Detection of Actinobacillus pleuropneumoniae ApxIV toxin antibody in serum and oral fluid specimens from pigs inoculated under experimental conditions

González, Wendy; Giménez-Lirola, Luis G.; Holmes, Ashley; Lizano, Sergio; Goodell, Christa; Poonsuk, Korakrit; Sitthicharoenchai, Panchan; Sun, Yaxuan; Zimmerman, Jeffrey

doi:10.1515/jvetres-2017-0021

Abstract

Introduction: The prevention and control of Actinobacillus pleuropneumoniae in commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety of A. pleuropneumoniae antigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique to A. pleuropneumoniae and produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.

Material and Methods: Four groups of pigs (six pigs per group) were inoculated with A. pleuropneumoniae serovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.

Results: All pigs inoculated with A. pleuropneumoniae serovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.

Conclusion: This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring for A. pleuropneumoniae.

References

1. Baarsch M.J., Foss D.L., Murtaugh M.P.: Pathophysiologic correlates of acute porcine pleuropneumonia. Am J Vet Res 2000, 61, 684–690.10.2460/ajvr.2000.61.684
Search in Google Scholar Back to article
2. Blackall P.J., Klaasen H.L.B.M., Van Den Bosch H., Kuhnert P., Frey J.: Proposal of a new serovar of Actinobacillus pleuropneumoniae: serovar 15. Vet Microbiol 2002, 84, 47–52.10.1016/S0378-1135(01)00428-X
Search in Google Scholar Back to article
3. Bochev I.:Porcine respiratory disease complex (PRDC): a review. I. Etiology, epidemiology, clinical forms and pathoanatomical features. Bulg J Vet Med 2007, 10, 131–146.
Search in Google Scholar Back to article
4. Bossé J.T., Janson H., Sheehan B.J., Beddek A.J., Rycroft A.N., Kroll J.S., Langford P.R.: Actinobacillus pleuropneunomiae: pathobiology and pathogenesis of infection. Microb Infect 2002, 4, 225–235.10.1016/S1286-4579(01)01534-9
Search in Google Scholar Back to article
5. Bossé J.T., Johnson R.P., Nemec M., Rosendal S.: Protective local and systemic antibody responses of swine exposed to an aerosol of Actinobacillus pleuropneumoniae serotype 1. Infect Immun 1992, 60, 479–484.10.1128/iai.60.2.479-484.1992
Search in Google Scholar Back to article
6. Bossé J.T., Li Y., Sárközi R., Gottschalk M., Angen Ø., Nedbalcova K., Rycroft A.N., Fodor L., Langford P.R.: A unique capsule locus in the newly designated Actinobacillus pleuropneumoniae serovar 16 and development of a diagnostic PCR. J Clin Microbiol 2017, doi:10.1128/JCM-02166.
Search in Google Scholar Back to article
7. Chiers K., Haesebrouck F., Van Overbeke I., Charlier G., Ducatelle R.: Early in vivo interactions of Actinobacillus pleuropneumoniae with tonsils of pigs. Vet Microbiol 1999, 68, 301–306.10.1016/S0378-1135(99)00048-6
Search in Google Scholar Back to article
8. Cho W.S., Chae C.:Expression of the ApxIV gene in pigs naturally infected with Actinobacillus pleuropneumoniae. J Comp Pathol 2001, 125, 34–40.10.1053/jcpa.2001.047411437514
Search in Google Scholar Back to article
9. Costa G., Oliveira S., Torrison J., Dee S.: Evaluation of Actinobacillus pleuropneumoniae diagnostic tests using samples derived from experimentally infected pigs. Vet Microbiol 2011, 148, 246–251.10.1016/j.vetmic.2010.08.02320884132
Search in Google Scholar Back to article
10. Costa G., Oliveira S., Torrison J.: Detection of Actinobacillus pleuropneumoniae in oral-fluid samples obtained from experimentally infected pigs. J Swine Health Prod 2012, 20, 78–81.
Search in Google Scholar Back to article
11. Desrosiers R., Moore C.: Indirect transmission of Actinobacillus pleuropneumoniae. J Swine Health Prod 1998, 6, 263–265.
Search in Google Scholar Back to article
12. Dreyfus A., Schaller A., Nivollet S., Segers R.P., Kobisch A.M., Mieli L., Sørensen V., Hüssy D., Miserez R., Zimmermann W., Inderbitzin F.: Use of recombinant ApxIV in serodiagnosis of Actinobacillus pleuropneumoniae infections, development and prevalidation of the ApxIV ELISA. Vet Microbiol 2004, 99, 227–238.10.1016/j.vetmic.2004.01.00415066725
Search in Google Scholar Back to article
13. Eamens G.J., Gonsalves J.R., Whittington A.M., Turner B.: Evaluation of serovar-independent ELISA antigens of Actinobacillus pleuropneumoniae in pigs, following experimental challenge with A. pleuropneumoniae, Mycoplasma hyopneumoniae and Pasteurella multocida. Aust Vet J 2012, 90, 225–234.10.1111/j.1751-0813.2012.00934.x22632286
Search in Google Scholar Back to article
14. Frey, J.: Detection, identification and subtyping of Actinobacillus pleuropneumoniae. Methods Mol Biol 2003, 216, 87–95.10.1385/1-59259-344-5:87
Search in Google Scholar Back to article
15. Giménez-Lirola L.G., Jiang Y.H., Sun D., Hoang H., Yoon K.J., Halbur P.G., Opriessnig T.: Simultaneous detection of antibodies against Apx toxins ApxI, ApxII, ApxIII, and ApxIV in pigs with known and unknown Actinobacillus pleuropneumoniae exposure using a multiplexing liquid array platform. Clin Vaccine Immunol 2014, 21, 85–95.10.1128/CVI.00451-13
Search in Google Scholar Back to article
16. Giménez-Lirola L.G., Xiao C.T., Zavala M., Halbur P.G., Opriessnig T.: Improving ante-mortem diagnosis of Erysipelothrix rhusiopathiae infection by use of oral fluids for bacterial, nucleic acid, and antibody detection. J Microbiol Methods 2013, 92, 113–121.10.1016/j.mimet.2012.11.014
Search in Google Scholar Back to article
17. Gottschalk M.: Actinobacillosis. In: Diseases of Swine, edited by J.J. Zimmerman, L. Kariker, A. Ramirez, K. Schwartz, G. Stevenson, Wiley-Blackwell, Hoboken, New Jersey, 2012, pp. 653–669.
Search in Google Scholar Back to article
18. Gottschalk M., De Lasalle F., Radacovici S., Dubreuil J.D.: Evaluation of long chain lipopolysaccharides (LC-LPS) of Actinobacillus pleuropneumoniae serotype 5 for the serodiagnosis of swine pleuropneumonia. Vet Microbiol 1994, 38, 315–327.10.1016/0378-1135(94)90150-3
Search in Google Scholar Back to article
19. Grøndahl-Hansen J., Barfod K., Klausen J., Andresen L.O., Heegaard P.M., Sørensen V.: Development and evaluation of a mixed long-chain lipopolysaccharide based ELISA for serological surveillance of infection with Actinobacillus pleuropneumoniae serotypes 2, 6 and 12 in pig herds. Vet Microbiol 2003, 96, 41–51.10.1016/S0378-1135(03)00208-6
Search in Google Scholar Back to article
20. Hensel A., Stockhofe-Zurwieden N., Ganter M., Petzoldt K.: Aerosol exposure of pigs to viable or inactivated Actinobacillus pleuropneumoniae serotype 9 induce antibodies in bronchoalveolar lining fluids and serum, and protects against homologous challenge. Vet Microbiol 1995, 47, 27–41.10.1016/0378-1135(95)00106-K
Search in Google Scholar Back to article
21. Huang H., Zhou R., Chen M.L., Liu J.J., Xu X.J., Chen H.C.: Cloning and expression of the ApxIVA gene of Actinobacillus pleuropneumoniae and development of an indirect ApxIVA-ELISA. Chin J Biotechnol 2005, 21, 131–136.
Search in Google Scholar Back to article
22. Inzana T.J.: Virulence properties of Actinobacillus pleuropneumoniae. Microb Pathog 1991, 11, 305–316.10.1016/0882-4010(91)90016-4
Search in Google Scholar Back to article
23. Inzana T.J., Fenwick B.: Serologic detection of Actinobacillus pleuropneumoniae in swine by capsular polysaccharide-biotin-streptavidin enzyme-linked immunosorbent assay. J Clin Microbiol 2001, 39, 1279–1282.10.1128/JCM.39.4.1279-1282.2001
Search in Google Scholar Back to article
24. Jacobsen M.J., Nielsen J.P., Nielsen R.: Comparison of virulence of differenct Actinobacillus pleuropneumoniae serotypes and biotypes using an aerosol infection model. Vet Microbiol 1996, 49, 159–168.10.1016/0378-1135(95)00184-0
Search in Google Scholar Back to article
25. Jobert J.L., Savoye C., Cariolet R., Kobisch M., Madec F.: Experimental aerosol transmission of Actinobacillus pleuropneumoniae to pigs. Can J Vet Res 2000, 64, 21–26.
Search in Google Scholar Back to article
26. Kamp E.M., Stockhofe-Zurwieden N., Van Leengoed L.A., Smits M.A.: Endobronchial inoculation with Apx toxins of Actinobacillus pleuropneumoniae leads to pleuropneumonia in pigs. Infect Immun 1997, 65, 4350–4354.10.1128/iai.65.10.4350-4354.19971756249317048
Search in Google Scholar Back to article
27. Kittawornrat A., Prickett J., Chittick W., Wang C., Engle M., Johnson J., Patnayak D., Schwartz T., Whitney D., Olsen C., Schwartz K., Zimmerman J.: Porcine reproductive and respiratory syndrome virus (PRRSV) in serum and oral fluid samples from individual boars: will oral fluid replace serum for PRRSV surveillance? Virus Res 2010, 154, 170–176.10.1016/j.virusres.2010.07.02520670665
Search in Google Scholar Back to article
28. Kittawornrat A., Prickett J., Wang C., Olsen C., Irwin C., Panyasing Y., Ballagi A., Rice A., Main R., Johnson J., Rademacher C., Hoogland M., Rowland R., Zimmerman J.: Detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in oral fluid specimens using a commercial PRRSV serum antibody ELISA. J Vet Diagn Invest 2012, 24, 262–269.10.1177/1040638711435679
Search in Google Scholar Back to article
29. Krejci J., Nechvatalova K., Kudlackova H., Faldyna M., Kucerova Z., Toman M.: Systemic and local antibody responses after experimental infection with Actinobacillus pleuropneumoniae in piglets with passive or active immunity. J Vet Med 2005, 52, 190–196.10.1111/j.1439-0450.2005.00844.x
Search in Google Scholar Back to article
30. Lacouture S., Mittal K.R., Jacques M., Gottschalk M.: Serotyping Actinobacillus pleuropneumoniae by use of monoclonal antibodies. J Vet Diagn Invest 1997, 9, 337–341.10.1177/104063879700900324
Search in Google Scholar Back to article
31. Leary S., Underwood W., Anthony R., Cartner S., Corey D., Grandin T., Greenacre C.B., Gwaltney-Bran S., McCrackin M.A., Meyer R., Miller D.: Methods of Euthanasia. In: AVMA Guidelines for the Euthanasia of Animals. American Veterinary Association, Schaumburg, Illinois (USA), 2013, pp. 18–41.
Search in Google Scholar Back to article
32. Loera-Muro V.M., Loera-Muro A., Morfín-Mata M., Jacques M., Avelar-González F.J., Ramírez-Castillo F., Ramírez-López E.M., Guerrero-Barrera A.L.: Porcine respiratory pathogens in swine farms environment in Mexico. Open J Anim Sci 2014, 4, 196–205.10.4236/ojas.2014.44025
Search in Google Scholar Back to article
33. Loftager M.K., Eriksen L., Nielsen R.: Antibodies against Actinobacillus pleuropneumoniae serotype 2 in mucosal secretions and sera of infected pigs as demonstrated by an enzyme-linked immunosorbent assay. Res Vet Sci 1993, 54, 57–62.10.1016/0034-5288(93)90011-4
Search in Google Scholar Back to article
34. Losinger W.C., Bush E.J., Smith M.A., Corso B.A.: Mortality attributed to respiratory problems among finisher pigs in the United States. Prev Vet Med 1998, 37, 21–31.10.1016/S0167-5877(98)00114-7
Search in Google Scholar Back to article
35. Markey B., Leonard F., Archambault M., Cullinane A., Maguire D.: General procedures in microbiology. In: Clinical Veterinary Microbiology. Mosby Elsevier, St. Louis, Missouri (USA), 2013, pp. 3–79.
Search in Google Scholar Back to article
36. Marois C., Gottschalk M., Morvan H., Fablet C., Madec F., Kobisch M.: Experimental infection of SPF pigs with Actinobacillus pleuropneumoniae serotype 9 alone or in association with Mycoplasma hyopneumoniae. Vet Microbiol 2009, 135, 283–291.10.1016/j.vetmic.2008.09.061
Search in Google Scholar Back to article
37. McFarland J.: The nephelometer: an instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. J Am Med Assoc 1907, 49, 1176–1178.10.1001/jama.1907.25320140022001f
Search in Google Scholar Back to article
38. Mur L., Gallardo C., Soler A., Zimmermman J., Pelayo V., Nieto R., Sánchez-Vizcaíno J.M., Arias M.: Potential use of oral fluid samples for serological diagnosis of African swine fever. Vet Microbiol 2013, 165, 135–139.10.1016/j.vetmic.2012.12.034
Search in Google Scholar Back to article
39. Nielsen R., van den Bosch J.F., Plambeck T., Sørensen V., Nielsen J.P.: Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae. Vet Microbiol 2000, 71, 81–87.10.1016/S0378-1135(99)00157-1
Search in Google Scholar Back to article
40. Opriessnig T., Hemann M., Johnson J.K., Heinen S., Giménez-Lirola L.G., O’Neill K.C., Hoang H., Yoon K.J., Gottschalk M., Halbur P.G.: Evaluation of diagnostic assays for the serological detection of Actinobacillus pleuropneumoniae on samples of known or unknown exposure. J Vet Diagn Invest 2013, 25, 61–71.10.1177/1040638712469607
Search in Google Scholar Back to article
41. Panyasing Y., Goodell C.K., Giménez-Lirola L., Kittawornrat A., Wang C., Schwartz K.J., Zimmerman J.J.: Kinetics of influenza A virus nucleoprotein antibody (IgM, IgA, and IgG) in serum and oral fluid specimens from pigs infected under experimental conditions. Vaccine 2013, 31, 6210–6215.10.1016/j.vaccine.2013.10.040
Search in Google Scholar Back to article
42. Perry M.B., Altman E., Brisson J.R., Beynon L.M., Richards J.C.: Structural characteristics of the antigenic capsular polysaccharides and lipopolysaccharides involved in the serological classification of Actinobacillus (Haemophilus) pleuropneumoniae strains. Serodiagn Immunother Infect Dis 1990, 4, 299–308.10.1016/0888-0786(90)90018-J
Search in Google Scholar Back to article
43. Pohl S., Bertschinger H.U., Frederiksen W., Mannheim W.: Transfer of Haemophilus pleuropneumoniae and the Pasteurella-like organism causing porcine pleuropneumonia to the genus Actinobacillus (Actinobacillus pleuropneumoniae comb. nov.) on the basis of phenotypic and deoxyribonucleic acid relatedness. Int J System Bacteriol 1983, 33, 510–514.10.1099/00207713-33-3-510
Search in Google Scholar Back to article
44. Pomorska-Mól M., Dors A., Kwit K., Kowalczyk A., Stasiak E., Pejsak Z.: Kinetics of single and dual infection of pigs with swine influenza virus and Actinobacillus pleuropneumoniae. Vet Microbiol 2017, 201, 113–120.10.1016/j.vetmic.2017.01.011
Search in Google Scholar Back to article
45. Prickett J.R., Johnson J., Murtaugh M.P., Puvanendiran S., Wang C., Zimmerman J.J., Opriessnig T.: Prolonged detection of PCV2 and anti-PCV2 antibody in oral fluids following experimental inoculation. Transbound Emerg Dis 2011, 58, 121–127.10.1111/j.1865-1682.2010.01189.x
Search in Google Scholar Back to article
46. Prickett J.R., Kim W., Simer R., Yoon K.J., Zimmerman J.: Oral-fluid samples for surveillance of commercial growing pigs for porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 infections. J Swine Health Prod 2008, 16, 86–91.
Search in Google Scholar Back to article
47. Rogers R.J., Eaves L.E., Blackall P.J., Truman K.F.: The comparative pathogenicity of four serovars of Actinobacillus (Haemophilus) pleuropneumoniae. Aust Vet J 1990, 67, 8–11.10.1111/j.1751-0813.1990.tb07382.x
Search in Google Scholar Back to article
48. Schaller A., Kuhn R., Kuhnert P., Nicolet J., Anderson T.J., MacInnes J.I., Segers R.P., Frey J.: Characterization of ApxIV, a new RXT determinant of Actinobacillus pleuropneumoniae. Microbiology 1999, 145, 2105–2116.10.1099/13500872-145-8-2105
Search in Google Scholar Back to article
49. Sebunya T.N., Saunders J.R., Osborne A.D.: Dose response relationship of Hemophilus pleuropneumoniae aerosol in pigs. Can J Vet Res 1983, 47, 54–56.
Search in Google Scholar Back to article
50. Shope R.E.:Porcine contagious pleuropneumonia. I. Experimental transmission, etiology and pathology. J Exp Med 1964, 119, 357–368.10.1084/jem.119.3.357
Search in Google Scholar Back to article
51. Straw B.E., Shin S.J., Yeager A.E.: Effect of pneumonia on growth rate and feed efficiency of minimal disease pigs exposed to Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae. Prev Vet Med 1990, 9, 287–294.10.1016/0167-5877(90)90074-R
Search in Google Scholar Back to article
52. Veterinary Diagnostic Laboratory (VDL) Iowa State University. Procedure for standard plate count; standard operative procedure (2^nd version). Document No. 9.344, Ames, IA, USA, 2014.
Search in Google Scholar Back to article
53. Tobias T.J., Bouma A., Daemen A.J., Wagenaar J.A., Stegeman A., Klinkenberg D.:Association between transmission rate and disease severity for Actinobacillus pleuropneumoniae infection in pigs. Vet Res 2013, 44, 2.10.1186/1297-9716-44-2357403623311865
Search in Google Scholar Back to article
54. Van Leengoed L.A., Kamp E.M.: Endobronchial inoculation of various doses of Haemophilus (Actinobacillus) pleuropneumoniae in pigs. Am J Vet Res 1989, 50, 2054–2059.
Search in Google Scholar Back to article
55. Velthuis A.G.J., De Jong M.C.M., Stockhofe N., Vermeulen T.M.M., Kamp E.M.: Transmission of Actinobacillus pleuropneumoniae in pigs is characterized by variation in infectivity. Epidemiol Infect 2002, 129, 203–214.10.1017/S0950268802007252
Search in Google Scholar Back to article

Detection of Actinobacillus pleuropneumoniae ApxIV toxin antibody in serum and oral fluid specimens from pigs inoculated under experimental conditions

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