Lysogenicity of atypical strains ofSalmonella paratyphi B

Summary Atypical strains ofS. paratyphi B have been described which caused gastroenteritis or were found by chance in healthy perons. These strains possessed atypical natural phages. The late fermentation of d-tartrate by some strains ofS. paratyphi B was examined.     

Acquisition of iron from transferrin bySalmonella paratyphi B

The ability ofSalmonella paratyphi B to obtain iron from human, bovine, and rabbit transferrin was studied. All three transferrins restored the growth potential for otherwise ironstarvedS paratyphi B producing enterobactin. No exogenous siderophore was needed for growth to occur. A non-enterobactin-producing derivative ofS. paratyphi B was unable to acquire iron from transferrin, and transferrin did not restore the growth potential of the organism. Sequestering the transferrin fromS. paratyphi B in dialysis tubing did not affect the ability of the transferrin to supply iron to the organism.     

The formation of bacteriophages in mixed cultures by recombination of genetic elements; characterisation of phage types ofSalmonella paratyphi B by phage reactions and lysogenic properties

Summary It was shown that bacteriophages, generated in mixed cultures of strains ofS. paratyphi B of different types are formed by recombination of elements from both strains. The characteristics of the bacteriophages found depend upon the “phage type” of the strains inoculated in the medium. Types ofS. paratyphi B can be characterised by a combination of phage reactions and lysogenic properties.     

Lysogenicity as a tool for bacteriophage typing ofSalmonella Paratyphi B

Summary and Conclusions The serological identity of the phage produced by a spontaneously lysogenic strain ofS. paratyphi B depends on the bacterial type of that particular strain. The frequency of the phenomenon of spontaneous lysogenicity is such, in the case ofS. paratyphi B, that it can be utilised as a tool for typing.It is clear, however, that the possibilities of the method are limited. Firstly, types exist that do not as a rule produce bacteriophages. Secondly, alysogenic strains may exist of types that as a rule are lysogenic.Therefore the method can be utilised only together with, and as a check upon, a system of phage reactions.In Holland the types Kampen and Leeuwarden are frequent types, that can readily be recognised by the identity of the phages they produce. This is sufficient to justify the use of the method.The relation shown between true lysogenicity and bacterial types may be of theoretical interest. Perhaps it will be possible in this way to relate every existing phage to a bacterial type. The theoretical aspects will, however, be discussed elsewhere.     

A sub-division ofSalmonella typhimurium into phage types based on the method of craigie and yen; Phages adaptable to species of the B and D group ofSalmonella; Phase adsorption as diagnostic aid

Summary and conclusions Phages were isolated which were adsorbed bySalmonella strains of groups B and D. These phages were adaptable to strains of species from these groups. The behaviour of some adaptations of these phages on strains ofS. paratyphi B has been described. A typing scheme forS. typhimurium has been established. So far, this consists of 32 types which have all been demonstrated with adaptations of a single phage.     

Growth of dimorphic human pathogenicfungi on media containing cycloheximide and chloramphenicol

Summary Studies of 24 strains ofBlastomyces dermatitidis confirmed previously published results that the yeast-phase of this fungus is more sensitive than the mycelial-phase to cycloheximide and chloramphenicol.Studies of 5 strains each ofHistoplasma capsulatum, Paracoccidioides brasiliensis andSporotrichum schenckii show that that these species also have a similar yeast-phase mycelial -phase sensitivity differential in regard to these antibiotics.A cycloheximide resistant strain ofB. dermatitidis was developed from a sensitive strain.The experimental results support the general practice of using 0.5 mg/ml cycloheximide and 0.05 mg/ml chloramphenicol in media for the isolation of the four fungi at 25° C. The results indicate, however, that some strains would not be recovered at 37° C with similar concentrations of these antibiotics.It is recommended that a concentration of not more than 0.2 mg/ml chloramphenicol should be used to preserve sputum which is subsequently to be cultured forB. dermatitidis, Histoplasma capsulatum, Paracoccidioides brasiliensis orS. schenckii.     

Incidence of resistance to ampicillin,chloramphenicol, kanamycin and tetracycline amongSalmonella species isolated in The Netherlands in 1972, 1973 and 1974

The resistance ofSalmonellae to drugs has been studied in the Netherlands since 1958. In 1972, 1973, and 1974 respectively, 14241, 13086, and 22927 strains were tested for resistance to ampicillin, chloramphenicol, kanamyci and tetracycline. From 1973 all strains were also tested for resistance to trimethoprim. In the period covered, the yearly incidence of resistance to at least one of the above drugs ranged from 39.2% to 45.6% of all strains obtained from various sources (humans, animals, animal products, sewage, etc.). A new finding in the period 1972 to 1974 was that many multiply resistant strains emerged inS. typhimurium and inS. dublin isolated from calves and cattle. In 1974, 64.4% of all strains ofS. typhimurium from these animals appeared to be resistant to ampicillin, tetracycline, chloramphenicol and kanamycin, and 25.5% of those ofS. dublin were found to be resistant to chloramphenicol and tetracycline. Of all strains ofSalmonellae examined in 1973 and 1974 respectively, 0.15 and 0.22% were resistant to trimethoprim, the main component of the twin-drug cotrimoxazol. Of the 142 strains ofS. typhi isolated in 1972 to 1974 two were resistant to tetracycline only, and one was resistant to all four antibiotics. The others had a normal susceptibility pattern.     

The pathway of betalain biosynthesis: Effect of cytokinin on enzymic oxidation and hydroxylation of tyrosine in Amaranthus tricolor seedlings

The effect of exogenously added tyrosine or l -3-(3,4-dihydroxyphenyl) alanine on the accumulation of betacyanin in response to cytokinin in Amaranthus tricolor (L.) var. tricolor half-seedlings depends on the age of the seedlings and the treatment of the seedlings prior to induction of pigment by benzyladenine. Neither extracted polyphenol oxidase, peroxidase or tyrosine hydroxylase activity, nor in vivo tyrosine hydroxylation is increased in response to exposure of seedlings to cytokinin. However a small percentage of the polyphenol oxidase activated or unmasked by Triton X-100 treatment of membrane fractions is increased after cytokinin treatment of half-seedlings for 22 h. It is concluded that cytokinin control may be on a multi-enzyme membrane-located complex involving part of the polyphenol oxidase activity of the tissue (possibly an isoenzyme), and that the majority of the polyphenol oxidase activity in Amaranthus is a constitutive membrane enzyme which is not involved in betacyanin synthesis. Although cytokinins do not affect in vivo tyrosine hydroxylation, this activity follows closely the accumulation of betacyanin which is first detectable about 6.5 h after the application of cytokinin. Only a very low level of in vivo hydroxylation can be demonstrated in half-seedlings treated for 6 h either with or without cytokinin but it begins to increase shortly after this time. A large increase in this activity by 16 h (independent of cytokinin) can be almost completely (79%) prevented by chloramphenicol (300 μM) although the drug increases accumulation of betacyanin. At this concentration about 30% of the protein synthesis in inhibited. In vitro tyrosine hydroxylation is, however, not reduced in half-seedlings treated with chloramphenicol during 16 h induction nor is extractable polyphenol oxidase reduced. It is concluded that chloramphenicol is inhibiting the synthesis of some protein essential for in vivo hydroxylation other than the activity measured during in vitro hydroxylation and that the inhibition of synthesis of 79% in vivo hydroxylation still leaves enough activity for maximum betacyanin synthesis.     

Pathogenic synergy: mixed intra-abdominal infections

In this article we review our researches into the pathogenesis of mixed infections. These may conveniently be divided into in vitro and in vivo studies. In vitro we confirmed that interference with the killing of aerobes by polymorphonuclear leucocytes (PMN’s) is a property of theBacteroides strains tested and appears to depend on competition for opsonins i.e. complement factors. Further studies are in progress to define which complement factors and which bacterial structures are involved. The influence ofB. fragilis on chemotaxis has also been studied. Our preliminary data suggest thatB. fragilis is itself poorly chemotactic and reduces the chemoattractivity ofProteus mirabilis. This observation is surprising when we consider that abscess formation is the hall-mark ofB. fragilis infections and needs clarification. In vivo we have developed a skin infection model in mice which is economical and gives reproducible and quantitative results. In this model we have demonstrated pathogenic synergy betweenEscherichia coli andB. fragilis. Further studies are planned to assess the role of complement and bacterial factors in this in vivo synergy.     

Isolation of Bdellovibrio sp. from Wastewater and Their Potential Application in Control of Salmonella paratyphi in Water

The problem of the increasing resistance of bacteria to conventional antibiotics gives the bacteria Bdellovibrio a great utility as a potential alternative source of antibiotics. Therefore, the preliminary goal of the present study was isolation and identification of antibiotic-resistant bacteria used as prey organisms for isolated Bdellovibrio sp., by xylose lysine desoxycholate (XLD) agar from different types of water in the Taif area, Saudi Arabia, and also to investigate water quality. Four antibiotic-resistant isolates of Salmonella sp. which were susceptible to Bdellovibrio were identified by morphological, biochemical and 16S rRNA characterization as Salmonella paratyphi. Seventeen strains of Bdellovibrio sp. were isolated from sewage wastewater using isolated S. paratyphi as prey bacteria by a double-layer plate. Only one of them causing a large plaque after 48 h of incubation at 37°C was designated Bdellovibrio AOA12. The shape of Bdellovibrio was confirmed by morphological characterization and electron microscopy. Bdellovibrio could lyse four antibiotic-resistant Gram-negative bacterial strains of Salmonella paratyphi but could not lyse Gram-positive bacteria such as Staphylococcus aureus and Bacillus cereus. The kinetic lysis of the Bdellovibrio as predator to four isolates of antibiotic-resistant Salmonella paratyphi as prey organisms was demonstrated. The results suggest that it may be possible to utilize Bdellovibrio to control antibiotic-resistant S. paratyphi in water.     

Sequences affecting the regulation of solvent production in<Emphasis Type="Italic"> Clostridium acetobutylicum</Emphasis>

The high solvent phenotype of Clostridium acetobutylicum mutants B and H was complemented by the introduction of a plasmid that contains either an intact or partially-deleted copy of solR, restoring acetone and butanol production to wild-type levels. This demonstrates that the solR open reading frame on pSOLThi is not required to restore solvent levels. The promoter region upstream of alcohol dehydrogense E (adhE) was examined in efforts to identify sites that play major roles in the control of expression. A series of adhE promoter fragments was constructed and the expression of each in acid- and solvent-phases of growth was analyzed using a chloramphenicol acetyl-transferase reporter system. Our results show that a region beyond the 0A box is needed for full induction of the promoter. Additionally, we show that the presence of sequences around a possible processing site designated S2 may have a negative role in the regulation of adhE expression.     

Incidence of resistance to chloramphenicol and tetracyclines among 13502Salmonella strains isolated in 1961

Summary The rate of resistance to tetracycline and chloramphenicol amongSalmonella strains isolated in the Netherlands in 1961 was found to be 3.96%, the corresponding figures for 1958/1959 and 1961 being 2.08 and 1.29 respectively. In this country the total number ofSalmonella types found to develop resistance to either tetracycline or chloramphenicol now amounts to 38. Almost 77% of all resistant strains isolated in 1961 were found among the human pathogenS.typhimurium. The relative frequency of resistance in this organism was 8.18%, as compared with 2.50% in 1958/1959 and 1.80% in 1960. In 1961 some cross infections caused by tetracycline resistant strains ofS.typhimurium were observed in man and on one occasion also in a herd of calves. A similar outbreak due to a tetracycline resistant strain ofS.bovis morbificans was seen in a hospital. As almost 87% of all antibiotic resistant strains found in 1961 originated from human patients, the resistance must be largely attributed to the therapeutic use of the drugs in question.     

Frequency of resistance to tetracycline and chloramphenicol amongSalmonella strains isolated in the Netherlands in 1962

Out of 12,472 strains ofSalmonella isolated in the Netherlands in 1962, 1365, or 10.94%, were found to be resistant to tetracycline or chloramphenicol or to both. Compared with the findings of the preceding years (1958/59:2.08%, 1960:1.29%, 1961:3.96%) this is a considerable increase. Of these 1365 strains, 1285, or 94.1%, were resistant to tetracycline and 46, or 3.4%, were resistant to chloramphenicol. The remaining 34 strains, or 2.5%, were resistant to both drugs.Among allSalmonella strains isolated in 1962, 5517 belonged to the speciesS. typhi murium. Of these, 1203 or 21.8%, were resistant to tetracycline. The resistance rates of strains originating from human patients, calves, pigs, other animals and other materials were 24.4%, 37.1%, 15.0%, 8.0% and 5.7% respectively.Factors which may possibly have contributed to the greatly increased frequency of drug resistance inS. typhi murium are: (1) the rapid spread of the use of tetracycline for therapeutic, prophylactic and nutritive purposes, and (2) the possibility of an episome-mediated transfer of drug resistance from relatively harmless intestinal bacteria, such asE. coli, toS. typhi murium.     

Physiological comparisons of transformed (crown gall) and nontransformedVinca rosea L. Cells

Summary In vitro growth rates of transformed (crown gall) and nontransformed cultures ofVinca rosea L. were greater at 32°C than at 25°C. The growth of transformed cells was significantly inhibited by kanamycin, neomycin, and chloramphenicol but not by cycloheximide. Nontransformed cells were inhibited by all four antibiotics., The relative growth rates of transformed cultures induced by four different strains, ofAgrobacterium tumefaciens did not correspond to the relative rates of tumor weight increase observed in vivo nor with the relative weights of tumor tissue in, plants 8 weeks after inoculation with the corresponding bacterial strains.     

Identification of a New Site for Ferrichrome Transport by Comparison of the FhuA Proteins of Escherichia coli, Salmonella paratyphi B, Salmonella typhimurium, and Pantoea agglomerans

The fhuA genes of Salmonella paratyphi B, Salmonella typhimurium, and Pantoea agglomerans were sequenced and compared with the known fhuA sequence of Escherichia coli. The highly similar FhuA proteins displayed the largest difference in the predicted gating loop, which in E. coli controls the permeability of the FhuA channel and serves as the principal binding site for the phages T1, T5, and 80. All the FhuA proteins contained the region in the gating loops required in E. coli for ferrichrome and albomycin transport. The three subdomains required for phage binding were contained in the gating loop of S. paratyphi B which is infected by the E. coli phages, whereas two of the subdomains were deleted in S. typhimurium and P. agglomerans which are resistant to the E. coli phages. Small deletions in a surface loop adjacent to the gating loop, residues 236 to 243 and 236 to 248, inactivated E. coli FhuA with regard to transport of ferrichrome and albomycin, but sensitivity to T1 and T5 was fully retained and sensitivity to 80 and colicin M was reduced 10-fold. Full-size FhuA hybrid proteins of S. paratyphi B and S. typhimurium displayed S. paratyphi B FhuA activity when the hybrids contained two-thirds of either the N- or the C-terminal portions of S. paratyphi B and displayed S. typhimurium FhuA activity to phage ES18 when the hybrid contained two-thirds of the N-terminal region of the S. typhimurium FhuA. The central segment of the S. paratyphi B FhuA flanked on both sides by S. typhimurium FhuA regions conferred full sensitivity only to phage T5. The data support the essential role of the gating loop for the transport of ferrichrome and albomycin, identified an additional loop for ferrichrome and albomycin uptake, and suggest that several segments and their proper conformation, determined by the entire FhuA protein, contribute to the multiple FhuA activities.     

Lytic effect ofVibrio cholerae elastase on Gram-positive and-negative bacteria

Elastase ofVibrio cholerae caused the lysis of freshly grown cells of Gram-negative (Pseudomonas aeruginosa, Proteus vulgaris, Salmonella paratyphi A andKlebsiella pneumoniae) bacteria. Gram-positive (Staphylococcus aureus andS. epidermidis) organisms were resistant to this enzyme. Heat killed and lyophilized Gram-positive and-negative bacteria (exceptS. aureus andS. epidermidis) showed higher sensitivity to elastase. Both Gram-negative and-positive bacteria were lyzed maximally by elastase at pH 8.0. At this pH, lytic activity of elastase was maximum in Tris-HCl and glycine-NaOH buffers followed by Tris-maleate and cacodylate buffers.     

On the occurrence ofSalmonella bareilly in man and animals and the mode of transmission of the salmonellose caused by this species

Summary In the Netherlands salmonellose due toS. bareilly was noted for the first time in 1948, initially only in man, but in the beginning of 1951 also in chicks. Between the end of March and the latter half of June the number of human cases has distinctly increased. Nine patients appeared to have had a recent contact with chicks which excretedS. bareilly or harboured this germ in their organs. The percentage of excreters of germs among chicks decreased after the first months. 481 eggs lain in autumn by young hens from poultry farms which had been found contaminated in spring 1951 did not containS. bareilly. Also a diseased person can spread the disease either by direct contact or by contaminating foodstuffs. In the course of our investigationS. bareilly has been incidentally cultivated from a rat and from material originating from a deer.     

Drug resistance in Salmonella strains isolated from domestic wastewater before and after treatment in stabilization ponds in an arid region (Marrakech,Morocco)

Some 118 Salmonella strains isolated before and after treatment in stabilization ponds were tested for antimicrobial resistance. In the treatment plant, which decreases the abundance of Salmonella by 99%, a significantly lower level of antibiotic resistance (P<0.01) was identified at the system's inflow point (19%) than at its outflow (29%). The serotypes most frequently identified as having multiple antibiotic resistance were Salmonella paratyphi B and S. typhimurium. High tetracycline resistance was observed at all sampling points, followed by resistance to ampicillin and streptomycin. Antibiotic resistance can be transferred from Salmonella to other members of the Enterobacteriaceae family, such as Escherichia coli K12; transfer frequencies in nutrient broth and filtered sewage water were 4.5×10^-4 and 7×10^-7, respectively.The authors are with the Université CADI AYYAD, Faculté des Sciences—Semlalia, Département de Biologie, Laboratoire de Microbiologie, Bd Le Prince My Abdallah, BP. S.15, Marrakech, Morocco.     

Comparative fluorescent antibody staining of histoplasma capsulatum and histoplasma duboisii with a specific anti-yeast phase H. Capsulatum conjugate

Summary The specific anti-yeast phaseHistoplasma capsulatum conjugate has been tested against 13 yeast phase strains ofH. capsulatum and 9 ofH. duboisii. The conjugate was specific forH. capsulatum, no yeast phase form ofH. duboisii obtained in vitro or in vivo reacted with it. The taxonomic implications of these results are discussed.