Listeriolysin O-based serodiagnosis of listeriosis and vis-à-vis cultural isolation of Listeria monocytogenes in slaughtered goats
DOI:
https://doi.org/10.48165/ijvsbt.19.4.05Keywords:
ELISA, Goats, Listeria monocytogenes, Listeriolysin O, SDS-PAGEAbstract
The present study was undertaken to ascertain prevalence, pathogenicity vis-a-vis seroprevalence of L. monocytogenes among goats slaughtered at Nagpur. The organism was isolated, identified, and characterized following USDA protocol from a total of 202 samples comprising blood samples (101) and chevon samples (101) revealing 1.98% prevalence of pathogenic L. monocytogenes. The virulent nature of pathogenic Listeria isolates was also assessed by in-vitro tests like haemolysis on sheep blood agar, CAMP test and PI-PLC assay. Listeriolysin O (LLO) protein produced by L. monocytogenes was purified employing ion-exchange chromatography. The purity of the LLO was confirmed using SDS-PAGE, which revealed it to be a homogeneous 58 kDa protein. Hyper immune serum was raised against purified LLO in healthy rabbits. An ELISA was standardise by the checker-board titration method and using LLO as an antigen and anti-LLO antibodies in raised hyper immune serum. The LLO-based indirect ELISA screening of streptolysin-O adsorbed sera from slaughtered goats demonstrated seropositivity for ALLO in 13.86% samples. Moreover, in this study, a positive correlation between seropositivity and isolation of pathogenic L. monocytogenes from the same animal was also noted endorsing the active functioning of ALLO in infection control. The results advocate use of LLO-based indirect ELISA on SLO adsorbed sera for rapid and reliable serodiagnosis of listeriosis in large populations where traditional isolation methods are not feasible.
Downloads
References
A Thesis: Barbuddhe, S.B. (1996). Studies on the kinetics and efficacy of listeriolysin-O for the sero-diagnosis of Listeria monocytogenes infection. Ph.D. Thesis submitted to Deemed University, IVRI, Izatnagar, U.P.
A Thesis: Chaudhari, S.P. (2001). Immunodiagnostic potential of PI-PLC, a virulence marker of Listeria monocytogenes, in listeric infection of buffaloes. Ph.D. Thesis submitted to Deemed University, IVRI, Izatnagar, (U.P.), India.
A Thesis: Shivaramu K.V. (2008). Studies on listeric infections in sheep by cultural, serological and molecular methods. MVSc thesis submitted to Deemed University Indian Veterinary Research Institute, Izatnagar, UP, India.
A Thesis: Thakur S (2000). Studies of Listeric infections on organized farms. M.V.Sc. Thesis submitted to Deemed University, Indian Veterinary Research Institute, Izatnagar, U.P. India.
Abd El-Rahim, I.H., Asghar, A.H., Fat'hi, S.M., and Ahmed, O.B. (2015). Serodiagnosis of Listeria monocytogenes infection in sheep. Int. J. Bioassays, 4, 3847-3850.
Aljicević, M., Beslagić, E., Zvizdić, S., Hamzić, S., and Mahmutović, S. (2006). Agglutination as screening test in routine diagnostic of listeriosis. Medicinski Arhiv., 60(2), 93-95.
Al–mashhadany, D. A., Ba-Salamah, H. A., Shater, A. R., and Al Sanabani, A. S. (2016). Prevalence of Listeria monocytogenes in red meat in Dhamar Governorate/Yemen. Prevalence, 2(12), 73-78.
Barbuddhe, S. B., Chaudhari, S. P. and Malik, S. V. S. (2002). The occurrence of pathogenic Listeria monocytogenes and antibodies against listeriolysin‐O in buffaloes. J. Vet. Med., Series B, 49(4): 181-184.
Barbuddhe, S. B., Malik, S. V. S., and Gupta, L. K. (2000). Kinetics of antibody production and clinical profiles of calves experimentally infected with Listeria monocytogenes. J. Vet. Med. Sci., Series B, 47(7), 497-502.
Barbuddhe, S. B., Malik, S. V. S., and Kumar, P. (1999). High seropositivity against listeriolysin O in humans. Ann. trop. med. parasitol., 93(5), 537-539.
Barbuddhe, S., & Chakraborty, T. (2008). Biotechnological applications of Listeria's sophisticated infection strategies. Microb. Biotechnol., 1(5), 361-372.
Berche P.A., Gaillard J.L., Geoffroy C.H. and Alouf J.E. (1987). T cell recognition of listeriolysin O is induced during infection with Listeria monocytogenes. J. Immunol. (Baltimore, Md.: 1950), 139(11), 3813-3821.
Berche, P., Bonnichon, M., Beretti, J. L., Raveneau, J., Gaillard, J. L., Veron, M., ... and Geslin, P. (1990). Detection of anti-listeriolysin O for serodiagnosis of human listeriosis. The Lancet, 335(8690), 624-627.
Bhanu Rekha, V., Malik, S.V.S., Chaudhari, S.P., and Barbuddhe, S.B. (2006). Listeriolysin O-based diagnosis of Listeria monocytogenes infection in experimentally and naturally infected goats. Small Rumin. Res., 66(1-3), 70-75.
Bouwer, H.G., Nelson, C.S., Gibbins, B.L., Portnoy, D.A., and Hinrichs, D.J. (1992). Listeriolysin O is a target of the immune response to Listeria monocytogenes. J. Exp. Med., 175(6), 1467-1471.
Christie, R., Atkins, N.E., and Munch-Petersen, E. (1944). A note on a lytic phenomenon shown by group B streptococci. Aust. J. Exp. Biol. Med. Sci., 22(3), 197-200.
Courtieu, A.L. (1991). Latest news on Listeriosis. Comp. Immunol. Microbiol. Infect. Dis., 14(1), 1-7.
Curtis, G.D.W., and Lee, W.H. (1995). Culture media and methods for the isolation of Listeria monocytogenes. Prog. J. Ind. Microbiol., 34, 63-75.
Das, S., Malik, S.V.S., Shrivastava, S., Gandhale, P., Kumar, S., Shoukat, S., Das, D.P., Barbuddhe, S.B. and Rawool, D.B. (2013). Comparative efficacy of Internalin C-based peptide and listeriolysin O-based enzyme linked immunosorbent assays for serodiagnosis of listeric infection in goats. Afr. J. Microbiol. Res, 7(48), 5471-5478.
Dhama, K., Karthik, K., Tiwari, R., Shabbir, M.Z., Barbuddhe, S., Malik, S.V.S., and Singh, R.K. (2015). Listeriosis in animals, its public health significance (food-borne zoonosis) and advances in diagnosis and control: a comprehensive review. Vet. Q., 35(4), 211-235.
Elezebeth, G., Malik, S.V.S., Chaudhari, S.P., and Barbuddhe, S.B. (2007). The occurrence of Listeria species and antibodies against listeriolysin-O in naturally infected goats. Small Rumin. Res., 67(2-3), 173-178.
Frece, J., Markov, K., Čvek, D., Kolarec, K., and Delaš, F. (2010). Comparison of conventional and molecular methods for the routine confirmation of Listeria monocytogenes in milk products produced domestically in Croatia. J. Dairy Res., 77(1), 112-116.
Geoffroy C.L., Gaillard J.L., Alouf J.E. and Berche P.A. (1987). Purification, characterization, and toxicity of the sulfhydryl-activated hemolysin listeriolysin O from Listeria monocytogenes. Infect. Immun., 55(7), 1641-1646.
Hernandez-Milian, A., and Payeras-Cifre, A. (2014). What is new in listeriosis? BioMed research international, 2014.
Husu, J.R. (1990). Epidemiological studies on the occurrence of Listeria monocytogenes in the feces of dairy cattle. J. Vet. Med., Series B, 37(1‐10), 276-282.
Islam, M.S., Husna, A.A., Islam, M.A., and Khatun, M.M. (2016). Prevalence of Listeria monocytogenes in beef, chevon and chicken in Bangladesh. Am. j. food sci. technol., 2(4), 39-44.
Jadhav, S., Bhave, M., and Palombo, E.A. (2012). Methods used for the detection and subtyping of Listeria monocytogenes. J. Microbiol. Methods, 88(3), 327-341.
Kaur, S., Malik, S.V.S., Bhilegaonkar, K.N., Vaidya, V.M., and Barbuddhe, S.B. (2010). Use of a phospholipase-C assay, in vivo pathogenicity assays and PCR in assessing the virulence of Listeria spp. Vet. J., 184(3), 366-370.
Kumar, P., Rao, J., and Haribabu, Y. (2014). Microbiological quality of meat collected from municipal slaughter houses and retail meat shops from Hyderabad Karnataka region, India. APCBEE procedia, 8, 364-369.
Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259), 680-685.
Larsen, S.A., and Jones, W.L. (1972). Evaluation and standardization of an agglutination test for human listeriosis. Appl. Microbiol., 24(1), 101-107.
Latha, C., Anu, C.J., Ajaykumar, V.J., and Sunil, B. (2017). Prevalence of Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus aureus, and Salmonella enterica Typhimurium in meat and meat products using multiplex polymerase chain reaction. Vet. World, 10(8), 927.
Lhopital, S., Marly, J., Pardon, P., and Berche, P. (1993). Kinetics of antibody production against listeriolysin O in sheep with listeriosis. J. Clin. Microbiol., 31(6), 1537-1540.
Lida T., Kanzaki M., Maruyama T., Inoue S., and Kaneuchi C. (1991). Prevalence of Listeria monocytogenes in intestinal contents of healthy animals in Japan. J. Vet. Med. Sci., 53: 873 – 875.
Lopez, A., and Bildfell, R. (1989). Neonatal porcine listeriosis. Can. Vet. J., 30(10), 828.
Low, J.C., Davies, R.C., and Donachie, W. (1992). Purification of listeriolysin O and development of an immunoassay for diagnosis of listeric infections in sheep. Can. Vet. J., 30(10), 2705-2708.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193, 265-275.
Malik, M. A., Sharma, J. K., and Neelesh, S. (2012). Prevalence of Listeria monocytogenes in different types of meat. Vet. Pract.,13(2), 204-205.
Miettinen A., Husu J.A., and Tuomi J.A. (1990). Serum antibody response to Listeria monocytogenes, listerial excretion, and clinical characteristics in experimentally infected goats. J. Clin. Microbiol., 28(2), 340-343.
Miettinen, A., and Husu, J. (1991). Antibodies to listeriolysin O reflect the acquired resistance to Listeria monocytogenes in experimentally infected goats. FEMS Microbiol. Lett., 77(2-3), 181-186.
Morandi, N., Canessa, A., and Rabagliati, A.M. (1981). Agglutinating antibodies against Listeria monocytogenes in healthy adult subjects. Bollettino Dell'istituto Sieroterapico Milanese, 60(5), 437-440.
Nass, W., and Ortel, S. (1977). Results and experience obtained with serodiagnosis of human listeriosis: an evaluation of analyses in the period from 1965-1975. Zeitschrift fur die gesamte Hygiene und ihre Grenzgebiete, 7(23), 482-485.
Negi M., Rawool D.B., Vergis J., Dhaka P., Vijay D., Mohan V., Suryawanshi R., Malik S.V.S., Kumar A., Barbuddhe S.B., and Ramteke P.W. (2014). Isolation and Identification of pathogenic Listeria monocytogenes from diarrhoeal cases in human infants and young animals. Adv. Anim. Vet. Sci., 2 (5S), 5 – 10.
Ojeniyi, B., Wegener, H.C., Jensen, N.E., and Bisgaard, M. (1996). Listeria monocytogenes in poultry and poultry products: epidemiological investigations in seven Danish abattoirs. J. Appl. Microbiol., 80(4), 395-401.
Osebold, J.W., and Aalund, O. (1968). Interpretation of serum agglutinating antibodies to Listeria monocytogenes by immunoglobulin differentiation. J. Infect. Dis., 118(2) 139-148.
Reiter R., Bowden M. and Palmer M. (1989). Ovine listeriosis in South coastal Western Australia. Aust.Vet. J. 66: 223–224.
Shakuntala, I., Das, S., Ghatak, S., Milton, A.A.P., Sanjukta, R., Puro, K.U., Pegu, R.K., Duarah, A., Barbuddhe, S.B. and Sen, A. (2019). Prevalence, characterization, and genetic diversity of Listeria monocytogenes isolated from foods of animal origin in North East India. Food Biotechnol., 33(3), 237-250.
Sibanda, T., and Buys, E. M. (2022). Listeria monocytogenes Pathogenesis: The Role of Stress Adaptation. Microorganisms, 10(8), 1522.
Singh, R., Kaur, S., and Bedi, J.S. (2019). Listeria contamination in chevon and mutton from retail meat shops and slaughter house environment of Punjab, India. FEMS Microbiol. Lett., 366(9), fnz111.
Suryawanshi, R.D., Jayarao, B., Barbuddhe, S.B., Chaudhari, S.P., Rawool, D.B., Mohan, V., Vergis, J., Negi, M. and Malik, S.V.S. (2014). Seropositivity of field veterinarians for listeric infection by indirect ELISAs employing recombinant and wild–type listeriolysin O. Adv. Anim. Vet. Sci., 2(5S), 17-21.
Suryawanshi, R.D., Malik, S.V.S., Jayarao, B., Chaudhari, S.P., Savage, E., Vergis, J., Kurkure, N.V., Barbuddhe, S.B. and Rawool, D.B. (2017). Comparative diagnostic efficacy of recombinant LLO and PI-PLC-based ELISAs for detection of listeriosis in animals J. Microbiol. Methods, 137, 40-45.
Tabouret, M., De Rycke, J., and Dubray, G. (1992). Analysis of surface proteins of Listeria in relation to species, serovar and pathogenicity. Microbiol., 138(4), 743-753.
Vaidya, G.R., Chaudhary, S.P., Zade, N.N., Khan, W.A., Shinde, S.V., Patil, A., and Kalambhe, D.G. (2018). Prevalence, virulence and antibiotic susceptibility of Listeria monocytogenes recuperated from slaughtered goats and pigs of Nagpur, Central India. Int. j. curr. microbiol. appl. sci., 7(4), 1566-1578.
Downloads
Published
Issue
Section
License
Copyright (c) 2023 Indian Journal of Veterinary Sciences & Biotechnology
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
