Ahmed Abdelaal, Magdy Elgioushy, Shaimaa Gouda, Mohamed El-Adl, Emad Hashish, Shimaa Elgaml, Mayar Ali, Attia El-Gedawy


In Egypt, limited studies were performed to investigate Jhon’s disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP). This study was concerned with diagnosis of MAP in cattle dairy farms using indirect-ELISA technique and amplification of IS900 using PCR technique for confirmation of infection and investigated adequate diagnostic markers for MAP through determining hemato-biochemical profile in cattle dairy farms. Screening of MAP was done by indirect ELISA between the periods from January 2017 to June 2018. A total of 200 cows suffered from persistent diarrhea (n=17) and apparently healthy cows in contact with them (n=183) were used in this study. Positive JD was reported in 62 cases (31%) and negative JD was reported in 138 cases (69%). Out of the positive JD, 17 cases were clinically affected by persistent diarrhea and weight loss (clinical JD), where 45 cases were apparently healthy (sub-clinical JD). Typical colonies of MAP were isolated from the fecal samples of clinically affected cows with positive staining of MAP bacilli. Amplification of IS900 using PCR technique confirmed the presence of MAP infection. Phylogenetic analysis revealed identical similarities with MAP strains in India and Egypt. Hemato-biochemical investigations revealed significant decrease in RBCs, PCV, total protein, globulin and glucose with non-significant change in oxidative stress markers in clinically affected cows. This study is considered one of the limited researches that highlight the role of MAP infection in dairy farms in Egypt.

Key words: dairy cattle; ELISA; IS900; Johne’s disease; MAP

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Norton S, Johnson WO, Jones G, Heuer C. Evaluation of Diagnostic Tests for Johne's Disease (Mycobacterium Avium Subspecies Paratuber-culosis) in New Zealand Dairy Cows. Journal of Veterinary Diagnostic Investigation 2010; 22 (3):341–51.

Pieper L, Sorge US, DeVries TJ, Godkin A, Lissemore K, Kelton DF. Evaluation of the Johne’s disease risk assessment and management plan on dairy farms in Ontario, Canada. Journal of dairy science 2015 ; 98 (10):6792–800.

Lombard JE. Epidemiology and economics of paratuberculosis. Veterinary Clinics: Food Animal Practice 2011; 27 (3):525–35.

Salem M, Heydel C, El-Sayed A, Ahmed SA, Zschöck M, Baljer G. Mycobacterium avium subspecies paratuberculosis: an insidious problem for the ruminant industry. Tropical Animal Health and Production 2013; 45 (2):351–66.

Tewari D, Hovingh E, Linscott R, Martel E, Lawrence J, Wolfgang D, Griswold D. Mycobacterium avium subsp. paratuberculosis (MAP) Antibody Response, Fecal shedding and Antibody Cross-reactivity to Mycobacterium bovis in MAP Infected Cattle Herds Receiving Johne's Vaccination. Clinical and Vaccine Immunology 2014; 00032–00014.

Beaudeau F, Belliard M, Joly A, Seegers H. Reduction in milk yield associated with Mycobacterium avium subspecies paratuberculosis (Map) infection in dairy cows. Veterinary Research 2007; 38 (4):625–34.

Chiodini RJ, Chamberlin WM, Sarosiek J, McCallum RW. Crohn’s disease and the mycobacterioses: a quarter century later. Causation or simple association? Critical Reviews in Microbiology 2012; 38 (1):52–93.

Gilardoni LR, Paolicchi FA, Mundo SL. Bovine paratuberculosis: a review of the advantages and disadvantages of different diagnostic tests. Rev Argent Microbiol 2012; 44 (3):201–15.

Bölske G, Herthnek D. Diagnosis of Paratuberculosis by PCR. Paratuberculosis: Organism, Disease, Control 2010; 267.

Sohal J, Singh S, Subodh S, Shoran N, Narayanasamy K, Singh P, Singh A, Maitra A. Mycobacterium avium subspecies paratuberculosis diagnosis and geno-typing: Genomic insights. Microbiological Research 2009; 164 (3):330–37.

Rosenberger G. Die Klinische Untersuchung des Rindes. 3rd ed. Paul Parey, Berlin,München. InTech 1990.

Manual OT. Paratuberculosis (Johne‟ s disease) Section 2.1. Chapter 2008.

Kalis C, Hesselink J, Barkema H, Collins M. Culture of strategically pooled bovine fecal samples as a method to screen herds for paratuberculosis. Journal of Veterinary Diagnostic Investigation 2000; 12: 547–51.

Fernández-Silva J, Abdulmawjood A, Bülte M. Diagnosis and molecular characterization of Mycobacterium avium subsp. paratuberculosis from dairy cows in Colombia. Veterinary Medicine International 2011; 352561.

Vary P, Andersen P, Green E, Hermon-Taylor J, McFadden J. Use of highly specific DNA probes and the polymerase chain reaction to detect Mycobacterium paratuberculosis in Johne's disease. Journal of Clinical Microbiology 1990; 28 (5):933–37.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 2013; 30: 2725–9.

Dacie JV. Dacie and Lewis practical haematology. Elsevier Health Sciences 2006.

Trinder P. Determination of glcose in blood using glucose oxidase with an alternative oxygen acceptor. Annals of Clinical Biochemistry 1969; 6 (1):24–27.

Allain CC, Poon LS, Chan CS, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clinical Chemistry 1974; 20 (4):470–75.

Fossati P, Prencipe L. Serum triglycerides determined colorimetrically with an enzyme that produces hydrogen peroxide. Clinical Chemistry 1982; 28(10):2077–80.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 1951; 193 (1):265–75.

Doumas B, Biggs H. Determination of Serum albumin: In standard methods of Clinical Chemistry (Cooper GA) Academic press Inc. New York 1972; 1:175

Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. American Journal of Clinical Pathology 1957; 28 (1):56–63.

Hausamen T-U, Helger R, Rick W, Gross W. Optimal conditions for the determination of serum alkaline phosphatase by a new kinetic method. Clinica Chimica Acta 1967; 5 (2):241–45.

Heinegård D, Tiderström G. Determination of serum creatinine by a direct colorimetric method. Clinica Chimica Acta 1973; 43 (3):305–10.

Marsh WH, Fingerhut B, Miller H. Automated and manual direct methods for the determination of blood urea. Clinical Chemistry 1965; 11 (6):624–27.

Koracevic D, Koracevic G, Djordjevic V, Andrejevic S, Cosic V. Method for the measurement of antioxidant activity in human fluids. Journal of Clinical Pathology 2001; 54 (5):356–61.

Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases the first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry 1974; 249 (22):7130––9.

Beutler E. Improved method for the determination of blood glutathione. J Lab Clin Med 1963; 61:882–88.

Frew JE, Jones P, Scholes G. Spectrophotometric determination of hydrogen peroxide and organic hydropheroxides at low concentrations in aqueous solution. Analytica Chimica Acta 1983; 155:139–50.

El-Bayomi K M, Saleh A A, Awad A, El-Tarabany M S, El-Qaliouby H S, Afifi M, El-Komy S, Essawi W M, Almadaly E A, El-Magd M A. Association of CYP19A1 gene polymorphisms with anoestrus in water buffaloes, Reproduction, Fertility and Development 2018; 30(3): 487–97.

Nims RW, Darbyshire JF, Saavedra JE, Christodoulou D, Hanbauer I, Cox GW, Grisham MB, Laval F, Cook JA, Krishna MC. Colorimetric methods for the determination of nitric oxide concentration in neutral aqueous solutions. Methods 1995; 7 (1):48–54.

Green E, Tizard M, Moss M, Thompson J, Winterbourne D, McFadden J, Hermon-Taylor J. Sequence and characteristics or IS 900, an insertion element identified in a human Crohn's disease isolate or Mycobacterium paratuberculosis. Nucleic Acids Research 1989; 17 (22):9063–73.

Harris NB, Barletta RG. Mycobacterium avium subsp. paratuberculosisin veterinary medicine. Clinical Microbiology Reviews 2001; 14 (3):489–512.

Tiwari A, VanLeeuwen JA, McKenna SL, Keefe GP, Barkema HW. Johne’s disease in Canada: Part I: Clinical symptoms, pathophysiology, diagnosis, and prevalence in dairy herds. The Canadian Veterinary Journal 2006; 47 (9): 874–82.

Clarke C. The pathology and pathogenesis of paratuberculosis in ruminants and other species. Journal of Comparative Pathology 1997; 116 (3):217–61.

Manning E, Collins M. Mycobacterium avium subsp. paratuberculosis: pathogen, pathogenesis and diagnosis [Johne's disease, paratuberculosis]. Revue Scientifique et Technique de l'OIE (France) 2001; 20(1):133–50.

Hulten K, Karttunen T, El-Zimaity H, Naser S, Collins M, Graham D, El-Zaatari F. Identification of cell wall deficient forms of M. avium subsp. paratuberculosis in paraffin embedded tissues from animals with Johne’s disease by in situ hybridization. Journal of Microbiological Methods 2000; 42: 185–95.

Bauerfeind R, Benazzi S, Weiss R, Schliesser T, Willems H, Baljer G. Molecular characterization of Mycobacterium paratuberc-ulosis isolates from sheep, goats, and cattle by hybridization with a DNA probe to insertion element IS900. Journal of Clinical Microbiology 1996; 34: 1617–21.

Motiwala AS, Strother M, Amonsin A, Byrum B, Naser SA, Stabel JR, Shulaw WP, Bannantine JP, Kapur V, Sreevatsan S. Molecular epidemiology of Mycobacterium avium subsp. paratuberculosis: evidence for limited strain diversity, strain sharing, and identification of unique targets for diagnosis. Journal of Clinical Microbiology 2003; 41: 2015–26.

Mahillon J, Chandler M. Insertion sequences. Microbiology and Molecular Biology Reviews 1998; 62: 725–74.

El-Deeb W, Fouda T, El-Bahr S. Clinico-biochemical investigation of paratuberculosis of Dromedary camels in Saudi Arabia: Proinflammatory cytokines, acute phase proteins and oxidative Stress Biomarkers. Pakistan Veterinary Journal 2014; 34 (4): 484–88.

Tharwat M, Al-Sobayil F, Ali A, Hashad M, Buczinski S. Clinical, ultrasonographic, and pathologic findings in 70 camels (Camelus dromedarius) with Johne’s disease. The Canadian Veterinary Journal 2012; 53 (5):543–48.

Weiss G, Goodnough LT. Anemia of chronic disease. New England Journal of Medicine 2005; 352 (10):1011–23.

Park BS, Park S, Jin K, Kim YM, Park KM, Lee J-N, Kamesaki T, Kim YW. Coombs-Negative Autoimmune Hemolytic Anemia in Crohn’s Disease. The American Journal of Case Reports 2014; 15:550–53.

Moore RE, Kirwan J, Doherty MK, Whitfield PD. Biomarker discovery in animal health and disease: the application of post-genomic technologies. Biomarker insights 2007; 2: 185–96.

Donat K, Erhardt G, Soschinka A, Brandt H. Decreased serum protein associated with Mycobacterium avium subspecies paratuberculosis shedding in German Holstein cows. Vet Rec 2014;

(16): 408.

Sweeney R, Collins M, Koets A, McGuirk S, Roussel A. Paratuberculosis (Johne's disease) in cattle and other susceptible species. Journal of Veterinary Internal Medicine 2012; 26 (6):1239–50.

Brady C, O'Grady D, O'Meara F, Egan J, Bassett H. Relationships between clinical signs, pathological changes and tissue distribution of Mycobacterium avium subspecies paratuberculosis in 21 cows from herds affected by Johne's disease. Veterinary Record 2008; 162 (5):147–52.

Elson CO, James SP, Graeff AS, Berendson RA, Strober W. Hypogammaglobulinemia due to abnormal suppressor T-cell activity in Crohn's disease. Gastroenterology 1984 ; 86 (3):569–6.

.Kamal AM. Some biochemical, heatological and clinical studies of selected ruminal and blood constituents in camels affected by various diseases. Res J Vet Sci 2008; 1 (1):16–27.

Salem M, El-Sayed A, Fayed A, Abo El-Hassan D. Subclinical infection of paratuberculosis among camels in Egypt. Journal of American Science 2012; 8:1141–47.

Cenesiz M, Ciftci G, Dalgin D, Kilic Y, Gul FY, Cenesiz S. Evaluation of Oxidant and Antioxidant Capacity in Paratuberculosis Positive Cattle. Pakistan Journal of Zoology 2016; 48 (5): :1603–06.

DOI: https://doi.org/10.26873/SVR-780-2019


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