We investigated the prevalence of virulent Rhodococcus equi in clinical isolates from 69 sporadic cases (60 men, 8 women, and 1 patient of unknown sex) in Chiang Mai, Thailand, between 1993 and 2001. Fifty were human immunodeficiency virus (HIV) positive, 3 were HIV negative, and HIV status was unknown for 16. Fifty-two (75%) of 69 isolates were strains of intermediate virulence that contained the virulence-associated 20-kDa antigen, and 17 isolates (25%) were avirulent. No virulent strains with the virulence-associated 15-17-kDa antigens were identified. R. equi was isolated from HIV-positive patients' houses and those of their neighbors: avirulent strains were widespread, but only 1 strain of intermediate virulence was isolated. R. equi strains of intermediate virulence were isolated from 4 (0.8%) of 500 submaxillary lymph nodes from apparently healthy pigs in Chiang Mai. The routes of R. equi acquisition should be investigated from the viewpoint of zoonosis and public health.

ABSTRACT. Rhodococcus equi was isolated from fecal and soil samples from four native Jeju horse farms and six Thoroughbred farms in Jeju, Korea. The isolates were examined for the presence of virulence-associated 15-17-kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and for the gene encoding VapA by PCR. R. equi was isolated from all 36 soil samples collected from the 10 farms with between 5.0 × 10 2 and 7.5 × 10 4 colony-forming units (cfu) per gram of soil, and from 37 of 40 fecal samples with between 5.0 × 10 1 and 1.1 × 10 5 cfu per gram of feces. Virulent R. equi was isolated from seven farms and appeared in 2.0% of isolates (10 of 508). Of the 10 virulent isolates, four contained a 90-kb type II plasmid, which has been found in isolates from the Kiso native horses of Japan, and the other six contained a new variant, which did not display the EcoRI and EcoT22I digestion patterns of the 10 representative plasmids already reported (85-kb types I, II, III, and IV; 87-kb types I and II; 90-kb types I, II, III, and IV). We designated the new variant as the "90-kb type V" plasmid, because its EcoRI digestion pattern is similar to that of the 90-kb type II plasmid. This is the first report of the prevalence of virulent R. equi in Jeju, Korea. The same virulence plasmid type is found in both Korean and Japanese isolates, providing insight into the origin, ancestry, and dispersal of native horses in Korea and Japan.

ABSTRACT. We recently demonstrated the presence of virulence-associated protein antigen (VapA)-positive Rhodococcus equi in Jeju Island, Korea. These bacteria contained one of two distinct plasmid types, a 90-kb type II plasmid, which has been found in isolates from the native Kiso horses of Japan, and a new variant, a 90-kb type V plasmid. However, the genotypic characters of the VapA-positive R. equi from Jeju native horses and their environments are poorly understood. Ninety-eight isolates from soil samples and 89 isolates from fecal samples were collected from five farms that breed or have bred Jeju native horses, and were tested for the presence of VapA by immunoblotting and PCR. Of the 98 soil isolates and 89 fecal isolates, seven and 13 were VapA-positive R. equi, respectively. In 2003, two Jeju foals died suddenly and were brought to the Faculty of Veterinary Medicine, Cheju National University, for postmortem examination. Pure cultures of R. equi were isolated from the lung lesions of both foals. Rhodococcus equi is one of the most important bacterial pathogens in one-to three-month-old foals. Infections caused by this organism are characterized by chronic, suppurative bronchopneumonia, which is often accompanied by ulcerative enteritis and mesenteric lymphadenitis [1,6,9]. Since the first isolation of Rhodococcus equi from the lung abscesses of Swedish foals by Magnusson in 1923 [4], the disease has been described in many areas of the world [1,6]. The discovery of virulence-associated antigens and virulence plasmids has clarified some aspects of the virulence of R. equi [14,18]. Virulent R. equi, which are characterized by a virulence-associated 15-17-kDa protein antigen (VapA) and a virulence plasmid of 85-90 kb, have been isolated from the lesions of infected foals and the disease has been reproduced experimentally [22,23]. However, avirulent R. equi, which have no virulence-associated antigens or plasmids and do not cause disease in foals, are distributed widely in horses and their environments [10,23]. Epidemiological studies have revealed that horse-breeding farms with endemic infections are heavily contaminated with virulent R. equi [9,17].The restriction-enzyme digestion patterns of virulent virulence plasmids from foals have allowed the plasmids of virulent isolates to be categorized into 12 closely related types and have revealed geographic differences in the distributions of these virulence plasmids, not only throughout the world, but also within Japan [7,11,15,16,19,20,22]. In a recent study, we isolated virulent R. equi containing the 90-kb type II plasmid that has been found in isolates from the native Kiso horses of Japan, from fecal and environmental samples of the native Jeju horse and thoroughbred horse farms on Jeju Island, Korea, and a new variant, the 90-kb type V plasmid [20]. However, only one virulent R. equi was isolated from soil samples collected from a Jeju native horse farm [20].The purposes of this study were to investigate the prevalence of virulent R. equi in Jeju native ...

Virulent Rhodococcus equi is an important pathogen in foals aged Key words: foal, Rhodococcus equi, soil, TennesseeRhodococcus equi is one of the most important bacterial pathogens in foals less than 3 months of age [1, 12,15,29]. Infections caused by this organism are c h a r a c t e r i z e d b y a c h r o n i c , s u p p u r a t i v e bronchopneumonia and/or enteritis [1, 12,15,29]. In horses, the disease is distributed worldwide, but little is known about the incidence of the infection in Tennessee, U. S. A.The discovery of virulence-associated antigens and virulence plasmids has permitted classification of the virulence of R. equi strains [23,30]. At least three virulence levels of R. equi have been identified: virulent, intermediate virulence, and avirulent [15]. Virulent R. equi is characterized by the presence of virulenceassociated 15-to 17-kDa antigens (VapA), and virulence plasmid DNA of 85-90 kb [23,30,33]. There are at least 11 virulence plasmid types (85-kb types I to IV, 87-kb types I and II, and 90-kb types I to V) present in VapA-positive R. equi from horses in the world by restriction fragment length polymorphism of plasmid DNAs [13,17,25,26,[31][32][33]. The virulent form has been found in pulmonary and/or intestinal lesions in foals and in the pulmonary lesions of AIDS patients (murine LD 50 =10 6 ) [28,30]. R. equi strains of intermediate virulence are identified by a virulence associated 20-kDa antigen (VapB) and virulence plasmid DNA of 79-100 kb [22]. The intermediate virulence form has been found in the submaxillary lymph nodes of pigs (murine LD 50 =10 7 ) and in the pulmonary lesions of AIDS patients [18,22]. In comparison, avirulent R. equi shows no evidence of either virulence-associated antigens or plasmid DNA (murine LD 50 >10 8 ), and is widespread in soil [15,29]. In recent studies, we have demonstrated geographic differences in the distributions of the virulenceassociated plasmids found in the Americas, Europe, Australia, Africa, Korea and Japan [2, 7-11, 13, 16, 17, 19, 25-27, 31-33, 35]. Most clinical isolates from the Americas, Australia, and Europe contain 85-kb type I or 87-kb type I plasmids [2, 7-11, 17, 19, 31]. The 85-kb type II plasmid is found only in French isolates and the 85-kb type III and type IV plasmids only in isolates from Texas [13,17,31]. These five types of virulence