Effects of task size on work-related and aberrant behaviors of youths with autism and mental retardation.

The effects of task size on rate of responding and on-task behavior as well as on nontask-related behaviors of students with autism and mental retardation on a repetitive task under conditions of no reinforcement for responding was analyzed. Task size, defined as the presence of either 36 or 250 beads in a container at the onset of the session, was compared in an alternating treatment design. The small-task condition resulted in higher on-task behavior for all participants and in higher work rate for four of the five participants. For the four participants who engaged in inappropriate use of task materials, higher levels of this behavior occurred in the large-task conditions. Other nontask-related behaviors were higher in the large-task condition for all participants with the exception of stereotypy, which was higher in the small-task condition for one participant. Better work-related behavior occurred for these participants in small- than in large-task conditions even though no tangible reinforcement was provided for task responding. Implications of these results are discussed in the context of arranging workplace environments to maximize productivity of persons with developmental disabilities.

Conjugational transfer of gentamicin resistance plasmids intra- and interspecifically in Staphylococcus aureus and Staphylococcus epidermidis.

We have previously reported the transfer of gentamicin resistance (Gmr) plasmids in a mixed culture inter- and intraspecifically between strains of Staphylococcus aureus and Staphylococcus epidermidis isolated at Michael Reese Hospital (Jaffe et al., Antimicrob. Agents Chemother. 21:773-779, 1982). We have now shown that representatives of these plasmids were transferred between apparently nonlysogenic strains of S. aureus either in mixed culture in broth or by filter-mating on agar medium. The mechanism of transfer appeared to be conjugation. A transferable Gmr plasmid (pSH8) mobilized or cotransferred a tetracycline R-plasmid and a chloramphenicol R-plasmid that were not independently transferable. The transfer of Gmr plasmids was accompanied by a high incidence of deletion mutations with varied loss of plasmid resistance determinants and, with some mutants, loss of the ability to effect self-transfer. Restriction endonuclease digestion of pSH8 and its deletion mutants made it possible to assign the property of self-transfer to a specific segment of the pSH8 genome and provided the basis for a physical and genetic map of that plasmid. Similar Gmr plasmids from S. aureus strains isolated in locations remote from Michael Reese Hospital had resistance determinants and transfer properties comparable to those of pSH8. Our observations provide evidence for the conjugal transfer of some staphylococcal plasmids, apparently independent of the presence of phage. This mechanism may be of significance in the intra- and interspecific dissemination of resistance to aminoglycosides and other antibiotics in Staphylococcus spp.

Structural and phenotypic varieties of gentamicin resistance plasmids in hospital strains of Staphylococcus aureus and coagulase-negative staphylococci.

We previously described a neonatal nursery epidemic of infections caused by a single strain of Staphylococcus aureus bearing a gentamicin resistance plasmid (Vogel et al., Antimicrob. Agents Chemother. 13:466-472, 1978). The same plasmid was present in two isolates of Staphylococcus epidermidis from the patients in this nursery and was transferable interspecifically from either S. aureus or S. epidermidis. During the ensuing 3 years, in the absence of further epidemics, we collected 162 gentamicin-resistant strains of S. aureus and coagulase-negative staphylococci from patients distributed throughout our hospital. Gentamicin resistance plasmids obtained from 41 representative S. aureus and coagulase-negative staphylococcal strains differed as determined by phenotypic and molecular analyses from the plasmid in the neonatal nursery epidemic. Nevertheless, these plasmids were structurally related to each other and to the plasmid of the original epidemic. Our results suggest an evolutionary relationship among these plasmids and support the hypothesis of a genetic reservoir of gentamicin resistance in coagulase-negative staphylococci transferable to S. aureus.

Gentamicin-resistant staphylococci as hospital flora: epidemiology and resistance plasmids.

In 1979, gentamicin-resistant (Gmr) Staphylococcus aureus was recovered from the clinical specimens of 22 epidemiologically unrelated hospitalized patients; 78% of the patients evaluated were also colonized with Gmr coagulase-negative staphylococci. Endonuclease fingerprinting was used to compare the Gmr plasmids within pairs of isolates of S. aureus and coagulase-negative staphylococci recovered from nine patients. Plasmids differed between pairs but were concordant within four pairs. Thus, in vivo interspecific plasmid transfer, although infrequent, may be important in the sporadic occurrence of Gmr S. aureus. To define the epidemiology of endemic Gmr coagulase-negative staphylococci, culture surveys were performed over two years. Gmr coagulase-negative staphylococci were acquired by 80% of the infants in a special-care nursery that previously had an outbreak of Gmr S. aureus. Among adult inpatients, a 48% colonization rate was related to prior exposure to antibiotics. In contrast, no colonization was found in outpatients or antepartum mothers.

Identity and interspecific transfer of gentamicin-resistance plasmids in Staphylococcus aureus and Staphylococcus epidermidis.

The hypothesis that emergence of gentamicin-resistant strains of Staphylococcus aureus and Staphylococcus epidermidis in a neonatal special care nursery was the result of transfer of a single plasmid between these two species was examined. In experiments with mixtures of staphylococci, either in mixed cultures or on human skin, isolates of S. aureus and S. epidermidis transferred their gentamicin-resistance plasmids both intra- and interspecifically. By electron microscopy, the molecular masses of the plasmids from S. aureus and S. epidermidis were the same, 12.2 +/- 0.36 (standard deviation) and 12.3 +/- 0.56 megadaltons, respectively. Restriction endonuclease analysis of the plasmids from five isolates of S. aureus and two isolates of S. epidermidis, with use of the enzymes HaeIII, EcoRI, XbaI, and HindIII, showed no differences in the digestion patterns of the seven gentamicin-resistance plasmids. The results supported the hypothesis that plasmid transfer between S. aureus and S. epidermidis occurs in nature.

Modulation of protein A formation in Staphylococcus aureus by genetic determinants for methicillin resistance.

Many methicillin-resistant (Mec(r)) strains of Staphylococcus aureus either produce no protein A or secrete it extracellularly (S. Winblad and C. Ericson, Acta Pathol. Microbiol. Scand. Sect. B 81:150-156, 1973). We found that methicillin resistance and protein A production were apparently lost coordinately from the natively Mec(r) strain A676. Restoration of the genetic determinant for methicillin resistance (mec) by transduction or transformation restored protein A production. In two other Mec(r) strains, loss of mec was accompanied by marked reduction in protein A formation. Genetic transfer of mec to derivatives of S. aureus 8325 affected protein A formation differently with different mec determinants. Those derived from strain A676 and two other Mec(r) strains reduced the scanty amount of protein A produced by strain 8325 to even lower or undetectable levels, whereas mec from two more Mec(r) strains increased its protein A content. This "mec-effect," i.e., stimulation or inhibition of protein A formation dependent on the combination of host strain and mec determinant, was reduced in methicillin-susceptible (Mec(s)) mutants produced by ethyl methane sulfonate treatment of Mec(r) strains. The mec-effect reappeared in spontaneous revertants to methicillin resistance. Phenotypic reduction of methicillin resistance in Mec(r) strains grown at 44 degrees C was accompanied by reduction of the mec-effect on protein A, but it had no effect on protein A formation in Mec(s) strains. Two independent mutants of strain 8325 produced large amounts of protein A at rates that were unaffected by growth at 44 degrees C or by the introduction of mec determinants.

Infections due to gentamicin-resistant Staphylococcus aureus strain in a nursery for neonatal infants.

An apparently homogeneous strain of Staphylococcus aureus resistant to gentamicin (Gmr), kanamycin, tobramycin, and sisomicin, but susceptible to amikacin and netilmicin, caused multiple infections in neonatal infants in a special care nursery. Nasal cultures revealed a high rate of carriage of the Gmr staphylococcus in infants without clinical infection. Segregating patients according to a modified cohort system and use of careful aseptic techniques led to apparent elimination of the Gmr strain. The resistance to aminoglycosides in this strain was mediated by an aminoglycoside 6'-N-acetyltransferase and a gentamicin phosphotransferase. Genetic determinants for these enzymes were borne on a circular covalently closed plasmid of approximately 11 megadaltons. These resistance determinants closely resemble those found in isolates of S. aureus that have caused nosocomial infections in patients in Europe.

Mutations in prophage phi11 that impair the transducibility of their Staphylococcus aureus lysogens for methicillin resistance.

Methicillin resistance (mec) is not transduced into Staphylococcus aureus 8325-4, but is transduced into this host after it has been lysogenized with phage phi11 and has acquired the penicillinase plasmid pI524 by a separate transduction (Cohen and Sweeney, 1970, 1973). Strain 8325-4 is competent for transformation of typical plasmid or chromosomal markers and for mec only if it is lysogenic for phi11 or a related prophage (Sjöström et al., 1974, 1975). A mutant strain of phi11 that was temperature sensitive (Ts) for vegetative multiplication did not mediate competence for transformation of its 8325-4 lysogen if the lysogen had been grown at a nonpermissive temperature (Sjöström and Philipson, 1974). We isolated four Ts mutants of phi11 that did not mediate transducibility of their 8325-4(pI524) lysogens for mec after growth at nonpermissive temperatures (40 to 42 degrees C). Transduction of typical plasmid or chromosomal markers was not affected. These phi11-Ts mutants mediated normal competence of their lysogens for transformation of a tetracycline resistance plasmid. Similarly, phi11-Ts mutants that rendered their lysogens temperature sensitive for transformation did not depress the frequency of transduction of mec. These two types of phi11-Ts mutants fell into two different genetic complementation groups that differed in the physiology of deoxyribonucleic acid synthesis and in the time of expression of the mutations during a single-burst growth cycle at a nonpermissive temperature. A virulent mutant of phi11, which plaqued with 100% efficiency on 8325(phi11), also failed to condition strain 8325-4 for transducibility of mec but retained the ability to confer competence for transformation of a tetracycline resistance plasmid. Different genetic loci and physiological functions are involved in phi11 mutations that affect transducibility of mec and those that affect competence for transformation of markers generally in S. aureus 8325-4.

Co-transduction of plasmids mediating resistance to tetracycline and chloramphenicol in Staphylococcus aureus.

Independent plasmids mediating resistance either to tetracycline (Tc) or chloramphenicol (Cm) were transduced successively into Staphylococcus aureus strain 8325. From this doubly resistant donor strain, Tc was co-transduced with a frequency of 40 to 50% when Cm was selected. Co-transduction of Cm was 5 to 10% with Tc selection. Plasmid elimination was infrequent and restricted to the Cm plasmid. A variant, doubly resistant strain gave 100% co-transduction of Tc and Cm and a high rate of joint elimination of both plasmid markers. Co-transduction of the plasmids from recombination-deficient donor strains was much reduced if the plasmids had been introduced separately into the donor strain, but occurred at the normal high rate if they had been introduced jointly. The plasmids were co-transformed at relatively low rates with closed circular deoxyribonucleic acid (DNA) from doubly resistant donors, but not with DNA from a mixed lysate of singly resistant strains. Our evidence favored a hypothesis of recombination-dependent, reversible linkage between the two plasmids as the basis for their co-transduction. Examination of plasmid DNA from the doubly resistant strains by ultracentrifugal and electron microscopic methods did not disclose any physical differences between singly and doubly resistant strains that might account for the observed co-transduction.

Isolation and characterization of a kanamycin resistance plasmid from Staphylococcus aureus.

A circular covalently closed duplex deoxyribonucleic acid plasmid carrying genes for resistance to kanamycin/neomycin has been identified in Staphylococcus aureus E419. The plasmid has a molecular size of 9.2 x 10(6) daltons and can be transduced into or can transform competent susceptible strains of S. aureus to kanamycin/neomycin resistance.

Absence of circular plasmid deoxyribonucleic acid attributable to a genetic determinant for methicillin resistance in Staphylococcus aureus.

Plasmid deoxyribonucleic acid was not detected by centrifugal analysis of lysates of penicillinase-negative strains of Staphylococcus aureus harboring a determinant of methicillin resistance derived from strain Villaluz. When these strains contained a penicillinase plasmid, the plasmid deoxyribonucleic acid of methicillin-resistant and methicillin-susceptible strains was indistinguishable by the methods employed. The results indicate that the genetic determinant for methicillin resistance in the strains examined was not associated with a circular plasmid comparable to those that have been shown to determine resistance to benzylpenicillin, tetracycline, and chloramphenicol in S. aureus.

Effect of the prophage and penicillinase plasmid of the recipient strain upon the transduction and the stability of methicillin resistance in Staphylococcus aureus.

Transduction of a methicillin-resistance determinant (mec) in Staphylococcus aureus RN450 was dependent on its prior lysogenization with an appropriate temperate phage. In addition, an appropriate transduced penicillinase plasmid was usually required. Some phage 80-resistant variants of RN450 or of its parental lysogenic strain, NCTC 8325, were also effective recipients for transduction of mec. Elimination of prophage from RN450 abrogated its effectiveness as a transductional recipient of mec. Elimination of prophage from a methicillin-resistant transductant of RN450 reduced resistance to undetectable levels in six of seven phage-eliminated strains. In four of these a variable number of clones again became phenotypically resistant after lysogenization alone or lysogenization combined with reintroduction of a penicillinase plasmid. In two prophage-eliminated strains, no evidence of residual mec could be adduced. The establishment, expression, or stability of the transduced mec in strain RN450 appeared to depend on some function determined by a prophage or a prophage and a penicillinase plasmid.

Phenotypic suppression of methicillin resistance in Staphylococcus aureus by mutant noninducible penicillinase plasmids.

Methicillin (intrinsic) resistance of Staphylococcus aureus was suppressed almost completely by regulatory gene (penI(1)) mutations of penicillinase plasmids that made penicillinase production strictly noninducible. Methicillin resistance was restored by secondary regulatory gene mutations that altered the noninducible phenotype or by complementation with a compatible plasmid that did not bear the noninducible mutation. No evidence was obtained for genetic linkage between a penicillinase plasmid and the gene for methicillin resistance. We suggest, therefore, that the mutant noninducible repressor acted in trans by binding to a site on the methicillin resistance determinant. This hypothesis would imply an appreciable degree of homology between penicillinase plasmids and methicillin resistance genes.

Transduction of Methicillin Resistance in Staphylococcus aureus Dependent on an Unusual Specificity of the Recipient Strain.

Resistance to methicillin was transduced by phage 80 or 53 from two naturally occurring methicillin-resistant strains of Staphylococcus aureus to methicillin-susceptible recipient strains at frequencies of 10(-7) to 10(-9). Ultraviolet irradiation of transducing phage and posttransductional incubation at 30 C were essential for useful frequencies of transduction. Effectiveness as a recipient for this transduction was highly specific. Strain NCTC 8325 (PS47) in its native state was an ineffective recipient but became effective after it had received by transduction one of several penicillinase plasmids. This acquired effectiveness was retained in most cases after elimination of the plasmid by ethidium bromide treatment. Like the donor strain, the progeny were heterogeneous in the degree of their resistance to methicillin, which was expressed by a higher proportion of cells as the temperature of incubation was lowered from 37 to 30 C. Separate transductants varied widely in the degree of resistance acquired by transduction. Methicillin resistance was stable in the donor and transductant strains. We favored the interpretation that methicillin resistance in our strains was determined by a single chromosomal gene, although the possibility that it was determined by two or more closely linked genes could not be excluded.

Differential depression of staphylococcal plasmid and chromosomal penicillinase genes by a class of unlinked chromosomal mutations (R2-).

We described previously a class of mutations (phenotypic designation R2(-)) in Staphylococcus aureus that were unlinked to penicillinase genes but nevertheless imposed constitutive synthesis of this enzyme upon wild-type plasmids. The plasmids affected by these mutations have been found to possess the same maintenance-compatibility type (I) and to form penicillinase of serotype A. Three plasmids that had a different maintenance-compatibility type (II) and formed penicillinase of serotype C were only slightly derepressed in R2(-) mutants. Three chromosomal penicillinase genes were variably derepressed, the range extending from negligible to nearly complete derepression. The differences in degree of derepression among the three types of penicillinase linkage groups were tentatively ascribed to position effects.

Evidence for genetic interaction between plasmid and chromosomal penicillinase linkage groups in Staphylococcus aureus.

Spontaneous reciprocal recombination was detected between the regulator genes of penicillinase plasmids and chromosomal penicillinase linkage groups of Staphylococcus aureus. The frequency of recombination varied with different penicillinase linkage groups.

Wild-type strain of Staphylococcus aureus containing two genetic linkage groups for penicillinase production.

Staphylococcus aureus strain 55C1, isolated from a patient in 1955, contained two genetic linkage groups for penicillinase formation. One was linked to genes that control resistance to cadmium and mercuric ions; it had properties of a plasmidborne gene. The other was not linked to resistance to these metal ions; it had properties of a chromosomal gene. Penicillinase formation by cells that contained either linkage group was inducible by penicillins. Induced penicillinase in cells that contained both linkage groups equalled the sum of that produced in cells containing each group singly. Exopenicillinase produced by cells containing either gene was serological type A. Constitutive penicillinase formation resulting from regulator gene mutations in either linkage group was repressed to differing extents by a wild-type determinant in the trans position. The genetic structure and the regulation of penicillinase formation in strain 55C1 resembled in general those for penicillinase linkage groups which Asheshov and Dyke described for diploid mutant strains of S. aureus PS 80. There were differences in detail, however.

Constitutive penicillinase formation in Staphylococcus aureus owing to a mutation unlinked to the penicillinase plasmid.

Previously described penicillinase-constitutive mutations in Staphylococcus aureus are caused by genetic lesions in a regulator gene (or genes) on the penicillinase plasmid in close linkage to the structural gene. This report describes a new class (R2(-)) of penicillinase-constitutive mutants of S. aureus unlinked to the plasmid. By transductional analysis, the penicillinase plasmids in these mutants were wild type. Wild-type plasmids transduced into penicillinase-negative (plasmid loss) derivatives of R2(-) mutants produced penicillinase constitutively in amounts comparable to a fully induced culture of the wild-type strain. Penicillinase production in R2(-) mutants was maximal at 30 to 32 C and was much reduced at 40 C.

Relation between iron uptake, pH of growth medium, and penicillinase formation in Staphylococcus aureus.

The uptake of iron and the formation of penicillinase was examined in cultures of wild-type Staphylococcus aureus. Uptake of iron was about twice as great at pH 4.7 as at pH 7.4 At pH 4.7, increase in iron uptake in the range of 1.0 to 4.0 mug per mg of bacterial protein was associated with a progressive increase in the rate of penicillinase formation, but a direct correlation between cellular iron content and rate of enzyme formation was not demonstrated. Addition of iron to deferrated medium enhanced penicillinase formation at pH 6.5 to 7.4 two- to fourfold in cultures induced with benzylpenicillin and in uninduced cultures. To demonstrate an effect on the uninduced cells, it was necessary to increase iron uptake by preliminary incubation of cells with iron in buffer. Calcium and certain other ions depressed iron uptake at acidic and at neutral pH, and, presumably as a result of this action, depressed the formation of penicillinase. Iron did not enhance penicillinase formation at pH 4.7 by two penicillinase constitutive mutants nor by wild-type cells undergoing induction at pH 6.5 by cephalosporin C or methicillin. After removal of cephalosporin C or methicillin during an early phase of induction, residual synthesis of enzyme was increased by prior uptake of iron. The results are considered compatible with the concept that uptake of iron, especially at acidic pH, interferes with the formation or function of penicillinase repressor.