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Mold infections

a technology of mold infections and fungi, applied in the field of mold infections, can solve the problems that milk production and a full spectrum of blood chemistry and biochemical assets have failed to reveal a consistent clinical correlate to jejunal hemorrhage syndrome, and achieve achieving the effect of reducing the risk of jhs

Inactive Publication Date: 2005-10-13
STATE OF OREGON ACTING BY & THROUGH THE STATE BOARD OF HIGHER EDUCATION OREGON STATE UNIV THE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, analyses of diets, ages of cows, levels of milk production and a full spectrum of blood chemistry and biochemical assets have failed to reveal a consistent clinical correlate to JHS.

Method used

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  • Mold infections
  • Mold infections
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Examples

Experimental program
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example 1

Methods

A. Sample Collection

[0100] Samples from nine cows which developed JHS or abomasal hemorrhage (AH) and from several negative control cows were collected. Details of the cases are included in Table 2. Due to the epidemiological nature of the study, consistent samples were not recovered from each farm where JHS was diagnosed. Instead, a mixture of blood, feed and tissues samples were submitted. Tissue samples included gut wall, mesenteric lymph node, cotyledon and liver. Samples from the gut region where hemorrhage occurred were also submitted. The negative control cow samples were blood samples taken from healthy cows (n=14) and three non-JHS dead cows at Pacific Northwest dairies.

B. DNA Extraction and SybrGreen Analysis

[0101] DNA was extracted from blood, tissues and feeds using the Qiagen QIAmp DNA mini kit. Percent recovery of DNA via the Qiagen method was 80%. A 1 μl sample of DNA was combined with SybrGreen PCR reagents, forward and reverse primers to bring the final...

example 2

Jejunal Hemorrhage Syndrome is Caused by Aspergillus fumigatus

[0129] Described herein is the finding that JHS is associated with high levels of A. fumigatus in the gut, gut wall and blood. In human studies, A. fumigatus is recognized for its invasive properties and pathogenicity in immunocompromised patients. These data demonstrate that it has a similar potential in ruminants. However, more than 100,000 fungal species are known, and there are more than 100 Aspergillus species. Only a very small proportion of their ITS domains have been published. Hence, detection of A. fumigatus could include unrecognized (non-sequenced) fungal species. In addition, other predisposing factor(s), for example stress-induced immuno-incompetence, may also contribute to the development of JHS.

A. Development of PCR Assay

[0130] To quantify A. fumigatus in feed, tissues and blood, a “real-time” quantitative PCR assay was developed. The assay was based on the sequences of the fungal ribosomal genes (FIG....

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Abstract

Isolated nucleic acids, kits and methods are disclosed for detecting, quantifying and / or identifying a fungal species in a sample or for diagnosing jejunal hemorrhage syndrome in a subject. The methods generally include amplifying a fungal nucleic acid sequence present in the sample using a primer selected from the group consisting of: SEQ. ID NO 5, SEQ. ID NO 6, SEQ. ID NO 7, SEQ. ID NO 8, SEQ. ID NO 9, SEQ. ID NO 10, SEQ. ID NO 11, SEQ. ID NO 12, SEQ. ID NO 13, SEQ. ID NO 14 or SEQ. ID NO 16; and observing or measuring the amplified nucleic acid sequence. The fungal species can thereby be detected, quantified, and / or identified. Fungal contamination also can be detected or measured in a biological or feed or food sample.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority from U.S. Application Ser. No. 60 / 446,367, filed on Feb. 10, 2003, and incorporated herein by reference for all purposes.BACKGROUND OF THE INVENTION [0002] The present disclosure relates to methods of detecting, quantifying, and identifying a fungal species in a sample, particularly for the detection or diagnosis of jejunal hemorrhage syndrome. [0003] Jejunal hemorrhage syndrome (JHS), also known as hemorrhagic bowel syndrome (HBS) and dead gut syndrome, is a newly-described disorder primarily affecting dairy cattle. Symptoms of JHS include point-source sub-mucosal hematomas, and can include ruptured hematomas with exsanguination into the lumen of the jejunum. The hematomas originate in the jejunal submucosa, and they dissect the mucosa from the underlying connective tissue. Despite the hemorrhage, clotting is unaffected. [0004] JHS is characterized by a sudden drop in milk production, abdominal pain due...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P19/34C12Q1/68G01N
CPCC12Q1/6895C12Q2600/16
Inventor FORSBERG, NEILPUNTENNEY, STEVEN
Owner STATE OF OREGON ACTING BY & THROUGH THE STATE BOARD OF HIGHER EDUCATION OREGON STATE UNIV THE
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