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October 2015 - The Maya Metabolomics EPI Research

 

  

The Maya Metabolomic Study

 in Honor of Maya, an esteemed member of Epi4Dogs

 Epi4Dogs is pleased to announce that we are collaborating with Dr. David A. Williams and Dr. Patrick Barko of the College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Champaign, Illinois. Together we are embarking upon a new research study, the Maya Metabolomic Studyhttp://vetmed.illinois.edu/cmmi/  in their Clinical Metabolomics and Microbiomics Initiative to further investigate possible environmental factors that may be involved in EPI. Metabolomics is an emerging field in which a very large number of small chemicals can be analyzed from samples of body fluids and tissues. This new technology has the potential to identify previously undetectable abnormalities associated with development of various diseases, including EPI, and it is hoped that this new information will provide clues as to why this disease develops and what we can do to prevent it.

We know that there are some genetic indications with EPI in dogs from the research done by Dr. Leigh Anne Clark of Clemson University Genetics and that it is strongly suspected that EPI is a combination of both genetics and environmental causes. The Maya Metabolomic Study will hopefully help to identify the environmental “trigger(s)” as to why EPI occurs and what causes the acinar cells in the pancreas to start destroying themselves.

To learn more about the complex field of metabolomics technology, interested readers can go to Metabolon’s website section that succinctly explains the study of metabolism and the metabolomics research process: http://www.metabolon.com/technology/about-metabolomics.aspx  and http://www.metabolon.com/technology/the-science-of-metabolism.aspx

Participation in this study will require one or two short visits to your veterinarian, or at a designated and authorized location, where samples of blood, feces, urine, and DNA will be collected by trained veterinary professionals. These samples will be shipped to or collected by Dr. Barko for analysis.

To find out if your dog qualifies for participation, you will need to fill out the EPI REGISTRY located here:  http://www.epi4dogs.com/epiregistry.htm (even if you filled it out a few years ago, please feel free to fill it out again, as i have recently included additional features to benefit the study).  Dr. Barko will sort the database in various ways as there are multi-levels of candidates needed for the study, and he will contact those via email who fit the criteria. This will be an on-going process. 

Never before has such an all inclusive/comprehensive EPI study been undertaken with every single body fluid collected. This will allow for further testing to be pursued with samples already on hand based on these study results. This was never possible before and a huge thank you to Dr. Patrick Barko for working diligently getting everything properly set-up for this extraordinarily comprehensive study!

I would like to especially and gratefully thank Maya’s family along with Drs. Williams and Barko. Because of each of them, this study is now possible.  And I would like to further extend a thank you to the University if Illinois, Toby's family, and the entire membership of Epi4Dogs for contributing much needed data for this study submitted on the Epi4Dogs “EPI Registry.” Again, Thank You!!!   

   The Research Investigators

 

Dr. Patrick Barko                                     Dr. David A. Williams
Veterinary Resident                                   MA VetMB PhD, Diplomate ACVIM, ECVIM-CA
Veterinary Clinical Medicine                      Professor in Small Animal Internal Medicine
College of Veterinary Medicine                  College of Veterinary Medicine
University of Illinois Urbana-Champaign           University of Illinois Urbana-Champaign
Champaign, Illinois                                Champaign, Illinois 

Dr. Williams Biography

Dr. David A Williams is a Professor of Veterinary Clinical Medicine at College of Veterinary Medicine, University of Illinois. He founded the "GI Lab" in 1985 at the University of Florida, when he introduced assay of serum trypsin-like immunoreactivity (TLI) in the dog to the United States. His research has been focused on the development and application of new tests for gastrointestinal diseases, particularly those affecting the pancreas, small intestine, stomach and liver of dogs and cats. Students and staff working in his subsequent GI Laboratories at Kansas State, Purdue, and most recently Texas A&M Universities established several other novel tests for gastrointestinal diseases for use by veterinarians internationally. These included feline serum TLI, canine and feline serum PLI, canine and feline fecal alpha 1 -proteinase inhibitor, unconjugated serum bile acids, canine TSH, and canine and feline serum cobalamin and folate assays. He received his veterinary degree from the University of Cambridge and his PhD from the University of Liverpool (where he first developed the canine TLI assay). He was an intern and resident at the University of Pennsylvania, and has held Faculty positions at the University of Florida, Kansas State University, Purdue University, Texas A&M University, and the University of Illinois, where he is presently Head of the Department of Veterinary Clinical Medicine. He continues to work as a consultant with the GI Lab, providing telephone consultations regarding management of patients diagnosed using the GI Lab services.

Research Interest

Dr. David A Williams's research interests are Development and application of new tests for gastrointestinal diseases, particularly those affecting the pancreas, small intestine, stomach and liver of dogs and cats.

 

THANK YOU !!!!        

Olesia C. Kennedy, President

Epi4Dogs Foundation, Inc.  

 

 

  Please consider supporting EPI Awareness the Maya Metabolomic Study on EPI by making a donation to Epi4Dogs Foundation Inc a 501c3 Non-Profit Public Charity  http://www.epi4dogs.com/donate.htm   or by participating in an Epi4Dogs fund raiser  http://www.epi4dogs.com/epiproducts.htm
 
 
 Canine EPI Research
CANINE EPI RESEARCH 
 

JUNE 2015ANIMAL GENETICS PUBLICATION
 

Association of DLA-DQB1 alleles with exocrine pancreatic insufficiency in Pembroke  Welsh Corgis

  1. J. M. Evans1
  2. K. L. Tsai1
  3. A. N. Starr-Moss1
  4. J. M. Steiner2 and
  5. L. A. Clark1,*

Article first published online: 19 JUN 2015

 
 
 
 
 
DOI: 10.1111/age.12317 
 
Exocrine pancreatic insufficiency (EPI) is a digestive disorder resulting from the insufficient secretion of enzymes from the pancreas. In dogs, this condition is often attributed to pancreatic acinar atrophy, wherein the enzyme-producing acinar cells are believed to be destroyed through an autoimmune process. Although EPI affects many diverse breeds, to date, molecular studies have been limited to the German Shepherd dog. A recent study of major histocompatibility genes in diseased and healthy German Shepherd dogs identified both risk and protective haplotypes. Herein, we genotyped DLA-DQB1 in Pembroke Welsh Corgis to determine whether dog leukocyte antigen alleles contribute to the pathogenesis of EPI across dog breeds. We evaluated 14 affected and 43 control Pembroke Welsh Corgis, which were selected based on an age of onset similar to German Shepherd dogs. We identified one protective allele (odds ratio = 0.13, P-value = 0.044) and one risk allele (odds ratio = 3.8, P-value = 0.047). As in German Shepherd dogs, the risk allele is a duplication of DLA-DQB1 (alleles DQB1*013:03 and 017:01); however, Pembroke Welsh Corgis have acquired a single polymorphism on DQB1*017:01. Thus, the DLA-DQB1 duplication is a risk allele for EPI in at least two breeds. 
 

February 2013   
Dr. Leigh Anne Clark and her staff at Clemson University in Clemson, South Carolina - "The Clemson Canine Genetics Research Laboratory http://www.clemsoncaninegenetics.com/ is expanding their EPI Research trying to find genetic markers to EPI. Through their EPI study in GSDs (German Shepherd Dog), they have found some interesting indications but are now looking to find the same indications in other breeds. 

Currently they are looking for cheek swabs from either purebred Corgi's or Labrador's that are afflicted with EPI. We would greatly appreciate it if you would be willing to participate in this research ... OR.... if you can pass this information on to someone that you might know that has either an EPI Corgi or Lab that would be willing to participate.... that would be GREAT!

Interested owners can directly contact Jacquelyn Evans in Dr. Leigh Anne Clark's lab at:[email protected]  and she will mail you cheek swabs, instructions, an informed consent form, and a pre-addressed and stamped envelope to return the samples and signed consent form to the Clark lab.

    

Dr. Clark's Contact Information  
Leigh Anne Clark, PhD.
Assistant Professor
Department of Genetics and Biochemistry
100 Jordan Hall
Clemson University
Clemson, South Carolina 29634

 

THANK YOU !!! 
  -----------------------------------------------------------------------------------------------------------
 
October 2012
Recent EPI Genetic Research (by Dr. Clark at Clemson Univ) can be read in it's entirety from 2ndchance info:
 
Topical Review
Current Status of Genetic Studies of Exocrine Pancreatic Insufficiency in Dogs
"Current Status of Genetic Studies of Exocrine Pancreatic Insufficiency in Dogs" 
by Leigh Anne Clark, PhD, and Melissa L. Cox, PhD
 
 ----------------------------------------------------------------------------------------------

October 2011Drs. Clark and Tsai have found an allele indication in some of the EPI population... this is the FIRST positive indication anyone has identified with EPI. Much work still needs to be done... but this is a huge breakthrough!  They have currently received a grant to further this study: "Pancreatic insufficiency in German Shepherd Dogs is caused by an autoimmune reaction that selectively destroys the pancreatic cells responsible for secreting digestive enzymes. Although once thought to be a simple recessive trait, recent work has shown that both genetic and environmental factors likely contribute to the disease. An initial whole-genome screen to identify genetic markers linked to pancreatic insufficiency yielded significant results on chromosome 12. This chromosomal region harbors the dog leukocyte antigen (DLA) system, the canine equivalent of the major histocompatibility complex (MHC), which is important in autoimmune and infectious diseases. Expression studies have shown that one MHC gene, DLA-88, is over-expressed in dogs with pancreatic insufficiency.

The objective of this work was to determine if alleles of the MHC gene, DLA-88, are genetic risk factors for exocrine pancreatic insufficiency (EPI) in the German Shepherd Dog (GSD). DLA-88 sequences were generated for 148 unrelated GSDs. Complete genotypes were determined for 132 dogs (69 cases and 63 controls). Statistical calculations were performed to assess differences in allele frequencies between cases and controls. Significant associations were found with three alleles. One allele confers an increased risk for EPI. Over 20% of EPI cases had at least one copy of this allele, whereas only one copy was observed in the control population. Two other alleles were found to confer protection against EPI. These data support a role for the immune system in the development of EPI in GDSs."


This information may not be copied and posted elsewhere without explicit permission from Dr. Clark
 
 
 
March 2010: Training for Dr. Clark and her staff on the SNP platform is scheduled for this month. As soon as all training is completed, the 200 SNP arrays collected last year for EPI will be tested. YEAH !!!

January 2010 Dr. Leigh Anne Clark has had the EPI Genetic SNP arrays ready to go to NIH for over a month, however, it has been difficult to secure a scheduled time to process this with NIH. Because of this delay, Dr. Keith E. Murphy, Professor and Chair of Genetics and Biochemistry at Clemson University has procured a SNP array platform for Clemson University (THANK YOU Dr. Keith Murphy !!!!) The platform has just arrived this week at Clemson allowing for the SNP array process to be performed in-house at Clemson. This is great news for our EPI Genetic Research effort and for all future genetic testing at Clemson University. Keep posted for genetic research updates soon.

July 24, 2009:  Dr. Kate Tsai, also previously from Texas A&M, has accepted a position as Research Assistant Professor at Clemson University and will be working on EPI Research with Dr. Leigh Anne Clark.

July 16, 2009: Clemson University is on schedule to order the SNP arrays this month. It should take approximately 4-6 weeks.

June 3, 2009:  Dr. Clark has concluded the collection of EPI and non-EPI GSDs samples. And she has already isolated the DNA from all the blood samples submitted. The next step is that the researchers need to prepare the DNA for use on the SNP array. In July 2009 they plan on starting the next phase which is to order the arrays and start running them. 

February 2009:  Previously EPI was suspected to be caused by autosomal recessive genes, but that is no longer thought to be the case. The following is a summary by lead researcher, Keith E. Murphy, PhD on the results of the November 2008 study:
          “To date, blood and serum samples have been collected from 31 unrelated German Shepherd Dogs (GSDs) having exocrine pancreatic insufficiency (EPI) and 45 unrelated GSDs that do not have EPI. In 2008, preliminary data were collected for 18 EPI-affected and 25 unaffected GSDs using the canine SNP array. This array is a newly-available technology that uses a comprehensive set of genetic markers, called single nucleotide polymorphisms (SNPs), to identify linkage with a trait. We were unable to identify a single region of interest but found that markers on multiple chromosomes were loosely linked with EPI. These data suggest that pancreatic insufficiency in the GSD is not inherited in an autosomal recessive fashion; but rather that it is likely a complex disorder, potentially having multiple genetic and environmental factors. To reduce the background (e.g., reduce false positives), it is necessary to collect SNP array data for additional EPI-affected and normal GSDs. To this end, we are presently collecting samples from additional dogs that will be used to probe the SNP array as part of a larger study.”
 
RESEARCHER COMMENTS  (Dr. Leigh Anne Clark's Corner)

June 2009: "EPI does have a genetic/heritable component, but it is likely more complex than autosomal recessive. In fact, a test breeding in Europe between
2 affected dogs resulted in a litter with NO affected puppies. Most likely there are environmental factors (stress, virus, etc.) or multiple genes contributing.
Unfortunately,
this means that there is no accurate way to identify carriers. Definitely do not breed affected dogs, and do not repeat matings that produced
EPI. It may also
be safest to not line breed dogs that are "carriers." 

   
THANK YOU! 

The following are the researchers involved in the EPI Research Study: 

Keith Murphy, Ph.D.
Professor and Chair, Dept of Genetics and Biochemistry, Clemson University
Grant for PAA from the CHF:
Murphy, K.E. and L.A. Clark (Co-Is). Analysis of a candidate gene for pancreatic acinar atrophy in the German Shepherd Dog. Canine Health Foundation. $22,680. 2004-2005.

Leigh Ann Clark, PhD in EPI
Ass't Professor, Dept of Genetics and Biochemistry, Clemson University
Dr. Clark studied under Dr. Murphy for her PhD and continues to work with him. She received the Texas A&M University College of Veterinary Medicine Fisher Institute Medical Research Award, 2004, for her dissertation, titled: Transmission genetics of pancreatic acinar atrophy in the German Shep Dog.

Kate Tsai, Ph.D.,                                                                                                        
Research Ass't Professor, Dept of Genetics and Biochemistry, Clemson University (previously from Dept of Pathobiology Texas A&M University, College of Veterinary Medicine and Biomedical Sciences)                          

Jörg M. Steiner, med.vet., Dr.med.vet., PhD, DACVIM, DECVIM-CA
Associate Professor with the Department of Small Animal Medicine and Surgery
Texas A&M University, College of Veterinary Medicine and Biomedical Sciences

in special collaboration with:                                       
David A. Williams MA VetMB PhD  
Diplomate ACVIM, ECVIM-CA
Developer of the TLI test  

 

 Pphoto is of Enzyme Diane's beloved Sarge - -Because of Sarge, many, many EPi dogs were given a 2nd chance at being properly managed because of Enzyme Diane's affordable generic enzymes, the cornerstone of treating EPI but brand name enzymes are extremely expensive that many could not afford....  thankfully Enzyme Diane's discounted and generic enzymes are affordable .... and we are forever indebted and grateful to one very special EPI pup...."Sarge".

EPI Genetic Research

 We are very grateful to have two excellent U.S. universities supporting the cause of EPI. When Dr. Keith Murphywent to Clemson University of SC, as Professor and Chair of Genetics and Biochemistry, Clemson stepped forward as a major contributor to the EPI Genetic Research along with Dr. Leigh Anne Clark who was still at Texas A&M University, as Research Assistant Professor in the Dept of Pathobiology conducting the EPI Genetic Research study.  May 2009, Dr. Leigh Anne Clark left Texas A&M University (TAMU) and accepted an Assistant Professor position at Clemson University in Clemson, South Carolina in the Genetics and Biochemistry Department. She now heads up her own Laboratory. Dr. Kate Tsai, also previously from Texas A&M, accepted a position in July 2009 at Clemson University as Research Assistant Professor and also works on EPI Research along with Dr. Leigh Anne Clark.  Genetic EPI Research has  been and continues to be conducted at Clemson University. Dr. Jorg Steiner, et al  at Texas A&M University continues to assist with Cobalamin & Folate testing and TLI testing for EPI.  Texas A&M University also continues to offer EPI consultations to vets through the TAMU GI lab.

Leigh Anne Clark, PhD.
Assistant Professor
Department of Genetics and Biochemistry
100 Jordan Hall
Clemson University
Clemson, South Carolina 29634 
Please feel free to visit Clemson Canine Genetics (CCG) website 
to see the various Genetic Research Projects taking place at Clemson Canine Genetics at Clemson University
in Clemson, South Carolina USA. We at epi4dogs are very pleased that CCG has selected
epi4dogs as their on-line source of information on EPI.

Dr. Leigh Anne Clark with daughter on the left with Dr. Kate Tsai on the right  

 RESEARCH UPDATES

Phase I of the EPI Research was concluded in November 2008, Phase II was completed the end of February 2009 with the collection of confirmed EPI German Shepherd Dogs GSDs) and Phase III, collecting from  normal-senior GSDs was completed the beginning of May 2009.  There has been a phenomenal outpouring of participation with the EPI Research Study... Thank you to everyone! 
  
DONATIONS ARE GREATLY NEEDED TO CONTINUE THE EPI GENETIC RESEARCH PROJECT
PLEASE CONSIDER MAKING A CONTRIBUTION
 AT:

Donations-Clemson Canine Genetics 

Enzyme EPI Clinical Trial 

THIS STUDY IS NOW CLOSED

....started June 17, 2012 / ended January 2013 due to depleted supplies of one of the controlled study enzymes...

 

June 17, 2012 and January 2013

I am very pleased to announce that epi4dogs will be assisting Texas A&M Gastrointestinal Laboratory  (TAMU) in finding candidates for a clinical trial for a possible new EPI canine enzyme

  •  All canine candidates must be over 1 year of age that are being or have been tested for EPI via the TLI blood test. It is preferable that the dog has not yet been started on any enzymes...but the health of the dog always comes first...so if the dog has been started on the powdered enzymes...they will still be considered. The dog will also need normal B12 levels and the absence of any other concurrent health condition.
  •  If you would like to participate, please have your vet (vet's only please!) contactDr. Joseph Cyrus Parambeth in Dr. Jorg Steiner's dept, theGastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843-4474 for details regarding the trial and/or to submit your case for consideration
    Phone979-862-2985 begin_of_the_skype_highlighting            979-862-2985      end_of_the_skype_highlighting    or    Email: [email protected]
  •  The study is for 45-50 days during which time you will be required to schedule 3 vet visits at which time your vet will distribute the enzymes to you, collect blood, and carry out a physical examination. You will have to collect fecal samples and complete the owner questionnaire during the course of this study.
  •  In appreciation, at the end of the trial, you will receive 6 additional weeks of pancreatic enzymes of your choice of the two used in the study for your pet.

More Details  

The purpose of this study is to evaluate the efficacy of a new pancreatic enzyme supplement in dogs with exocrine pancreatic insufficiency (EPI) by comparing the treatment response to that for another product that has already been successfully used in dogs. The study is for a period of 45-50 days. Briefly, the dog would be placed on a 21-day course of one pancreatic enzyme supplement and then on another 21-day course of another supplement. During the study period, the participant will need to be seen by his/her veterinarian for a total of 3 visits (21 days apart, i.e. when starting or changing the enzyme supplements). During each visit, the veterinarian will perform a physical examination and collect blood for assessment of blood and serum parameters. The owner, would need to collect three sets of three fecal samples at each time point of the study, and fill out a daily questionnaire. These fecal samples, collected over different periods of time would be used to evaluate the efficacy of the enzymes in helping your dogs digest their food. A six week free enzyme supplement of your choice (either the first or the second one used in the study) would be provided on successful completion of the study. Your dog must be over 1 year of age, have clinical signs and a diagnostic confirmation of EPI using the Trypsin Like Immunoreactivity (TLI) test) to be enrolled in the study. Other inclusion criteria include a normal serum cobalamin level, and absence of any other concurrent diseases. During the entire course of study, no other concurrent medication or changes in the diet are permissible. Owner and patient confidentiality would be maintained at all times.

Upon acceptance of participation, there will be the following forms to review and/or sign:

  • Instructions
  • Instruction Chart
  • Consent Form
  • Owner Questionnaire
  • Owner Statement

 

 If you are interested in possibly helping further benefiting the treatment of EPI dogs (and hopefully cats too in the near future) please consider participating.....THANK YOU!!

Brief Communication - J Vet Intern Med 2010;24:450–452
Heritability of Exocrine Pancreatic Insufficiency in
German Shepherd Dogs

E. Westermarck, S.A.M. Saari, and M.E. Wiberg

Brief Communication - J Vet Intern Med 2010;24:450–452
Heritability of Exocrine Pancreatic Insufficiency in
German Shepherd Dogs

E. Westermarck, S.A.M. Saari, and M.E. Wiberg


Background: Several studies have revealed that exocrine pancreatic insufficiency (EPI) is an inherited disease in German
Shepherd Dogs (GSDs). Pedigree analyses have suggested an autosomal recessive inheritance model.
Objective: Test mating of 2 dogs with EPI.
Animals: A sire and dam purebred GSD both with EPI and a litter of 6 puppies.
Methods: Test mating and long-term follow-up of offspring. The pancreas was biopsied via laparotomy on 26 occasions.
Serum trypsin-like immunoreactivity was measured. Study was approved by Animal Ethics Committee.
Results: During the 12-year study period only 2 of the 6 offsprings developed pancreatic acinar atrophy (PAA). In 1 puppy,
end-stage PAA and in the other puppy partial PAA was diagnosed.
Conclusions and Clinical Importance: PAA is not a congenital disease in GSDs. This study provided evidence that PAA is
not inherited in a simple autosomal recessive fashion.
Key words: Biopsy; Pancreatic acinar atrophy; Polygenic inheritance; Test mating; Trypsin-like immunoreactivity.

Abbreviations:
EM electron microscopy
EPI exocrine pancreatic insufficiency
GSD German Shepherd Dog
PAA pancreatic acinar atrophy
TLI trypsin-like immunoreactivity

     Canine exocrine pancreatic insufficiency (EPI) is a disease characterized by inadequate production of digestive enzymes by pancreatic acinar cells. Affected dogs typically show such clinical signs as polyphagia, weight loss, yellowish poorly digested loose and pulpy feces, increased fecal volume, and frequent defecation.1. EPI has been reported in many breeds, but it is most commonly seen in German Shepherd Dogs (GSDs). In GSDs, the underlying cause of EPI is pancreatic acinar
atrophy (PAA). A characteristic of PAA is selective destruction of the digestive enzyme producing acinar cells.1, PAA has features of autoimmune disease, including genetic suspectibility to disease. 1.2.
    
Several studies have indicated that EPI is an inherited disease in GSDs, and most research of the inheritance model by pedigree analysis suggests that EPI is inherited in an autosomal recessive fashion.3–5 The prevalence of affected dogs in most reports has, however, been lower (!15%) than expected 25% for a simple autosomal recessive trait; thus, a polygenic mode of inheritance has been proposed.4 The mutation or candidate gene has not been identified.6.

Materials and Methods
     This study was performed during 1987 to 2000. Both parents were purebred GSDs (dam 4 years, sire 6 years), with different progenitors. EPI diagnosis for both parents was documented on the basis of results of the serum trypsin-like immunoreactivity (TLI) assay 7. (dam 0.9 mg/L; sire 0.8 mg/L; control 5.2–35.0 mg/L). The dogs were fed regular commercial dog food twice daily, and 50 g of raw pig pancreas was added to every meal.

     At the time of mating, both dogs were clinically in good condition. The dam delivered 9 puppies, but 3 died immediately or a short time after birth. Necropsy was performed on these puppies, and histologic examination of the pancreas did not reveal abnormalities.

     Of the 6 remaining puppies, 4 were females and 2 males. The puppies were weaned at 6 weeks of age and released to their owners.

     The puppies were raised in private homes. The owners were informed orally about EPI and were also provided with a written information package. They were told that the dogs would likely be afflicted with EPI. The investigators and the dog owners maintained close ties throughout the study. All of the dogs lived in the Helsinki area.

     Two clinically healthy Beagles were included as control dogs. The dogs were sacrificed for an unrelated research project at the age of 3
years. The experimental protocol was approved by the Ethics Committee for Animal Experiments of the College of Veterinary Medicine, Helsinki, Finland.

Tests Performed on the Puppies
     Serum TLI was measured by radioimmunoassay.7. The tests were performed for the 1st time at 6 weeks of age.

     The biopsies from the pancreas were taken via laparotomy under general anesthesia. The gross appearance of pancreas was evaluated. One biopsy specimen was obtained from the right duodenal limb of the pancreas at each time point by ligation of a piece of pancreatic tissue (ca. 4.0mm in diameter) through a ventral midline abdominal incision. From all puppies the 1st biopsy was taken at 6 weeks of age and subsequently at approximately 0.5, 1.2, 2.0 years of age. Besides puppy no. 2 at 5.5 and 7.0 years of age. The biopsy specimens for histologic an electron microscopy (EM) examinations were processed in a routine manner.8. The EM findings of acinar cells were classified into 4 stages.8.

     Necropsy was performed on all dogs, and several samples of the pancreas were examined. The pancreas of 2 control dogs was also studied.

Results
    The dam developed a fatal mesenteric torsion 6 months after parturition. The sire was euthanized at the age of 12 years. Necropsy of both the dogs revealed a severely atrophic pancreas typical of an end-stage PAA.

    The results for puppy no. 1 have been published in detail elsewhere.8 Briefly, the female puppy was clinically healthy at birth and throughout adolescence, and grew normally without signs of maldigestion. The 1st biopsy from the pancreas at the age of 6 weeks was histological normal, but EM examination revealed mild stage 1 abnormalities. The dog was clinically normal at 22 months of age, but serum TLI was decreased to 0.3 mg/L (o2.5 mg/L indicating EPI; normal reference range 5.2–
35.0 mg/L). Examination of the pancreas indicated no gross or histologic abnormalities, but EM revealed widespread degenerative changes (stages 2–4). One month
later, the dog developed typical clinical signs of EPI, and at 25 months of age the gross and histologic examination of the pancreas revealed typical features of PAA.2 Treatment with powdered pancreatic extracta and antibiotics was introduced, with good treatment response.

     Puppy no. 2 (female) was clinically healthy during the 1st 5 years. Laparotomy was performed 4 times (at 1.5, 5,14, and 24 months of age). Gross and histologic examination was normal every time, and EM studies revealed stage 1 changes. Serum TLI was measured 12 times, and on 2 occasions the TLI value was subnormal (3.4 and 4.1 mg/L).

     At the age of 5.5 years, the dog was still clinically healthy and of normal weight, but serum TLI was abnormally low (2.1 mg/L) and at laparotomy, the gross
examination showed partial atrophy of the pancreas. There were scattered areas of pancreas that had lost their glandular appearance and changes typical for partial
PAA resulting from atrophic lymphocytic pancreatitis.1,2 Lymphocytic inflammation was most extensive in the border zone areas of normal and partially affected tissue.
EM showed evidence of atrophy, mainly type 3 and 4 degenerative changes. The islet cells were normal.

     The dog was treated with immunosuppressives (prednisolon  and azathioprined).9 Prednisolon treatment was discontinued after 1 year and azathioprine treatment
after 1.5 years.

     The dog lived another 6 years after diagnosis of partial AA. No signs of EPI were present. Serum TLI was measured 11 times; the values were always o5.2 mg/L, 5
times being abnormally low, o2.5 mg/L (range 0.2–2.4 mg/L).

     The dog died of internal hemorrhage at the age of 12 years. At necropsy the pancreas was severely diminished in size and histologic examination showed mild lymphocytic pancreatitis, especially in the border zone regions. In areas of more advanced tissue destruction, the findings were similar to those found in end-stage PAA.

     Puppy nos. 3 to 6 showed no clinical signs typical of EPI during their lifetime. A laparotomy was performed 4 times on each dog (between 6 weeks and 24 months of age), and the pancreas on every occasion was grossly normal, and a biopsy was taken. Histologic examinations revealed a normal pancreas. EM of the biopsies often
showed type 1 changes in acinar cells. After each laparotomy, recovery was uncomplicated and postoperative assays of serum amylase and lipase activities provided
no indication of pancreatitis or pancreatic tissue damage. Serum TLI was measured 7 to 15 times, and in each dog the value was at least once subnormal. The lowest values in dog nos. 3 to 6 were 2.7, 3.4, 3.4, and 4.9 mg/L, respectively. The dogs died due to different causes at ages 8 to 13 years. At necropsy, the pancreas was grossly and histologically normal in each dog.

     The TLI value of the 2 control dogs (7.5 and 8.4 mg/L) was within the normal range, and necropsy of the dogs revealed a normal pancreas both grossly and histologically. EM study of the control samples revealed evidence of type 1 changes.

Discussion
     Several previous studies have suggested that EPI is inherited in an autosomal recessive fashion. Here, when 2 affected EPI dogs were mated, only 2 of the 6 offspring
revealed atrophic changes in the exocrine part of the pancreas. This is much less than expected if a disease is inherited by Mendelian genetics. A polygenic mode of
inheritance is therefore the only explanation for this outcome. This is, however, a logical result as PAA in GSDs has been shown to be an autoimmune disease.2
     Performing test mating in dogs is laborious, particularly in late-onset diseases such as EPI. Test matings have been used more often in early-onset diseases, but also in these cases the keeping of affected animals purely for test purposes is ethically problematic. Fortunately, EPI is a treatable disease, and with medication an affected dog can live a more or less normal life. In our study, all dogs, including the two with degenerative changes in the pancreas, lived a normal life.

     Our previous report on puppy no. 1 described the rapid progression from EM findings to the gross appearance of a completely atrophied pancreas. We noted also
that mild stage 1 changes were found in acinar cells at the age of 6 weeks.8 To check whether the stage 1 changes were fixation artifacts instead of real degenerative changes, the pancreas of 2 healthy control dogs was examined. These dogs revealed similar changes. In conclusion, the changes earlier reported to be stage 1 changes are artifacts, thereby confirming that previous evidence indicating that PAA is a congenital disease in GSDs is not reliable.

     To halt the degenerative process on puppy no. 2, immunosuppressive medication was initiated. Whether the treatment was responsible for arresting the destruction process remains, however, unknown. Some dogs have been reported to stay in the subclinical phase of EPI for Exocrine Pancreatic Insufficiency 451
years, or sometimes for life, without treatment.9 Currently, no treatment is recommended when dogs with partial PAA have no clinical signs.9

    The other 4 siblings showed neither clinical signs typical of EPI nor degenerative or inflammatory changes in the pancreas. During their lifetime a pancreatic biopsy
was taken by laparotomy 4 times, and the pancreas was also examined at necropsy.

     To take serial biopsies from pancreas was the only reliable way to gain information about the pathological processes involved with the early stages of PAA before
the clinical signs of EPI appear. The 1st biopsies were taken after the weaning of the puppies to find out if EPI is a congenital disease. In an earlier study it was shown
that in about every 2nd EPI dog the clinical signs appear before 2 years of age.1 This is why we took almost all the biopsies during the 1st 2 years. During the study, the recovery from biopsy samplings was uneventful and no complications were detected.

      Measurement of serum TLI is the most commonly used test of pancreatic function, being both a sensitive and practical method for detecting severe EPI. We have reported that repeatedly low serum TLI values (o5.2 mg/L) in clinically healthy dogs can be a valuable marker of subclinical EPI and suggestive of partial PAA in GSDs.10 The results of puppy no. 2 are in agreement with this conclusion. During the last 6 years of its life the serum (TLI value was constantly low, fluctuating between subnormal (2.5–5.2 mg/L) and abnormally low (o2.5 mg/L). However, we have also concluded that diagnosing partial PAA may be problematic with an indirect pancreatic function test because of overlapping results in normal and partially affected dogs.10 This was also seen in these puppies, as each of the 4 healthy puppies had on at least some occasion subnormal serum TLI values.

     Our findings revealed that PAA is not a congenital disease in GSDs, and in this canine family PAA was not inherited by a simple autosomal recessive fashion of a
single gene. The study also showed that the atrophic process can destroy the entire exocrine part of the pancreas in a very short time, but the process can be halted, remaining in this position for the rest of the dog’s life. The function test helped to detect the stage of the exocrine pancreas, but it was not completely reliable.

From the Department of Equine and Small Animal Medicine (Westermarck, Wiberg) and the  Department of Basic Veterinary Sciences (Saari), Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland. Data for one of the dogs have previously been published in Am J Vet Res 1993;54:1088-1094. Corresponding author: Elias Westermarck, Department of Equine and Small Animal Medicine, Faculty of Veterinary Medicine, University of Helsinki, P.O. Box 57, 00014 Helsinki, Finland; e-mail: [email protected]
Submitted June 2, 2009; Revised September 3, 2009; Accepted
November 18, 2009.
Copyright r 2010 by the American College of Veterinary Internal
Medicine
10.1111/j.1939-1676.2009.0461.x

Footnotes
a Viokase V, Fort Dodge Laboratories, Fort Dodge, IA
b Tylan, Elanco, Geneva, Switzerland
c Prednisolon, Leiras, Tammisaari, Finland
d Azamun, Leiras

References
1. Westermarck E, Wiberg M. Exocrine pancreatic insufficiency
in dogs. Vet Clin Small Anim 2003;33:1165–1179.
2. Wiberg ME, Saari SAM, Westermarck E. Exocrine pancreatic
atrophy in German Shepherd Dogs and rough-coated Collies: An
end result of lymphocytic pancreatitis. Vet Pathol 1999;36:530–541.
3. Weber W, Freudiger U. Erbanalytische Untersuchungen uber
die chronisce exocrine Pankreasinsuffizienz beim Deutshen Scha¨ ferhund.
Schweiz Arch Tierheilkd 1977;119:257–263.
4. Westermarck E. Hereditary nature of canine pancreatic degenerative
atrophy in the German Shepherd Dog. Acta Vet Scand
1980;21:389–394.
5. Moeller ME, Steiner JM, Clark LA, et al. Inheritance of pancreatic
acinar atrophy in German Shepherd Dogs. Am J Vet Res
2002;10:1429–1434.
6. Clark LA, Wahl JM, Steiner JM, et al. Linkage analysis and
gene expression profile of pancreatic acinar atrophy in the German
Shepherd Dog. Mammalian Genome 2005;16:955–962.
7. Williams DA, Batt RM. Sensitivity and specificity of radioimmunoassay
of serum trypsin-like immunoreactivity for the
diagnosis of canine exocrine pancreatic insufficiency. J Am Vet
Med Assoc 1988;192:195–200.
8. Westermarck E, Batt RM, Vaillant C, Wiberg M. Sequential
study of pancreatic structure and function during development of
panreatic acinar atrophy in German Shepherd Dog. Am J Vet Res
1993a;54:1088–1094.
9. Wiberg ME, Westermarck E. Subclinical exocrine pancreatic
insufficiency in dogs. J Am Vet Med Assoc 2002;220:1183–1187.
10. Wiberg ME, Nurmi AK, Westermarck E. Serum trypsin like
immunoreactivity measurement for the diagnosis of subclinical exocrine
pancreatic insufficiency. J Vet Intern Med 1999;13:426–432.
452 Westermarck et al

Previous Research

2005

Linkage analysis and gene expression profile of pancreatic acinar atrophy in the German Shepherd Dog

Clark, LA., Wahl, JM., Steiner, JM., Zhou, W., Ji, W., Famula, TR., Williams, DA., Murphy, KE.:

http://www.ncbi.nlm.nih.gov/pubmed/16341675 

 

 

2003                                                                                                                                                                                                                  Pancreatic acinar atrophy in German shepherd dogs and rough-coated Collies                                         Etiopathogenesis and response to long-term enzyme replacement treatment                                                                                                                      Department of Clinical Veterinary Sciences, Section of Medicine - Faculty of Veterinary Medicine-University of Helsinki, Finland https://oa.doria.fi/bitstream/handle/10024/749/pancreat.pdf?sequence=1

2002

Inheritance of pancreatic acinar atrophy in German Shepherd Dogs

E. Michael Moeller , BS Jörg M. Steiner , Dr med vet, PhD Leigh Anne Clark , BS Keith E. Murphy  PhD Thomas R. Famula , PhD David A. Williams , VetMB, PhD Mary E. Stankovics , DVM Amy S. Vose , BA

http://avmajournals.avma.org/doi/abs/10.2460/ajvr.2002.63.1429  

 

 

1999
Exocrine Pancreatic Atrophy in German Shepherd Dogs and Rough-coated Collies:
An End Result of Lymphocytic Pancreatitis
M. E. WIBERG, S. A. M. SAARI, AND E. WESTERMARCK
http://www.vetpathology.org/cgi/reprint/36/6/530.pdf 

 

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June 2009 Research update

June 3, 2009     

     Dr. Leigh Anne Clark has completed collecting samples from EPI afflicted German Shepherd Dogs and non-EPI GSDs...THANK YOU EVERYONE!!!!   
     As of May 18, 2009 Dr. Leigh Anne Clark has taken accepted a position at Clemson University in South Carolina as Ass't Professor, Dept of Genetics and Biochemistry. She has moved her lab and will continue her EPI Genetics Research at Clemson.
    For the latest on the EPI Research Study, please go to the"EPI Research" tab.

Thank YOU!

4-22-09 THOSE WITH SENIOR GERMAN SHEPHERD DOGS - - THANK YOU FOR YOUR HELP !

BLOOD SAMPLES are no longer NEEDED FROM  OLDER-NORMAL GSDs!

As soon as those who have recently contacted Dr. Leigh Anne Clark and is sending in their samples please be sure to send in your samples by May 15, 2009....Texas A&M University (TAMU) will have completed collecting blood samples from senior (8 years old or older) normal (dogs that DO NOT have EPI) German Shepherd Dogs (GSDs) for Phase III of the EPI Research Study with SNP technology in an effort to identify the complex genetic markers to EPI. Shiloh Shepherds and GSD-mixes will be tested at a later date.

 

For future collections to be announced at a later date: INSTRUCTIONS FOR  EPI  RESEARCH
To participate in contributing a blood & serum sample please share these instructions with your vet. Most vets will draw blood free if it is for research. The Fed Ex shipping costs will be paid at this time and the Fed Ex # will be given to you once you contact Dr. Leigh Anne Clark

Elaine's "Boomer" 

 1. Dog will need to fast 12 hours before the draw.
2. Draw approximately 10ml blood into syringe
3. Fill purple-top with 5-6mls
4. Invert the tube several times after filling
5. Fill red-top tube with remainder of blood
6. Spin blood to separate serum
7. After separation, transfer serum to second red-top tube
8. Please make sure tubes are clearly labeled
9. Return samples in a Styrofoam cooler with a freezer pack.
10. Do not ship on a Friday.
11. Ship standard overnight via Fed Ex:

Blood Samples (with clear identification!) are to be shipped to:

Clemson University
51 New Cherry Rd
319 BRC Bldg
Clemson, SC 29634

Saving these instructions for possible future blood collections

INSTRUCTIONS (for next phase collections):

If in the future additional BLOOD SAMPLES are needed (Shiloh Shepherds and GSD-mixes) at a later date - - it will be announced. 
The blood contribution instructions will remain the same, but sent to Clemson Univ instead of Texas A&M. 
1. Dog will need to fast 12 hours before the draw.  
2. Draw approximately 10ml blood into syringe
3. Fill purple-top with 5-6mls
4. Invert the tube several times after filling
5. Fill red-top tube with remainder of blood
6. Spin blood to separate serum
7. After separation, transfer serum to second red-top tube
8. Please make sure tubes are clearly labeled
9. Return samples in a Styrofoam cooler with a freezer pack.
10. Do not ship on a Friday.
11. Ship standard overnight via Fed Ex
: Ship to: Clemson University, 51 New Cherry Road, 319 BRC Bldg, Clemson SC 29634

 

 

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