---

Introduction: Brucellosis is one of the most important zoonotic diseases that causes miscarriage and infertility in animals and causes human fever. The use of the common SS9 strain of Brucella abortus has several side effects for livestock. Brucella P39 protein is one of the plasma peripheral space proteins that is considered as one of the important immunogenic indicators. With the production of the new protein combination of P39, more studies can be done on the ability of this protein to stimulate immune responses against Brucella. Therefore, in this research, the production and purification of this protein in Escherichia coli bacteria has been done as a new compound.
method: In this experimental study, using the polymerase chain reaction, the P39 gene was propagated by the bacterium Brucella abortus. After purifying the P39 gene, it was cloned into plasmid carriers pSK+ and pGEX4T1. Therefore, pSK+-P39 and pGEX4T1-P39 structures were prepared. To produce the recombinant protein P39, the plasmid structure pGEX4T1-P39 first entered the Escherichia coli bacterium BL21. The protein was then produced by IPTG by induction of pGEX4T1-P39 plasmid. The resulting protein was purified using the orderly purification protein glutathione S-transferase. The amount of purified protein was measured using the Brad Ford method.
Results: The nucleotide sequence of the gene propagated by the cloned PCR in the plasmid carrier  pSK+ was exactly the same as the P39 gene of Brucella abortus. Production of P39 protein was performed by induction of pGEX4T1-P39 plasmid. The purified protein content was 200 micrograms per milliliter.
Conclusion: The production of the new protein P39 compound Brucella Abortus, which is unstable in the cytoplasm of the Escherichia coli bacterium, is possible using carriers with additive proteins such as pGEX4T1 in the host of Escherichia coli strain BL21.

---

Introduction: In many studies, immunogenicity of Brucella proteins such as P39 in animals is investigated. In this study, we evaluated antigenicity of recombinant P39 from Brucella abortus in patients with Brucellosis. Materials and Methods: In this experimental study, at first recombinant P39 was produced in Escherichia coli. Sera reactivity of six infected individuals against the recombinant P39 protein was analysed by Western Blot. Results: Data indicated that P39 protein from Brucella abortus was recognized by patients, sera antibodies. Conclusion: Our data showed that recombinant P39 protein can be detected as an antigen by sera in infected human. Therefore, recombinant P39 have same epitopes with natural form of this antigen.

---

Background: Brucellosis is one of the most common bacterial zoonotic infections in the world. The incidence of this infection is quite high and is endemic in several countries. According to WHO report, the prevalence of zoonotic and human brucellosis is on the rise in the Mediterranean region, the Middle East, and west Asian countries. The aim of this study was to investigate the usage of silver nanoparticles in treatment of brucellosis. Materials and Methods: In this experimental study, the activity of silver nanoparticles against Brucella meltensis 16M was determined by agar well diffusion method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of silver nanoparticles were determined by macrodilution method. Also, the antibacterial effect of silver nanoparticles was studied in mouse model. Results: The results showed that silver nanoparticles in low concentrations can kill Brucella melitensis 16M in laboratory conditions. MIC and MBC of silver nanoparticles were 4 ppm and 6 ppm in macrodilution method, respectively. The anti-brucella effect of silver nanoparticles was also observed in mouse model. Conclusion: This study demonstrated that silver nanoparticles can be used against brucellosis.

---

Background: Brucellosis is caused by brucella which is a facultative intracellular pathogen invading both professional and nonprofessional phagosytic cells. Eucalyptus globulus is one of the most widely used medicinal plants in folk medicine throughout the world. The aim of this study was to evaluate the antibacterial effects of Eucalyptus globulus extracts on intramacrophage Brucella melitensis 16M. Materials and Methods: In this experimental study, after preparing aquatic, ethanolic, and acetonic extracts of Eucalyptus globules, the effect of the extracts on intramacrophge survival of B. melitensis 16M obtained from cell culture of Balb/c mice peritoneal macrophages was studied. In order to do this, after lysis of macrophages, through preparation of serial dilutions and culture on Mueller Hinton agar medium, the number of colonies grown was counted. Results: The maximum antimicrobial activity of Eucalyptus globulus extracts on intramacrophage B. melitensis 16 M were in 1:40 dilution (21.62 mg/ml) of the aquatic extract, 1: 640 dilution (1.26 mg/ml) of the ethanolic extract, and 1:320 dilution (2.59 mg/ml) of the acetonic extract after 24h. Conclusion: Aquatic, acetonic, and ethanolic extracts of Eucalyptus globulus possess antimicrobial properties against intramacrophage B. melitensis 16M and ethanolic extract has the most effective antimicrobial activity on intramacrophage Brucella melitensis therefore, these extracts can be useful in treatment of brucellosis.

---

Background: Brucellosis is a debilitative disease that imposes heavy costs on the economy and society. Therefore, using the most accurate and efficient method to diagnose this disease is essential. In Iran, Brucella melitensis is the common causative agent for brucellosis and BP26 protein of this bacterium has a good level of antigenicity. Thus, the aim of this study is to produce Brucella melitensis recombinant BP26 protein with a PET28a expression vector.

Materials and Methods: In this applied-fundamental study, genomic DNA was isolated from bacterial culture through proteinase K (pK) and phenol/chlorophorm protocol. Then, two pairs of primers were designed based on the known sequence in the gene bank for amplification of Brucella melitensis bp26 gene and PCR reaction was set up and optimized. The PCR product was cloned first into PTZ57R/T vector and accessed on the PET28a vector and sequenced. The recombinant vector was transformed and expressed into E. coli BL21 (DE3). Then, the recombinant protein was purified with Ni-NTA column of chromatography against His tag.

Results: The size of PCR product was in accordance with the part of bp26 gene size in the gene bank. The bp26 gene without adding IPTG had little expression and 3 hours after adding IPTG with a 1 Mm concentration to culture media, extreme expression was observed.

Conclusion: The production of recombinant BP26 protein from isolated Brucella melitensis native to Markazi province was done. Noticing the importance of BP26 protein and its significance for future studies on providing brucellosis diagnosis kits, its production was made possible.

---

Background: Brucellosis is one of the most important diseases among humans and animals. Clinical management of brucellosis due to an increased rate of treatment failure and recurrence is extremely worrying. The aim of this study was to determine the antimicrobial susceptibility pattern of the brucella isolates.

Materials and Methods: From April to September 2014 a total of 30 brucella isolates that were cultured on brucella agar has been studied. The species identification was carried out and to determine the effect of antibiotics on bacteria antibiogram testing was performed by disk diffusion.

Results: In this study, 30 brucella strains were isolated from cultured specimens and antibiogram testing was performed. All microbial positive specimens were sequenced by PCR. All isolates were Brucella melitensis. According to the tests, suceptibility to tetracycline, minocycline, gentamicin, tigecyclin was 100%, to doxycycline 93.3%, co-amoxiclave 66.7%, rifampin 44.7%, streptomycin 86.7%, ciprofloxacin 80%, cotrimoxazole 76.7% and ceftriaxone 73.3%.

Conclusion: This study shows that the predominant strain in our patients was Brucella melitensis. Also, due to high levels of resistance to rifampin to use the other effective drugs like gentamicin, streptomycin, ciprofloxacin or cotrimoxazole in combination with doxycycline or tetracycline.

---

Background: Brucella is an intracellular bacterium that causes chronic infection in humans and domestic animals. The underlying mechanisms that cause prolonged illness are complex and not fully understood. Immune responses may have an important role in the chronicity of infection. Here, we evaluated the lymphocyte proliferation responses in patients with chronic and acute brucellosis.

Materials and Methods: This descriptive - analytical study was performed on 22 patients with acute brucellosis, 21 patients with chronic brucellosis and 21 healthy people with the similar age, sex and genetic background as control group. Peripheral lymphocytes were isolated using Ficoll and the cellular proliferation was quantified in presence of antigen and phytohemaglutinin-A by MTT method.

Results: The brucella antigen-specific stimulation index in patients with chronic brucellosis was significantly lower than the acute brucellosis patients (p=0.001). Also, stimulating the lymphocytes with phytohemaglutinin-A has shown that proliferative response in patients with chronic brucellosis was lower than the other groups (p=0.04).

Conclusion: The results indicated that chronic brucellosis inhibits lymphocyte proliferation. This inhibition of lymphocyte proliferation may be due to the induction of anergy.