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Pathogenic fungi can affect
livestock via infection of the gastrointestinal tract and
hematogenous distribution to tissues (1). In humans, nosocomial
disseminated mycoses are becoming more prevalent (2). Immunoincompetence
is the main predisposing factor for nosocomial infection in
humans. Mortality rates in immunocompromised patients are
high (>50%; 2). The most common mycotic infections
in humans are mediated by Aspergillus fumigatus and Candida
albicans (2).
Livestock are also susceptible to mycotic infections (3). A recent survey found
mycotic GI lesions in 5% of slaughter cattle with the most common infecting pathogens
being A. fumigatus and C. albicans (4). Predisposing factors
to mycotic infections included feeding of moldy feed, immunocompromising diseases,
acidosis, anti-microbial therapy, reflux of abomasal contents, metabolic disturbances,
post-partum stress, viral erosive disearses such as IBR, anti-inflammatory treatment
and abortion (4). Primary portals for mycotic invasion included the omasum and
the Peyer's patches. Infection is typically associated with GI hemorrhage. Recent
studies have linked jejunal hemorrhage syndrome (JHS, hemorrhagic bowel syndrome,
acute hemorrhagic enteritis) and abortion to A. fumigatus infection
(3). However, it appears that A. fumigatus may not be the sole cause.
Its feeding may require other predisposing conditions, such as stress or immunosuppression,
for manifestation of pathogenicity (5).
Research at Oregon State University has determined that JHS (HBS) dairy cows
and cows with mycotic abortions have significant amounts of A. fumigatus DNA
in their GI tracts, mesenteric lymph nodes, liver, blood and cotyledon (3). Some
asymptomatic cows harbor A. fumigatus DNA in their blood, however, these
levels of DNA are typically far lower (1/20th to 1/ 50,000th ) levels detected
in sick cows.
Interpretation of your results
Oregon State University's methods for quantitation of A. fumigatus required
almost two years to develop. DNA from your blood / tissue sample(s) was extracted
using a standard DNA extraction method (Qiagen) and then purified. The DNA sample
was then assayed for the presence of A. fumigatus genomic DNA using A.
fumigatus -specific primers and a BioRad MyiQ thermocycler.
Primers were designed to hybridize to unique A. fumigatus sequences
in the fungal internally-transcribed spacer (ITS-1) domain. A Sybr-Green real-time
polymerase chain reaction (PCR) method was then used to amplify the DNA in your
sample(s) and determine the concentration of A. fumigatus DNA. A standard
curve using purified genomic A. fumigatus DNA was run with all assays.
Results are provided using both a "+" system and by providing actual concentration
of A. fumigatus DNA in your sample(s).
Score |
Result |
GU
X 10000/ml |
|
|
| --- |
negative |
0 |
no detectable A.fumigatus |
| + |
low |
>0-1 |
trace |
| ++ |
borderline |
1-3 |
positive |
| +++ |
positive |
3-10 |
positive |
| ++++ |
strong positive |
10-25 |
strong positive |
| +++++ |
very strong
positive |
>25 |
very strong positive |
Value > 3X10000 GU/ml
are rarely observed in healthy (asymptomatic) cows or in non-HBS
downer cows.
Caution: Use results only as an adjunct to other diagnostic
indicators. Detection of A. fumigatus alone may not be taken as a
definitive diagnosis. Values are expressed in "genomic units" (GU). One hundred GUs are
the equivalent genomic DNA present in one A. fumigatus organism (
i.e., one spore). In blood, however, not all GUs are viable. A significant
proportion of GUs may arise from digestion of A. fumigatus genomic
DNA and liberation of intact ITS-1 DNA fragments.
References: 1.
Jensen H, Shoenheyder H, Basse A. Acute disseminated aspergillosis
in a cow with special reference to penetration and spread.
J. Comp Path 1991; 104: 411-417.
2.
Chen SCA, Halliday CL, Meyer W. A review of nucleic acid-based
diagnostic tests for systemic mycoses with an emphasis
on polymerase chain reaction-based assays. Med Mycol.
2002; 40: 333-357.
3. Puntenney S, Wang Y, Forsberg NE. Mycotic infections
in livestock: recent insights and studies on etiology,
diagnostics and prevention of hemorrhagic bowel syndrome.
Proc. SW. Nutr Manag Conf. p.49-62, 2003.
4. Sarfati J, Jensen H, Latge J. Route of infections in
bovine aspergillosis. J Med Vet Mycol 1996; 34: 379-383.
5. Kirkpatrick MA, Timms LL, Kersting KW, et al .
Case Report - jejunal hemorrhage syndrome of dairy cattle.
Bov Prac 2001; 35: 104-115.
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