMONISINS
 GH. SADEGHI
University of Kurdistan
 GH. SADEGHI
University of Kurdistan
                                                                   GH. SADEGHI
                                                                  University of Kurdistan




Clinical guide to major mycotoxins in poultry
  Mycotoxin      Type of bird       Dietary      Main effect
                                     level
  Aflatoxin      Broiler chicken     3 ppm       No detrimental effect after 2
                                                 weeks of exposure
                  Laying hens       2.5 ppm      Minimal dietary concentration can
                                                 affecting performance when given
                                                 for 3 weeks or more
Ochratoxin A     Broiler chicken   1.5 -2 ppm    Minimal dieary level capable of
                                                 reducing weight gain
                                   2 -4 ppm      Increased relative weight of
                                                 kidney, liver; decreaseed relative
                                                 weight of thymus and bursa
Trichotecenes:   Broiler chicken   1 -4 ppm      Oral lesions, decrease FI and
                                                 WG, abnormal feathering
 T2 toxin and
     DAS                            1-2 ppm      Oral lesions, decrease FI and egg
                                                 production

                                                Adapted from Lesson et al. 2001
                                             GH. SADEGHI
                                            University of Kurdistan




      Prevention of mycotoxicosis
•Transgenic Strategies to Reduce Mycotoxins in
 Crops:
          Reducing infection by the fungus
          Degrading the mycotoxin in planta
          Disrupting mycotoxin biosynthesis
          Inducing insect resistance
           • Bt CORN HYBRIDS
            Transgenic corn with the Bt gene for insect
            resistance is a promising solution to the
            fumonisin problem
                                               GH. SADEGHI
                                              University of Kurdistan




Prevention of Microbial Growth in Feed
• Microorganisms require unbounded water
  to growth in feeds.
• Feeds containing more than 12-14 % moisture
  can support bacterial, mold, and Yeast growth
  unless they are frozen.
• In semi- moist feeds at room temperature
   (22oC),mold growth is visible within 3 days.
• There are 3 approach to control microbial growth:
     • Use of antimicrobial agents
     • Use of special packaging
     • Use of water absorbents material
                                                          GH. SADEGHI
                                                         University of Kurdistan




                Antimicrobial Agents
• Over 20 compounds are used in the feed industry
  to inhibit fungal or microbial growth.
• Some have general and some specific application
   in feed product:
   • The Benzoates and Parabens are wide spectrum
     antimicrobials
   • propionate are used primarily to inhibit yeasts
   and molds, but are also effective against bacteria,
   fungi and yeast.
• An intelligent choice of an antimicrobial include:
   • study of specific microbial problem
   • solubility and ease of application of the agent
   • pH requirement
   • effect of antimicrobial agent on palatability
                                              GH. SADEGHI
                                             University of Kurdistan




        Antimicrobial Agents used in feeds

Name                      Limit or Restriction
Benzoic acid              0.1 %
Formic acid               2.5 %
Propionic acid            None
Sorbic acid               None
Calcium propionat         None
Calcium sorbate           None
Potassium sorbate         None
Potassium bisulfate       Not for use in B1 sources
Potassium metabisulfate   Not for use in B1 sources
Methyl paraben            0.1 %
Propyl paraben            0.1 %
                                            GH. SADEGHI
                                           University of Kurdistan



    Antimicrobial Agents used in feeds Cont.

Name                    Limit or Restriction

Sodium benzoate         0.1 %

Sodium bisulfate        Not for use in B1 sources

Sodium metabisulfate    Not for use in B1 sources

Sodium nitrite          0.002 %

Sodium propionate       None

Sodium sorbate          None

Sodium sulfite          Not for use in B1 sources
                                                   GH. SADEGHI
                                                  University of Kurdistan




                Especial Packaging
• Control of spoilage in semi-moisture product (18-36% moisture)
  is not simply a matter of adding antimicrobial agent.
• If feed microbial load is higher than 500,000 CFU, it is
  difficult to control spoilage with antimicrobial agents.
• A second means of controlling spoilage is by the use of
  special packaging .
•The aim of this process is to reduce O2 tension in the feed
  package during storage to restrict the growth of aerobic
  microorganisms.
• Thus, an inert gas, such as N2 or CO2 is introduce into the
  bag before it sealed.
• this method is more applicable in fish feed industry
                                                          GH. SADEGHI
                                                         University of Kurdistan




           Water Absorbents Material
• Microorganisms require an abundant supply of water to
  growth, therefore, reducing the supply of water in a feed
  is an effective way to limit their growth.
• A third approach to controlling spoilage in feeds is by
  controlling the water activity (Aw).
   • Aw: the ratio of the water vapor in the feed to the vapor
          pressure of pure water at the same temperature
   • Aw of pure water is 1.0 and n feeds is less than 1.0
   • The Aw for microorganisms growth are:
                • 0.91 for bacteria
                • 0.88 for yeasts
                • 0.80 for molds
                                                       GH. SADEGHI
                                                      University of Kurdistan




           Water Absorbents Material
• water exist in feeds in 3 forms (liquid, vapor and chemically bound
• Control of Aw in feeds involve increasing the proportion
 of bounded water, thereby decreasing the proportion
 present as a liquid (free).
• It is possible to achieve Aw values in suitable range by
 the addition of compounds such as:
                NaCl
                Propylene glycol
                Other dry feedstuffs (grains)
                                                GH. SADEGHI
                                               University of Kurdistan




   Detoxification of mycotoxin-contaminated
                      feeds
• Different strategis developed to control of mycotoxicosis
  include :
           • Physical Methods
           • Chemical methods
• Physical methods include:
               • Extraction with organic solvents
               • Heat inactivation
               • Irridation
               • Adsorption by adsorbents
• Chemical methods include:
               • Ammoniation
               • treatment with oxidizing agents
               • treatment with organic acids
                                                GH. SADEGHI
                                               University of Kurdistan




             Organic solvent extraction
• Several solvents or mixture of solvents can effectively
 extract mycotoxins( especially aflatoxins) from feeds
 include:
            • 95% Ethanol
            • 90% aqueous acetone
            • 80% isopropanol
            • hexane – ethanol
            • hexane – methanol
            • hexane – methanol – water
            • methylene chloride (CH2Cl2 )
                                                 GH. SADEGHI
                                                University of Kurdistan




                   Heat inactivation
•Aflatoxins are very resistant to thermal inactivation, but
 some research works showed that roasting significantly
 reduced the concentration of aflatoxin in peanuts that
 AFB1 have been reduced by 50 – 70 %

                        Irradiation
•A marked reduction in the concentration of aflatoxins
 in contaminated products has been showed.
• Peanut oil exposed to short wave and long wave
  UV light showed a significant reduction in aflatoxin level
• A 14 hour exposure to sunlight destroyed 50% of AFB1
  in peanut flakes
                                               GH. SADEGHI
                                              University of Kurdistan




                     Adsorbents
• Adsorbents are compounds that are not absorbed from
 the GIT and have ability to bind physically with
 chemical compounds, precluding their absorption.
• Adsorption therapy has been one of the most important
 method of preventing the absorption of ingested toxins.
• However, adsorbents will also bind concurrently
 administered drugs used for therapeutic purposes.
• The use of adsorbents have been extensively studied in
 livestock exposed to dietary aflatoxins.
                                               GH. SADEGHI
                                              University of Kurdistan




                       Adsorbents

• Different adsorbents are used include:
      • Activated charcoal
             • 200ppm and or 0.2 - 0.5 g/kg
      • Hydrated sodium calcium aluminosilicate (HSCAS)
             • 0.5 %
      • Sodium bentonite
             • 0.5 %
                                             GH. SADEGHI
                                            University of Kurdistan




         Detoxification by ammoniation
• Ammonia or ammonia-related compounds have the
 most practical application
• Ammoniation of contaminated corn, cottonseed,
 cottonseed meal and peanut meal has been shown to
 reduce the aflatoxin level by greater than 99%
• Primarily 2 procedure are used:
   • HP/HT: used in feed mill
             use 0.2-2% ammonia, 20-60min
             35-50 psi, 80-120 oC
   • AP/AT: used in farm
             • Use 1-5% ammonia, 2-3 week
                                                 GH. SADEGHI
                                                University of Kurdistan




         Detoxification by oxidizing agent
• Other chemical method is Hydrogen peroxide (H2O2)
• It is an oxidizing agent that destroy up to 97% of
 aflatoxins

          Detoxification by organic acids
• Treatment of maize with following organic acids has
reduced the infection with Aspergillus flavus fungi and
decreased aflatoxin production after 6 months.
              • Isobutyric acid
              • Propionic acid
              • Acetic acid
                                                GH. SADEGHI
                                               University of Kurdistan




                  CONCLUSIONS
The ultimate solutions to the global feed toxins, especially
mycotoxin problem are:
 Reduction of fungal infection and mycotoxin levels in
  crop plants in the field and in storage.
 Novel strategies need to be developed such as the
  reduction of fumonisin levels in genetically modified
 Bt corn.
 Detoxification of contaminated feedstuffs by suitable
  methods
And finally by inform poultry producers about the health
 hazards of mycotoxins
              GH. SADEGHI
             University of Kurdistan




     ???




questions?
‫سنندج‬   ‫?‪Questions‬‬




                ‫از توجه شما سپاسگزارم‬
                                                GH. SADEGHI
                                               University of Kurdistan




• growing poultry should not receive more than 20 ppb
  aflatoxin in the diet. However, feeding levels lower than
  20 ppb may still reduce their resistance to disease,
  decrease their ability to withstand stress and bruising,
  and generally make them unthrifty
 • Laying hens generally can tolerate higher levels than
   young birds, but levels should still be less than 50 ppb
• Aflatoxin contamination can reduce the birds’ ability to
   withstand stress by inhibiting the immune system.
• This malfunction can reduce egg size and possible lower
   egg production.
• In addition, one must pay special attention to the use of
  contaminated corn in layer rations because eggs are
                                                GH. SADEGHI
                                               University of Kurdistan




              Prevention of mycotoxosis
• The best way to prevent aflatoxicosis is to
  prevent the infection of the feed by Aspergillus
 if moisture content is kept below 14% and temperature is
  kept below 25 degrees Celsius, then mold will have a
  difficult time surviving.
• Unfortunately, there are problems with chemical
  treatment of infected crops.
  What ever treatment is used adds to the cost of
   production and none of the processes are 100%
   effective.
  Some treatments may decrease the food value of the
  crop, and some chemical treatments may produce
  secondary biologically active agents that may also be
                                                 GH. SADEGHI
                                                University of Kurdistan




•` Mixing of corn with dry grain has shown to help decrease
   moisture content of the stored material below the level
   that mold can begin to develop
• Also, high CO2 levels along with decreased humidity have
  been shown to decrease aflatoxin levels

• The most effective and practical method of detoxification
  of contaminated grain is ammoniation.
    This can kill up to 99% of mold growth
    Proprionic acid has very little effect
• Also, the pretreatment of food with Phenobarbitals or
  diethyl stilbestrol, used as dietary supplements, will help
  reduce the effects of aflatoxins by binding to them and
  preventing their breakdown and absorption.
                                               GH. SADEGHI
                                              University of Kurdistan




• Clay minerals and hydrated sodium calcium alumiosilicate
  at levels as low as 0.5% of dietary intake, can also help
  reduce the effects of aflatoxins in the body.

								
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