Good Hygiene Practices along the coffee chain

Module 1.1

Fungal Overview
Background: yeasts and moulds (fungi) in food
 Eukaryotic cell structure
 More complex than prokaryotic (bacteria)
 Yeasts
 Unicellular (3 – 5μm)
 Can divide rapidly (but slower than bacteria -
2-3h)
 Moulds
 Tubular cells (30 - 100μm) (hyphae)
 Grow by apical extension (can grow very long -
filamentous fungi)
 Reproduce by sexual and asexual production of
spores
 Adapted to lower moisture conditions than
most bacteria

Slide 2 Module 1.1 – Fungal Overview

 Background: fungi in food
 ‘Useful’ fungi
 Edible mushrooms
 Used in processing / preservation

 Spoilage fungi
 Can grow on foods with lower available water than
most bacteria (some as low as aw = 0.65)
 Typically spoil semi-moist foods – cheeses, cured
meats, bread, cakes, fruit preserves etc
 Cereals, grains, nuts, coffee, cocoa that are
incorrectly stored (damp, moist conditions) – huge
food and feed losses annually

 Toxigenic fungi

Slide 3 Module 1.1 – Fungal Overview

 Toxigenic fungi: Overview of mycotoxins

 Fungal metabolites
 When ingested, inhaled or absorbed through skin cause lowered
performance, sickness or death in man or animals, including birds.
 Acute effects
• Headache, fever, nausea, diarrhœa, vomiting, weakness, tremors,
convulsions
• In some cases death
 Chronic or long-term effects
• Cancer
• Genetic or birth defects
 Over 200 kinds of mycotoxin, produced by about 150 different fungi
 Certain crops are commonly associated with certain mycotoxins
 Ecological associations of mould with crop plants
 Certain post-harvest conditions can favour certain moulds

Slide 4 Module 1.1 – Fungal Overview

 Mycotoxins of major significance

Mould species Mycotoxins

Aspergillus parasiticus Aflatoxins B1, B2, G1, G2

Aspergillus flavus Aflatoxins B1, B2
Fusarium sporotrichiodes T-2 toxin
Fusarium graminearum Deoxynivalenol, Zearalenone
Fusarium moniliforme Fumonisin B1
Penicillium verrucosum Ochratoxin A
Aspergillus ochraceus Ochratoxin A
Penicillium expansum Patulin

Slide 5 Module 1.1 – Fungal Overview

 Aflatoxins

 Commonly associated with maize,

groundnuts, tree nuts, spices, CYA
CBS
dried fruit etc.
 Carry-over from animal feed to CZ

foods of animal origin for humans:

e.g. Aflatoxin M1 in milk
 International guidelines exist for
prevention and control

Slide 6 Module 1.1 – Fungal Overview

 Other important mycotoxins

 Trichothecenes – Fusarium spp

 Associated with a variety of cereals and wet harvest conditions
 Zearalenone – Fusarium spp
 Associated with maize grown in temperate climates
 Fumonisins – Fusarium spp
 Primarily associated with maize
 Patulin - Penicillium spp, Aspergillus spp
 Associated with apple products
 Ochratoxin – Aspergillus spp, Penicillium spp
 Associated with cereals, wine, grape juice, dried fruit, coffee
and cocoa

Slide 7 Module 1.1 – Fungal Overview

 OTA contamination in coffee
 OTA long known as a renal toxin and carcinogen which is also
teratogenic (produces birth defects)
 Evidence of genotoxicity published in the early 1990’s - if true,
categorizes OTA with aflatoxin
 Studies in Europe on dietary exposure concluded the most
significant sources are grain and grain products; beer; wine; dried
fruit; coffee
 Several countries have already adopted maximum levels of
contamination in coffee
 Some importers have rejected contaminated batches
 EU harmonised limits for roasted and soluble coffees - in force
from January 2005

Slide 8 Module 1.1 – Fungal Overview

 OTA producers in coffee
 OTA producers in coffee:
 Aspergillus ochraceus (and related)
 Aspergillus carbonarius A. Colonies of A. flavus from
Aspergillus flavus group. B. & C. C
 Aspergillus niger complex
Typical colonies of Penicillium spp. A B
 Elsewhere:
 Penicillium verrucosum
 Penicillium nordicum

 These organisms interact with other coffee-associated organisms,

and not just Coffee Berry Borer (CBB) and Colletotrichum etc. The
fungi include:
•Cladosporium spp.
 Fusarium stilboides
•Penicillium brevicompactum
 Candida edax •Auriobasidium pululans
 Cryptococcus album •Eurotium repens

 Additional context are the conditions man’s activities impose in the

orchard and during processing and trading

Slide 9 Module 1.1 – Fungal Overview

 Conditions for activity of OTA producers

 Not all isolates of a species that is known to produce a mycotoxin will do

so:
 A. niger complex  5% usually weak
 A. carbonarius  80% often strong
 A. ochraceus and similar  80% often strong
 The range of conditions over which a mycotoxin producer can grow is
broader than those over which it can produce mycotoxin:
 A. niger complex: Aw and temperature limits n.a.
 A. carbonarius: Aw limits  0.92 and 0.85 temperature limits  35˚C and 37˚C
 A. ochraceus: Aw limits  0.82 and 0.78 temperature limits  40˚C and 42˚C
 The interaction of physiological and ecological properties is too complex -
thus laboratory studies are only indicative
 At this stage of our understanding, only field studies can clarify the
limiting conditions for OTA contamination in coffee production

Slide 10 Module 1.1 – Fungal Overview

 Effect of pH and Aw on mould growth

Xerophile Mesophile Hydrophile

pH 3.0 4.0 5.0 7.0 pH 3.0 4.0 5.0 7.0 pH 3.0 4.0 5.0 7.0
Aw Aw Aw
0.99+ 0.99+ 0.99+

0.98 X
H 0.98 0.98

0.94 0.94 0.94

0.905 0.905 0.905

Slide 11 Module 1.1 – Fungal Overview

 Factors controlling mould growth

 Initial contamination?
 Oxygen / gaseous environment?
 Nutrients?
 Temperature?
 Water activity? Aw =
 What is it?
 How do we measure it?

Slide 12 Module 1.1 – Fungal Overview

 Moisture content (m.c.) and Aw

Cherry robusta

 m.c. describes the

1.0
sample; Aw predicts

0.9
microbial growth
potential

0.8
++ +

 In commerce, m.c. is

0.7
measured but the

aw
0.6
microbial stability is only
predicted by Aw so we

0.5
need to inter-convert
0.4
 So we need to understand
0.3

the precision of this inter-

conversion 10 20 30 40 50 60
mc

Slide 13 Module 1.1 – Fungal Overview

 Evaluating moisture in commodities
Moisture content - dry or wet basis?
 Chemical methods
 Oven method
 Temperature?
 Time?
 Air circulation?
 Vacuum?
 Electrical methods
 Capacitance
 Conductance
 Other gravimetric methods
 Empirical / traditional sensory methods

Slide 14 Module 1.1 – Fungal Overview

 Evaluating moisture in commodities

Water activity

 Internal equilibration?
 Equilibration with chamber air?

Slide 15 Module 1.1 – Fungal Overview

 Precision and accuracy of measurement

 Uniformity of
commodity
 Sampling
One type of low-cost
 Calibration moisture meter
investigated under the
 Methodology ‘global coffee project’ SINAR moisture meter

 Frequency
 Quality of standards
 Instrument stability
 Robustness
 Kind of use ‘EDABO’ distillation method of moisture
determination developed in Brazil

Slide 16 Module 1.1 – Fungal Overview

 Moisture and Aw in complex systems

 The husk is more

hygroscopic than the 100 y = -0.0585x2 + 3.7691x + 30.236
bean - it forms a R2 = 0.9774
90
barrier that slows
OTA 80 bean
water loss during prod husk
drying and slows

e.r.h.
70
hsk
water ingress during limit
60 bn
re-wetting. y = -0.0459x2 + 3.2896x + 26.158
50
 From the perspective R2 = 0.979

of mould growth, the 40

0 10 20 30 40 50
significance of a given
m .c. (db)
moisture content of
bean and cherry is
quite different. BEAN LIMIT- -HUSK LIMIT

Slide 17 Module 1.1 – Fungal Overview