High protection of mice against Brucella abortus by oral immunization with recombinant probiotic Lactobacillus casei vector vaccine,expressing the outer membrane protein OMP19 of Brucella species

Brucellosis is a zoonotic disease threatening the public health and hindering the trade of animals and their products, which has a negative impact on the economic development of a country. Vaccination is the most effective way to control brucellosis. The recombinant vector vaccines are promising candidates for immunization in humans and animals. In this study, the gene encoding OMP19 antigen was primarily amplified and cloned into an expression vector called pT1NX, and then transformed to L. casei cell via electroporation technique. The expression was confirmed using specific antibody against the recombinant protein via immunological screening tests such as western blot and immunofluorescence assay. Finally, recombinant L. casei was orally fed to mice and the results were further recorded, indicating that the mice group which received OMP19 through L. casei based vaccine represented a very good general and mucosal immune responses protective against challenges with virulent B. abortus 544 strain compared with negative control recipient groups. Therefore, the vaccine produced in this research plan can be a very good candidate for protection against brucellosis.     

The protoxin Cry1Ac of Bacillus thuringiensis improves the protection conferred by intranasal immunization with Brucella abortus RB51 in a mouse model

Brucellosis is a zoonotic disease affecting many people and animals worldwide. Preventing this infection requires improving vaccination strategies. The protoxin Cry1Ac of Bacillus thuringiensis is an adjuvant that, in addition to increasing the immunogenicity of different antigens, has shown to be protective in different models of parasitic infections. The objective of the present study was to test whether the intranasal co-administration of pCry1Ac with the RB51 vaccine strain of Brucella abortus confers protection against an intranasal challenge with the virulent strain B. abortus 2308 in BALB/c mice. The results showed that co-administration of pCry1Ac and RB51, increased the immunoprotection conferred by the vaccine as evidenced by the following: (1) decrease of the splenic bacterial load when challenged intranasally with the virulent strain; (2) greater in vivo cytotoxic activity in response to the transference of previously infected cells; (3) further proliferation of cytotoxic TCD8+ cells in response to stimulation with heat-inactivated bacteria; (4) increased production of TNF-α and IFN-γ; and (5) significant IgG2a response. These results indicate that the use of the Cry1Ac protein as a mucosal adjuvant via the intranasal route can be a promising alternative for improving current RB51 vaccine against brucellosis.     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

布鲁氏菌病的研究进展

布病,尤其是由羊种布鲁氏菌引起的布病,目前是世界上最常见的一种人兽共患传染病,全世界每年报道的布病病例超过500万。布鲁氏菌已经被CDC列为B类病原体,成为潜在的生物武器。布鲁氏菌也是最常见的实验室源性病原体。布病的流行地域在不断变化,每年都有新发地和复发地的报道。除了那些牛布病已经根除的国家,布病仍然在许多国家的人和动物中存在。全世界由于布病给动物产品及公共卫生安全而带来的经济损失是巨大的。虽然许多有效的疫苗正在用于动物免疫,但是还没有理想的人用疫苗可以使用。动物布病的控制将显著降低人布病的发生。     

布鲁菌介导细胞自噬的研究进展

布鲁菌是布鲁菌病的病原体,在世界范围内给养殖业带来巨大损失。自噬是细胞的一种代谢方式,细胞在自噬相关基因的调控下清除细胞内病原微生物和吞噬降解受损、衰老细胞器及大分子物质,以维持机体内环境平衡。布鲁菌入侵细胞后,能够诱发细胞自噬,而这一复杂的过程需要很多细胞因子的参与,探究布鲁菌引发的细胞自噬已成为揭示布鲁菌致病机制的新热点,现就这一新热点的研究进展进行综述。     

布鲁氏菌病的研究与防控进展

布鲁氏菌病（Brucellosis,简称“布病”）是由布鲁氏菌（Brucella）引起的危害严重的人畜共患病,主要危害人畜生殖系统,人与人之间几乎不传播,患病动物是主要的传染源,近几年疫情急骤回升。作者综述了布病的研究现状及其流行趋势,初步探讨布病综合防控面临的主要困境。     

奶牛布鲁氏菌病的病原分离鉴定及血清学研究

通过细菌分离、动物试验、细菌学与血清抗体之间相互关系、温度与感染动物之间相互关系等分离一例奶牛布鲁氏菌。发现细菌学与血清抗体呈平行关系，试管凝集反应与补体结合反应存在一定差异，感染动物14天出现波浪热，持续到剖杀前(31天)。     

布鲁菌S2疫苗株免疫牛与牛种、羊种布鲁菌自然感染牛ELISA鉴别诊断方法的建立

为建立区分猪种布鲁菌S2疫苗株接种奶牛与布鲁菌自然感染奶牛,BLAST比对分析羊种、牛种、猪种、犬种、沙林鼠种和绵羊种6种布鲁菌基因序列,发现repA—related基因是猪种布鲁菌与牛种及羊种布鲁菌的差异基因。设计引物PCR扩增获得repA-related基因片段,克隆并原核表达得到了布鲁菌repA—related融合蛋白,以repArelated蛋白建立间接EI．IsA检测方法。用repA—related蛋白间接ELISA检测猪种s2疫苗株接种动物血清为阳性,检测牛种和羊种布鲁菌自然感染动物血清为阴性。repA—related蛋白间接EusA能从试管凝聚实验（SAT）及常规ELIsA检测阳性的奶牛血清样本中,区分出s2疫苗接种牛与牛种布鲁菌感染牛。     

奶牛四种流产性疾病的流行现状及疫苗研究进展

奶牛布氏杆菌病、传染性鼻气管炎、病毒性腹泻-黏膜病及新孢子虫病是导致我国奶牛流产发病比例升高的主要疾病,有效防控这四种疾病一直是世界关注的重点之一。本文对上述四种疾病的流行现状及疫苗研究进展进行了综述。     

Impact of geographical factors on the spread of animal brucellosis in the Republic of Kazakhstan

In Latin and Central America and in most Asian countries, brucellosis remains an insufficiently studied disease. This study aims to determine the national and regional incidence of brucellosis among cattle (cows) and small ruminants (sheep, goats) in the Republic of Kazakhstan, as well as to identify the effect of climatic and geographical factors on the incidence rates. Thematic maps were created in an open geographic information system QGIS version 2.8. in order to identify the natural and socio-economic factors that influence the spread of the disease overlay method was used. Local cluster analysis was used in order to identify additional causes of the disease. Findings show the following values of Pearson correlation between the overall population and the number of animals infected: 0.68 for cows, p ≤ 0.005, and 0.56 for sheep and goats, p ≤ 0.03. Thus, the larger the heard in a given area, the greater likelihood of having brucellosis. Data processing reveals that Kazakhstan has almost twice as many regions good for cattle breeding as regions that are good for the small ruminants farming. The correlation variables for cattle and small ruminants are approximately the same. On the basis of the performed research the author proposes to amend the accepted methodology of epidemiology surveillance by the methods based on spatial (geographical) analysis. It is also proposed to adjust the process of breeding cattle and small ruminants considering the additional health recommendations that take into account the geographical aspects of the spread of the disease.