Isolation and molecular identification of Brucella spp. in bovine herds kept at householders in the Delta region of Egypt by MALDI-TOF and AMOS-PCR

Supplementary Files

Table I
Table II


Cattle and buffaloes
Brucella spp.

How to Cite

Abd Eltawab, A., El-Hofy, F., Hamdy, M., Moustafa, S., Soliman, E., Ahmed, W., Ramadan, M., & Wareth, G. (2020). Isolation and molecular identification of Brucella spp. in bovine herds kept at householders in the Delta region of Egypt by MALDI-TOF and AMOS-PCR. Veterinaria Italiana, 56(4), 297-300.


Brucellosis is a widespread disease in Egypt which cause huge economic losses in the dairy industry. The present study aims at isolating and identifying Brucella (B.) spp. circulating in bovine and buffalo dairy herds kept at farmers houses in four districts of the Delta region of Egypt. One hundred and five tissue specimens were collected from seropositive cattle and buffaloes. The samples included 10 vaginal swabs, 3 placentas, 3 uteri and 86 supra-mammary lymph nodes from dams, as well as 3 stomach contents from aborted fetuses. Matrix-assisted laser desorption ionization (MALDI) and the conventional biotyping techniques were used for preliminary identification of isolates into the genus level. AMOS-PCR was applied to differentiate Brucella isolates into species level. Nineteen Brucella strains have been identified, four B. abortus strains were recovered from cattle and 15 B. melitensis strains were isolated from both cattle (n = 8) and buffaloes (n = 7). The predominant occurrence of B. melitensis in bovines raises the fact that B. melitensis clone can cross species barriers and can establish a permanent reservoir in cattle and buffaloes. Presence of culture-positive animals at householders represent a high-risk factor for human infection. This knowledge is of significant importance in the control of brucellosis in bovines.


Abd-El Halim M., Mohamed A. & Shalaby N. 2017. Prevalence of brucellosis in buffaloes and its control measures. Journal of Veterinary Medical Research, 24, 161-170.

Al Dahouk S., Tomaso H., Prenger-Berninghoff E., Splettstoesser W.D., Scholz H.C. & Neubauer H. 2005. Identification of Brucella species and biotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Crit Rev Microbiol, 31, 191-196.

Alton G.G., Jones L.M., Angus R. & Verger J. 1988. Techniques for the brucellosis laboratory. Institut National de la recherche Agronomique (INRA).

Alvarez J., Saez J.L., Garcia N., Serrat C., Perez-Sancho M., Gonzalez S., Ortega M.J., Gou J., Carbajo L., Garrido F., Goyache J., & Dominguez L. 2011. Management of an outbreak of brucellosis due to B. melitensis in dairy cattle in Spain. Research in veterinary science,90, 208-211.

Bricker B. & Halling S. 1994. Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR. Journal of clinical microbiology, 32, 2660-2666.

Chiliveru S., Appari M. & Suravajhala P. 2015. On Brucellapathogenesis: looking for the unified challenge in systems and synthetic biology. Systems and synthetic biology,9, 73-75.

Croxatto A., Prod'hom G. & Greub G. 2012. Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology. FEMS microbiology reviews,36, 380-407.

Ducrotoy M., Bertu W., Ocholi R., Gusi A., Bryssinckx W., Welburn S. & Moriyón I. 2014. Brucellosis as an emerging threat in developing economies: lessons from Nigeria. PLoS neglected tropical diseases,8, e3008. DOI:3010.1371/journal.pntd.0003008.

El-Diasty M., Wareth G., Melzer F., Mustafa S., Sprague L.D. & Neubauer H. 2018. Isolation of Brucella abortus and Brucella melitensis from seronegative cows is a serious impediment in brucellosis control. Vet Sci, 5 (1), 28. doi: 10.3390/vetsci5010028.

Fretin D., Whatmore A.M, Al Dahouk S., Neubauer H., Garin-Bastuji B., Albert D., Van Hessche M., Ménart M., Godfroid J., Walravens K. & Wattiau P. 2008. Brucella suis identification and biovar typing by real-time PCR. Veterinary microbiology, 131, 376-385.

Galinska E.M. & Zagorski J. 2013. Brucellosis in humans - etiology, diagnostics, clinical forms. Annals of agricultural and environmental medicine: AAEM, 20, 233-238.

Godfroid J., Scholz H., Barbier T., Nicolas C., Wattiau P., Fretin D., Whatmore A., Cloeckaert A., Blasco J. & Moriyon I. 2011. Brucellosis at the animal/ecosystem/human interface at the beginning of the 21st century. Preventive veterinary medicine, 102, 118-131.

Hassanain N.A. & Ahmed W.M. 2012. Sero-prevalence of brucellosis in Egypt with emphasis on potential risk factors. World Journal of Medical Sciences, 7, 81-86.

Hosein H.I., Zaki H.M., Safwat N.M., Menshawy A.M.S., Rouby S., Mahrous A. & Madkour B.E. 2018. Evaluation of the General Organization of Veterinary Services control program of animal brucellosis in Egypt: an outbreak investigation of brucellosis in buffalo. Veterinary world, 11, 748-757.

Matope G., Bhebhe E., Muma J.B., Skjerve E. & Djønne B. 2009. Characterization of some Brucella species from Zimbabwe by biochemical profiling and AMOS-PCR. BMC research notes, 2, 261.

Murugaiyan J., Walther B., Stamm I., Abou‐Elnaga Y., Brueggemann‐Schwarze S., Vincze S., Wieler L., Lübke‐Becker A., Semmler T. & Roesler U. 2014. Species differentiation within the Staphylococcus intermedius group using a refined MALDI‐TOF MS database. Clinical Microbiology and Infection,20, 1007-1015.

Pappas G., Papadimitriou P., Akritidis N., Christou L. & Tsianos E.V. 2006. The new global map of human brucellosis. Lancet Infect Dis, 6, 91-99.

Radostits O., Gay C., Blood D. & Hinchcliff K. 2000. A textbook of the diseases of cattle, sheep, pigs, goats, and horses. WB Saunders, London, 1329-1337.

Refai M. 2002. Incidence and control of brucellosis in the Near East region. Veterinary microbiology,90, 81-110.

Wareth G., Hikal A., Refai M., Melzer F., Roesler U. & Neubauer H. 2014a. Animal brucellosis in Egypt. J Infect Dev Ctries,8, 1365-1373.

Wareth G., Melzer F., El-Diasty M., Schmoock G., Elbauomy E., Abdel-Hamid N., Sayour A. & Neubauer H. 2017. Isolation of Brucella abortus from a dog and a cat confirms their biological role in re-emergence and dissemination of bovine brucellosis on dairy farms. Transbound Emerg Dis,64, e27-e30.

Wareth G., Melzer F., Elschner M.C., Neubauer H. & Roesler U. 2014b. Detection of Brucella melitensis in bovine milk and milk products from apparently healthy animals in Egypt by real-time PCR. J Infect Dev Ctries,8, 1339-1343.