Aquariums as reservoirs for multidrug-resistant Salmonella Paratyphi B.
Multidrug-resistant Salmonella enterica serovar Paratyphi B dT+ isolates from patients with gastroenteritis gastroenteritis: see enteritis.
Acute infectious syndrome of the stomach lining and intestines. Symptoms include diarrhea, vomiting, and abdominal cramps. were identical with isolates from their home aquariums. Matched isolates had identical phage phage: see bacteriophage.
phage - A program that modifies other programs or databases in unauthorised ways; especially one that propagates a virus or Trojan horse. See also worm, mockingbird. The analogy, of course, is with phage viruses in biology. types, Xbal and IS200 profiles, and Salmonella genomic island 1 (SGI (SGI, Sunnyvale, CA, www.sgi.com) A manufacturer of workstations and servers, founded in 1982 by Jim Clark. The company was founded as Silicon Graphics, Inc., but changed to its acronym in 1999. 1). Ornamental fish tanks are reservoirs for SGIl-containing S. Paratyphi B dT+.
Strains of Salmonella enterica serovar Paratyphi B that use d-tartrate as a carbon source (S. Paratyphi B dT+, formerly S. enterica serovar Java)primarily cause gastroenteritis (1). Since the late 1990s, multidrug-resistant S. Paratyphi B dT+ has been increasingly isolated from infected persons in different parts of the world. One type, which is resistant to streptomycin streptomycin (strĕp'tōmī`sĭn), antibiotic produced by soil bacteria of the genus Streptomyces and active against both gram-positive and gram-negative bacteria (see Gram's stain), including species resistant to other , spectinomycin spectinomycin /spec·ti·no·my·cin/ (spek?ti-no-mi´sin) an antibiotic derived from Streptomyces spectabilis, used as the hydrochloride salt in the treatment of gonorrhea. , trimethoprim trimethoprim /tri·meth·o·prim/ (-meth´o-prim) an antibacterial closely related to pyrimethamine; almost always used in combination with a sulfonamide, primarily for the treatment of urinary tract infections. , and sulfonamides Sulfonamides Definition
Sulfonamides are medicines that prevent the growth of bacteria in the body.
Sulfonamides are used to treat many kinds of infections caused by bacteria and certain other microorganisms. , carries a chromosomally located class 2 integron with the dfrA1-sat1-aadA1 (Tn7) array of gene cassettes (2). This clone is predominantly associated with poultry and poultry products in Germany and the Netherlands (2,3). Human cases of gastroenteritis caused by S. Paratyphi B dT+ with the resistance phenotype ApCmSmSpSuTc (Ap, ampicillin ampicillin (ăm'pĭsĭl`ĭn), a penicillin-type antibiotic that is effective against both gram-negative microorganisms and gram-positive microorganisms such as Escherichia coli. ; Cm, chloramphenicol chloramphenicol (klōr'ămfĕn`əkŏl'), antibiotic effective against a wide range of gram-negative and gram-positive bacteria (see Gram's stain). It was originally isolated from a species of Streptomyces bacteria. ; Sm, streptomycin; Sp, spectinomycin; Su, sulfonamides; Tc, tetracycline tetracycline (tĕ'trəsī`klēn), any of a group of antibiotics produced by bacteria of the genus Streptomyces. They are effective against a wide range of Gram positive and Gram negative bacteria, interfering with protein ) have also been found in Canada (4), the United Kingdom (5), France (6), and Australia (7), and their incidence is increasing. In most of the studied isolates, the resistance genes blaP1, floR, aadA2, sull n. 1. A plow. , and tetG are located in a complex class 1 integron recently designated In104 (7) (Figure 1). This integron is located within the Salmonella genomic island 1 (SGI1) that was first identified in S. enterica serovar Typhimurium DT 104 strains with the same phenotype (8). However, the source of the SGI1-containing S. Paratyphi B dT+ has not been identified. Whether isolates obtained in different countries are clonally related is also not known.
Although a few epidemiologic studies suggest that antimicrobial drug-susceptible S. Paratyphi B dT+ may be linked to aquacultural practices (9,10), no molecular data confirm this. However, the first reported SGI1-containing S. Paratyphi B dT+ isolate with drug-resistance phenotype ApCmSmSpSuTc was isolated in 1997 from a tropical fish in Singapore (11), raising the possibility that tropical fish and aquariums are a reservoir. The aim of this study was to determine if domestic aquariums are reservoirs for SGI1-containing, multidrug-resistant S. Paratyphi B dT+ that infect humans.
S. Paratyphi B dT+ with the resistance phenotype ApCmSmSpSuTc had been isolated sporadically in various states of Australia since 1997, and initial surveys showed a potential association with ownership of home aquariums (D. Lightfoot, unpub, data). In 2000, multidrug-resistant S. Paratyphi B dT+ with an identical phage type (reaction does not conform [RDNC RDNC Resource Discovery Network Centre (UK) ]), designated here as Aus2, and the same drug-resistance profile (ApCmSm SpSuTc) was isolated from humans with gastroenteritis and from fish tanks in the homes of 2 infected patients (Table). In 2003 and 2004, 13 cases of ApCmSmSpSuTc S. Paratyphi B dT+ were investigated by state and commonwealth health departments, and all were associated with home aquariums containing tropical fish (J. Musto et al., unpub, data). Of these, 11 cases were phage type RDNC Aus3, 1 was phage type 1 var 15, and 1 was phage type 3b var. Water and gravel were collected from the domestic aquariums of 5 patients with RDNC Aus3-type infections, and identical isolates were recovered from each fish tank. Four matched sets of isolates, 2 from 2000 and 2 from 2003, were further examined (Table). One isolate (SRC (SouRCe) Contrast with DST, which is an abbreviation of "destination." 50) characterized previously (7) was used as a control (Table).
To determine if the resistance phenotype of these strains was due to SGI1 (4,5,7,8,11), polymerase chain reaction polymerase chain reaction (pŏl`ĭmərās') (PCR), laboratory process in which a particular DNA segment from a mixture of DNA chains is rapidly replicated, producing a large, readily analyzed sample of a piece of DNA; the process is (PCR PCR polymerase chain reaction.
polymerase chain reaction
Polymerase chain reaction (PCR) ) with primer pairs shown in Figure 1 was used as previously described (7). The left and right junctions of SGI1 with the chromosome and of In104 with SGI1 were present in all cases. Regions containing the gene cassettes were amplified by using standard primers (L1 and R1) in the 5'- and 3'-conserved segments of class 1 integrons. Fragments of 1.0 and 1.2 kb were amplified from all isolates, and digestion of these amplicons with RsaI generated a profile (data not shown) that was indistinguishable from the pattern for the 2 amplicons containing the aadA2 and blaP1 cassettes found in In104 and S. Paratyphi B dT+ isolates SRC49 and SRC50 from 2001 (7). The aadA2 gene cassette was linked to SO26 in the SGI1 backbone, which indicates that it is on the left, as in In104, and the expected 1.8-kb PCR fragment was generated by using primers in groEL and blaP1 (Figure 1), which places the blaP1 cassette on the right. Southern hybridization hybridization /hy·brid·iza·tion/ (hi?brid-i-za´shun)
1. crossbreeding; the act or process of producing hybrids.
2. molecular hybridization
3. of XbaI-digested whole-cell DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. with a probe for the floR gene as described previously (7) identified a band of [approximately equal to] 12 kb, which is consistent with an SGI1 structure identical to that reported previously (7,8,11) and the groEL-blaP1 amplicon linked this 12-kb XbaI fragment with the adjacent 4.3-kb XbaI fragment (Figure 1).
To obtain further evidence for the identity of the matched human and fish tank isolates, macrorestriction analyses of XbaI-digested whole-cell DNA by pulsed-field gel electrophoresis (PFGE PFGE Pulsed-Field Gel Electrophoresis ) were performed as previously described (12). Several studies (3-6,13) suggest that S. Paratyphi B dT+ isolates possess considerable genetic heterogeneity. However, the SGI1-containing isolates appear to be homogeneous. The band patterns for all SGI1-containing S. Paratyphi B dT+ were identical from humans and fish tanks with phage type RDNC Aus3 (Figure 2A) and Aus2 (data not shown). IS200 profiles were also analyzed by hybridization of an IS200 probe with Pst I-digested whole-cell DNA as described elsewhere (6). Again, all strains showed identical profiles (Figure 2B and data not shown) that differed by 1 band from profile IP1 recently described (6). Thus, matched isolates from humans and their fish tanks were indistinguishable from each other.
An unusual observation in this study was that isolates with different phage types showed identical PFGE and IS200 profiles, indicating that they represented a clonal cluster. The control strain SRC50 (RDNC) also displayed the same patterns, demonstrating that it also is a member of the same clone. Thus, variation in phage type (Table) appears to have occurred within a single clone. Variation in phage type has also been reported in other studies of multidrug-resistant S. Paratyphi B dT+ strains (4-6), although a number of related but slightly different XbaI PFGE patterns were observed in those studies. This finding suggests that all multidrug-resistant S. Paratyphi B dT+ found globally have a single origin, but that variations, possibly because of acquisition of other temperate phages or plasmids, have arisen over time. However, direct comparisons of strains from different countries will be needed to confirm this hypothesis.
This is the first definitive report showing that ornamental fish tanks are a reservoir for multidrug-resistant S. Paratyphi B dT+ (ApCmSmSpSuTc phenotype) containing SGI1 that causes severe disease in humans, particularly young children. In addition to containing SGI1, the matched isolates from humans and their fish tanks had the same phage type and the same XbaI macrorestriction digest pattern and IS 200 profile. These findings identify home aquariums containing tropical fish as the most important, although not necessarily the only, source of multidrug-resistant S. Paratyphi B dT+. The fact that 12%-14% of Australian households have ornamental fish (14) and as many as 12 million American and 1 million Canadian families own domestic aquariums (9), together with the young age of most affected patients, indicate that multidrug-resistant S. Paratyphi B dT+ in home aquariums is a risk factor for Salmonella infection and thus becomes a public health issue.
We thank Linda Falconer for skillful skill·ful
1. Possessing or exercising skill; expert. See Synonyms at proficient.
2. Characterized by, exhibiting, or requiring skill. technical assistance.
Renee S. Levings is supported by a grant from the New South Wales Department of Primary Industries New South Wales Department of Primary Industries (NSW DPI) is part of the Government of New South Wales and is responsible for agriculture, fisheries and aquaculture, state forests, and minerals and petroleum in the state. The Minister for Primary Industries is Ian Macdonald. and a University of Wollongong History
The University of Wollongong was founded in 1951 when a Division of the then New South Wales University of Technology (re-named the University of New South Wales in 1958) was established in Wollongong. Postgraduate Research Award.
Ms Levings is a PhD candidate at the Elizabeth Macarthur Agricultural Institute. Her research interests are the analysis of clustered antimicrobial drug-resistance genes and their spread among members of the Enterobacteriaceae.
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Address for correspondence: Steven P. Djordjevic, Microbiology and Immunology Section, New South Wales Department of Primary Industries, Elizabeth Macarthur Agricultural Institute, Private Mail Bag 8, Camden, New South Wales Camden in New South Wales, Australia, is a historic town and Sydney Suburb 65 km southwest of the Sydney CBD and near the city of Campbelltown.
Camden is now a smaller, yet well known town lying on the fringe of the Sydney Metropolitan area. 2570, Australia; fax: 61-2-4640-6384; email: firstname.lastname@example.org
Renee S. Levings, * ([dagger]) Diane Lightfoot, ([double dagger]) Ruth M. Hall, ([section]) and Steven P. Djordjevic *
* Elizabeth Macarthur Agricultural Institute, Camden, New South Wales, Australia; ([dagger]) University of Wollongong, Wollongong, New South Wales Wollongong is the 3rd largest city in the state of New South Wales, Australia, after Sydney and Newcastle. It is also a Local Government Area administered by the Wollongong City Council. , Australia; ([double dagger]) University of Melbourne
In 2006, Times Higher Education Supplement ranked the University of Melbourne 22nd in the world. Because of the drop in ranking, University of Melbourne is currently behind four Asian universities - Beijing University, , Melbourne, Victoria, Australia; and ([section]) University of Sydney The University of Sydney, established in Sydney in 1850, is the oldest university in Australia. It is a member of Australia's "Group of Eight" Australian universities that are highly ranked in terms of their research performance. , Sydney, New South Wales New South Wales, state (1991 pop. 5,164,549), 309,443 sq mi (801,457 sq km), SE Australia. It is bounded on the E by the Pacific Ocean. Sydney is the capital. The other principal urban centers are Newcastle, Wagga Wagga, Lismore, Wollongong, and Broken Hill. , Australia
Table. Genetic characteristics of Salmonella enterica serovar Paratyphi B dT+ isolates used in this study Isolate no. * Source ([dagger]) Phage type ([double dagger]) Set 1 SRC229 H Aus2 SRC230 H Aus2 SRC231 FT Aus2 Set 2 SRC232# H Aus2 SRC233# H Aus2 SRC233A FT Aus2 Set 3 SRC145 H Aus3 SRC142 FT Aus3 SRC143 FT Aus3 Set 4 SRC149 H Aus3 SRC147 FT Aus3 SRC148 FT Aus3 Control SRC50 H RDNC Isolate no. * State ([section]) Date of isolation Set 1 SRC229 ACT 2000 SRC230 ACT 2000 SRC231 ACT 2000 Set 2 SRC232# Vic 2000 SRC233# Vic 2000 SRC233A Vic 2000 Set 3 SRC145 Vic 2003 SRC142 Vic 2003 SRC143 Vic 2003 Set 4 SRC149 Vic 2003 SRC147 Vic 2003 SRC148 Vic 2003 Control SRC50 Vic 2001 Isolate no. * Age, y/Sex SGI1 ([paragraph]) Set 1 SRC229 <1/F + SRC230 1/M + SRC231 -- + Set 2 SRC232# 11/F + SRC233# 11/F + SRC233A -- ND Set 3 SRC145 74/F + SRC142 -- + SRC143 -- + Set 4 SRC149 12/M + SRC147 -- + SRC148 -- + Control SRC50 14/M + ** * All isolates were resistant to ampicillin, chloramphenicol, streptomycin, spectinomycin, sulfonamides, and tetracycline. ([dagger]) H, human isolates; FT, fish tank isolates. ([double dagger]) Determined by using standard procedures and designations (http://www.geocities.com/avinash_abhyankar/typing.htm). RDNC, reaction does not conform. RDNC Aus2 and RDNC Aus3 are 1 var and 3b var phage-typing variants, respectively, and are identifiable and reproducible phage-typing patterns awaiting formal assignment by the World Health Organization-designated International Reference Laboratory, Colindale, UK. ([section]) ACT, Australian Capital Territory; Vic, Victoria. ([paragraph]) SGI1, Salmonella genomic island 1; ND, strain not available for molecular analysis. (#) Isolates are from the same person. ** Data from Levings et al. (7).