Mycobacterium avium subspecies paratuberculosis – an important food borne pathogen of high public health significance with special reference to India: an update

Figures & data

Table 1. Global bio-load of MAP in milk: Country-wise.

Species	Countries	Sample type	Tests	Bioload (%)	References
Bio-load world-wide
Cattle	USA	Raw milk	Culture	2.4–28.6	Sweeney et al. 1992a; Streeter et al. 1995; Stabel 2002; Pillai & Jayarao 2002
	UK			6.9	Grant et al. 2002a
	Argentina			8.3	Paolicchi et al. 2003
	Czech Republic			2.0	Ayele et al. 2005
	Northern Ireland			0.3	O'Reilly et al. 2004.
	Australia			35.0	Taylor et al. 1981
	USA		PCR	33.0	Pillai and Jayarao 2002
	Iran			14.7	Soltani et al. 2008
	Iran		PCR	32.0	Anzabi et al. 2013
			Microscopy	20.0
	USA	Individual milk	ELISA	3.3–82.4	Sweeney et al. 1994; Lombard et al. 2006a, 2006b
	Denmark			15.4	Nielsen et al. 2002
				10.7–33.7	Klausen et al. 2003; Nielsen & Ersboll 2006
	Canada			1.7–11.2	Hendrick et al. 2005a; b
				2.5	Hendrick et al. 2006.
	USA	Bulk tank milk	Culture	2.8–20.6	Jayarao et al. 2004
	USA		PCR	13.4–39.0	Jayarao et al. 2004; Wilson et al. 2010.
	UK			22.0	Botsaris et al. 2013.
	Chile			49.0	Kruze et al. 2013
	Iran			8.6–23.0	Haghkhah et al. 2008
	Switzerland			23.0	Muehlherr et al. 2003.
	Chile		ELISA	9.4	Salgado et al. 2007
	Denmark			70.0	Nielsen et al. 2000
	USA	Pasteurized milk	Culture	2.8	Ellingson et al. 2005; Stabel 2002
	UK			1.8–6.7	Millar et al. 1996; Grant et al. 2002a, 2005
	Brazil			2.7	Carvalho et al. 2012a
	Argentina			2.8	Paolicchi et al. 2012.
	Canada			15.0	Grant et al. 2002b.
	N. Ireland			6.9	O'Reilly et al. 2004.
	Czech Republic			1.6	Ayele et al. 2005
Human	USA	Human milk		–	Naser et al. 2000, 2009; Bannantine et al. 2014
Bio-load in India
Cattle	North India	Raw milk	Culture	67.0	Shankar et al. 2010
			PCR	6–30.8	Singh et al. 2007a; Shankar et al. 2008, 2010; Kaur et al. 2010
		Individual milk	PCR	16.0	Sharma et al. 2008
			ELISA	28.0
		Pasteurized milk	Culture	67.0	Shankar et al. 2010
			PCR	33.0–38.8	Shankar et al. 2008, 2010
Buffaloes	Rajasthan	Individual milk	ELISA	31.1	Vinodh Kumar et al. 2014
Cattle & buffaloes	Mathura, Agra & New Delhi	Individual milk		38.0	Singh et al. 2016
			d-ELISA	46.2
		Un-pasteurized	PCR	100.0	Singh et al. 2007b
		Pasteurized		100.0
Goats	CIRG, Mathura	Raw milk	Culture	20.0–69.8	Singh & Vihan 2004; Kumar et al. 2008
			PCR	7.6–37.7	Kumar et al. 2008; Raghuvanshi et al. 2010
			ELISA	30.7–70.1
			Microscopy	13.8	Raghuvanshi et al. 2010
		Individual milk	PCR	8.3	Raghuvanshi et al. 2010; Singh et al. 2016
			ELISA	37.5–38.0
			Microscopy	16.6
			d-ELISA	54.9	Singh et al. 2016
		Pooled milk	PCR	5.8	Raghuvanshi et al. 2010
			ELISA	11.7
			Microscopy	5.8

Table 2. Presence of MAP in milk products from domestic livestock.

	Milk products from domestic livestock
S. No.	Animal species	Product Details	Country	Bio-load (%)	Reference
1	Cattle	Semi hard cheese	Switzerland	4.0	Stephan et al. 2007
2		Cheese	USA	23.0	Clark Jr. et al. 2006
3		Curd		23.0
4		Milk powder	Czech Republic	35.0	Hruska et al. 2005
5		Milk products	India	22.0–100.0	Shankar et al. 2010
6		Ice cream & flavoured milk		56.0–78.0
7	Goats and sheep	Hard cheese	Czech Republic	5.4	Ikonomopoulos et al. 2005
8		Semi hard cheese		5.0
9	Goats	Milk paneer	India	0.0–16.6	Raghuvanshi et al. 2013

Table 3. Presence of MAP in meat, organs and secretory products of livestock species.

S. No.	Product details	Country	Reference
(1)	Beef (cattle)	USA	Jaravata et al. 2007
		Denmark	Okara et al. 2010
		Italy	Savi et al. 2015
(2)	Raw meat products	Czech Republic	Lorencova et al. 2014
(3)	Meat & meat products	UK	Jaravata et al. 2007
(4)	Semen (bull)	UK	Larsen and Kopecky 1970
(5)	Epididymis (bulls)	USA	Collins 1997
(6)	Foetus (in-utero)	USA	Sweeney et al. 1992b
(7)	Intestinal tract, pulmonary lymph nodes, mammary gland, milk, uterus, amniotic fluid, testicles	Czech Republic	Kopecna et al. 2008
		India	Vohra et al. 2008a
(8)	Udder	USA	Collins 1997
		India	Vohra et al. 2008a
(9)	Kidneys, lungs, liver	USA	Collins 1997
(10)	Supra-mammary lymph nodes	Czech Republic	Pavlik et al. 2000; Vohra et al. 2008a
(11)	Intestine	India	Kumar et al. 2007; Singh et al. 2008b; 2008c; 2008d; Yadav et al. 2008; Chaturvedi et al. 2017
(12)	Mesenteric Lymph Nodes	India	Kumar et al. 2007; Singh et al. 2008c; 2008d; Yadav et al. 2008; Chaturvedi et al. 2017
(13)	Vaginal Secretion	India	Vohra et al. 2008b

Table 4. In-activation of MAP bacilli inoculated into milk during Pasteurization temperature.

	Pasteurization					Presence of MAP
Pasteurization Equipments	Temperature (°C)	Time	Detection level (CFU/ml)	Maximum Inoculums (log CFU/ml)	No. of samples (n)	No. (n)	%	Minimum reduction (log CFU)	Reference
Laboratory Test tubes	63	30 min	2	5	36	36	100	<1	Chiodini and Hermon-Taylor 1993
	72	15 sec			36	36	100	<1
	63.5	30 min	2	7	66	63	95	2	Grant et al. 1996
	71.7	15 sec			66	60	91	2
	65–76	30 min	5	8	16	5	31	2	Stabel 1997
Pasteurizer	65–75	15 sec	NA	6	12	0	NA	>6
Capillary tubes	63	30 min	4	5	30	0	NA	>5	Keshwani and Frank, 1998
	72	15 sec	NA	NA	30	0	NA	>5
Pasteurizer	72	15 sec	0.04	3	117	7	6	<1	Grant et al. 1998
Pasteurizer	≤90	15 sec	2	6	54	18	33	6	Grant et al. 1999
	72	25 sec	NA	NA	9	0	NA	>6
Laboratory test tubes	63	30 min	3	7	7	0	NA	7	Gao et al. 2002
	72	15 sec	NA	NA	11	2	18	5
Pasteurizer	63	30 min	0.04	8	36	4	11	4	Stabel and Lambertz 2004
	66	16 sec	NA	NA	36	34	94	1
	72–74	16 sec	NA	NA	108	8	7	1
Local pasteurization unit	71.7	15 sec	NA	NA	244	4	1.6	NA	Ayele et al. 2005
			NA	NA	10	1	10.0	NA
			NA	NA	66	13	19.7	NA
Slug flow pasteurization unit	62.7	30 min	NA	10^8	3	1	33.3	7	Stabel et al. 2004
	65.5	16 sec	NA	10^8	3	3	100.0	4
	71.7	15 sec	NA	10^5	3	1	33.3	4
	71.7	20 sec	NA	10^8	3	2	66.6	>8
	74.4	15 sec	NA	10^8	3	1	33.3	7
HTST pasteurization unit	62.7	30 sec	NA	10^8	3	0	0.0	>8	Stabel et al. 2004
	65.5	16 sec	NA	10^8	3	3	100.0	>5
	71.7	15 sec	NA	10^8	3	1	33.3	>8
	71.7	20 sec	NA	10^8	3	1	33.3	> 8
	74.4	15 sec	NA	10^8	3	0	0.0	> 8

Table 5. Mycobacterium avium subspecies paratuberculosis (MAP) in water supplies and environment.

Sn.	Environmental samples	Country	Reference
1.	Environmental and domestic aerosols	UK	Rhodes et al. 2014
2	River water	UK	Pickup et al. 2006
		India	Singh et al. 2012b
3	Drinking water supply system	UK	Pickup et al. 2006
		USA	Pierce et al. 2009b; Beumer et al. 2010
		Czech Republic	Klanicova et al. 2013
		USA	Falkinham et al. 2013
		UK	Aboagye et al. 2011
		Canada	Gill et al. 2011
		Czech Republic	Hruska et al. 2012
4	Domestic showers	USA	Feazel et al. 2009
		Australia	Thomson et al. 2013
5	Dam water and sediment	Australia	Whittington et al. 2005
6	Soil	USA	Reizman et al. 2011
		Chile	Salgado et al. 2011
		UK	Pickup et al. 2005
		Australia	Dhand et al. 2009
		India	Singh et al. 2012b
7	Distilled water	USA	Collins et al. 2001
8	Surface water	USA	Grewal et al. 2006
		UK	Pickup et al. 2006
		Ireland	Whan et al. 2005

Table 6. Percent bio-load of Mycobacterium avium subspecies paratuberculosis in the four domestic livestock species from different regions of India in past 32 years (1985–March 2017).

	Period A			Period B		Period C#
Species	1985–1990	1991–1995	1996–2000	2001–2005	2006–2010	2011-March, 2017	Cumulative (1985 – March, 2017)
Goats	11.4 (464 /4057)	13.1 (292 / 2221)	11.1 (443 / 3974)	21.5 (533/2479)	29.7 (443/1490)	39.8 (2394/ 6010)	22.5 (4569/20 231)
Sheep*	ND	ND	ND	25.0 (5 /20)	25.9 (118/454)	75.8 (154/203)	40.9 (277/ 677)
Sub total	11.4 (464 /4057)	13.1 (292/2221)	11.1 (443/3974)	21.52 (538/2499)	28.8 (561/1944)	41.0 (2548/ 6213)	23.1 (4846/ 20 908)
Cattle*	ND	ND	ND	47.0 (80/170)	31.0 (558/1799)	53.0 (1116/2104)	42.7 (1741/ 4073)
Buffaloes*	ND	ND	ND	41.9 (70/167)	22.1 (115/520)	53.0 (181/341)	35.6 (366/1028)
Sub total	11.4 (464 /4057)	13.1 (292/2221)	11.1 (443/3974)	44.5 (150/337)	29.0 (673/2319)	52.5 (1284/ 2445)	41.3 (2107/ 5101)
Grand Total	11.4 (464 /4057)	13.1 (292/2221)	11.1 (443 /3974)	24.2 (688 /2836)	28.9 (1234 /4263)	44.2 (3832/8658)	26.8 (6976/26 009)
	11.6 (1199/10 252)			27.0 (1922/7099)

Note: Figures in parenthesis are numbers (Positive/Total); #Up to March 2017; ND = Not done; * Based on 4 tests (microscopy, ELISA, PCR, Culture)

Table 7. Percent bio-load of MAP* in Human Population: 9 years (2008–March 2017): CIRG studies.

	Period A	Period B	Period C	Period D
Tests	2008–2013	2013–2014	2014–2015	2015-March, 2017	Cumulative (2008-March, 2017)
Serum ELISA	(34.0) 7887/23 196	(49.2) 190/386	(31.7) 235/740	(14.8) 73/492	(33.7) 8385/24 814
Blood PCR	(8.4) 260/3093	(18.1) 4/22	(17.4) 19/109	(10.7) 9/84	(8.8) 292/3308
Stool PCR	(5.9) 6/101	(55.5) 10/18	(44.7) 17/38	(41.6) 5/12	(22.4) 38/169
Total (%) n	(30.2) 8153/26 390	(47.8) 204/426	(30.5) 271/887	(14.7) 87/588	(30.8) 8715/28 291

*Figures in parenthesis are percent.

Table 8. Association of Mycobacterium avium subspecies paratuberculosis (MAP) infection with different kinds of human diseases/ailments.

Sn	Diseases/ailments	References
1	Crohn's disease	Hermon-Taylor et al. 2009; Naser et al. 2009; Singh et al. 2012a; MCNeese et al. 2015
2	Inflammatory bowel disease	Abubakar et al. 2008; Momotani et al. 2012; Nazareth et al. 2015; Timms et al. 2016
3	Ulcerative colitis	Pierce et al. 2010; Singh et al. 2011; Tuci et al. 2011
4	Irritable bowel syndrome(IBS)	Scanu et al. 2007; Singh et al. 2008a
5	Sarcoidosis	Reid and Chiodini 1993
6	Type-1 diabetes	Naser et al. 2013; Kawasaki et al. 2014; Singh et al. 2014c
7	Autoimmune thyroiditis	Pinna et al. 2014
8	Blau syndrome	Dow & Ellingson 2010
9	Multiple sclerosis	Mameli et al. 2014
10	Autism	Dow 2011
11	Autoimmune arthritis	Blau 1985; Moudgil et al. 1997
12	Auto-immune hepatitis	Miyata et al. 1995
13	Primary biliary cirrhosis	Vilagut et al. 1997
14	Scleroderma	Danieli et al. 1992
15	Kawasaki disease	Yokota et al. 1993
16	Alzheimer's disease	Cossu et al. 2011
17	Behcet's disease	Direskeneli & Saruhan-Direskeneli 2003
18	Takayasu's arteritis	Aggarwal et al. 1996

Table 9. Bio-type profile (Molecular Epidemiology) of MAP strains originating from different geographical regions of India in last 12 years (2004–March 2017) using IS1311 PCR_REA methodology.

			MAP bio-type profile (IS1311 PCR_RE)
Species	State	IS900 positive DNA (n)	Indian Bison Type (n)	Cattle Type (n)
Goats	1. Uttar Pradesh	141	141	NIL
	2. Himachal Pradesh	5	5	NIL
	3. Madhya Pradesh	7	7	NIL
	4. Assam	3	3	NIL
	5. Rajasthan	6	6	NIL
Sheep	1. Uttar Pradesh	15	15	NIL
	2. Tamil Nadu	20	20	NIL
Cattle	1. Uttar Pradesh	123	119	4
	2. Punjab	13	11	2
	3. Himachal Pradesh	6	6	NIL
Buffaloes	1. Uttar Pradesh	29	28	1
	2. Punjab	13	11	2
	3. Tamil Nadu	2	2	NIL
Sub-total A		383	377 (98.4)	6 (1.5)
Wild Ruminants
Deer	Uttar Pradesh	3	2	1
Wild Bison	Tamil Nadu	4	4	0
Rabbits	Tamil Nadu	6	6	0
Sub-total B		13	12 (92.3)	1 (7.7)
Human and milk samples
Human	Multiple states	49	43	6
Commercial milk	Multiple states	26	26	NIL
Sub-total C		75	69 (92.0)	6 (8.0)
Environmental samples
Soil	Uttar Pradesh	15	15	-
Yamuna river water	Uttar Pradesh	2	2	-
Sub-total D		17	17 (100%)	-
Grand total (A+ B + C + D)		488	472 (96.7%)	16 (3.2%)

*Figures in parenthesis are percent.

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