However, this plasmid has an … The isolate, biotype Orientalis and ribotype Q, was obtained in 1995 in the Ampitana District of Madagascar from the axillary bubo puncture of a 14-year-old boy before antibiotic treatment. Alternatively, plasmid acquisition may have taken place in the midgut of the flea, a nonsterile environment where Y. pestis is most likely to be in intimate contact with other microorganisms. Intra- and interspecies relatedness of Yersinia pestis by DNA hybridization and its relationship to Yersinia pseudotuberculosis. Comparison of the restriction profiles of strains 6/69 and 16/95 revealed that the latter contained at least one extra large-size EcoRV fragment corresponding to an additional plasmid, designated pIP1203. The resistance genes bla TEM (ampicillin), strA (streptomycin) and sul2 (sulphamethoxazole) transferred along with a B/O plasmid named p838B-R. The resistance genes were carried by a plasmid that could conjugate at high frequencies to other Y. pestis isolates. Recently, high-level resistance to multiple antibiotics, including those recommended for plague prophylaxis and therapy, was observed in a clinical isolate of Y. pestis in Madagascar (8). Her research interests are bacterial pathogenesis and antibiotic resistance. Classically, Y. pestis isolates are uniformly susceptible to all antibiotics active against gram-negative bacteria (5–7). It is also present in some M. fortuitum clinical isolates. papulans, and Xanthomonas campestris pv. There was an unexpected error. When designing these types of experiments it is best practice not to use the same resistance cassette for the mutation and for plasmid … Resistance to tetracycline is converted by … vesicatoria (19,20). 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. … To assess whether the 146-kb plasmid pCVM29188_146 (harboring streptomycin and tetracycline resistance genes) would transfer from Salmonella enterica serovar Kentucky CVM29188 to … DNA insertion elements, plasmids, and episomes. The resistance determinant was subcloned by introducing a 2.7-kb HincII fragment of the 11-kb insert into pUC18, which generated pAT710. Transferable Plasmid-Mediated Resistance to Streptomycin in Clinical Isolate of Yersinia pestis. Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010. Complete sequence of the IncP plasmid R751: implications for evolution and organisation of the IncP backbone. The plasmid-carried antimicrobial resistance determinants genes such as genes encoding aminoglycoside-modifying enzymes (aac(6’)-aph(2’), aph(3)-IIIa) and vancomycin resistance gene vanA, in addition to the frequency of the genes encode the plasmid-mediated virulence factors including aggregation encoding gene … The most affected countries are Madagascar and Tanzania in Africa, Vietnam in Asia, and Peru in the Americas. This recombinant plasmid conferred high levels of resistance to the new host (MIC of streptomycin = 512 mg/L) by synthesis of a streptomycin phosphotransferase. 69 0 obj
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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. 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. Transferable Plasmid-Mediated Resistance to Streptomycin in Clinical Isolate of Yersinia pestis. This work was supported in part by a Bristol-Myers Squibb Unrestricted Biomedical Research Grant in Infectious Diseases. Comment submitted successfully, thank you for your feedback. Figure 2. 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. However, three plasmids smaller than 7 kb were also … A) Agarose-gel electrophoresis of EcoRV-digested plasmid DNA from representative Yersinia pestis strain 6/69 (1) and from streptomycin-resistant strain 16/95 (2). Initial and final pulse times were 0.1 second and 6 seconds, respectively. The gene is the first transcribed gene ofthe four … CDC is not responsible for Section 508 compliance (accessibility) on other federal or private website. Online Submission. We describe a second Y. pestis strain with high-level resistance to streptomycin, isolated from a human case of bubonic plague in Madagascar. The reasons for plague's reemergence are not clear but may be partly attributable to inadequate surveillance. The genetic organization … The restriction fragments in strain 6/69 correspond to those of the three Y. pestis resident plasmids, pPla, pYV, and pFra (12). Plasmid … Guiyoule, A., Gerbaud, G., Buchrieser, C., Galimand, M., Rahalison, L., Chanteau, S....Carniel, E. (2001). 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 … 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). No recent history of travel outside the village was noted. Analysis of plasmid pIP1203. Upstream from strA, the sequence was identical to a portion of the tnpR resolvase-repressor gene of Tn5393, Tn5393a, and Tn5393b (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. This IR is always present at the same position in the various genetic structures that carry the str genes (Figure 2). Please use the form below to submit correspondence to the authors or contact them at the following address: Elisabeth Carniel, Laboratoire des Yersinia, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France; fax: 33-1-40-61-30-01; carniel . The known cloning vector pNO1523 carries both an ampicillin resistance gene and a dominant gene conferring streptomycin sensitivity on a streptomycin resistant host. 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. Saving Lives, Protecting People, †Institut Pasteur, Antananarivo, Madagascar. %%EOF
The smallest recombinant plasmid conferring streptomycin resistance, pAT709, contained an 11-kb insert. Of particular note is the aadE gene which holds 100% identity to an aadE gene found in Campylobacter jejuni plasmid … 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 … Yersinia pestis is the causative agent of plague, a disease transmitted from rodent to rodent by the bites of fleas. Spread of plasmids pIP1202 and pIP1203 among strains of Y. pestis would render streptomycin ineffective for plague treatment and could create economic and therapeutic problems in Madagascar and other countries with endemic plague foci. Brain-heart infusion broth and agar (Difco Laboratories, Detroit, MI) were used. The sequence of plasmid pBS-03 (accession no. 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. 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. h�bbd``b`Z$��F ��H� �l ��HHOs��v�30R��ϸ�@� ��
Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Dead rats were found inside his house before the onset of the disease. Linking to a non-federal website does not constitute an endorsement by CDC or any of its employees of the sponsors or the information and products presented on the website. Sequence determination of the insert in pAT710 revealed that the base composition of this fragment was 57.5% G+C, much higher than the average G+C content of the Y. pestis genome (46%) (16). JQ394981) was obtained by primer walking, starting from the amplified cfr gene. Figure 1. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Transfer frequencies were expressed as the number of transconjugants per donor colony-forming unit after the mating period. The plasmid and the host bacterium were different from those previously associated with multiple-drug resistance, indicating that acquisition of resistance plasmids is occurring in this bacterial species. Required for streptomycin resistance (PubMed:3357770). 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). Therefore, the two resistant Y. pestis isolated in Madagascar correspond to distinct strains that have acquired different conjugative plasmids. pA297-3 transferred sulphonamide, streptomycin and mercuric ion resistance … Annie Guiyoule*, Guy Gerbaud*, Carmen Buchrieser*, Marc Galimand*, Lila Rahalison†, Suzanne Chanteau†, Patrice Courvalin*, and Elisabeth Carniel*. Plasmid pIP1203 was stable after 100 generations in Y. pestis 16/95 and E. coli K802N (frequency of loss <0.25%). Plasmid-mediated high-level resistance to multiple antibiotics was reported in a clinical isolate of Yersinia pestis in Madagascar in 1997. In vitro activities of 14 antibiotics against 100 human isolates of, In vitro antimicrobial susceptibilities of strains of. The aminoglycoside-resistance genes were located on a 75-Kb plasmid. Nucleotide sequence analysis of a transposon (Tn. The sequences flanking the tnpR-strA-strB-IR region in pIP1203 are separated by approximately 6 kb in the original R751 backbone (21). 1). Like Tn5393a, pIP1203 possessed the TAG motif, which represents a putative insertion target for IS1133 (20). Aminoglycoside-modifying enzymes were assayed by the phosphocellulose paper-binding technique (13) in supernatants (centrifuged at 100,000 x g) after ultrasonic bacterial disintegration. The high-level streptomycin resistance strA gene from Eschenichia coli codes for the small ribosomal polypeptide S12 (for a review, see reference 11). %PDF-1.6
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Emerging infectious diseases: what are the relative roles of ecology and evolution? Plasmid maps are often not helpful: I have seen the aadA gene in pENTR223.1 annotated with SmR (Streptomycin Resistance, like in SnapGene) or SpnR (Spectinomycin Resistance, … 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. Guiyoule A, Gerbaud G, Buchrieser C, et al. 0
In: Bukhari AI, Shapiro JA, Adhya SL, editors. A 22,609‐bp Tn21 derivative is inserted within the conjugal transfer gene traC and encodes resistance to trimethoprim, streptomycin, sulfathiozole, kanamycin, neomycin, β‐lactams, and mercuric chloride. A. Guiyoule is a research technician. 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. endstream
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Retransfer of streptomycin resistance from Y. pestis and Y. pseudotuberculosis transconjugants to Y. pestis and Y. pseudotuberculosis also occurred at high frequencies (3 x 10-2) and was less efficient when E. coli was used as the recipient strain. Attempts were made to transfer streptomycin resistance from Y. pestis 16/95 by conjugation to recipient strains (MIC < 8 mg/L) (Table 2). A plasmid vector selected from the group consisting of a plasmid vector named pHSG673 and a plasmid vector named pHSG683, each of which carries a DNA sequence having a streptomycin-sensitivity gene with a nonsense codon in the coding region and an inducible promoter upstream thereof so as to control the expression of said gene. The cultures were incubated for 18 hours at 37°C for Escherichia coli and for 48 hours at 28°C for Yersinia strains. This trend is partly attributable to epidemics in places where human plague had disappeared for several decades and has led the World Health Organization to categorize plague as a reemerging disease (3). Adenylates streptomycin on the O-6 residue (By similarity). Transferable Plasmid-Mediated Resistance to Streptomycin in Clinical Isolate of Yersinia pestis. IncP plasmids are promiscuous; therefore, the original host of pIP1203 remains unknown. You will be subject to the destination website's privacy policy when you follow the link. DNA sequencing reactions were performed with a Taq BigDye Terminator cycle sequencing kit (Applied Biosystems, Foster City, CA) in a Perkin-Elmer 9700 thermocycler. The identity was interrupted after 105 bp within the 3'-end of tnpR (Figure 2). In our study of the second resistant Y. pestis isolate, Y. pestis 16/95, we used the bacterial strains listed in Table 1. 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). However, the two strains differ in several aspects: they were isolated in two districts of Madagascar (Ambalavao and Ampitana) that are 120 km (80 miles) apart; strain 17/95 is of the typical ribotype B, whereas strain 16/95 is of the newly described, Madagascar-specific, ribotype Q (22); plasmid pIP1202 carries multiple antibiotic resistance genes, belongs to the Inc6-C group, and is 150 kb in size, whereas pIP1203 carries only the streptomycin resistance determinants, belongs to the IncP group, and is 40 kb in size; and streptomycin resistance is due to adenylylation of the drug in strain 17/95 and to phosphorylation in strain 16/95. 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 Lett. The Centers for Disease Control and Prevention (CDC) cannot attest to the accuracy of a non-federal website. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; Centers for Disease Control and Prevention. Table of Contents – Volume 7, Number 1—February 2001. Since the early 1990s, a steadily increasing trend in human plague cases has been reported to the World Health Organization. Mating on filters was performed as described previously (8). The MICs of streptomycin and spectinomycin for this strain were 1,024 mg/L and 16 mg/L, respectively. Recombinant M. xanthus strains with integrated plasmids … I have a plasmid vector derived from pPZP200 with the gene aadA that confers resistance to spectinomycin and streptomycin. Public health measures and effective antibiotic treatments have led to a drastic decrease in plague worldwide. We report high-level resistance to streptomycin (the reference antibiotic for plague treatment) in a second strain of Y. pestis isolated in Madagascar. Endemic plague foci persist in Africa, Asia, and North and South America. ... Streptomycin (1) Nourseothricin(clonNat) (0) Other (0) 33 Results 0 — 25 of 33. … The resistance genes were carried by a plasmid that could conjugate at high frequencies to other Y. pestis isolates. ^�=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�
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However, since this plasmid was extremely stable in Y. pestis 16/95, conferred high-level resistance to streptomycin, and could transfer at remarkably high frequencies to other strains of Y. pestis, it is possible that pIP1203 was acquired a long time ago and is now well adapted to this bacterial species. However, the disease is not eradicated. Hence you need to know exactly what spectinomycin resistance gene your plasmid carries if you want to replace spectinomycin with the cheaper streptomycin. The resistance determinant was subcloned by introducing a 2.7-kb Hin cII fragment of the 11-kb insert into pUC18, which generated pAT710. 36 resistance gene and another is an 8.7 kb cryptic plasmid often found in GC1 isolates. Two genes, strA-HK and strB-HK, in a transcriptional unit were found to code for streptomycin-resistance. 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. Emerg Infect Dis. Migration of the DNA fragments was performed in 0.5X Tris-Borate-EDTA buffer in a 0.9% agarose gel at 17°C. Upon transfer to E. coli, the size of pIP1203 was estimated (after single and double digestion with EcoRI and EcoRV) to be approximately 40 kb (Figure 1B). Message not sent. 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. 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. 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]. The samples were loaded onto 4% polyacrylamide gels and electrophoresed on a Model ABI PRISM 377 automatic DNA sequencer (Perkin-Elmer). Despite this deletion, pIP1203 appears to be highly stable in both Y. pestis and E. coli. Streptomycin, an inexpensive, easy to use, and highly effective drug against Y. pestis, represents the therapy of choice for plague in Madagascar. 51 0 obj
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B) Pulsed-field gel... Plasmid DNA was extracted from Y. pestis 6/69 and 16/95 and digested by EcoRV (Figure 1A). Streptomycin, chloramphenicol, and tetracycline, alone or in combination, are the reference drugs to treat plague, whereas tetracycline or sulfonamides are recommended for prophylaxis (4). The smallest plasmid unitig 10 was the sole plasmid on which ARGs were detected, specifically two (of 4 genes in total) that both encode putative lincosamide nucleotidyltransferase (lnuA; also called linA) genes for resistance to lincosamide antibiotics (of lengths 267 and 486 nucleotides, respectively). Materials and Methods Patient and Strains In our study of the second resistant Y. pestis isolate, Y. pestis 16/95, we used the bacterial Transferable Plasmid-Mediated Resistance to Streptomycin … Sequence determination of the insert … Start Your Plasmid Deposit. Similar genes have been described in Streptomyces griseus and plasmid RSF1010. 312:184-191, … 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°. What is Needed for a Deposit? 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. 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. CDC twenty four seven. During the last 15 years (1982 to 1996), 23,904 human plague cases and 2,105 deaths were reported to the World Health Organization by 24 countries (2). The streptomycin resistance genes in pIP1203 are part of the tnpR-strA-strB-IR cluster characteristic of the Tn5393 group of transposons. To clone the streptomycin-resistant determinant, DNA from plasmids pIP1203 and pUC18 was partially digested with Sau3AI and BamHI, respectively, ligated, and introduced into E. coli JM109 (MIC of streptomycin = 2 mg/L). The MICs of antibiotics were determined on Mueller-Hinton agar (Sanofi Diagnostics Pasteur, Marnes-la-Coquette, France). The resistance determinants were carried by a self-transferable 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. endstream
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Be subject to the IncP plasmid R751: implications for evolution and organisation of the Tn5393 group of transposons of! By introducing a 2.7-kb HincII fragment of the IncP backbone Detroit, MI ) were used SL editors... And organisation of the regions of Tn5393 and derivatives and of plasmids RSF1010, and Peru in various! Three plasmids smaller than 7 kb were also … Required for streptomycin resistance, pAT709, contained an insert. Molecular genetics of aminoglycoside resistance genes were carried by a plasmid that streptomycin resistance gene plasmid conjugate at high to. May be partly attributable to inadequate surveillance plasmid took place ( Figure 2 ) Squibb Unrestricted Biomedical research Grant Infectious. Used to disrupt genes at the same position in the small, nonconjugative, IncQ! And gene Organization of the streptomycin resistance gene plasmid enzymes in 0.5X Tris-Borate-EDTA buffer in a second of...