PCR–RFLP of outer membrane proteins gene of 
Dichelobacter nodosus: a new tool in the epidemiology of 
footrot

Currently only phenotypic epidemiological markers, serogrouping and virulence testing of Dichelobacter nodosus, are available for investigating footrot outbreaks in small ruminants. These methods have limitations in tracing the source of infection. In this study, a genotypic marker, PCR–RFLP of outer membrane protein gene, was used to characterize D. nodosus. The technique was evaluated in a controlled experiment involving two strains of bacteria. PCR–RFLP was found to be highly specific in differentiating isolates obtained from recipient animals infected with different strains. Subsequently, this technique was used to characterize isolates obtained from field cases of footrot in Nepal. A total of 11 patterns was recognized among 66 Nepalese D. nodosus isolates representing four different serogroups. PCR–RFLP also discriminated isolates with similar phenotypic characteristics. However, all isolates which, phenotypically, were virulent were represented by only two patterns irrespective of their serogroups. It is suggested that PCR–RFLP described here could be a useful epidemiological marker in the study of footrot.

Download Full-text

Isolation of an ompC-like outer membrane protein gene from Salmonella typhi

Gene ◽

10.1016/0378-1119(87)90366-0 ◽

1987 ◽

Vol 61 (1) ◽

pp. 75-83 ◽

Cited By ~ 30

Author(s):

José Luis Puente ◽

Valia Flores ◽

Marcos Fernández ◽

Yolanda Fuchs ◽

Edmundo Calva

Keyword(s):

Membrane Protein ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Protein Gene ◽

Salmonella Typhi ◽

Membrane Protein Gene

Download Full-text

Outer membrane characteristics of Salmonella enteritidis phage type 4 growing in chickens

Epidemiology and Infection ◽

10.1017/s0950268800057174 ◽

1993 ◽

Vol 111 (3) ◽

pp. 449-454 ◽

Cited By ~ 13

Author(s):

H. Chart ◽

D. Conway ◽

B. Rowe

Keyword(s):

Outer Membrane ◽

Salmonella Enteritidis ◽

Outer Membrane Proteins ◽

Phage Type ◽

Virulence Plasmid ◽

Uptake System ◽

Ferric Ions ◽

Phenotypic Characteristics ◽

A Subunit

SummaryStrains of Salmonella enteritidis belonging to phage type 4 (SE4) were grown in the peritoneal cavities of chickens, and without subculture on laboratory media examined for inducible in vivo phenotypic characteristics. These bacteria expressed three major outer membrane proteins (OMPs) of 33, 35 and 36 kilodaltons (kDa), and iron regulated OMPs of 74, 78 and 81 kDa. Bacteria growing in vivo did not express flagella, or fimbriae with a subunit molecular mass of 14 kDa (14 kDa fimbriae). Two OMPs of 55 and 23 kDa, expressed during culture in nutrient broth, were repressed during growth in chickens. Possession of a 38 MDa ‘mouse virulence’ plasmid did not influence the expression of OMPs, flagella or fimbriae. It was concluded that strains of SE4 growing in chicken tissues, use an enterobactin mediated iron uptake system to obtain ferric ions, do not express flagella or 14 kDa fimbriae and appear not to express novel OMPs involved in survival in vivo.

Download Full-text

Anaplasma phagocytophilum Has a Functional msp2 Gene That Is Distinct from p44

Infection and Immunity ◽

10.1128/iai.72.7.3883-3889.2004 ◽

2004 ◽

Vol 72 (7) ◽

pp. 3883-3889 ◽

Cited By ~ 31

Author(s):

Quan Lin ◽

Yasuko Rikihisa ◽

Suleyman Felek ◽

Xueqi Wang ◽

Robert F. Massung ◽

...

Keyword(s):

Outer Membrane ◽

Anaplasma Phagocytophilum ◽

Ixodes Scapularis ◽

Outer Membrane Proteins ◽

Gene Families ◽

Single Copy ◽

The United States ◽

Granulocytic Anaplasmosis ◽

The United Kingdom ◽

Membrane Protein Gene

ABSTRACT The msp2 and p44 genes encode polymorphic major outer membrane proteins that are considered unique to the intraerythrocytic agent of Anaplasma marginale and the intragranulocytic agent of Anaplasma phagocytophilum, respectively. In the present study, however, we found an msp2 gene in A. phagocytophilum that was remarkably conserved among A. phagocytophilum strains from human granulocytic anaplasmosis (HGA) patients, ticks, and a horse from various regions in the United States, but the gene was different in a sheep isolate from the United Kingdom. The msp2 gene in the A. phagocytophilum strain HZ genome was a single-copy gene and was located downstream of two Ehrlichia chaffeensis omp-1 homologs and a decarboxylase gene (ubiD). The msp2 gene was expressed by A. phagocytophilum in the blood from HGA patients NY36 and NY37 and by A. phagocytophilum isolates from these patients cultured in HL-60 cells at 37°C. The msp2 gene was also expressed in a DBA/2 mouse infected by attaching ticks infected with strain NTN-1 and in a horse experimentally infected by attaching strain HZ-infected ticks. However, the transcript of the msp2 gene was undetectable in A. phagocytophilum strain HZ in SCID mice and Ixodes scapularis ticks infected with strain NTN-1. These results indicate that msp2 is functional in various strains of A. phagocytophilum, and relative expression ratios of msp2 to p44 vary in different infected hosts. These findings may be important in understanding roles that Msp2 proteins play in granulocytic ehrlichia infection and evolution of the polymorphic major outer membrane protein gene families in Anaplasma species.

Download Full-text