Identification of Brucella melitensis from camel’s blood by vitek2 and real time polymerase chain reaction
DOI:
https://doi.org/10.15584/ejcem.2024.1.17Keywords:
Brucella melitensis, brucellosis, camels, conventional PCR, RT-PCR, serological assaysAbstract
Introduction and aim. Brucellosis is a zoonotic disease. Experimental clinical and laboratory diagnosis is still facing problems in identifying the organism. The present study will diagnose a Brucella infection in camel blood in Qatar using serological assays. Isolation and identification were performed on a camel blood sample. Brucella in bacterial isolates was determined by real-time polymerase chain reaction (RT-PCR) as a gold standard test.
Material and methods. A total of 220 samples, 200 random serum samples, and 20 EDTA blood samples were selected among the above-mentioned random samples, and 20 serum samples from camel handlers were collected from Al Shahaniya prov ince, Qatar. The Rose Bengal test (RBT), buffered antigen plate agglutination test (BAPAT), and enzyme linked immunosorbent assay (cELISA) for the monoclonal antibody in serum samples were performed using commercially available kits. For the molecular detection of Brucella, conventional PCR and real-time PCR (GPS kit) were used for the genus-specific insertion sequence IS711. Brucella melitensis (MICROBOSS Hightech GmbH kit) was used to identify subspecies.
Results. The results identified by vitek2 compact (30%) showed B. melitensis in 6 samples out of 20 isolates. Both conventional (66.67%) and RT-PCR (83.33%) analyses supported this, demonstrating the presence of Brucella. These tests also showed that Brucella species were present in Rose Bengal 182/200 (91%), BAPAT 182/200 (91%), and cELISA (90%) 180/200 in camel serum.
Conclusion. To conclude, the prevalence of brucellosis in dromedary camels is higher in this region, and as a matter of urgency, measures should be taken to control the disease.
Downloads
References
Kirk MD, Pires SM, Black RE, et al. Correction: World Health Organization Estimates of the Global and Regional Disease Burden of 22 Foodborne Bacterial, Protozoal, and Viral Diseases: A Data Synthesis. PLoS Med. 2010;12(12):e1001940. doi: 10.1371/journal.pmed.1001940
Wernery U, Kinne J, Joseph M, Johnson B, Nagy P. Where do Brucella organisms hide in serologically positive lactating dromedaries. In Proc. International Camel Conference, College of Veterinary and Animal Sciences, Rajasthan Agricultural University, Bikaner, India. 2007;68-70.
OIE. Brucellosis (Brucella abortus, B. melitensis and B. suis) (Infection with B. abortus, B. melitensis and B. suis). In: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. France: Paris; 2016.
Zhang Z, Yu S, Wang X, et al. Human brucellosis in the People’s Republic of China during 2005-2010. Int J Infect Dis. 2013; 17(5):e289-e292. doi: 10.1016/j.ijid.2012.12.030
Xiao P, Yang H, Di D, et al. Genotyping of Human Brucella melitensis Biovar 3 Isolated from Shanxi Province in China by MLVA16 and HOOF. PLoS ONE. 2015;10(1):e0115932. doi: 10.1371/journal.pone.0115932
Shimol SB, Dukhan L, Belmaker I, et al. Human brucellosis outbreak acquired through camel milk ingestion in southern Israel. Isr Med Assoc J. 2012;14:475-478.
Ahmadi A, Mohsenpour B, Doroudian P, et al. A seroprevalence and relationship survey of brucellosis between pregnant women and women with spontaneous abortion in Iran. Med J Islam Repub Iran. 2017;31(1):42. doi: 10.14196/mjiri.31.42
Alkahtani AM, Assiry MM, Chandramoorthy HC, Al-Hakami AM, Hamid ME. Seroprevalence and risk factors of brucellosis among suspected febrile patients attending a referral hospital in southern Saudi Arabia (2014-2018). BMC Infect Dis. 2020;20:1-8. doi: 10.1186/s12879-020-4763-z
Hull NC, Schumaker BA. Comparisons of brucellosis between human and veterinary medicine. Infect Ecol Epidemiol. 2018;8:1500846. doi: 10.1080/20008686.2018.1500846
Norman FF, Monge-Maillo B, Chamorro-Tojeiro S, Perez-Molina J-A,Lopez-Velez R. Imported brucellosis: a case series and literature review. Travel Med Infect Dis. 2016;14:182-199. doi: 10.1016/j.tmaid.2016.05.005
Wang Z, Wang SS ,Wang GL, Wu TL, Lv YL, Wu QM. A pregnant mouse model for the vertical transmission of Brucella melitensis. Vet J. 2014; 200(1):116-121. doi: 10.1016/j.tvjl.2013.12.021
Franco MP, Mulder M, Gilman RH, Smits HL. Human brucellosis. Lancet Infect Dis. 2007; 7(12):775-786. doi: 10.1016/S1473-3099(07)70286-4
Dadar M, Alamian S, Behrozikhah, AM, et al. Molecular identification of Brucella species and biovars associated with animal and human infection in Iran. Vet Res Forum. 2019;10:315-321. doi: 10.30466/vrf.2018.89680.2171
Gwida MM, El-Gohary AH, Melzer F, et al. Comparison of diagnostic tests for the detection of Brucella spp. in camel sera. BMC Research Notes. 2011;4:525. doi: 10.1186/1756-0500-4-525
Ghorbani A, Rabbani Khorasgani M, Zarkesh-Esfahani H, et al. Comparison of serology, culture, and PCR for detection of brucellosis in slaughtered camels in Iran. Comp Clin Pathol. 2013;22:913-917. doi: 10.1007/s00580-012-1499-1
Marei A, Boghdadi G, Abdel-Hamed N, et al. Laboratory diagnosis of human brucellosis in Egypt and persistence of the pathogen following treatment. J Infect Dev Ctries. 2011;5:786-791. doi: 10.3855/jidc.1538
Sayour AE, Elbauomy EM, El-Kholi MK, Abeer AE Shehata. Brucellosis Prevalence and Serologic Profile of Male One-Humped. Global Veterinaria. 2015;14(1):67-76. doi: 10.5829/idosi.gv.2015.14.01.9260
Corbel MJ. Brucellosis in humans and animals. Geneva, Switzerland: World Health Organization; 2006.
Gwida, MM, El-Gohary AH, Melzer F, Khan I, Rosler U, Neubauer H. Brucellosis in camels. Res Vet Sci. 2012;92:351-355. doi: 10.1016/j.rvsc.2011.05.002
Bricker BJ. PCR as a diagnostic tool for brucellosis. Vet Microbiol. 2002; 90:435-446. doi: 10.1016/s0378-1135(02)00228-6
Jeffrey T, Foster Faith M, Walker Brandy D, et al. African Lineage Brucella melintensis Isolates from Omani Livestock. Front Microbiol. 2018;8:2702. doi: 10.3389/fmicb.2017.02702
Hamdy MER, Abdel-Haleem MH, Dawod RE, et al. First seroprevalence and molecular identification report of Brucella canis among dogs in Greater Cairo region and Damietta Governorate of Egypt. Vet World. 2023;16(1):229-238. doi: 10.14202/vetworld.2023.229-238
Rahil AI, Othman M, Ibrahim W, Yahya M. Brucellosis in Qatar: A retrospective cohort study. Qatar Med J. 2014;2014(1):25-30.
Ahmed ME, Musa MT. Characterization of Bacteria Isolated from Dromedary Camels Affected with Pneumonia for the First Time in Sudan. Annu Res Rev Biol. 2015;7(1):61-67. doi: 10.9734/ARRB/2015/16744
Wakjira BS, Jorga E, Lakew M, et al. Animal Brucellosis: Seropositivity rates, Isolation and Molecular Detection in Southern and Central Ethiopia. Vet Med. 2022;13:201-211. doi: 10.2147/VMRR.S372455
Aidaros H. Global perspectives – the Middle East: Egypt. Rev Sci Tech. 2005;24:589-596.
McDermott J, Arimi S. Brucellosis in sub-Saharan Africa: epidemiology, control and impact. Vet Microbiol. 2002;90:111-134. doi: 10.1016/s0378-1135(02)00249-3
Amin M, Ahmed S, Zaki H, Ismail R. Serological and molecular studies on the diagnosis of bovine brucellosis. Nature and Science. 2012;10:68-76.
Hu J, Zhang X, Yang H, et al. Brucellosis screening and follow-up of seropositive asymptomatic subjects among household members of shepherds in China. Eur J Clin Microbiol Infect Dis. 2021;40(6):1325-1328. doi: 10.1007/s10096-020-04115-z
Li F, Du L, Zhen H, et al. Follow-up outcomes of asymptomatic brucellosis: a systematic review and meta-analysis. Emerg Microbes Infect. 2023;12(1):2185464. doi: 10.1080/22221751.2023.2185464
Probert WS, Schrader KN, Khuong NY, Bystrom SL, Graves MH. Real-time multiplex PCR assay for detection of Brucella spp., B. abortus, and B. melitensis. J Clin Microbiol. 2004;42(3):1290-1293. doi: 10.1128/JCM.42.3.1290-1293.2004
Spicic S, Zdelar-Tuk M, Ponsart C, et al. New Brucella variant isolated from Croatian cattle. BMC Vet Res. 2021;17(1):126. doi: 10.1186/s12917-021-02833-w
Enright FM. The pathogenesis and pathobiology of Brucella infection in domestic animals. Chapter 12 in Animal Brucellosis. K. Nielsen and J. R. Duncan, ed. CRS Press, Boca Raton, FL. Perry, MB, and Bundle D R. Antigenic relationships of the lipopolysaccharides of Escherichia hermannii strains with those of Escherichia coli O157:H7, Brucella melitensis, and Brucella abortus. Infect Immun. 1990; 58:1391–1395.
Salisu U, Kudi C, Bale J et al. Seroprevalence of Brucella antibodies in camels in Katsina State, Nigeria. Trop Anim Health Prod. 2017;49(5):1041-1046. doi: 10.1007/s11250-017-1297-5
Dawood HA. Brucellosis in camels (Camelus dromedorius) in the south province of Jordan. Am J Agric Biol Sci. 2008;3(3):623-626.
Omer MM, Musa MT, Bakhiet MR, Perrett L. Brucellosis in camels, cattle and humans: Associations and evaluation of serological tests used for diagnosis of the disease in certain nomadic localities in Sudan. Res Vet Sci. 2010;29:663-669. doi: 10.20506/rst.29.3.2003
Anvarinejad M, Pouladfar GR, Pourabbas B, et al. Detection of Salmonella spp. with the BACTEC 9240 Automated Blood Culture System in 2008-2014 in Southern Iran (Shiraz): Biogrouping, MIC, and antimicrobial susceptibility profiles of isolates. Jundishapur J Microbiol. 2016;9(4):e26505.
Hassanzadeh P, Hassanzadeh Y, Mardaneh J, Rezai E, Motamedifar M. Isolation of Methicillin-Resistant Staphylococcus aureus (MRSA) from HIV Patients Referring to HIV Referral Center, Shiraz, Iran, 2011-2012. Iran J Med Sci. 2015;40(6):526-530.
Alshaikh AM, Al-Haidary A, Aljumaah RS, Al-Korashi MM, ElNabi GRA, Hussein MF. Camel brucellosis in Riyadh region, Saudai Arabia. J Camel Pract Res. 2007;14:113-117. doi: 10.20506/rst.11.3.612
Ducrotoy M, Bertu WJ, Matope G, et al. Brucellosis in Sub-aharan Africa: Current challenges for management, diagnosis and control. Acta Trop. 2017;165:179-193. doi: 10.1016/j.actatropica.2015.10.023
Al-Majali AM, Talafha AQ, Ababneh MM, Ababneh MM. Seroprevalence and risk factors or bovine brucellosis in Jordan. J Vet Sci. 2009;10(1):61-65. doi: 10.4142/jvs.2009.10.1.61
Baily GG, Kraahn J B, Drasar BS, Stoker, NG. Detection of Brucella melitensis and Brucella abortus by DNA amplification. J Trop Med Hyg. 1992;95:271-275.
Herman L, De Ridder H. Identification of Brucella spp. By using the polymerase chain reaction. Appl Environ Microbiol. 1992;58:2099-2101. doi: 10.1128/aem.58.6.2099-2101.1992
Hamdy MER, Amin AS. Detection of Brucella species in the milk of infected cattle, sheep, goats and camels by PCR. Vet J. 2002;163:299-305. doi: 10.1053/tvjl.2001.0681
O’Leary S, Sheahan M, Sweeney T. Brucella abortus detection by PCR assay in blood, milk and lymph tissue of serologically positive cows. Res Vet Sci. 2006;81:170-176. doi: 10.1016/j.rvsc.2005.12.001
Tsegay A, Tuli G, Kassa T, Kebede N. Seroprevalence and risk factors of brucellosis in abattoir workers at Debre Zeit and Modjo export abattoir, Central Ethiopia. BMC Infect Dis. 2017;17:101. doi: 10.1186/s12879-017-2208-0
Lignereux L, Chaber AL, Fretin D, Godfroid J, Saegerman C. Brucella melitensis biovar 1 isolation in a captive wildlife population in the United Arab Emirates. First isolation in the scimitar-horned Oryx (Oryx dammah). Vet Microbiol. 2022;266:109360. doi: 10.1016/j.vetmic.2022.109360
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2024 European Journal of Clinical and Experimental Medicine

This work is licensed under a Creative Commons Attribution 4.0 International License.
Our open access policy is in accordance with the Budapest Open Access Initiative (BOAI) definition: this means that articles have free availability on the public Internet, permitting any users to read, download, copy, distribute, print, search, or link to the full texts of these articles, crawl them for indexing, pass them as data to software, or use them for any other lawful purpose, without financial, legal, or technical barriers other than those inseparable from having access to the Internet itself.
All articles are published with free open access under the CC-BY Creative Commons attribution license (the current version is CC-BY, version 4.0). If you submit your paper for publication by the Eur J Clin Exp Med, you agree to have the CC-BY license applied to your work. Under this Open Access license, you, as the author, agree that anyone may download and read the paper for free. In addition, the article may be reused and quoted provided that the original published version is cited. This facilitates freedom in re-use and also ensures that Eur J Clin Exp Med content can be mined without barriers for the research needs.




