Source: CLEMSON UNIVERSITY submitted to NRP
LEPTOSPIROSIS IN PET DOGS IN SOUTH CAROLINA 2002-2012: INCIDENCE, ZOONOTIC POTENTIAL AND IMPLEMENTATION OF A DIAGNOSTIC PCR ASSAY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
0213246
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Nov 1, 2007
Project End Date
Aug 31, 2009
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
CLEMSON UNIVERSITY
(N/A)
CLEMSON,SC 29634
Performing Department
LIVESTOCK-POULTRY HEALTH
Non Technical Summary
Leptospirosis is a disease of zoonotic potential and is thus important from a public health standpoint. It is a reportable disease in people. Humans are susceptible to all of the pathogenic serovars that are found in domestic animals, and are considered an incidental host. In addition to indirect transmission through contaminated water or soil, people may also become infected by direct contact with urine or tissues from infected animals. The disease in people is often associated with occupation, which allows for direct exposure to infected animals, or recreational activities that allow for exposure to contaminated water and soil. Since dogs can have more direct contact with people than do livestock species and wildlife, dogs can be an important potential source of infection in people. The disease in people can vary from subclinical to severe and can potentially result in death from meningitis, renal failure or hepatic failure. Accurate, rapid diagnostic testing capabilities to detect the disease in animals are thus warranted to help eliminate human exposure as much as possible. In addition, such testing will allow for more rapid implementation of appropriate treatment for affected dogs and potentially exposed people. In addition to the zoonotic potential that exists between dogs and people, infected dogs could also potentially serve as a source of infection to other domestic animals, such as livestock species. If a dog becomes infected with a serovar that is potentially pathogenic to other domestic animals, the dog may develop persistent infection in the kidneys. The result is a carrier state in which the organisms may be shed in the urine for ≥1 year. The shed organisms serve are a source of infection for other animals. Impacts: The information obtained from this project is intended to provide a better perspective of the incidence of leptospirosis in pet dogs in South Carolina, which has not been previously reported. Investigation and implementation of more sophisticated diagnostic techniques within the CVDC laboratory will lead to more rapid, accurate diagnosis which could have direct impact on animal and human health.
Animal Health Component
45%
Research Effort Categories
Basic
20%
Applied
45%
Developmental
35%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3113830110012%
3113830116016%
3114010110012%
3114010116012%
7223830110012%
7223830116012%
7224010110012%
7224010116012%
Goals / Objectives
1) To investigate the incidence of leptospirosis in the pet dog population in South Carolina. This will be a retrospective and prospective investigation over the time period of 2002-2012. This information will be based on laboratory submissions to the CVDC. 2) To implement a Polymerase Chain Reaction (PCR) assay for detection of leptospirosis at the Clemson Veterinary Diagnostic Center (CVDC). 3) To compare the PCR method to currently utilized diagnostic techniques. 4) To obtain epidemiologic information on reported human leptospirosis cases in uth Carolina over the same time period, and to investigate a possible correlation with pet dog infection.
Project Methods
The retrospective portion of this study will involve searching the VETLIMS database to determine how many samples from dogs were submitted to CVDC for leptospirosis testing since 2002, and how many of these samples fit the criteria for being positive or suspect positive based on the testing performed at CVDC. These samples may include urine submitted for dark field microscopy, serum submitted for microagglutination (MAT) testing, kidney and/or liver submitted for histopathology with silver stain, and in-house or field necropsies in which any of these tests were requested and performed. Information obtained from this search would be summarized into a table indicating the accession number, tests performed and results of the tests. In addition, dog breed, age, sex, vaccination status, type/brand of vaccination, and location (county) will be included if such information was/is provided by the submitting veterinarian. The gathering of information from the VETLIMS database will be facilitated by Mike Martin, DVM, MPH, Veterinary Informatics Specialist, Clemson University, Livestock Poultry Health, Animal Health Programs. Samples from dogs are continuously being submitted to CVDC for leptospirosis testing, and test results will continuously be assessed. In addition to the tests currently available at CVDC and the implementation of PCR testing, other diagnostic methods can be explored, tested and compared to the current methods using these submitted samples as new methods become available. When possible, serum, urine and fresh tissues may be frozen and stored until new test methods are available for use within the laboratory. Formalin fixed, processed tissues are maintained in paraffin embedded blocks for a minimum of 7 years, and can be used for special staining or potentially, immunohistochemistry, should this become commercially available. For this study, all of the diagnostic testing on dogs will be performed at CVDC. A positive leptospirosis test will be defined as a MAT titer of ≥800 in a single serum sample, a fourfold increase in MAT titers in paired serum samples (samples submitted from the same animal approximately 2 weeks apart), detection of spirochetes in urine samples with dark field microscopy, and/or histopathologic lesions compatible with leptospirosis with detection of Leptospira-like organisms using silver stain in liver or kidney. A MAT titer of ≥800 will be used because titers this high are considered unlikely with vaccination. A positive PCR test will be defined as a positive leptospirosis infection after validation of the test is complete. PCR validation and testing will be supervised by Adam Leaphart, PhD, Microbiologist, Clemson Veterinary Diagnostic Center. Information on reported human leptospirosis infections during the same time period will be obtained through collaborative efforts with personnel at DHEC, and a possible correlation with a pet dog infection will be investigated when possible. Information on the test method(s) used to determine positive infection in humans will be gathered when possible.

Progress 11/01/07 to 08/31/09

Outputs
OUTPUTS: A PCR test for Leptospirosis which can be performed on urine and fresh tissue was implemented at the Clemson Veterinary Diagnostic Center in 2008. The laboratory information management systems have been searched to identify the numbers of canine submissions that were tested for Leptospirosis during the period of 2002 to current. Included in this search were those cases in which darkfield microscopy on urine, serum microagglutination test, Steiner stains on tissues and/or PCR tests were performed. This is an ongoing process as samples continue to be submitted. The numbers of positive results obtained and comparisons between test results if multiple tests were requested on the same animal are still in the process of being determined. The practicing veterinarians in the community that are interested in testing for Leptospirosis in dogs are made aware of the new PCR test. If veterinarians choose to submit urine, they are encouraged to request the PCR test instead of darkfield microscopy. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The retrospective searches through laboratory information management systems at CVDC have indicated that since 2002, the microagglutination test has been performed on approximately 430 dogs, darkfield microscopy on urine has been performed on approximately 20 dogs, and PCR has been performed on urine from 1 dog. There has thus far been some difficulty in assessment of numbers of Steiner stains performed on formalin fixed kidney and liver from dogs between 2002 and June 2008. This has been due to some limitations in the searchable database of the previously used laboratory information management system at CVDC. A new laboratory management information system was implemented in June of 2008, and since then, Steiner stains have been performed on kidney and/or liver from 4 dogs.

Publications

  • No publications reported this period


Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: A PCR test for Leptospirosis which can be performed on urine and fresh tissue was implemented at the Clemson Veterinary Diagnostic Center in 2008. The laboratory information management systems have been searched in attempt to identify the numbers of canine submissions that were tested for Leptospirosis during the period of 2002 to current. Included in this search were those cases in which darkfield microscopy on urine, serum microagglutination test, Steiner stains on tissues (kidney and liver) and/or PCR tests on fresh tissue and/or urine were performed. This is an ongoing process as samples continue to be submitted. The numbers of positive results obtained and comparisons between test results if multiple tests were requested on the same animal are still in the process of being determined. The practicing veterinarians in the community that are interested in testing for Leptospirosis in dogs are made aware of the new PCR test. If veterinarians choose to submit urine from live dogs, they are encouraged to request the PCR test, as this test allows for more rapid and accurate results. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Serum microagglutination test was discontinued at Clemson Veterinary Diagnostic Center (CVDC) in 2008. These samples are now being sent to the Texas Veterinary Medical Diagnostic Laboratory for testing. The decision was made due to shortage of personnel in the CVDC serology department, and the difficulty in maintaining the positive controls for the test.

Impacts
The implemetation of the new PCR test will allow for more rapid and accurate detection of infected dogs. This benefits not only the dogs that are infected, but also the veterinarians and pet owners who have been in contact with these dogs and are therefore at potential risk for infection themselves. Results of the retrospective searches through the laboratory information management systems at the Clemson Veterinary Diagnostic Center have indicated that between 2002 and March 2008, the microagglutination test has been performed on approximately 230 dogs, darkfield microscopy on urine has been performed on approximately 17 dogs, Steiner stain has been performed on tissue(s) from 73 dogs and PCR has been performed on urine from 1 dog.

Publications

  • No publications reported this period