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The genetic basis of antimicrobial resistance in Ontario isolates of Actinobacillus (Haemophilus) pleuropneumoniae was studied. Two Ontario isolates of A. pleuropneumoniae were found to be resistant to sulfonamides (Su), streptomycin (Sm) and ampicillin (Amp). Resistance to Su and Sm was specified by a 2.3 megadalton (Mdal) plasmid which appeared to be identical to pVM104, which has been described in isolates of A. pleuropneumoniae from South Dakota. Southern hybridization showed that the 2.3 Mdal Su Sm plasmid was highly related to those Hinc II fragments of RSF1010 known to carry the Su Sm genes, but was unrelated to the remainder of this Salmonella resistance plasmid. Resistance to Su and Amp was specified by a 3.5 Mdal plasmid and appeared identical to pVM105 previously reported. The beta-lactamase enzyme had an isoelectric point of approximately 9.0. Southern hybridization showed no relationship to the TEM beta-lactamase. A third isolate of A. pleuropneumoniae was found to be resistant to chloramphenicol (Cm), Su and Sm by virtue of a 3.0 Mdal plasmid which specified a chloramphenicol acetyl transferase. We conclude that resistance to Su, Sm, Amp and Cm is mediated by small plasmids in A. pleuropneumoniae. Although the Su and Sm resistance determinants are highly related to those found in Enterobacteriaceae, the plasmids themselves and the beta-lactamase determinant are different.

pFM739, an R plasmid from Neisseria sicca that encodes penicillin, streptomycin and sulphonamide resistance, and the enterobacterial IncQ(P-4) plasmid RSF1010, which encodes streptomycin and sulphonamide resistance, were incompatible, and were mobilized by the same conjugative plasmids. Restriction mapping confirmed a high degree of similarity between both R plasmids; pFM739 carried DNA fragments corresponding to the known replication and resistance regions of RSF1010. pFM739 also carried an extra segment with the same restriction map as that described for the beta-lactamase-coding region of transposon Tn3. It is suggested that the R plasmids isolated from commensal Neisseria sp. could have resulted from transposition of a Tn3-like genetic element to an RSF1010-like plasmid, and that they contain deletion derivatives of transposon Tn3.

Of 7 plasmids we tested, the plasmid pORF2 was eliminated in vitro with the most efficiency by treatment with subinhibitory concentrations of novobiocin, coumermycin and 10 quinolones. It showed a cure rate of 43% by enoxacin; 12% by novobiocin, pefloxacin, ciprofloxacin and CI-934; 7% by coumermycin and ofloxacin; 9% by amifloxacin; and 4% by AM-833. On the other hand, pSC194, pBR322 and pMH612 were poorly cured in vitro by quinolones, except pSC194 which was cured 33% by enoxacin. R1, pP1603, and pUB110 were unaffected by the treatment. Mice were challenged intraperitoneally with a 2XLD50 of Escherichia coli carrying the ORF2 plasmid and were treated per os with 1 X or 1/2 X ED50 of either enoxacin or CI-934. The frequency of loss of ampicillin resistance determined 3 h after treatment shows curing effects of 92% for CI-934, 89% for enoxacin and 20% for untreated control.

The pnd gene of R plasmid R483, like the srnB gene of the F plasmid, increases the degradation of stable RNA in Escherichia coli. The nucleotide sequence of the pnd locus was determined and compared with that of the srnB locus. The genes have open reading frames that are 54% homologous, and both have an upstream inverted repeat sequence. The pnd gene expression seems to decrease the osmotic barrier of the cytoplasmic membrane, since no plasmolytic vacuoles were formed in the cells carrying the gene when the cells were exposed to hypertonic sucrose solution. This result suggests that RNase I in the periplasm passes through the altered membrane to degrade stable RNA in the cytoplasm.

Trimethoprim resistance mediated by the Staphylococcus aureus multi-resistance plasmid pSK1 is encoded by a structure with characteristics of a composite transposon which we have designated Tn4003. Nucleotide sequence analysis of Tn4003 revealed it to be 4717 bp in length and to contain three copies of the insertion element IS257 (789-790 bp), the outside two of which are flanked by directly repeated 8-bp target sequences. IS257 has imperfect terminal inverted repeats of 27-28 bp and encodes for a putative transposase with two potential alpha-helix-turn-alpha-helix DNA recognition motifs. IS257 shares sequence similarities with members of the IS15 family of insertion sequences from Gram-negative bacteria and with ISS1 from Streptococcus lactis. The central region of the transposon contains the dfrA gene that specifies the S1 dihydrofolate reductase (DHFR) responsible for trimethoprim resistance. The S1 enzyme shows sequence homology with type I and V trimethoprim-resistant DHFRs from Gram-negative bacteria and with chromosomally encoded DHFRs from Gram-positive and Gram-negative bacteria. 5' to dfrA is a thymidylate synthetase gene, designated thyE.

The Pseudomonas aeruginosa isolates in Taiwan were grouped with antisera against O antigens and tested with 13 different antimicrobial agents. Group E (28%) lead in serogrouping, followed by Group G (19%), Group F (16%), Group B and Group H (12%, respectively), and there were no significant differences in distribution of serogroups among the isolates from different areas in Taiwan. Amikacin (75%) and piperacillin (74%) were the most effective antibiotics, followed by tobramycin (63%), carbenicillin (62%), ticarcillin (57%) and gentamicin (56%). Group F strains were relatively more susceptible to most of the antibiotics tested than strains of other serogroups. All strains tested were multiple drug resistance. Thirty percent (67/224) of P. aeruginosa carried plasmids. The average number of resistant markers per strain was 8.7 for plasmid-carrying strains as compared to 7.6 for plasmid-free strains. The plasmid-carrying strains showed higher drug resistance than plasmid-free strains. Twenty-seven percent (13/48) of plasmid-carrying strains contained R plasmids which were detected by transformation (6/23) and conjugation (7/25). These R plasmids totally encoded at least seven different resistant markers. The R plasmid from P. aeruginosa 119 was not modified after transforming into Escherichia coli RR1. The R plasmids of P. aeruginosa transferred into E. coli RR1 or Serratia marcescens 73 caused some changes in the outer membrane of transformants or transconjugants. Some R plasmid-carrying strains and their transformants or transconjugants exhibited the ability to produce acetyltransferase.

It has been shown that the plasmid R89S derivatives can be used as integrative vectors for bacteria in which the plasmid is unable to replicate autonomously. The chromosomal and plasmid fragments of phototrophic bacterium Rhodobacter sphaeroides have been cloned in plasmid pVZ365, a SmRKmR-derivative of R89S. The obtained recombinant plasmids were mobilized into R. sphaeroides cells by the I pcP-group conjugative plasmid R751. The frequencies of the SmR-transconjugants formation are 3.7.10(-5) to 5.6.10(-3) per recipient cell. The formation of the SmR-transconjugants has not been revealed in case of the plasmid pVZ365 mobilization. The recombinant molecules containing R. sphaeroides plasmid fragments have been shown to integrate into endogenous plasmids and form cointegrates with them.

In 1983, a small outbreak of infections caused by a previously unrecognized multiply-drug-resistant Shigella flexneri 3a strain occurred on the Hopi Indian reservation. The index patient, a diabetic woman with recurrent Escherichia coli bacteriuria on prophylactic trimethoprim/sulfamethoxazole (TMP/SMZ) therapy, was hospitalized with concurrent E. coli urinary tract infection and shigellosis. Both E. coli isolated from her urine and S. flexneri isolated from her stool were resistant to ampicillin, carbenicillin, streptomycin, sulfisoxazole, tetracycline, and TMP/SMZ. Both isolates contained a 35-MDa plasmid transferrable to recipient E. coli, and transconjugates acquiring the plasmid from either donor strain also acquired resistance to the same agents. The number and size of fragments generated by plasmid digestion with DNA restriction endonuclease ClaI were similar. A review of clinical microbiology records showed that an E. coli strain isolated from the patient 3 w before the onset of shigellosis had identical antimicrobial resistance to the E. coli and Shigella isolated during the outbreak. These studies indicate that the index patient receiving prophylactic TMP/SMZ was the likely source of the R-plasmid for the outbreak strain of Shigella.

Six strains of S. typhimurium were isolated from water samples of Dal Lake, Srinagar (India). These strains could survive and multiply in the Lake water and its filtrate. Viable count of this bacterium increased abruptly on 6th day of incubation with a peak level on 7th and 9th day, followed by sharp decline on 12th-14th day. Viable count was higher at 37 degrees C than at room temperature. At 4 degrees C this bacterium survived for longer time than at 22 degrees C or 37 degrees C. Viable count was higher and survival longer in the filtrate than in Lake water. Drug resistance could be transferred to Escherichia coli recipient strains from S. typhimurium, both in filtrate and Dal Lake water at 4 degrees C, 22 degrees C and 37 degrees C. The prolonged survival of pathogenic bacteria and transfer of R-plasmid in Dal Lake water could be a health hazard.

The kanamycin resistance determinant of the broad-host-range plasmid RP4 encodes an aminoglycoside 3'-phosphotransferase of type I. The nucleotide sequence of the kanamycin resistance gene (Kmr) and the right end of the insertion element IS8 of plasmid RP4 has been determined. The gene (816 bp) is located between IS8 and the region (Tra 1) encoding plasmid factors mediating bacterial conjugation. Kmr and Tra 1 are transcribed toward each other. The nucleotide sequence has been compared to five related aphA genes originating from gram-negative and gram-positive organisms and from antibiotic producers. Among these that of Tn903 shares the highest degree of similarity (60%) with the RP4 gene. Significant similarities were also detected between the amino acid sequences of the six enzymes. The C-terminal domains of six different aminoglycoside 3'-phosphotransferases (APH(3'] are highly conserved. They are substantially similar to segments of a variety of enzymes using ATP as cofactor. The role of the C-terminal sequences of APH(3') as potential domains for ATP recognition and binding is discussed.

Haemophilus species have been implicated as the source of plasmid-mediated ampicillin resistance in Neisseria gonorrhoeae. Previous attempts to transfer conjugally the resistance plasmids from Haemophilus species to N. gonorrhoeae have met with limited success. Using both biparental and triparental mating systems, it was found that transfer will occur if the commensal Neisseria species, Neisseria cinerea, is used as a transfer intermediate. This organism stably maintains resistance plasmids of Haemophilus and facilitates transfer of these plasmids to N. gonorrhoeae, in a triparental mating system, at a transfer frequency of 10(-8). Both Haemophilus ducreyi and N. gonorrhoeae carry mobilizing plasmids capable of mediating conjugal transfer of the same resistance plasmids. However, restriction endonuclease mapping and DNA hybridization studies indicate that the mobilizing plasmids are distinctly different molecules. Limited homology is present within the transfer region of these plasmids.

To develop a highly efficient means for generating methotrexate resistant (MTXr) hematopoietic cells in vivo, a recombinant retroviral genome was constructed that encodes a MTXr dihydrofolate reductase (DHFRr). Cell lines producing high titers of virus capable of transmitting the DHFR gene were generated and used to infect mammalian cells in vitro. Analysis of infected fibroblasts indicated that the DHFRr gene was transmitted intact and conferred a high level of MTXr upon cells. Based on these findings, DHFRr-containing virus was used to infect murine bone marrow cells in vitro. Following infection, the transduced cells were introduced into lethally irradiated recipients via bone marrow transplantation techniques. The presence of the proviral sequences in cells of the spleen and bone marrow of engrafted recipients was associated with significantly increased survival of mice treated with otherwise lethal doses of MTX.

The paper presents data on the nature of the conjugation-like process (CLP) of genetic marker transfer on the membrane filters in group A streptococci. Studied are the CLP development depending on the duration of the donor-recipient contact and the influences of various physiological conditions on the degree of CLP manifestation and frequency. The effect of sub-inhibiting erythromycin concentrations on the frequency of the erythromycin marker transfer is first found. The original experimental evidence and literature data suggest a role played by transposing elements in CLP determination on the membrane filters in group A streptococci.

Present status of the problem of origin of extra-chromosomal inheritance elements is reviewed in the light of literary data and some results obtained by the author. The role of plasmids in the ecology of bacteria, in their adaptive variability and evolution is discussed. Extra-chromosomal inheritance is regarded as a particular case of biological inheritance.

The aim of the present study was to determine which types of beta-lactamases codified by plasmids are transferred by conjugation from several species of enterobacteria. To this end, 352 strains of ampicillin-resistant enterobacteria from clinical samples from the Hospital Civil of Bilbao were evaluated. Their beta-lactamase activity and their capacity to transfer this capacity by conjugation were evaluated. The several types of plasmidic beta-lactamases in the strains that conjugated and in their respective transconjugants were characterized by analytic isoelectric approach, and also the sensitivity of these stains to 20 beta-lactamic antibiotics and the size of their plasmids. Twenty different types were detected, with a clear predominance of TEM 1. Type TEM 2 was found in 19% of the strains which conjugated, and much less commonly the types SHV 1, HMS 1 and a beta-lactamase of an approximate pl of 4.9 were found. The transfer of these beta-lactamases is mediated by a great variety of plasmids and is associated with variable levels of resistance to penicillins and unstable cephalosporins. The presence of betalactamases with activity on the more stable cephalosporins has not been detected.

Expression of plasmid-encoded resistances in regenerating protoplasts of Bacillus subtilis occurs only after wall synthesis has been resumed. This is observed for protoplasts obtained from cells already containing plasmids (pC194, pT127, pK545) or for plasmid-bearing cells coming from a PEG-mediated transformation. Recovery of expression needs a 2-h incubation of protoplasts, previously washed to get rid of their lysozyme content, in rich hypertonic medium (SMMP). A longer incubation (24-h) results in the obtention of regenerants; however, most of them have lost their resistant phenotype in contrast to those obtained from the usual solid regeneration plates. This finding suggests either a high curing effect or some kind of gene inactivation phenomenon. Discussion is focused on the critical points that have to be considered when polyethylenglycol-mediated transformation of protoplasts is applied to recombinant DNA technology.

Lactose fermenting Salmonella typhimurium are endemic in São Paulo, but not in Rio de Janeiro Two isolations are described from the latter city. These Rio de Janeiro strains have a plasmid of 7.4 megadaltons. These plasmids were not auto-transferable, were thermostable and were not eliminated by acridine orange. One of these strains arose from a plasmid that had the lactose operon repressed, leading us to speculate about the evolution of the lactose fermenting character in Brazilian Salmonella.

DNA of S. fradiae B-45 partially cleaved by Sau3A restrictase was cloned in S. lividans TK64 in the plasmid vector pIJ702. Three recombinant plasmids pVG251, pVG262, and pVG253 with tlr1, tlr2 and tlr3 genes were isolated from the transformed clones of S. lividans TK64 with higher inducible resistance to tylosin as compared to the plasmid-free strain. DNA-DNA blot hybridization was performed between the total DNA cleaved by several restrictases from S. fradiae B-45 and some other strains and the DNA probes containing the tlr genes. It was shown that tlr1 and tlr3 genes were unique in S. fradiae B-45. Sequences homologous to tlr2 gene were present both in DNA of S. fradiae B-45 in 7 copies and in strains of S. antibiotics and S. hygroscopicus producing respectively oleandomycin and turimycin.

Cointegrate plasmids were formed in vivo between the broad-host-range R-plasmid RP4 and two catabolic plasmids derived from Pseudomonas putida HS1. One of these was the wild-type plasmid pDK1 encoding the complete inducible toluene/xylene (TOL) catabolic pathway and one was pDKT1, a deletion derivative of pDK1 selected after growth of HS1 on benzoate and supporting growth on only toluene. The two plasmids formed, pDK2 and pDKT2 respectively, each consisted of a complete RP4 replicon in which was an insert of the parent plasmid DNA respectively 40 and 20 kbp in size. The detailed restriction maps of the two plasmids were determined and many of the catabolic genes were located by subcloning and enzyme assay of recombinant plasmids in Escherichia coli and Pseudomonas hosts. The insert in pDK2 contained both operons of the catabolic pathway, the 'upper pathway' operon (xylCAB) and the meta pathway operon (xylDLEGF(I,J,K)H), and a region identified as having the function of the regulator gene xylS. The insert in pDKT2 contained only the upper pathway operon and the regulatory region. Within each of the three coding regions there was great similarity with the same regions on TOL plasmids pWW0 and pWW53-4 apparent (a) by the same order of the genes, (b) by a similar pattern of restriction sites and (c) by hybridization studies. However, the order and orientations of the three coding regions differed from those previously described for both pWW0 and pWW53-4. The significance of these findings to the evolution of TOL plasmids is discussed.

The MerR protein mediates the induction of the mercury resistance phenotype in bacteria; it has been isolated in order to study the effects of metal-ion induced changes in the metabolism of prokaryotic cells at the molecular level. After DNA sequences responsible for negative autoregulation were removed, the 16-kilodalton protein was overproduced and purified to more than 90 percent homogeneity by a salt extraction procedure that yields about 5 milligrams of protein per gram of cells. Complementation data, amino terminal analysis, gel filtration, and deoxyribonuclease I protection studies demonstrate that the purified merR gene product is a dimer under nondenaturing conditions and that it binds specifically to DNA, in the presence and absence of mercury, at a palindromic site which is directly between the -10 and -35 regions of the structural genes and adjacent to its own promoter. These initial results indicate that MerR is a DNA-binding metalloregulatory protein that plays a central role in this heavy metal responsive system and they delineate an operator site in the mer operon.

By the use of an Hfr strain made by integrative suppression of a derepressed R factor, R100-1 in a dnaA-ts mutant of Escherichia coli and several other appropriate control strains with and without the R factor, a screening system was established which can be used to detect agents which are either inhibitory to the replication of the R factor and lead to its "curing" or are preferentially inhibitory to the growth of the R-carrying bacteria. The validity of this system was shown with several agents already known to be experimentally effective.

Bacterial resistance to mercury has been studied in two different strains from animal origin, Salmonella typhimurium 9205 and Escherichia coli 467. These two strains are resistant to mercuric chloride but sensitive to phenylmercury, and thus belong to the group of bacteria that possess a "narrow" spectrum resistance. The presence of plasmids within the cells has been demonstrated through conjugation experiments and direct detection of extrachromosomal DNA in transconjugants. These plasmids, termed p9205-1 and p467-30, differ from each other by both their size (100 and 50 Kbp, respectively) and structure, as shown by the restriction patterns arising from digestion by nucleases BamHI, HindIII, PstI and EcoRI. Each plasmid has been treated with enzyme SalI to yield DNA fragments that have been cloned into pBR322. Two recombinant plasmids, p9205-1/Sal and p467-30/Sal, have thus been constructed, each of them harboring a 2 Kbp fragment that appears to contain the merA gene coding for mercuric reductase. From the analysis of the restriction maps of these recombinant plasmids as well as the functional behaviour of the bacteria that they are able to transform, it can be concluded that they are identical.

Plasmid pBP11 contains a sequence homologous to Tn21-like element Tn2410 encoding dihydropteroate synthetase and beta-lactamase OXA-2. The nucleotide sequence of a 1.5 kb segment of this region has been determined including the bla gene. It reveals strong sequence homology with the OXA-2 operon of plasmid R46. The implications of an additional 319 bp segment in pBP11 for the different evolution of R46/pKM101 and pBP11 are discussed.

A total of 62 clinical strains of Staphylococcus aureus collected from Kaohsiung Medical College were examined for susceptibility of several antimicrobial agents and underwent plasmid analysis. The results of the antibiotic susceptibility test revealed that 33.9% were resistant to methicillin and 9.7% were resistant to gentamicin, whereas, resistance to heavy metal ions such as arsenic and cadmium were as high as 95% and 76% respectively. Antibiotic multiresistant isolates of S. aureus were also found. Multiresistance to nalidixic acid-tetracycline, ampicillin-nalidixic acid-tetracycline, and ampicillin-chloramphenicol-kanamycin-streptomycin- tetracycline were most frequently encountered. The phage typing results indicated that type 52 was predominant in 27%, followed by 11% of 52/83A and most strains fell into group I and a mixed group of I & III. In addition, these types expressed were highly correlated with a resistance to ampicillin, chloramphenicol, streptomycin, nalidixic acid, kanamycin and methicillin. Plasmid DNA profiles from these isolates exhibited different patterns. The plasmid of mol wt. 27 x 10(6) was associated with resistant to ampicillin and kanamycin, whereas, a plasmid of mol wt. 30 x 10(6) effected resistance to ampicillin, kanamycin and streptomycin. The determinant which conferred resistance to chloramphenicol was carried by a plasmid of mol wt. 3 x 10(6), and tetracycline resistance was encoded on a plasmid of mol wt. 2.8 x 10(6) or 28 x 10(6) in different isolated strains. Analysis of plasmids by HindIII digestion showed that plasmids of different molecular weight but with similar phenotypic characteristics often share common fragments.

A pR351 plasmid (Tc Ap Cb) conferring drug resistance of naturally occurring E. coli strain was examined. Conjugation and transduction experiments have indicated that this plasmid is R plasmid aggregate consisting of three independent plasmids: a) conjugative plasmid pR351 A (SuTc) fi- (F) belonging to incompatibility group L, b) conjugative plasmid pR351 B (SuApCb) fi- (F) belonging to incompatibility O, and c) non-conjugative plasmid pR351 C (ApCb). The existance of these plasmids in pR351 plasmid aggregate was confirmed by the agarose gel analysis of plasmid DNA isolated from the lysates of E. coli J53 transconjugants carrying pR351 A or pR351 B plasmids and from E. coli K12 C600 transductant carrying pR351 C plasmid. Molecular mass of these plasmids was found to be 55.60 and 3 Mdal respectively. The non-conjugative pR351 C plasmid could be mobilized by Col B and F factors. Our findings have indicated that two fi- (F) R plasmids can stably coexist in R plasmid aggregate.

The region of the antibiotic resistance plasmid R100 that encodes the plasmid-specific transfer gene traM has two tandemly aligned promoters separated by 145 nucleotides. The principal transcripts are 705 and 562 nucleotides long. Minor transcripts are 1550 and 1700 nucleotides long. The 705-base transcript appears to encode an 11 kD traM protein. The 562-base transcript does not encode a detectable protein. When subcloned on short fragments, the promoter for the 562-base transcript initiates efficiently but that for the 705 site does not. The 3' ends of the 705 and 562 base transcripts end inside the traJ ORF. Thus they provide additional sense RNA to compete with traJ for finP, the antisense translational regulator of traJ. A model is proposed for the participation of these sense and antisense transcripts in the control of expression of the traJ gene.

Two hundred and fifty Shigella strains isolated in Japan were examined for their drug-resistances and their conjugal transferability. Drugs used and the isolation frequency of resistant strains were: sulfanilamide (SA) 96%, streptomycin (SM) 85%, tetracycline (TC) 82%, fosfomycin (FOM) 76%, chloramphenicol (CP) 51%, trimethoprim (TMP) 46%, ampicillin (ABPC) 45%, nalidixic acid (NA) 6%, kanamycin (KM) 5%, cephaloridine (CER) 4%, gentamicin (GM) 0%, amikacin (AMK) 0%, colistin (CL) 0%, norfloxacin (NFLX) 0%. All of the KM-resistances and none of the FOM-resistances were transferable. Each 20 to 30% of CP, TC, SM, SA or ABPC-resistant isolates transferred each drug-resistance. Two thirds of the isolates were brought to Japan by travellers mostly from South-East Asia. The isolation frequency of resistant strains was not different between domestic and foreign strains except that NA-resistance was not found in foreign strains. The frequency of isolates carrying transferable R plasmids were higher in domestic strains than foreign strains. Most of the isolated had multiple drug-resistance. Major resistance types were: (CP, TC, SM, SA, ABPC) 22%; (TC, SM, SA, TMP) 20%; (CP, TC, SM, SA, ABPC, TMP) 16%; (TC, SM, SA) 8%. Strains showing different resistance types were different among each other in their serum type, conjugal transferability, and derived country. Stepwise evolution of multiple resistance in one country was not demonstrated. The resistance types of Shigella isolates in Japan have been influenced by yearly change of imported strains.

Many plasmids affect the host cells. Their effects cannot be explained only by the expression of the well-known genes coding for antibioticresistance, bacteriocinogeny and hemolysis or the analogous genes (side-effects). The side effects are not characteristic of all plasmids operating under similar conditions. Forecasting of the side-effects inducikility by any definite plasmid is impossible now. Sometimes the same functions exert the contrary effects on the bacterial cell. The connection between the presence of plasmids, especially R-plasmids and the complex cellular property, virulence, is of great interest. Often, bacteria become less virulent obtaining the plasmids. Two possible reasons causing such an effect are discussed. The first one is a direct effect of plasmids on cellular physiology. The second reason is connected with population shifts caused by the fact that the cells with initial low virulence possess the recipient ability predominantly. The decreased virulence of bacteria harbouring R-plasmids, in authors opinion, is quite a natural phenomenon based on plasmid host cells adaptation to the existence in "the realm of antimicrobial agents".

Tetracycline resistance determinants of R plasmids from the fish pathogens Aeromonas hydrophila, Edwardsiella tarda, and Pasteurella piscicida were classified as class D by their expression of resistance to tetracycline and minocycline and through their DNA structure.