Sub-Theme I

Fungal pathogenesis

Introduction
Fungi and fungal spores are widely distributed in air, dust and fomites. Fungi may exist as
commensals forming part of normal flora at certain body sites. Of the more than 250000
described fungal species, only a few are linked to disease in animals and humans. Fungi are
most commonly found as plant pathogens. Humans and animals are relatively resistant due
in part to their relatively high body temperature. Reptiles, fish and insects are less resistant
since their body temperature is the same as environmental temperature.
Fungal invasion of tissues
A mycosis is a fungal infection. A mycotoxicosis is a disease caused by the intake of
fungal toxins.
Mycoses are caused by either true fungal pathogens that cause primary infections or
opportunistic fungal pathogens that cause secondary infections. Opportunistic fungal
pathogens are often part of the normal flora.
Mycoses can be divided in three types: Cutaneous or superficial, subcutaneous and
systemic.
Fungi that only grow at room temperature cause superficial mycoses (cutaneous &
subcutaneous) since they cannot survive at body temperature. Dermatophytosis is a
superficial mycosis of the skin involving the keratinised layers of the epidermis.
A subcutaneous mycosis involves the deeper layers of the skin and other subcutaneous
structures, e.g., the dermis and lymphatics.
Systemic mycoses in warm-blooded animals involves the internal organs and are caused
by fungi that are able to grow at the host’s body temperature. In other words, at 37°C and
higher.
Mycotoxicosis is a systemic disease caused by secondary metabolites of filamentous fungi.
Examples are aflatoxin produced by Aspergillus species and fumonisin B1 produced by
Fusarium species. Mycotoxins target and damage organs such as the liver, kidneys or brain.

Pathogenesis, virulence factors and mechanisms of tissue damage by fungi

Epidemiology of mycoses
Most fungi, even pathogens, do not require a host to complete their life cycle. So, infections
are not transmitted from one animal to the next. Exceptions are dermatophytes and Candida
species that naturally inhabit a host. These are transmissible. Mycoses are often
undiagnosed or misdiagnosed as resistant bacterial infections.
Primary fungal pathogen
A true primary fungal pathogen can invade and grow in a healthy, non-compromised host.
They usually come from and environmental reservoir and infect animals that have been
exposed to a large dose of spores or that are immunologically naïve to the fungus. For
example, the dermatophytes. The dermatophytes most commonly encountered are the
Trichophyton species and Microsporum species.
A potential host is more susceptible to fungal invasion when natural physical barriers are
broken down, for example skin abrasions open the way for invasion of dermatophytes and
development of dermatophytosis. Dermatophytes also produce keratolytic substances that
break down keratinocytes. Suppression or deficiency in the cell mediated immune response
(CMI) leads to increased susceptibility to fungal invasion. Increased progesterone levels
increase the susceptibility of female animals to candidiasis (the disease caused by Candida
species). Candida species are yeasts (single celled fungi) that often form part of the normal
flora on mucosal surfaces.
Opportunistic (Secondary) fungal infections
Opportunistic fungal pathogens take advantage of hosts that are debilitated or
immunocompromised due to a primary condition to cause infection. These fungi may be from
an environmental reservoir for example Cryptococcus or Aspergillus. They may also be
commensals (normal flora) in healthy hosts for example Candida species or Malassezia
species. However, when it comes to fungi the distinction between primary pathogens and
opportunists is a grey area. Sometimes opportunists appear to cause disease in healthy
hosts.
Predisposing factors
A very important predisposing factor for opportunistic fungal infections is antibiotic therapy.
The overuse of broad-spectrum antimicrobials kills off the bacterial normal flora that keep
yeasts under control and so pave the way for yeast overgrowth. A good example is otitis
externa in dogs due to Malassezia pachydermatis. Long term corticosteroid treatment is
another factor as it suppresses the immune system. Diabetes also suppresses the immune
system. Catheters can also carry fungi into the blood stream.

Pathogenesis
Portals of entry are inhalation, skin damage (abrasions, cuts, scratches, penetration wounds)
or ingestion.
Cutaneous (Superficial) mycoses
Primary mycoses tend to be cutaneous and superficial. Fungal spores on blankets, halters,
brushes or saddles contaminate the skin surface or hair follicles. The spores germinate and
hyphae penetrate the keratinised layers of the skin. An example is dermatophytosis
(ringworm) which is an infection of the superficial layers of the skin by fungi of the genera
Microsporum, Trichophyton or Epidermophyton.
Subcutaneous
Subcutaneous infection results when fungal spores are inoculated into the deeper layers of
tissue below the skin. Lymphocutaneous sporotrichosis (Sporothrix schenckii) develops
when the spores are inoculated into the skin by a thorn or wood splinter. Opportunistic fungi
can also sometimes cause such infections.
Systemic
The respiratory tract can also be a portal of entry as some spores are so small that they can
be inhaled and settle in the lungs or airsacs. The spores attach to and are taken up by
epithelial cells and phagocytes. A good example is aspergillosis in birds. The intestine can
also serve as a portal of entry as spores can be ingested with mouldy feed such as hay,
peanuts and parrot seeds. Once again aspergillosis is a good example. The spores
germinate inside the phagosome and escape into the cytosol of the host cell. Such infections
can develop from localised to systemic when fungi erode blood vessels to gain access to the
blood stream.

Virulence factors
Fungi produce virulence factors that enhance their ability to cause disease and the severity
of disease they can cause. A fungus can have a number of virulence factors that work in
combination. These virulence factors evolved to aid the survival of fungi in the environment,
to resist ingestion by amoebae or to survive inside amoebae, but many are also useful when
the fungus has to survive in an animal.
Virulence factors are proteins, polysaccharides, phospholipases and structures (capsules)
that help the fungus to colonise and survive the immune system attacks of the host. Some of
these factors are: The ability to adhere to host cells by means of cell wall glycoproteins,
capsules that resist phagocytosis or aid in survival inside the host cell.
Fungi are primarily saprophytes that digest their substrate by secreting enzymes such as
keratinase, elastase and collagenase. They then absorb the nutrients that are released from
damaged cells. The ability to secrete these enzymes is a virulence factor that enables
dermatophytes to colonise and survive on skin.
Another virulence factor is the ability to grow at higher temperatures including the fever
range. Opportunistic fungi can acquire this virulence factor. Non-pathogenic fungi and fungi
that cause superficial infections do not have this factor and so cannot survive in a warm-
blooded animal. The ability to exhibit thermal dimorphism improves the chances of survival
of a fungus in a warm-blooded host. A fungus is thermally dimorphic when it can grow as a
mould at 20 to 30°C, and as a yeast at 37°C. The yeast form can survive and reproduce at
temperatures up to 40°C, low oxygen tension and low nutrient availability. It more resistant
to killing by phagocytes as well.

Tissue damage
Fungi that cause mycoses can damage tissues in several ways. They can produce
enzymes (proteins) such as keratinases, elastases and collagenases that digest
keratinocytes, collagen or blood vessel endothelium. The primary purpose is often to free the
nutrients inside the cells for absorption by the fungus. Fungal growth is invasive producing
masses or balls of hyphae that lead to physical obstruction of bronchi and arteries as well as
subsequent necrosis of tissues. These fungal masses can also displace or destroy vital
structures. The infection elicits an inflammatory response that also causes tissue damage.
The response is pyogranulomatous. Hypersensitivity can develop to particular fungal spores.
Some filamentous fungi produce mycotoxins at certain developmental stages in response to
environmental stressors as they grow on a substrate such as peanuts or maize. Mycotoxins
serve various purposes in the environment such as suppressing competing microorganisms.
However they can cause mycotoxicosis in animals. The portal of entry for these toxins is
ingestion. When ingested in large amounts, they can have cytotoxic effects in animals such
as cellular disorders, oxidative stress, inhibition of translation of mRNA, DNA damage and
apoptosis. Many are carcinogenic. The most well-known of these toxins is aflatoxin,
produced by Aspergillus flavus, that causes liver damage. Other producers of mycotoxins
are Fusarium and Penicillium.
The host’s most important defence mechanisms are cell-mediated immunity which includes
phagocytosis and inflammation. Immunity tends to be short term, for example only a year for
the dermatophytes. Immunity can lead to delayed hypersensitivity reaction following
inhalation of spores of Aspergillus fumigatus.
In summary, there is a complex interplay between fungal virulence factors and host defence
factors which determines whether disease will develop or not. The two important determining
factors are inoculum size and general immunity of the host.

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