winter season in South Korea, several outbreaks of highly pathogenic avian infl uenza virus (H5N1) were confi rmed among domestic poultry and in migratory bird habitats. Phylogenetic analysis showed that all isolates were closely related and that all belong to the A/ bar-headed goose/Qinghai/5/2005-like lineage rather than the A/chicken/Korea/ES/2003-like lineage.

ABSTRACT:The prevalence of brucellosis was determined in the ruminants (buffaloes, cattle, sheep and goats) of five different districts viz. Bagerhat, Bogra, Gaibangha, Mymensingh and Sirajgonj of Bangladesh. A total of 550 sera samples of 105 buffaloes, 188 cattle, 127 goats and 130 sheep were screened by RBT and were further confirmed with I-ELISA. A structured questionnaire was used to collect epidemiological information on the animals. The overall serological prevalence derived from the samples was 2.87% in buffaloes, 2.66% in cattle, 3.15% in goats, and 2.31% in sheep. The prevalence was relatively higher in females than that in males in cattle, goats and sheep but, an insignificantly higher prevalence was observed in males than that in females in the case of buffalo. A significant association was found between abortion or age and occurrence of brucellosis (P < 0.01). The results of the study provide (a) a comparison of the prevalence of brucellosis in different livestock species in Bangladesh, (b) constitute baseline data for further study of Brucella infections, and (c) are a starting point for the control of brucellosis.

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Two new primer sets of a 766-and a 344-bp fragment were introduced into the conventional Bruce-ladder PCR assay. This novel multiplex PCR assay rapidly and concisely discriminates Brucella canis and Brucella microti from Brucella suis strains and also may differentiate all of the 10 Brucella species.The alphaproteobacterial genus Brucella consists of 10 species: B. abortus, B. canis, B. suis, B. ovis, B. neotomae, B. melitensis, B. ceti, B. pinnipedialis, B. microti, and B. inopinata (3, 5, 16, 17). Brucella species show a host preference, but some strains can be transmitted among a variety of animals, including humans (11,12,14,18).To accelerate effective prevention and control of brucellosis, a fast and accurate identification method is necessary. Many studies have developed PCR-based assays for the discrimination of Brucella species (2,4,7,8,10). Recently, López-Goñi et al. reported that the Bruce-ladder PCR assay could differentiate most Brucella species, including marine mammal and vaccine strains B. abortus S19, B. abortus RB51, and B. melitensis Rev.1 (6, 10, 13). However, these assays did not solve the problem of erroneous identification of B. canis isolates (such as B. suis) and basic differentiation between two marine mammal Brucella species (B. ceti and B. pinnipedialis) (9, 13, 15). Therefore, the aim of this study was to develop a fast, simple, and accurate one-step multiplex PCR assay to differentiate 10 Brucella species using 22 reference strains and 106 field isolates in South Korea.To discriminate between B. canis and B. suis, alignments of their whole-genome sequence or each biovar short-gun

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BackgroundA Brucella eradication program has been executed in Korea. To effectively prevent and control brucellosis, a molecular method for genetic identification and epidemiological trace-back must be established. As part of that, the MLVA typing assay was evaluated and applied to B. abortus isolates for analyzing the characteristics of the regional distribution and relationships of foreign isolates.ResultsA total of 177 isolates originating from 105 cattle farms for the period 1996 to 2008 were selected as representatives for the nine provinces of South Korea. A dendrogram of strain relatedness was constructed in accordance with the number of tandem repeat units for 17 loci so that it was possible to trace back in the restricted areas. Even in a farm contaminated by one source, however, the Brucella isolates showed an increase or decrease in one TRs copy number at some loci with high DI values. Moreover, those 17 loci was confirmed in stability via in-vitro and in-vivo passage, and found to be sufficiently stable markers that can readily identify the inoculated strain even if minor changes were detected. In the parsimony analysis with foreign Brucella isolates, domestic isolates were clustered distinctively, and located near the Central and Southern American isolates.ConclusionThe MLVA assay has enough discrimination power in the Brucella species level and can be utilized as a tool for the epidemiological trace-back of the B. abortus isolates. But it is important to consider that Brucella isolates may be capable of undergoing minor changes at some loci in the course of infection or in accordance with the changes of the host.

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