Article Index "Johne's Disease in Sheep and Goats" Article Index

JOHNE'S DISEASE IN SHEEP AND GOATS

By: William P. Shulaw, DVM, MS
Extension Veterinarian, Cattle and Sheep, Ohio State University Extension
About the Author

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Etiology
Johne's disease, or paratuberculosis, is caused by Mycobacterium avium subspecies paratuberculosis, an acid-fast bacterium recently renamed because DNA studies have shown it to be very similar to M. avium. Mycobacterium paratuberculosis is capable of causing disease (Johne's disease) in all ruminants, and paratuberculosis has been described in several wild and captive exotic species. It may survive for extended periods in the environment in soil, water, and manure, and it is resistant to many common disinfectants. It may be killed by cresylic acid compounds and sodium orthophenylphenol (i.e., One Stroke EnvironT - available in many farm outlets). Worldwide, various "strains" have been isolated from cattle, sheep, goats, and from farmed red deer in New Zealand. The existence of "strains" is now well documented in the scientific community as is the difficulty in growing some of them in the laboratory.

Transmission and Pathogenesis
Contaminated feed, water, bedding, and soiled udders are thought to be the major routes of spread of the organism. Young animals less than six months of age are thought to be the most susceptible to infection. Older animals are more resistant to infection and development of disease but can develop delayed hypersensitivity and antibody responses which may interfere with some diagnostic tests. Intrauterine infection in cattle has been well documented, but the ultimate disease status of the offspring is unknown. A confirmed isolation of M. paratuberculosis from a sheep fetus in our laboratory occurred in 1989 and again in 1990. Another report has demonstrated antibodies to M. paratuberculosis in the serum of colostrum-deprived lambs. It appears likely that fetuses so infected in utero could develop clinical disease as adults or young adults, but this is unknown at this time.

Once organisms enter the small intestine, they are engulfed by macrophages - the cells involved in killing foreign bacteria - located in the intestinal wall. Macrophages generally are unable to kill them. The bacteria multiply inside the macrophage which eventually dies.

The body's immune responses to the intracellular bacteria create the typical thickening and microscopic distortion of the intestinal wall. Hypersensitivity reactions may increase the cellular response.

Evidence suggests that, at times, M. paratuberculosis may be found in the blood and lymph fluid which allows the bacteria access to other parts of the body. Organisms have been isolated from the uterine wall and placenta in the cow. The bacteria are also known to be present in the colostrum and milk of some infected cows, and it is likely that this occurs in other species. This method of transmission has not been documented in sheep and goats but is likely to occur.

Clinical Signs and Differential Diagnosis
Emaciation, or wasting disease, unresponsive to dewormers and antibiotics is the usual sign. Appetite is often good, in spite of weight loss, until the animal is near death. Normal stools are the usual observation even in clinically diseased animals, but intermittent diarrhea or softened, pasty stools are occasionally observed. The profuse watery diarrhea seen in cattle is not a common feature of the disease in sheep and goats. Intermittent, low-grade fever may be observed. Intermandibular edema (bottle jaw), lethargy, and depression are sometimes seen. The disease may be confused with intestinal parasitism, chronic malnutrition, caseous lymphadenitis (especially animals with internal abscesses), ovine progressive pneumonia (OPP), environmental toxins, and cancer.

Diagnostic Tests
Culture of feces (manure) and tissue is the "gold standard" for diagnosis but may be unreliable in sheep and some other ruminant species when using standard laboratory techniques. It is the generally recognized "official" test and may require 8-24 weeks of incubation for colonies to be observed. Several reports have described difficulties in growing the bacteria from known infected sheep tissues and feces. In many laboratories, M. paratuberculosis has been readily culturable from cattle and goats, but not from sheep. Recent work in Australia and Spain suggests that broth culture methods followed by DNA-based techniques may be successful in identifying the organism from sheep feces and tissues. In goats, fecal and tissue culture using cattle-adapted techniques seems to work satisfactorily; however, our state diagnostic laboratory has encountered a goat flock with classic paratuberculosis from which M. paratuberculosis could not be grown. Similar problems have also been reported in cattle from Australia, suggesting that strain differences can make culture difficult in many species.

Gross and microscopic examination of tissues taken from dead animals showing signs of disease are reliable techniques for diagnosis; however, sheep and goats may exhibit some gross and microscopic lesions not typical of those seen in cattle. These include: (1) ridges and thickening of the small intestine and cecum are not always seen as is common in cattle; (2) the colon and the rectum are not regularly involved - disease is more often confined to the small intestine; (3) cording, or prominence of the mesenteric lymphatics, is common along with swollen mesenteric lymph nodes; and (4) microscopically, a great amount of variation can exist in the type of lesions seen. This may be a result of strain differences in the infecting organisms and of differences in the ability of the animal to mount an effective immune response.

Acid-fast staining of intestinal mucosa or lymph node smears, often performed in cattle, usually reveals the presence of typical clumps of acid-fast bacteria within macrophages. Smears of rectal mucosa biopsies are difficult to get and may not be very helpful because the disease process is less likely to involve the rectum in sheep and goats. Intradermal Johnin testing (skin testing) may yield many false positive and false negatives. It has been suggested that the absence of any positives in a screening test of all animals may suggest that the herd or flock is not infected. However, this test is not widely practiced in sheep and goats.

Many serologic tests, or blood tests, have been used to diagnose paratuberculosis in cattle and small ruminants. Complement fixation is the official test in many countries for cattle. It is sometimes used in sheep and other small ruminants destined for export from the United States. Cross reactions with other bacteria have been reported for this test in cattle, and it has a relatively low sensitivity. It is no longer considered sufficiently accurate for diagnostic purposes.

Enzyme-linked immunosorbent assay (ELISA) may be useful in the future. However, this assay lacks standardization and validation across laboratories at the present time, and the difficulty of using culture as the "gold standard" complicates validation of tests. The currently licensed test as formatted for cattle has not yet been shown to be useful in sheep. One report examining its usefulness in goats looks promising, but only relatively small numbers of animals were used, and the test method was modified. Evidence exists that infections in sheep with Corynebacterium pseudotuberculosis, the cause of contagious abscesses in sheep and goats, may cause cross reactions and false positives in ELISA.

Agar gel immunodiffusion (AGID) testing has been used to assist in the diagnosis of clinical Johne's disease in sheep, goats, and llamas. In cattle, goats, and sheep, it correlates well with heavy shedding of M. paratuberculosis in the feces. This test may be the most sensitive and specific serologic test we have for sheep; however, it probably lacks sufficient sensitivity in subclinical animals to use in a test-and-cull program. One report describing culture and AGID test use in a large infected goat flock suggested that culture was able to detect more animals at a single test than AGID tests. However, AGID testing may be useful in some control efforts and in certification programs for goats.

A DNA probe kit available from IDEXX, Inc., will detect the organism reliably, but the format and processing steps are only approved for cattle. It certainly should be useful for other species, but it is currently only formatted for use with cattle feces. Research has indicated that current technology requires that about 1,000 or 10,000 organisms per gram of feces need to be present before the test will be positive. The fecal culture techniques routinely used for cattle and goats can detect fewer numbers of bacteria in feces. The probe technique may be useful for small ruminants, but is currently not widely available and usually quite expensive.

Control in Infected Flocks
Control of the disease in infected flocks is currently difficult because of the lack of a live animal test that will reliably detect animals that are not shedding M. paratuberculosis or which are shedding very low numbers. In goat flocks, AGID testing and culture often work well if the strains are culturable. Some suggestions for control include:

  • Necropsy (autopsy) as many sick or cull animals as possible to confirm the diagnosis and get an idea of how bad the problem is. Culture several representative clinical animals (both tissue and feces) until you can determine if the strain in the flock is culturable. If feasible, culture all animals over 1-1.5 years at least annually. Cull infected animals immediately.
  • Because the AGID test seems to correlate well with bacterial load and fecal shedding, you may chose to test an infected flock with AGID prior to lambing/kidding to remove the heaviest shedders.
  • Maintain a sanitary environment at lambing/kidding time. Use deep, fresh bedding or sunny, clean pastures.
  • Clean the udders of dams before nursing if feasible. Move young animals and their mothers to "clean" pasture as soon as possible.
  • Identify weight-loss ewes/does and isolate them from the flock until a diagnosis is made or until they are culled. Do not place these animals in contact with young animals "to give them more attention."
  • Consider depopulation; however, it may be difficult to repopulate with stock that can be verified as free of infection. It would be well to let the premises stand idle for at least a year before repopulation with clean animals.
  • Some flocks may be able to raise valuable lambs and kids as colostrum-deprived orphans, using milk replacer, in a separate, clean area in order to establish a new flock. In other words, establish two flocks and gradually phase out the old flock. This has been successful for control of OPP and CAE. Even though it is labor-intensive in the first few years, it may be the best way to salvage valuable genetic material. The possibility of in utero infection exists and might cause this technique to fail. As a precaution, progeny from known infected dams should be culled.
  • Vaccination has been shown in cows, goats, and sheep to lower the number of clinical cases in the herd or flock, but it will not prevent all animals from becoming infected. Some vaccinates shed bacteria, and a proportion will develop clinical signs. Much work needs to be done with vaccines to evaluate their usefulness because reports from around the world are somewhat contradictory. Currently available vaccines may interfere with ELISA, and other serology, and frequently cause reactivity in tuberculin testing. There are no vaccines currently available for sheep and goats in the United States.

Prevention
If the flock or herd is believed to be not infected, use biosecurity measures to prevent introduction of M. paratuberculosis. Some things to consider include these.

  • Most flocks and herds get infected by the purchase of infected animals. Maintain a closed flock if possible. Ask whether paratuberculosis has been observed in the seller's herd. Consider using the AGID test prior to purchase, or on arrival, and during quarantine. Submit fecal material for culture and consider culturing purchased animals every six months until they leave the herd/flock.
  • Be cautious when using cow colostrum for newborn lambs or kids. Infected cows may shed M. paratuberculosis in their colostrum and milk, and many dairy herds in the United States are infected. Pasteurization of colostrum may render it physically undesirable and will not reliably destroy M. paratuberculosis (145 degrees F. for 30 minutes is not always effective). Also be aware that many goat flocks appear to be infected so another flock's colostrum supply may be infectious.
  • Recipient animals in embryo transfer programs have been a source of infection in many cattle herds, and recipient animals of unknown status may introduce the infection in small ruminant flocks and herds.
  • Wildlife are not currently thought to be important carriers of M. paratuberculosis, but exposure to other captive exotic ruminants may be a risk factor. Grazing sheep on pastures with infected cattle, or on pastures to which manure from infected cattle has been applied, may spread the infection to sheep.

Currently, control of Johne's disease in flocks and herds of all species is difficult. Because of the nature of the organism and the slow progress of the infection, it is likely to remain so. However, we can make progress using the tools and the knowledge we have. It is important that we understand this disease and take steps to prevent further spread of the infection in our livestock species.

Reviewers
Kent H. Hoblet, DVM, MS Extension Veterinarian, Dairy The Ohio State University 241 Sisson Hall 1900 Coffey Road Columbus, OH 43210

Suelee Robbe, DVM, MS Veterinary Diagnostic and Production Animal Medicine 1701 Veterinary Medicine Iowa State University Ames, IA 50011

Susan M. Stehman, VMD, MS Senior Extension Veterinarian Diagnostic Laboratory P.O. Box 5786 College of Veterinary Medicine Cornell University Ithaca, NY 14852-5786

Figure 1 Figure 1. Johne's disease may be confused with other wasting diseases. The ewe in the foreground has Johne's disease. The black-faced ewe has caseous lmyphadenitis.
Figure 2 Figure 2. This sample of small intestine from a sheep with Johne's disease shows the classic thickening commonly seen in cattle with the disease.
Figure 3 Figure 3. This sample of small intestine from a different sheep shows relatively little thickening. The apparent ridges and thickening are a shrinkage artifact.
Figure 4 Figure 4. Positive results on agar gell immunodiffusion testing are an indication of heavy shedding of Mycobacterium paratuberculosis.

About the author: Ohio State University Fact Sheet
Veterinary Preventive Medicine
1900 Coffey Road, Columbus, OH 43210
VME-0003-01
Originating Web Site: http://ohioline.osu.edu/vme-fact/0003.html

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