Technologies of suspension cultivation in bioreactor of the Chlamydophila abortus strain on McCoy cell culture
Abstract
The purpose of the study was to investigate the cultural method of the diagnosis of Chlamydophila abortus. For this purpose, laboratory diagnosis of pathological material from the farm was carried out at the LLP Research and Production Enterprise “Antigen”, where, based on the dThe purpose of the study was to investigate the cultural method of the diagnosis of Chlamydophila abortus. For this purpose, laboratory diagnosis of pathological material from the farm was carried out at the LLP Research and Production Enterprise “Antigen”, where, based on the data of anamnesis, data of occurrence of similar epizootic picture for the last two years, examination and diagnostic manipulations (clinical symptoms, autopsy data, epizootological anamnesis), the preliminary diagnosis “Enzooticata of anamnesis, data of occurrence of similar epizootic picture for the last two years, examination and diagnostic manipulations (clinical symptoms, autopsy data, epizootological anamnesis), the preliminary diagnosis “Enzootic abortion of ewes caused by Chlamydophila abortus” was made. Samples from the organs of aborted foetuses and swabs with vaginal contents were taken to confirm this diagnosis. Complement fixation tests with chlamydia antigen were found to be positive in 7 (23%) of the 30 animals that were examined. To isolate the pathogen, the technology of suspension cultivation in a bioreactor of the Chlamydophila abortus strain on McCoy cell culture was used. Isolated pathological material was grown in McCoy cell cultures that were suspended in a bioreactor in Dulbecco’s Modified Eagle’s Medium (DMEM) nutrient medium with 10% foetal bovine serum and antibiotics at a concentration of 2×105 cells/mL. Enzyme immunoassay was a confirmation of the successful result of cultivation. Enzyme immunoassay confirmed the presence of the pathogen in 60.9% of cases. 25 samples were received, which were positive. Thus, the accuracy of the cultural diagnostic method turned out to be almost 3 times higher than the serological diagnostic method. Serological examination revealed 23% presence of Chlamydophila abortus in the samples, and the cultural diagnostic method revealed 60.9%. Unfortunately, in practice, the use of the cultural diagnostic method is associated with a time-delayed result, which is a disadvantage of this diagnostic method compared to the use of polymerase chain reaction
Keywords
laboratory diagnostics; small cattle; enzootic abortion of ewes; intracellular parasitism; cultural diagnostic method; enzyme immunoassay
[1] Adesiyun, A.A., Knobel, D.L., Thompson, P.N., Wenzel, J., Kolo, F.B., Kolo, A.O., Conan, A., & Simpson, G.J. (2020). Sero-epidemiological study of selected zoonotic and abortifacient pathogens in cattle at a wildlife-livestock interface in South Africa. Vector Borne and Zoonotic Diseases, 20(4), 258-267. doi: 10.1089/vbz.2019.2519.
[2] Akter, R., El-Hage, K.M., Sansom, F.M., Carrick, J., Devlin, J.M., & Legione, A.R. (2021). Metagenomic investigation of potential abortigenic pathogens in foetal tissues from Australian horses. BMC Genomics, 22(1), article number 713. doi: 10.1186/s12864-021-08010-5.
[3] Al-Ahmed, T.A., & Salman, S.S. (2020). Seroprevalence of enzootic abortion and border disease in small ruminants in Al-Basra province, Iraq. Plant Archives, 20(2), 2722-2727.
[4] Alzuguren, O., Domínguez, L., Chacón, G., Benito, A.A., & Mencía-Ares, O. (2023). Infectious abortions in small domestic ruminants in the Iberian Peninsula: Optimisation of sampling procedures for molecular diagnostics. Frontiers in Veterinary Science, 10, article number 1152289. doi: 10.3389/fvets.2023.1152289.
[5] Bommana, S., & Polkinghorne, A. (2019). Mini review: Antimicrobial control of Chlamydial infections in animals: Current practices and issues. Frontiers in Microbiology, 10, article number 113. doi: 10.3389/fmicb.2019.00113.
[6] Braun, C., Alcázar-Román, A.R., Laska, A., Mölleken, K., Fleig, U., & Hegemann, J.H. (2019). CPn0572, the C. pneumoniae ortholog of TarP, reorganizes the actin cytoskeleton via a newly identified F-actin binding domain and recruitment of vinculin. PloS One, 14(1), article number e0210403. doi: 10.1371/journal.pone.0210403.
[7] Büyük, F., Özgen, E.K., Karakurt, E., Coşkun, M.R., Büyük, E., Özmen, M., Dağ, S., Çelik, E., Gülmez Sağlam, A., Karadeniz Pütür, E., Ulucan, M., Nuhoğlu, H., & Şahin, M. (2020). Accomplished management of chlamydophila abortus-induced enzootic sheep abortions: The case of Şavşat (Turkey). Kafkas Universitesi Veteriner Fakultesi Dergisi, 26(6), 777-785. doi: 10.9775/kvfd.2020.24380.
[8] Campbell, L.A., & Hahn, D. (2020). Chlamydia pneumoniae infections. In M. Tan, J.H. Hegemann & C. Sütterlin (Eds.), Chlamydia biology: From genome to disease (pp. 31-58). London: Caister Academic Press. doi: 10.21775/9781912530281.02.
[9] Cheok, Y.Y., Lee, C.Y., Cheong, H.C., Looi, C.Y., & Wong, W.F. (2020). Chronic inflammatory diseases at secondary sites ensuing urogenital or pulmonary chlamydia infections. Microorganisms, 8(1), article number 127. doi: 10.3390/microorganisms8010127.
[10] Cheong, H.K., Lee, K.Y., Cheok, Y.Y., Tan, G.M., Looi, K.Y., & Wong, W.F. (2019). Chlamydiaceae: Diseases in primary hosts and zoonosis. Microorganisms, 7(5), article number 146. doi: 10.3390/microorganisms7050146.
[11] Cingolani, J., McCauley, M., Lobley, A., Bryer, A.J., Wesolowski, J., Greco, D.L., Lokareddy, R.K., Ronzone, E., Perilla, J.R., & Pomet, F. (2019). Structural basis for homotypic fusion of chlamydial inclusion bodies by the SNARE-like protein IncA. Nature Communications, 10, article number 2747. doi: 10.1038/s41467-019-10806-9.
[12] Cross, A.R., Baldwin, V.M., Roy, S., Essex-Lopresti, A.E., Prior, J.L., & Harmer, N.J. (2019). Zoonoses under our noses. Microbes and Infection, 21(1), 10-19. doi: 10.1016/j.micinf.2018.06.001.
[13] Esmaeili, H., Bolourchi M., Mokhber-Dezfouli, M.R., Khaltabadi Farahani, R., & Teimourpour, A. (2021). Detection of Chlamydia abortus and risk factors for infection in small ruminants in Iran. Small Ruminant Research, 197, article number 106339. doi: 10.1016/j.smallrumres.2021.106339.
[14] European Commission. (2019). Bovine and swine diseases. Retrieved from https://food.ec.europa.eu/system/ files/2020-12/la_annual-situation_2019.pdf.
[15] Fayez, M., Elmoslemany, A., Alorabi, M., Alkafafy, M., Qasim, I., Al-Marri, T., & Elsohaby, I. (2021). Seroprevalence and risk factors associated with Chlamydia abortus infection in sheep and goats in Eastern Saudi Arabia. Pathogens, 10(4), article number 489. doi: 10.3390/pathogens10040489.
[16] Filardo, S., Di Pietro, M., & Sessa, R. (2022). Better in vitro tools for exploring chlamydia trachomatis pathogenesis. Life (Basel, Switzerland), 12(7), article number 1065. doi: 10.3390/life12071065.
[17] Gojam, A., & Tulu, D. (2020). Infectious causes of abortion and its associated risk factor in sheep and goat in Ethiopia. International Journal of Veterinary Science & Technology, 4(1), 7-12.
[18] Gu, W., Deng, X., Lee, M., Sucu, Y.D., Arevalo, S., Stryke, D., Federman, S., Gopez, A., Reyes, K., Zorn, K., Sample, H., Yu, G., Ishpuniani, G., Briggs, B., Chow, E.D., Berger, A., Wilson, M.R., Wang, C., Hsu, E., Miller, S., DeRisi, J.L., & Chiu, C.Y. (2021). Rapid pathogen detection by metagenomic next-generation sequencing of infected body fluids. Nature Medicine, 27(1), 115-124. doi: 10.1038/s41591-020-1105-z.
[19] Hahn, D.L. (2021). Chlamydia pneumoniae and chronic asthma: An updated systematic review and populationbased risk meta-analysis. PLoS One, 16(4), article number e0250034. doi: 10.1371/journal.pone.0250034.
[20] Imkamp, F., Albini, S., Karbach, M., Kimmich, N., Spinelli, C., Herren, S., Sprecher, R., Meier, K., & Borel, N. (2022). Zoonotic Chlamydiae as rare causes of severe pneumonia. Swiss Medical Weekly, 152, article number w30102. doi: 10.4414/smw.2022.w30102.
[21] Kirimbayeva, Z., Abutalip, A., Mussayeva, A., Kuzembekova, G., & Yegorova, N. (2023). Epizootological monitoring of some bacterial infectious diseases of animals on the territory of the Republic of Kazakhstan. Comparative Immunology, Microbiology and Infectious Diseases, 102, article number 102061. doi: 10.1016/j. cimid.2023.102061.
[22] Komar, A., Kozerecka, O., Besarab, O., & Galkin, A. (2019). Development and validation of a highly informative immuno-enzymatic analysis for the determination of free prostat-specific antigen. Innovative Biosystems and Bioengineering, 3(4), 220-231. doi: 10.20535/ibb.2019.3.4.185877.
[23] Kvapil, P., Račnik, J., Kastelic, M., Marková, J., Murat, J.B., Kobédová, K., Pittermannová, P., Budíková, M., Sedlák, K., & Bártová, E. (2021). Biosurveillance of selected pathogens with zoonotic potential in a zoo. Pathogens (Basel, Switzerland), 10(4), article number 428. doi: 10.3390/pathogens10040428.
[24] Longbottom, D., Livingstone, M., Ribecca, P., Beckman, D., van der Ende, A., Pannekoek, J., & Vanrompuy, D. (2021). Whole genome de novo sequencing and comparative genomic analyses suggests that Chlamydia psittaci strain 84/2334 should be reclassified as Chlamydia abortus species. BMC Genomics, 22(1), article number 159. doi: 10.1186/s12864-021-07477-6.
[25] Mussayeva, A., Yegorova, N., Namet, A., Kozhabayev, M., & Syrym, N. (2023). Salmonella sheep abortion: Distribution, diagnosis, and control measures. Journal of Applied Animal Welfare Science. doi: 10.1080/10888705.2023.2214272.
[26] Mussayeva, A., Yegorova, N., Yerishov, M., Dossanova, A., Suchshikh, V., Namet, A., Siyabekov, S., Nussupova, S., Yespembetov, B., & Syrym, N. (2021). Molecular-biological properties of the attenuated strain of salmonella abortus-equi E-841, used in the creation of a vaccine against abortion of mares. American Journal of Animal and Veterinary Sciences, 16(2), 144-150. doi: 10.3844/ajavsp.2021.144.150.
[27] Nogarol, C., Marchino, M., Scala, S., Belvedere, M., Renna, G., Vitale, N., & Mandola, M.L. (2024). Seroprevalence and risk factors associated with chlamydia abortus infection in sheep and goats in North-Western Italy. Animals, 14(2), article number 291. doi: 10.3390/ani14020291.
[28] Nyzhnyk, B., Valchuk, O., Kataieva, T., Dreval, D., & Derkach, I. (2024). Common causes of abortion in cows. Scientific Reports of the National University of Life and Environmental Sciences of Ukraine, 20(1), 1-18. doi: 10.31548/ dopovidi.1(107).2024.020.
[29] Origlia, J.A., Cadario, M.E., Frutos, M.S., Lopez, N.F., Corva, S., Unzaga, M.F., Piscopo, M.V., Cuffini, C., & Petruccelli, M.A. (2019). Detection and molecular characterization of Chlamydia psittaci and Chlamydia abortus in psittacine pet birds in Buenos Aires province, Argentina. Revista Argentina de Microbiología, 51(2), 130-135. doi: 10.1016/j.ram.2018.04.003.
[30] Özgür, K. (2022). Molecular and histopathologic investigation of Pestivirus, Chlamydophila abortus and Listeria monocytogenes infections in aborted sheep foetuses. Journal of the Hellenic Veterinary Medical Society, 73(1), 3889-3896. doi: 10.12681/jhvms.26289.
[31] Rennick, J.J., Johnston, A.P., & Parton, R.G. (2021). Key principles and methods for studying the endocytosis of biological and nanoparticle therapeutics. Nature Nanotechnology, 16, 266-276. doi: 10.1038/s41565-02100858-8.
[32] Şevik, M. (2023). Border disease virus and chlamydophila abortus co-infection in aborted sheep foetuses. Journal of the Hellenic Veterinary Medical Society, 74(3), 5961-5964. doi: 10.12681/jhvms.30570.
[33] Tyrunskiy, V., Bogdanova, N., & Lyutskanov, P. (2023). Protective properties of the fleece of Taurian ewesof the Askanian fine fleece breed depending on the breeding differentiation rank. Animal Science and Food Technology, 14(2), 76-88. doi: 10.31548/animal.2.2023.76.
[34] Verzhykhovsky, O., & Nedosekov, V. (2024). Key aspects of biosafety in modern animal husbandry. Ukrainian Journal of Veterinary Sciences, 15(3), 41-54. doi: 10.31548/veterinary3.2024.41.
[35] Zhang, X., Yang, Q., Lang, Y., Jiang, X., & Wu, P. (2020). Rationale of 3,3’,5,5’-tetramethylbenzidine as the chromogenic substrate in colorimetric analysis. Analytical Chemistry, 92(18), 12400-12406. doi: 10.1021/acs. analchem.0c02149.