Paratuberculosis, diagnosis and

control

Dr. Jubeda Begum

 Introduction

 Johne’s disease (JD) is a chronic progressive granulomatous enteritis of ruminants

(mostly) caused by Mycobacterium avium subspecies paratuberculosis (MAP).

 Order: Actinomycetales, Family: Mycobacteriaceae and a member of Mycobacterium

avium intracellulare complex.

 MAP is a 0.5x1.5 microns size acid fast bacillus that occurs in clumps (Lilenbaum et al.,
2007).

 The family currently counts more than 130 established and validated species and
subspecies (Turenne et al., 2007).
 MAP genome size 4.4 to 4.7 Mb, G+C % = 66-67%.
 Dependency on mycobactin and presence of 15-20 copies of insertion element IS 900
(Bannantine et al., 2002).
 Disease occurrence

 Johne and Frothingham first recognized the disease in Germany (1895) (Lilenbaum et al.,
2007). It was first diagnosed in India in 1913 at Hissar followed by reports of JD
throughout the country with incidence around 2% and is endemic in India (Singh et al.,
2005; Ronald et al., 2012).

 Twort isolated the organism and experimentally produced the disease in cattle in 1910
(Hermon-Taylor, 2000). OIE listed B disease and has trade restrictions (Singh et al.,
2007).

 MAP is not limited to ruminants but expansion of hosts from polygastric to monogastric
and from monogastric to carnivores and wild ruminants reveals its wide host range and
complex epidemiology.
 First recognized in cattle, then in sheep and
later in goats.

 Transmitted horizontally via fecal oral route,

milk and vertically via placental route.
 Signs

 Primarily there are two clinical signs: cachexia and chronic diarrhea (less common in sheep
and goat).

 Infection of MAP in animals and human beings lead to inflammation of intestines and
mesenteric lymph nodes (Sechi et al., 2005).

 MAP is known to persist in harsh microenvironment inside activated macrophages (Ghosh et

al., 2013) causing chronic granulomatous enteropathy characterized by persistent diarrhoea,
emaciation and death in large ruminants.
 Economic impact

 High economic impact on animal industry resulting in estimated annual loss of USD 250
million in US alone (Ott et al., 1999; Hasonova et al., 2006).

 Indian studies showed consistently high bio-load of the disease in large ruminants.

 Over 70 percentage of the rural households in India depend on livestock.

 Therefore a direct or indirect interference with livestock production due to premature

culling, reduced milk production, decreasing fertility, emaciation and diarrhea will directly
hinder the economy of the country.
 Vaccines
 Currently there is no cure for Johne’s disease. Current vaccine preparations for
paratuberculosis do not prevent infection but do reduce fecal shedding and clinical disease
thereby slowing the spread of disease (Uzonna et al., 2003; Johnston et al., 2009).

 Initially, vaccines to prevent paratuberculosis were commercially available only in some

regions of North America and Europe where the economic losses are a serious issue
(Lilenbaum et al., 2007)

 At present, no commercial JD vaccine is available which can provide solid protection and
the vaccine available is not cost effective especially to small farmers who have minimum
holdings for their livelihood.

 In order for the potential benefit of new paratuberculosis vaccine candidates to be assessed,
experimental infection should lead to measurable shedding of the bacterium in the feces, as
it is the only antemortem quantitative assessment of infection status (Stabel et al., 2009).
 Zoonotic potential

 Recovery of MAP from human breast milk (Naser, 2000) and gut (Singh, 2014)
increased the concern for the control of disease in animals.

 Zoonotic potential not fully established, seems related to the etiology of Crohn’s
disease in humans (Coussens et al., 2004).

 Isolation of MAP from intestinal biopsies of Crohn’s disease patients led to the
concern that it may be a potential zoonosis (Mishina et al., 1996).
 Diagnosis

 Early and accurate diagnosis of JD is the prerequisite for the control of infection thereby
improving the sustainability and productivity of Indian dairy herds (Rawat et al., 2014).

 Main problem of diagnosis is the detection of infection in apparently healthy animals

(Shin et al., 2004).

 Diagnosis is difficult due to low sensitivity of the test developed so far, prolonged
incubation, intermittent shedding of MAP, variability in host immune response, fastidious
nature, and high cost of imported kits (Whitlock and Buergelt , 1996).

 Gold standard test is culture and isolation of MAP from feces (OIE, 2008).
 Diagnosis

 Diagnostic test for MAP infection divided into 2 types (OIE, 2008)

 A) Bacteriology and molecular approach (direct microscopy, conventional culture,

Radiometric culture (BACTEC); PCR - IS900 PCR , most widely used molecular
test to diagnose MAP infection).

 B) Immunological approach (CMI: Gamma interferon assay, DTH reaction, LTT,

NO estimation test; HI: AGID, ELISA, CFT).

 Fecal culture, although technically difficult and time consuming, will detect infected
animals in 6 months or more before they could develop clinical signs, which is very
important since this disease has a slow progression and many animals are non-
clinical carriers of disease.
 Prevention and Control Practices

 There are some vaccines for this disease; however they are used only in very well
defined situations and under strict regulatory control.

 Vaccination of young calves has shown a reduction in disease incidence but it does not
prevent shedding or subsequent new cases in the herd.

 Control involves good sanitation and management practices including screening tests for
new animals to identify and eliminate infected animals and ongoing surveillance of adult
animals.
Prevention and Control Practices

 In suspected herd calves, kids, or lambs should be birthed in areas free of manure removed
from the dam immediately after birth

 Bottle-fed pasteurized colostrum (or tested disease free colostrum), and raised separate
from adults until at least one year old.

 Prevent infection by closing the herd or securing additions from Johne’s free or Johne’s
test negative herds.

 Purchase replacements from a herd that has individual cow/calf records, good
management practices and is currently herd test negative or from a herd that has had no
evidence of Johne’s disease for 5 years as a second choice
Prevention and control option for beef
herds

 Management changes  Two herd programme

 Test and cull  Embryo transfer

 Partial depopulation  Vaccinate

 Limitations in the control of JD

 Control of disease in India is hindered by the presence of large number of cases of disease
in domestic livestock species, lack of information on strain diversity and non-availability of
“indigenous kits” for early detection.

 For diagnosis, culture (feces, milk, and intestinal tissues), ELISA and PCR have been
routinely used. Long incubation (12–16 weeks) and low sensitivity (>50%) limit the use of
culture (Motiwala et al., 2003).

 Utility of serological tests is limited due to low specificity and sensitivity, as immune
response may not be detectable either due to anergy or late appearance in pathogenesis.
 Limitations in the control of JD

 For the control of disease in dairy herds it is essential to know frequency and distribution
of MAP infection.

 Vaccination may interfere with eradication programmes that are based on detection and
subsequent elimination of infected animals and vaccination against.

 Paratuberculosis can also interfere with tests for bovine tuberculosis.

 Future perspective

 As there is no single reliable test for the confirmatory diagnosis of JD at present,

development of single lab on chip diagnostic method or DIVA vaccine should be
encouraged
Thank You