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Background: Brucellosis is one of the most prevalent and common diseases between humans and animals.Prompt diagnosis and timely treatment of this disease can prevent many complications. In this regard, this study aims to comparatively evaluate ELISA, PCR and serological methods to identify Brucella abortus. Methods: In this study, the serum of 100 patients referred to Tonekabon private laboratory from July 2020 to January 2021 was examined by PCR, ELISA and Wright, Coombs Wright, and 2ME methods for the detection of Brucella abortus. Results: In this study, the mean age of the sample was 43.3 ± 18.2 of which 21% were infected with Brucella abortus according to the above serological methods. According to ELISA test, 22% of the samples were IgM, 6% of the samples were IgG and 16% were PCR positive. Kappa agreement coefficient in Wright and Coombs Wright test and 2me were significant (P

Brucellosis is one of the most common zoonoses in the Middle East. It is causing economic losses to the livestock industry and has a great public health concern. Little is known about the genetic diversity and distribution of brucellae in Iran. Therefore, forty Brucella spp. strains (B. abortus and B. melitensis) isolated from animals and humans were analyzed by whole genome sequencing (WGS) technology using single nucleotide polymorphism (SNP) analysis and core genome multilocus sequence typing (cgMLST). Brucella isolates were obtained from lymph nodes (cows and camels), milk (cows, camels and sheep), and aborted foetus samples (sheep and goats), as well as cerebrospinal fluid and blood of humans. The isolates were originating from thirteen provinces of Iran and isolated between 2015 and 2020. According to in-silico MLST, ST8 and ST2 were the most frequent sequence types in B. melitensis and B. abortus, respectively. Based on phylogeographic reconstruction using cgSNP analysis, the investigated Iranian B. melitensis strains belonged to the American and Mediterranean lineages of the B. melitensis phylogeny. Furthermore, cgSNP analysis revealed a similarity between Iranian B. abortus isolates and strains from Iraq and Egypt. Therefore, the origin of the Iranian strains can be suggested to be strains from neighboring and Middle East countries. Moreover, cgMLST analysis showed that the Iranian B. melitensis strains were closely relative to strains recovered from sheep and humans in Iraq, Afghanistan, Syria, Turkmenistan, and Pakistan. In the current panel of strains, cgMLST and cgSNP analysis provided an appropriate and accurate tool for effective traceback analyses for Brucella spp. from Iran. The results of cgSNP and cgMLST helped to understand the geographic distribution and interspecies transmission of Iranian strains and highlight the importance of specific brucellosis control measures in Iran with regard to the One-Health approach.

Abstract Background Brucellosis is an endemic zoonotic disease with rising health and economic concerns in many areas worldwide. Musculoskeletal pains are among the main complications of human brucellosis, which are often difficult to diagnose due to the variability of clinical symptoms. Brucellar discitis is a very disabling problem in some chronic forms of the disease which may lead to serious vertebral and neurological consequences. Case presentation In this case report, we reported the isolation of Brucella abortus from lumbar disc bulging in a woman who had rheumatoid arthritis and diabetes mellitus as underlying conditions. The patient had several negative brucellosis serological tests and dorsolumbar pains with urinary incontinence over a 2-month period. The diagnosis was confirmed by magnetic resonance imaging (MRI) examination of lumbar spine as well as disc culture. MRI examination was performed without intravenous contrast and revealed the presence of disc bulging, left foraminal narrowing at L5-S1, left foraminal narrowing, anterolisthesis grade II at L4-L5. The diagnosis was also confirmed by isolation of B. abortus biovar 1 from bulging disc culture. The isolate was characterized by AMOS PCR, Bruce-ladder PCR and biotyping, resulting in the identification of B. abortus from L4-L5 and L5-S1 disc bulging regions. The patient was treated with two drugs i.e. doxycycline and rifampin for 3 months. In the follow-up, in addition to improving the patient’s general condition, low-back pain was also significantly reduced. Conclusions MRI, serology, cultural and molecular test along with patient history are important to make a rapid diagnosis of brucellosis’ discitis, thereby decreasing the delay for the brucellosis treatment. The present report suggests that the infection by Brucella spp. should be fundamentally considered among the causative agents of back pain especially in the endemic areas of Brucella infections.

Bovine brucellosis, an infectious disease transmitted by Brucella melitensis and Brucella abortus, presents a significant zoonotic risk for agricultural economics and animal health. The primary objective of this study was to present a comprehensive understanding of the prevalence and features of Brucella strains within the industrial dairy farming sector in Iran. Rose Bengal plate test, standard agglutination test, and indirect enzyme linked immunosorbent assay tests were used to confirm all seropositive animals. A total number of 1,311 bovine samples from seropositive animals including were collected from 224 farms in 21 provinces of different regions of Iran and examined. The discovered Brucella isolates were phenotyped and molecularly characterized. The isolates were all B. abortus or B. melitensis. Bacteria analysis revealed that 70.53% of seropositive farms were tested positive for Brucella strains, predominantly B. melitensis biovar 1 (43.42%) and B. abortus biovar 3 (27.11%). Geographical distribution revealed that B. melitensis biovar 1 was the most common in dairy cow farms (16 provinces), followed by B. abortus biovar 3 (six provinces). Also, the prevalence of B. melitensis biovar 2, B. melitensis biovar 3, B. abortus biovar 1, B. abortus biovar 2 and RB51 vaccine were restricted to certain provinces. AMOS (abortus melitensis ovis suis)- polymerase chain reaction and Bruce-ladder PCR confirmed species identification. These results highlighted the complexity of bovine brucellosis in Iran and illustrated that B. melitensis was spread from small ruminants to cattle. This study provided important epidemiological insights for targeting future brucellosis control programs in the Iranian dairy farms. [ABSTRACT FROM AUTHOR]

Brucellosis is a neglected zoonotic disease with significant public health impacts. It is primarily associated with classic Brucella species in mammals, but recent research has shown atypical Brucella strains in various hosts, including reptiles. Despite the known physiological differences between reptiles and humans, the rise in invasive reptile populations and other factors make reptiles and the pathogens they carry a potential public health concern. This study collected 90 venomous snakes from different regions in Iran and examined their liver samples, buccal, and cloaca cavities for Brucella spp. The samples underwent bacteriological analysis and DNA extraction. PCR amplification of the Omp28 gene and IS711-based PCR were used for the molecular identification of Brucella species. Subsequently, the Omp28 gene sequences were determined, and a phylogenetic tree was created. Brucella abortus was detected in the liver tissue of two snake species, namely Pseudocerastes persicus and Vipera albicornuta, at a prevalence rate of 2.2%. This finding is exceptional, as it has not been previously documented in venomous snake populations in the wild. The study confirms the presence of Brucella spp. in venomous snakes, suggesting the potential for zoonotic transmission. This discovery raises questions about the evolutionary forces shaping Brucella populations in reptiles. Understanding the prevalence and impact of Brucella in snakes is essential for wildlife conservation and assessing zoonotic risks. Proper safety measures are crucial when handling reptiles that may carry Brucella. Further research is needed to fill knowledge gaps regarding Brucella in these snake species and its potential zoonotic implications. [ABSTRACT FROM AUTHOR]

Berberis integerrima Bonge. (Syn: Berberis densiflora Boiss. & Buhse) is a shrub widely distributed in Middle East and central part of Asia. An ethnobotanical study revealed that indigenous and tribal people in Iran use B. integerrima root decoction for treatment of brucellosis. Therefore, the aim of this study was bioassay directed isolation of antibacterial compounds from this plant based on their in vitro bactericidal activity against Brucella abortus. Briefly, the ethanol extract of B. integerrima was fractioned and subjected to preliminary antibacterial screening tests against Brucella. The more active fraction (Fr.3) was subjected to purification by repeated chromatography systems. Quaternary benzylisoquinoline alkaloids including columbamine, palmatine, berberine, and jatrorhizine were four main components identified in the selected active fraction. Except for berberine which is reported before, palmatine, columbamine and jatrorhizine are isolated for the first time from this plant. Anti-brucellosis properties of isolated compounds 1-4 were studied against B. abortus under different test conditions. In minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) results, jatrorhizine (4) showed more antibacterial activity with MIC and MBC of 0.78 and 1.56 μg/mL, respectively. In both agar well diffusion and disk diffusion ANOVA results showed that there were statistically significant differences between compounds 1-4 versus placebo in all of the tested concentration (P

Background: Brucellosis is one of the most prevalent and common diseases between humans and animals.Prompt diagnosis and timely treatment of this disease can prevent many complications. In this regard, this study aims to comparatively evaluate ELISA, PCR and serological methods to identify Brucella abortus. Methods: In this study, the serum of 100 patients referred to Tonekabon private laboratory from July 2020 to January 2021 was examined by PCR, ELISA and Wright, Coombs Wright, and 2ME methods for the detection of Brucella abortus. Results: In this study, the mean age of the sample was 43.3 ± 18.2 of which 21% were infected with Brucella abortus according to the above serological methods. According to ELISA test, 22% of the samples were IgM, 6% of the samples were IgG and 16% were PCR positive. Kappa agreement coefficient in Wright and Coombs Wright test and 2me were significant (P <0.001). Serological diagnostic indices and ELISA sensitivity were 68.75% and 68.75%, respectively. The lowest prediction rate of Brucella abortus among diagnostic methods was related to Elisa (IgM). Based on Fisher's exact test, there was no significant relationship between the percentage of Brucella abortus positive PCR cases and age, sex, previous history of infection with Brucella, fever, body aches and dairy consumption. Conclusion: Based on the results of our study, the accuracy of all methods is comparative and the lowest accuracy is related to Elisa (IgM) which has a lower level of predictability than other methods. The highest level of prediction belonged to Wright and 2me tests. [ABSTRACT FROM AUTHOR]

Brucellosis is a zoonotic infectious disease in western regions of Iran, especially in Hamedan province. Following the Famenin brucellosis cohort study, the main aim of the current study was the molecular detection of Brucella species (spp.) in sheep and goats from Famenin, Hamedan, Iran. A total of 23 Brucella-seropositive samples (sheep=21 and goats=2), which had been screened from 1,660 animals in the Famenin cohort study, were used to detect Brucella-DNA using the BCSP31 target gene and IS711 locus. In total, 20 of 23 samples were positive for Brucella infection by using specific primers. Additionally, Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) were confirmed in 90% (n=18) and 10% (n=2) of positive samples, respectively. There was no sample with the co-infection of B. abortus and B. melitensis. In this study, B. abortus was isolated from one of the goat samples. This is the first report on Brucella spp. in animals in the region. It was found that B. melitensis is the dominant spp. responsible for brucellosis in animals from Famenin. Molecular techniques are reliable tools to detect Brucella infection, especially in cases without serology findings and conclusive results. [ABSTRACT FROM AUTHOR]

Brucellosis is a zoonotic bacterial disease that causes enormous economic losses in livestock populations and severe debilitation in humans globally. This study analyzes the seroprevalence of human brucellosis in Iran from 1970 to 2023, revealing key epidemiological trends based on data from 20,046 individuals. Through serological, culture, and molecular tests, it identifies the Brucella species affecting humans over five decades. The findings emphasize the need for ongoing monitoring and improved disease control efforts. Subsequently, we cultured specimens from seropositive individuals who exhibited suspected symptoms of brucellosis. The specimens included blood (n = 9017), placentas (n = 203), bone marrow (n = 221), aborted fetuses (n = 70), cerebrospinal fluid (CSF) (n = 62), synovial fluid (n = 5), urine (n = 4), sperm (n = 3), lumbar discs (n = 3), abscesses (n = 2), milk (n = 2), and vertebral bodies (n = 1). The overall seroprevalence of brucellosis in Iran was 44.9 % based on RBPT, 20.7 % by SAT, and 8.7 % by 2-ME tests. The highest SAT prevalence occurred in the 2000s (25.8 %), followed by the 1990s (23.8 %), 1980s (22.2 %), 1970s (14.8 %), and 2010s (10.4 %). Two main Brucella species were identified in infected humans as Brucella melitensis and Brucella abortus. Positive culture rates varied by sample type, with 9.9 % for blood, 10.4 % for bone marrow, and up to 100 % for fluids like urine and milk. The most common strain was B. melitensis biovar 1 (89.2 %), while B. abortus biovar 1 had a much lower incidence (0.3 %). This investigation showed that the highest rates of culture positivity were in samples of urine, synovial fluid, abscess, and milk. These findings suggest that B. melitensis continues to be an important public health concern and that recent years of decline may be due to better control measures of the disease. The combined use of serology and culture is recommended for diagnosing brucellosis. Increased awareness among farmers and consumers of raw dairy products, coupled with nationwide control measures, is crucial for combating zoonotic brucellosis. • A comprehensive seroprevalence rate of 44.9 % by RBPT, 20.7 % by SAT, and 8.7 % by 2-ME. • The highest seroprevalence occurred in the 2000s at 25.8 %, with a subsequent decline to 10.4 % in the 2010s. • Positive culture rates of 9.9% in blood, with higher detection rates in CSF (14.5%) and synovial fluid (60%) was reported. [ABSTRACT FROM AUTHOR]

Background: Brucellosis is an important zoonotic disease in Iran. There are several reservoirs for this disease in nature. There is little information in this regard in Iran. Aims: The present study investigated the prevalence and diversity of Brucella species in ovine reticuloendothelial organs including liver, spleen, intestine (mesenteric lymph nodes), and lung. This help to address the ability of the reservoir species in disease transmission to other animals through these organs. Methods: A total of 200 ovine visceral organs including 44 intestines, 51 lungs, 52 spleens, and 52 livers were collected. The presence of different Brucella species was studied using the polymerase chain reaction (PCR). Results: The results demonstrated the prevalence of Brucella species in 56 (28.4%) samples. In addition, liver and intestine were the most and the least infected organs, respectively. In general, 48 out of 56 positive samples were infected with Brucella abortus, while 10 samples were infected with Brucella melitensis. Conclusion: Eventually, the results approved the possible presence of B. abortus among domestic animals, especially sheep and cattle, and highlighted the role of sheep as reservoir hosts for its potential transfer to cattle. [ABSTRACT FROM AUTHOR]

Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Berberis integerrima Bonge. (Syn: Berberis densiflora Boiss. & Buhse) is a shrub widely distributed in Middle East and central part of Asia. An ethnobotanical study revealed that indigenous and tribal people in Iran use B. integerrima root decoction for treatment of brucellosis. Therefore, the aim of this study was bioassay directed isolation of antibacterial compounds from this plant based on their in vitro bactericidal activity against Brucella abortus. Briefly, the ethanol extract of B. integerrima was fractioned and subjected to preliminary antibacterial screening tests against Brucella. The more active fraction (Fr.3) was subjected to purification by repeated chromatography systems. Quaternary benzylisoquinoline alkaloids including columbamine, palmatine, berberine, and jatrorhizine were four main components identified in the selected active fraction. Except for berberine which is reported before, palmatine, columbamine and jatrorhizine are isolated for the first time from this plant. Anti-brucellosis properties of isolated compounds 1-4 were studied against B. abortus under different test conditions. In minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) results, jatrorhizine (4) showed more antibacterial activity with MIC and MBC of 0.78 and 1.56 μg/mL, respectively. In both agar well diffusion and disk diffusion ANOVA results showed that there were statistically significant differences between compounds 1-4 versus placebo in all of the tested concentration (P <0.001). In conclusion, all of four alkaloids showed potent antibacterial activity against B. abortus but jatrorhizine and columbamine with free hydroxyl group on C-3 or C-2 showed more activity than palmatine and berberine without any free hydroxyl group on their structures. The antibacterial effects of columbamine (15 μg/mL) and jatrorhizine (15 μg/mL) were comparative to streptomycin (10 μg/mL) as standard drug which candidate them for more pharmacological researches to find new antibacterial agents against brucellosis. [ABSTRACT FROM AUTHOR]

Objectives: Brucellosis is a major zoonotic disease that poses a significant public health threat worldwide. The classical bacteriological detection process used to identify Brucella spp. is difficult and time-consuming. This study aimed to develop a novel molecular assay for detecting brucellosis. Methods: All complete sequences of chromosome 1 with 2.1-Mbp lengths were compared among all available Brucella sequences. A unique repeat sequence (URS) locus on chromosome 1 could differentiate Brucella abortus from Brucella melitensis. A primer set was designed to flank the unique locus. A total of 136 lymph nodes and blood samples were evaluated and classified by the URS-polymerase chain reaction (PCR) method in 2013-2014. Results: Biochemical tests and bacteriophage typing as the golden standard indicated that all Brucella spp. isolates were B. melitensis biovar 1 and B. abortus biovar 3. The PCR results were the same as the bacteriological method for detecting Brucella spp. The sensitivity and specificity of the URS-PCR method make it suitable for detecting B. abortus and B. melitensis. Conclusion: Quick detection of B. abortus and B. melitensis can provide the most effective strategies for control of these bacteria. The advantage of this method over other presented methods is that both B. abortus and B. melitensis are detectable in a single test tube. Furthermore, this method covered 100% of all B. melitensis and B. abortus biotypes. The development of this URS-PCR method is the first step toward the development of a novel kit for the molecular identification of B. abortus and B. melitensis. [ABSTRACT FROM AUTHOR]

Brucellosis is a world prevalent endemic illness that is transmitted from domestic animals to humans. Brucella spp. exploits urease for survival in the harsh conditions of stomach during the gastrointestinal infection. In this study, we examined the immune response and the protection elicited by using recombinant Brucella urease ( rUrease) vaccination in BALB/c mice. The urease gene was cloned in pET28a and the resulting recombinant protein was employed as subunit vaccine. Recombinant protein was administered subcutaneously and intraperitoneally. Dosage reduction was observed with subcutaneous ( SC) vaccination when compared with intraperitoneal ( IP) vaccination. rUrease induced mixed Th1-Th2 immune responses with high titers of specific IgG1 and IgG2a. In lymphocyte proliferation assay, splenocytes from IP and SC-vaccinated mice displayed a strong recall proliferative response with high amounts of IL-4, IL-12 and IFN-γ production. Vaccinated mice were challenged with virulent Brucella melitensis, B. abortus and B. suis. The SC vaccination route exhibited a higher degree of protection than IP vaccination (p value ≤ 0.05). Altogether, our results indicated that rUrease could be a useful antigen candidate for the development of subunit vaccines against brucellosis. [ABSTRACT FROM AUTHOR]

• The first comprehensive molecular typing of Brucella spp. infecting humans and livestock in Iran is presented. • Twenty –three Brucella isolates were characterized by phenotypic typing, AMOS-PCR, Bruce-ladder and MLST. • B. melitensis isolates belonged to 3 MLST-9 genotypes ST8, ST7, and ST102 – the latter previously reported only once, from a traveler with links to Iran. • Two widely dispersed MLST-9 genotypes were identified from B. abortus isolates (ST1 and ST2). • The study provides the first data describing how Brucella isolates from Iran relate to the global population structure. Brucellosis is an endemic infection in Iran and represents a serious health problem in humans and livestock causing important economic losses. The objective of this study was to undertake molecular characterization of Brucella spp. isolated from humans and livestock in several provinces of Iran including by multi-locus sequence typing (MLST), in order to understand the genotypes circulating in Iran and their relationship to genotypes globally. A total of 23 Brucella isolates were isolated from eight milk samples (seven cows, and one camel), human blood samples (seven), bovine lymph nodes (two), and samples from aborted fetuses (three sheep, two cows, and one goat). Phenotypic and molecular identification of Brucella isolates was performed on all isolated bacteria and showed that all were either Brucella melitensis or Brucella abortus. B. melitensis was associated with ovine/caprine and camel samples, most human isolates, and a significant minority of cattle isolates. In contrast B. abortus from livestock was associated only with isolations from bovine samples, as well as a single human sample. These results indicate that both B. melitensis and B. abortus contribute to the human brucellosis burden in Iran. B. melitensis isolates comprised three MLST-9 genotypes, the common and globally distributed ST8, a single representative of ST7, and several additional examples of ST102, a genotype previously only reported in a single isolate from a human brucellosis case believed to be acquired through travel to Iran. B. abortus isolates represented two globally common MLST-9 genotypes (ST1 and ST2), with relationships to biotype and other PCR-based typing methods consistent with previous observations. The results provide the basis for further studies examining the molecular epidemiology of Brucella circulating in Iran and the relationships of local isolates to those present globally. [ABSTRACT FROM AUTHOR]

Brucella spp. commonly infect humans in various regions worldwide. Human brucellosis mainly spreads through the consumption of contaminated raw dairy products and meat from domestic livestock (water buffalo, goats, sheep, cattle, pigs and camels). In this regard, the origin and routes of transmission of this bacterium should be carefully determined in order to control the source of infection. This study aimed to evaluate the rate of Brucella spp. contamination of camel milk samples sent for analysis to the national brucellosis laboratory during 2018 in Iran. For this purpose, 96 milk samples from 96 dairy camel herds were randomly collected from two provinces and investigated for the presence of Brucella spp contaminations by both bacterial culture method and polymerase chain reaction (PCR). No clinical manifestation of brucellosis was reported in camels from which milk samples were collected. Using the culture method, three milk samples (3%) originating from two camels of Isfahan province (4%) and one camel from the Semnan province (2%), were contaminated with Brucella abortus. According to PCR analyses, B. abortus gene was detected in 14 (14.5%) milk samples, including 9 and 5 samples from Isfahan (18%) and Semnan (11%) province, respectively. PCR method revealed significant differences (p = 0.02) in the level of contamination with B. abortus between milk samples collected from two regions. These results represent the first report regarding the isolation of B. abortus from raw camel milk in Iran and highlight the importance to screen apparent healthy camels. Therefore, the consumption of raw camel milk may contribute to the spread of human brucellosis in endemic regions. [ABSTRACT FROM AUTHOR]

A zoonotic disease called brucellosis can cause flu-like symptoms and heart inflammation. The bacteria responsible for this disease can also enter the brain, causing a condition called neurobrucellosis that can result in long-term neurological problems. In this study, researchers aimed to determine the changes in the hippocampal cells of rats infected with Brucella. For the study, 24 adult male albino rats were inoculated with 1 × 106 CFU Brucella abortus 544. The rats were then deeply anesthetized, and their hippocampus samples were taken for stereological, histological, and molecular studies. The results showed that the infected rats had increased microgliosis and astrogliosis. Furthermore, a high level of caspase-3 in their hippocampal tissue indicated their susceptibility to apoptosis. Additionally, there was a decrease in expression of Ki67, which further supported this. Sholl's analysis confirmed a significant failure in glial morphology. The study demonstrated that the pathogen has the ability to destroy the hippocampus and potentially affect its normal physiology. However, more research is needed to clarify various aspects of neurobrucellosis. • Brucella increases astrogliosis and changes the morphological characteristics of astrocytes in the hippocampus. • Brucella Increases microgliosis in rats' hippocampus and alters microglia cells' morphological characteristics. • Brucella infection induces the expression of caspase-3 in Pyramidal cells and reduces neurogenesis in the hippocampus. [ABSTRACT FROM AUTHOR]

Introduction: Brucella spp. are gram-negative, facultative intracellular bacteria pathogens responsible for brucellosis, a zoonotic disease that can cause abortion, fetal death, and genital infections in animals and undulant fever in humans. Lipopolysaccharide (LPS) is known as a major virulence factor of Brucella spp. The wboA gene is capable of encoding a glycosyltransferase that appears to play a major role in LPS biosynthesis. Hence, the characterization of this gene can help in the clarification of the pathogenicity of Brucella spp. Methods: This study was carried out at Razi Vaccine and Serum Research Institute in 2011. Briefly, the wboA gene in B. abortus biovar 3 and B. melitensis biovar 1, the predominant biovars in Iran, were amplified by using two pairs of specific primers. Polymerase chain reaction (PCR) products were cloned into a thymine-adenine (TA) cloning vector and transformed into an E. coli DH5α before being sequenced. Multiple alignments of identified sequences were performed, with all wboA sequences deposited in the GenBank sequence database. Results: This study showed that a mismatch has occurred in B. melitensis biovar 1; this biovar is predominant in Iran. In contrast, the wboA gene from B. abortus biovar 3 was similar to that of other B. abortus variations. Conclusion: The comparison and alignment of the wboA gene of native Brucella strains in Iran to all wboA sequences deposited in GenBank revealed that the wboA gene has changed in the long term; hence, because of its unique nucleotide pattern, the gene can be used for specific diagnosis of B. abortus and B. canis. [ABSTRACT FROM AUTHOR]