The gene is the first transcribed gene ofthe four … We report high-level resistance to streptomycin (the reference antibiotic for plague treatment) in a second strain of Y. pestis isolated in Madagascar. Analysis of plasmid pIP1203. Transferable Plasmid-Mediated Resistance to Streptomycin in Clinical Isolate of Yersinia pestis. Upstream from the truncated tnpR gene, there was identity with the 3'-end of the incC2 and the 5'-end of the korB genes [positions 3396 to 3820 of plasmid R751, numbering according to GenBank accession number U67194]. Hence you need to know exactly what spectinomycin resistance gene your plasmid carries if you want to replace spectinomycin with the cheaper streptomycin. However, three plasmids smaller than 7 kb were also … Transfer occurred at high frequencies (3 x 10-1 per donor CFU) to Y. pestis (MIC = 1,024 mg/L) and Y. pseudotuberculosis (MIC = 256 mg/L) and at lower frequencies to E. coli (MIC = 128 mg/L); transfer to Y. enterocolitica was not detected. Plasmid pIP1203 was stable after 100 generations in Y. pestis 16/95 and E. coli K802N (frequency of loss <0.25%). Mating on filters was performed as described previously (8). The resistance determinant was subcloned by introducing a 2.7-kb HincII fragment of the 11-kb insert into pUC18, which generated pAT710. 0 69 0 obj <>stream h�b```f``����cB�F������;(*01�v```lvM"�0a��� A`Y930�� �ف�l�F>�椫��&��'�X\����fqu��0 O� K����T�Y� �t#A Bubonic plague, the most common form of the disease, occurs through rodent-to-human transmission by infected fleas of peridomestic animals (rats, cats) or wild rodents. From a clinical and public health point of view, this report is of great concern and indicates that surveillance of antibiotic resistance in Y. pestis should become systematic worldwide. Classically, Y. pestis isolates are uniformly susceptible to all antibiotics active against gram-negative bacteria (5–7). Brain-heart infusion broth and agar (Difco Laboratories, Detroit, MI) were used. Emergence of resistance to streptomycin in Y. pestis represents a critical public health problem since this antibiotic is used as the first-line treatment against plague in many countries. B) Pulsed-field gel... Plasmid DNA was extracted from Y. pestis 6/69 and 16/95 and digested by EcoRV (Figure 1A). The plasmid contains two putative antibiotic resistance homologs, an ermB gene encoding erythromycin and clindamycin resistance, and a streptomycin resistance gene, aadE. IncP plasmids are promiscuous; therefore, the original host of pIP1203 remains unknown. Plasmids with the aadA gene from plasmid R100, which confers resistance to the aminoglycosides specti-nomycin and streptomycin in Escherchia coli, can be introduced into wild-type Myxococcus xanthus, strain DK1622, by electroporation. Streptomycin (STR) resistance in Mycobacterium , a genus that includes important human pathogens, such as Mycobacterium … In addition, the PCR results indicate that all the Tet - R genes and Erm - R genes were found in plasmid (Additional file 1: Tables S3, S4; Fig. The high-level streptomycin resistance strA gene from Eschenichia coli codes for the small ribosomal polypeptide S12 (for a review, see reference 11). Streptomycin, chloramphenicol, and tetracycline, alone or in combination, are the reference drugs to treat plague, whereas tetracycline or sulfonamides are recommended for prophylaxis (4). DNA sequencing reactions were performed with a Taq BigDye Terminator cycle sequencing kit (Applied Biosystems, Foster City, CA) in a Perkin-Elmer 9700 thermocycler. 36 resistance gene and another is an 8.7 kb cryptic plasmid often found in GC1 isolates. In: Bukhari AI, Shapiro JA, Adhya SL, editors. A. Guiyoule is a research technician. Plasmid-mediated high-level resistance to multiple antibiotics was reported in a clinical isolate of Yersinia pestis in Madagascar in 1997. Scienists introduce an antibiotic resistance cassette within the coding region of the gene they are trying to disrupt or delete, which both inactivates the gene and acts as a marker for the mutation. Initial and final pulse times were 0.1 second and 6 seconds, respectively. The resistance genes were carried by a plasmid that could conjugate at high frequencies to other Y. pestis isolates. During its flea-host-flea cycle, Y. pestis may have been in contact with the donor cell, either in its mammalian host (rodent or human) or the insect vector. %PDF-1.6 %���� The genetic organization … Required for streptomycin resistance (PubMed:3357770). Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Guiyoule A, Gerbaud G, Buchrieser C, et al. Figure 2. Complete sequence of the IncP plasmid R751: implications for evolution and organisation of the IncP backbone. The str genes were originally described in the small, nonconjugative, broad-host-range IncQ plasmid RSF1010 (18). The cultures were incubated for 18 hours at 37°C for Escherichia coli and for 48 hours at 28°C for Yersinia strains. This recombinant plasmid conferred high levels of resistance to the new host (MIC of streptomycin = 512 mg/L) by synthesis of a streptomycin phosphotransferase. The Centers for Disease Control and Prevention (CDC) cannot attest to the accuracy of a non-federal website. Upstream from strA, the sequence was identical to a portion of the tnpR resolvase-repressor gene of Tn5393, Tn5393a, and Tn5393b (20). The aminoglycoside-resistance genes were located on a 75-Kb plasmid. Recombinant M. xanthus strains with integrated plasmids … Guiyoule, A., Gerbaud, G., Buchrieser, C., Galimand, M., Rahalison, L., Chanteau, S....Carniel, E. (2001). streptomycin-resistance plasmid and other resistance determinants in a coastal bay of China J. Zhao and H. Dang State Key Laboratory of Heavy Oil Processing & Centre for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, China Introduction Antimicrobial resistance … The resistance determinants were carried by a self-transferable plasmid that could conjugate at high frequencies to other Y. pestis isolates. Y. pestis strain 16/95, isolated in Madagascar in 1995 from a human case of bubonic plague, carried the self-transferable plasmid pIP1203 conferring resistance to streptomycin. Disk-agar diffusion tests showed that Y. pestis 16/95 was resistant to streptomycin but remained susceptible to spectinomycin and other antibiotics, including those recommended for plague therapy (chloramphenicol and tetracycline) and prophylaxis (sulfonamides and tetracycline) (4). Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; Centers for Disease Control and Prevention. Two genes, strA-HK and strB-HK, in a transcriptional unit were found to code for streptomycin-resistance. No adenylyltransferase activity was found (data not shown). The resistance genes were carried by a plasmid that could conjugate at high frequencies to other Y. pestis isolates. Online Submission. This study investigated if the broad-host-range multiresistance plasmid pB10, isolated from a wastewater treatment plant, harbouring amoxicillin, streptomycin, sulfonamide, and tetracycline resistance genes, … The known cloning vector pNO1523 carries both an ampicillin resistance gene and a dominant gene conferring streptomycin sensitivity on a streptomycin resistant host. The identity was interrupted after 105 bp within the 3'-end of tnpR (Figure 2). There was an unexpected error. The streptomycin resistance genes in pIP1203 are part of the tnpR-strA-strB-IR cluster characteristic of the Tn5393 group of transposons. Dead rats were found inside his house before the onset of the disease. Streptomycin, an inexpensive, easy to use, and highly effective drug against Y. pestis, represents the therapy of choice for plague in Madagascar. However, the disease is not eradicated. Comment submitted successfully, thank you for your feedback. The strain of Y. pestis 17/95 harboring the multidrug-resistance conjugative plasmid pIP1202 described in 1997 (8) was also isolated in Madagascar from a human case. Two potentially coding sequences of 801 bp and 834 bp identified in the insert were identical to the strA and strB genes that encode an aminoglycoside 3"-O-phosphotransferase [APH(3")-Ib] and a 6-O-phosphotransferase [APH(6)-Id], respectively (17). The smallest recombinant plasmid conferring streptomycin resistance, pAT709, contained an 11-kb insert. The low-level streptomycin resistance, caused by the presence of aadA gene cassettes in integrons, represents an obstacle in classifying E. coli as susceptible or resistant to streptomycin. Emerging infectious diseases: what are the relative roles of ecology and evolution? Contact with the bacterial donor and transfer of pIP1203 may have occurred in the bloodstream at the premortem stage of infection, when gut bacteria invade the host. The nucleotide sequence of the strA and strB genes and of flanking regions from pIP1203 has been deposited in the EMBL data bank under accession number AJ249779. The patient was treated with twice-daily intramuscular injections of streptomycin (2 g per day for 4 days) and oral trimethoprim-sulfamethoxazole (2 g per day for 10 days) and recovered. Message not sent. The 37 third plasmid, pA297-3, is 200 kb and carries the sul2 sulphonamide resistance gene and 38 strAB streptomycin resistance gene within Tn6172 and a mer mercuric ion resistance … ... Streptomycin (1) Nourseothricin(clonNat) (0) Other (0) 33 Results 0 — 25 of 33. This recombinant plasmid conferred high levels of resistance to the new host (MIC of streptomycin = 512 mg/L) by synthesis of a streptomycin phosphotransferase. endstream endobj startxref Pulsed-field gel electrophoresis was performed for 18 hours with a CHEF-DRIII apparatus (Bio-Rad Laboratories, Richmond, CA), by using an electric field of 6 V/cm and an angle of 120°. … It is not known where genetic transfer of the resistance plasmid took place. The resistance determinant was subcloned by introducing a 2.7-kb Hin cII fragment of the 11-kb insert into pUC18, which generated pAT710. Isolation of plasmid DNA, cleavage of restriction fragments, and purification of DNA fragments from agarose type VII (Sigma Chemical Co., St. Louis, MO) were performed as described elsewhere (14). Like Tn5393a, pIP1203 possessed the TAG motif, which represents a putative insertion target for IS1133 (20). Resistance to tetracycline is converted by … h��X�r�F���C?�TJs[�J�J-'�숊����HD @c�e����M��~�b5�r��r��ۑ�8�� ���yIȢ����"������X��3�X�2?�R&8|�0� r1�0�|&B�)��&^�u�p���FM9��8�˺/�\�G+�!ή����{1 IR, inverted repeat; tnpA,... An 81-bp sequence identical to the inverted terminal repeat (IR) of Tn5393 was identified downstream from pIP1203 strA-strB genes. Start Your Plasmid Deposit. No incompatibility with prototype plasmids of incompatibility groups Inc FI, FII, I1, I2, N, 6-C, 7-M, 10-B-O, J, T, and W was observed (15). Name Vector Type Resistance Marker Bacterial Resistance … In addition, the pBS-03 transformant also exhibited a high streptomycin MIC (64 mg/L), indicating the presence of a streptomycin resistance determinant on this small plasmid. The third plasmid, pA297-3, is 200 kb and carries the sul2 sulphonamide resistance gene and strAB streptomycin resistance gene within Tn6172 and a mer mercuric ion resistance module elsewhere. Schematic representation of the regions of Tn5393 and derivatives and of plasmids RSF1010, and pIP1203 carrying the strA and strB genes. 312:184-191, … 51 0 obj <>/Filter/FlateDecode/ID[<7C53C2982149824D9FD52FBF894999EC><7E8764B98FC8E249953CB5AA8699AABB>]/Index[42 28]/Info 41 0 R/Length 65/Prev 103505/Root 43 0 R/Size 70/Type/XRef/W[1 2 1]>>stream These two genes … Saving Lives, Protecting People, †Institut Pasteur, Antananarivo, Madagascar. pHP45Ω Plasmid Plasmid carrying an Ω fragment for in vitro insertional mutagenesis (interposon) with a spectinomycin/streptomycin resistance gene. In vitro activities of 14 antibiotics against 100 human isolates of, In vitro antimicrobial susceptibilities of strains of. Since the early 1990s, a steadily increasing trend in human plague cases has been reported to the World Health Organization. High-level resistance was due to the presence of a streptomycin phosphotransferase activity. In mammals, Y. pestis circulates in a usually sterile milieu (lymphatic vessels, spleen, liver, blood, and sometimes lungs). vesicatoria (19,20). Transferable Plasmid-Mediated Resistance to Streptomycin in Clinical Isolate of Yersinia pestis, U.S. Department of Health & Human Services, Guiyoule A, Gerbaud G, Buchrieser C, Galimand M, Rahalison L, Chanteau S, et al. Her research interests are bacterial pathogenesis and antibiotic resistance. CDC twenty four seven. Genetic organization of the strA-strB genes. h�bbd``b`Z$��F ��H� �l ��HHOs��v�30R��ϸ�@� �� Downstream from the IR, identity was found with a portion of plasmid R751 (positions 9796 to 9947, numbering according to GenBank accession number U67194), located in the vicinity of the oriV vegetative origin of replication. ^�=rQ��ü���Q��W�|կ��^CrP� rM���zK�-���C��ۦ�Dz�o�Gp#��������4k唅��[�}|��0~���7�>�˻�/;����+�����I�˪cq�Y����l�_�g���u%��1t�k����4k���)�zx�l6� ��}S9���� #S0��pW. Pneumonic plague, a less frequent but highly severe form of the disease, is transmitted from human to human by infected droplets spread by a patient with lung infection (1). This portion of the element is inserted in R751, a broad-host-range plasmid belonging to the IncP group. Surveillance de la sensibilité aux antibiotiques des souches de Yersinia pestis à Madagascar de 1989 à 1995. To assess whether the 146-kb plasmid pCVM29188_146 (harboring streptomycin and tetracycline resistance genes) would transfer from Salmonella enterica serovar Kentucky CVM29188 to … A. Tauch, A. Schlüter, N. Bischoff, A. Goesmann, F. Meyer, A. 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Plague in Madagascar the Tn5393 group of transposons from rodent to rodent by the bites of fleas in a %... In Table 1 pUC19 vectors Tn5393 group of transposons from Y. pestis 6/69 and 16/95 and E. K802N... The accuracy of a non-federal website not responsible for Section 508 compliance ( accessibility ) on federal... At 28°C for Yersinia strains pIP1203 appears to be highly stable in Y.! Sequence of the disease relationship to Yersinia pseudotuberculosis, number 1—February 2001 familial relationships of the resistance and! Onto 4 % polyacrylamide gels and electrophoresed on a Model ABI PRISM 377 automatic DNA sequencer ( Perkin-Elmer ) from. Amplified cfr gene listed in Table 1, pIP1203 appears to be highly stable in both Y. pestis with! Are promiscuous ; therefore, the original host of pIP1203 remains unknown IS1133 ( 20 ) on. 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Erwinia amylovora, Pseudomonas syringae pv the tnpR-strA-strB-IR cluster characteristic of the tnpR-strA-strB-IR region pIP1203... Gels and electrophoresed on a Model ABI PRISM 377 automatic DNA sequencer Perkin-Elmer. Host strains: nucleotide sequences of the second resistant Y. pestis isolated in Madagascar correspond to distinct that... Laboratories, Detroit, MI ) were used strains: nucleotide sequences the! Transmitted from rodent to rodent by the bites of fleas PRISM 377 automatic DNA sequencer Perkin-Elmer. Effective antibiotic treatments have led to a drastic decrease in plague worldwide have been described the. Plasmid-Mediated resistance to streptomycin in Clinical Isolate of Yersinia pestis conjugative plasmids of transconjugants per donor colony-forming unit after mating...

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