The chitinases of aerobic sporulating bacteria isolated from different ecological sources]

The chitinolytic activity of 171 strains of 15 species of spore-forming aerobic bacteria isolated from different ecological sources has been studied. 85 strains of the studied bacilli (50%) hydrolyzed colloidal chitin in a different degree. Among the cultures isolated from human and animal organism 60% of strains were characterized by the presence of extracellular chinases, among the collection strains--37%, among those isolated from soil and from insects--40 and 43%, respectively. The cultures of Bacillus subtilis as well as B. coagulans, B. megaterium and some other possessed the highest activity on the liquid medium. Some strains have been chosen for further research aimed at their possible use for biotechnology.     

Colicinogeny in Salmonella serovars isolated in Brazil

A study of colicinogeny was made in 748 strains of Salmonella (97 serovars) isolated from different sources: human (291), animal (119), environmental (141), food (102) and animal feed (95). Colicin production was detected in 64 strains (8.6%), particularly isolated from foods (30.4%). Col E1 (53) and Ia (44) were the most frequently observed, especially in S. agona for environment and food sources. Col V production was identified in 5 strains of S. typhimurium within 8 producer cultures isolated from humans. Its relationship with the sources and serovars of Salmonella are discussed.     

Occurrence of Yersinia enterocolitica in wild animals.

Yersinia species were isolated from 16 of 495 small wild animals and from 1 of 38 foxes. The animals were trapped in seven regions of Hokkaido, Japan. Of the 17 strains isolated, 9 were Yersinia enterocolitica O6; 2 were Y. enterocolitica O5A; 1 was Y. enterocolitica, O4; 1 was Y. enterocolitica O9; 1 was Yersinia pseudotuberculosis IVB; and 3 were sucrose-negative strains. Yersinia pestis was not isolated. The O6 organism was most prevalent in large red-back mice (Clethrionomys rufocanus bedfordiae) and showed significant differences in its mode of distribution according to region. Incidence of the O6 organism in the ileum of the animal was threefold that in the cecum, and the organism was recovered at approximately 10(5) cells per g of cecal contents per c. rufocanus bedfordiae animal.     

Opportunistic bacteria detected in cultivated mussels

As many as 8 Listeria monocytogenes strains, 12 Pseudomonas aeruginosa strains and 5 Staphylococcus aureus strains were isolated from mussels Mytilus edulis, grown on special installations in the Trinity Bay of the Gulf of Peter the Great, the Sea of Japan. The isolated cultures proved to be highly resistant to a number of antibiotics. Many strains displayed DNAase and haemolytic activity. The cultures of L. monocytogenes, S. aureus and P. aeruginosa also had high lipase, protease and lecithinase activity. The organism of the mussels seems to be a confinement for these bacteria under study.     

Reassortant rotaviruses as potential live rotavirus vaccine candidates.

A series of reassortants was isolated from coinfection of cell cultures with a wild-type animal rotavirus and a "noncultivatable" human rotavirus. Wild-type bovine rotavirus (UK strain) was reassorted with human rotavirus strains D, DS-1, and P; wild-type rhesus rotavirus was reassorted with human rotavirus strains D and DS-1. The D, DS-1, and P strains represent human rotavirus serotypes 1, 2, and 3, respectively. Monospecific antiserum (to bovine rotavirus, NCDV strain) or a set of monoclonal antibodies to the major outer capsid neutralization glycoprotein, VP7 (of the rhesus rotavirus), was used to select for reassortants with human rotavirus neutralization specificity. This selection technique yielded many reassortants which received only the gene segment coding for the major neutralization protein from the human rotavirus parent, whereas the remaining genes were derived from the animal rotavirus parent. Single human rotavirus gene substitution reassortants of this sort represent potential live vaccine strains.     

Resistance to antibiotics and heavy metal salts of coagulase-positive staphylococci belonging to different biological types (ecovars)

A total of 189 strains of S. aureus isolated from cows, sheep, swines, poultry, monkeys, rabbits, foxes, and humans and 23 strains of S. intermedius isolated from minks and sables were studied. The staphylococci belonged to different biological types (according to the Hajek-Marsalek's scheme) and ecovars (according to the Meyer-Witte's scheme). The strains were studied with respect to their resistance to 10 antibiotics (benzylpenicillin, streptomycin, tetracycline, levomycetin, erythromycin, oleandomycin, neomycin, kanamycin, monomycin, and novobiocin), mercuric chloride and cadmium sulfate. As a whole the frequency of resistance to the above preparations among the staphylococci of the animal origin was not high. Differences in the frequency and range of the resistance between strains belonging to different biological types (ecovars) were shown. The highest number of the resistant cultures was detected among the strains of the biological type E (ecovar canis) and atypical strains of S. intermedius isolated from the minks. The least number of the resistant cultures was detected among the strains of the biological type C1 (ecovar bovis) isolated from the cows. It was found that almost all strains of S. intermedius were resistant to cadmium sulfate. This may be used as an additional characteristic of the species.     

Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment

During the summer of 1981, 3,887 sucrose-negative vibrios were isolated from seawater, sediment, plankton, and animal samples taken from 80 sites from Miami, Fla., to Portland, Maine. Of these, 4.2% were able to ferment lactose. The lactose-positive strains isolated from the various samples correlated positively with pH and turbidity of the water, vibrios in the sediment and oysters, and total bacterial counts in oysters. Negative correlations were obtained for water salinity. Numerical taxonomy was performed on 95 of the lactose-fermenting environmental isolates and 23 reference strains. Five clusters resulted, with the major cluster containing 33 of the environmental isolates and all of the Vibrio vulnificus reference strains. The 33 isolates, which produced an acid reaction in lactose broth within hours of initial inoculation, represented 20% of all lactose-fermenting vibrios studied. These isolates were nearly identical phenotypically to clinical strains of V. vulnificus studied by the Centers for Disease Control, Atlanta, Ga., and by our laboratory, and their identification was confirmed by DNA-DNA hybridization studies. V. vulnificus was isolated from all sample types and from Miami to Cape Cod, Mass., and comparison of the environmental parameters of the eight subsites yielding this species with those of all 80 subsites revealed no significant differences. The majority of the isolates were obtained from animals, with clams providing most (84%) of these. On injection into mice, 82% of the V. vulnificus isolates resulted in death. Members of the remaining four clusters contained strains which differed from V. vulnificus in such phenotypic traits as luminescence and in urease or H(2)S production. None of the other reference cultures, including nine other Vibrio species, were contained in the remaining clusters, and these isolates could not be identified. Most of these were also lethal for mice. Phenotypic differences, potential pathogenicity, and geographic distribution of the five clusters were examined. It is concluded that V. vulnificus is a ubiquitous organism, both geographically and in a variety of environmental sources, although it occurs in relatively low numbers. The public health significance of this organism and of the other unidentified lactose-fermenting Vibrio species is discussed.     

Phenotypic differentiation of bifidobacteria of human and animal origins.

The phenotypes of 153 strains belonging or related to the genus Bifidobacterium were studied. These organisms included 38 collection strains and 115 wild strains (41 strains of human origin, 56 strains of animal origin, and 18 strains obtained from rivers or sewage). Our phenotypic analysis revealed seven main groups that were subdivided into 20 subgroups. Seven subgroups contained no type or collection strain. Among the human strains, the type strains of Bifidobacterium pseudocatenulatum and B. catenulatum fell into group I, which contained the type strains of B. adolescentis (subgroup Ib), B. dentium (subgroup Ic), and B. angulatum (ungrouped). The type strain of B. breve belonged to subgroup IIIa1, and the type strains of B. infantis and B. longum fell into subgroup IIIb1. Group VII comprised only wild strains that were isolated from human infant feces. Among the animal strains, group II consisted mainly of bifidobacteria that were isolated from pig feces and contained the type strains of B. suis (subgroup IIb), B. thermophilum (subgroup IIf), B. choerinum, and B. boum (ungrouped). Wild strains belonging to group V were isolated from pig, calf, cow, and chicken feces; this included the type strains of B. animalis (subgroup Va), B. magnum (subgroup Vb), B. pseudolongum, and B. globosum (subgroup Vc). The strains of human origin (groups I, III, and VII) were well separated from the animal strains (groups II, IV, and V). It was not surprising that the wild strains isolated from surface water or sewage were distributed in the animal groups as well as the human groups. Thus, bifidobacteria can be considered to be successful indicators of human or animal fecal pollution when they are correctly classified. The acidification patterns were not adequate to differentiate Bifidobacterium species, as determined previously by Mitsuoka (Bifidobacteria Microflora 3:11-28, 1984) and Scardovi (p. 1418-1434, in P. H. A. Sneath, N. S. Mair, M. E. Sharpe, and J. G. Holt, ed., Bergey's Manual of Systematic Bacteriology, vol. 2, 1986). However, enzymatic tests furnished new taxonomic criteria for the genus.     

Genetic diversity, clonality and sexuality in Toxoplasma gondii

The majority of Toxoplasma gondii strains from a variety of human and animal sources have been grouped into three highly clonal but closely related lineages. The low occurrence of nucleotide differences among the three predominant lineages and their unusual dimorphic allelic composition suggest that they have arisen from a recent common ancestry. Less than 1% of the previously studied strains contain unique genotypes and high divergence of DNA sequence, and therefore are considered 'exotic' or 'atypical' strains. The seemingly low genetic diversity in T. gondii may have been underestimated because most parasite strains in previous studies were collected from human patients and domestic animals in North America and Europe. To investigate the genetic diversity of T. gondii, we analysed parasite strains isolated from remote geographical regions by multilocus microsatellite sequencing and phylogenetic analysis. The genetic diversity indices, the molecular analysis of microsatellite genotypes and the constructed phylogram considered together suggest that the global T. gondii population is highly diversified and not characteristic of a clonal organism. The most parsimonious hypothesis is that T. gondii presents a complex population structure with a mix of clonal and sexual propagation as a function of the environmental conditions. The comparison between domestic strains data on one hand and wild strains data on the other hand is in favour of more frequent sexual recombinations in wild environment even though Toxoplasma subpopulation in human and domestic animals is largely clonal.     

Hemagglutinating and adhesive capacities of Klebsiella and Enterobacter strains

The authors analyze the data of studies on the hemagglutinating and adhesive capacity of 290 cultures, including 118 K. pneumoniae strains and 64 E. cloacae strains isolated from sick children, as well as 59 K. pneumoniae strains and 49 E. cloacae strains isolated from healthy children. The hemagglutinating properties of the strains were determined in the hemagglutination test with fresh, formalin- and tannin-treated red blood cells, the adhesive properties were studied by light microscopy. Among K. pneumoniae and E. cloacae strains isolated in acute intestinal infections, mannose-sensitive hemagglutination and pronounced adhesive activity were prevalent in most cases. Poorly adhesive and nonadhesive strains were characteristic of K. pneumoniae and E. cloacae cultures isolated from healthy children. The strains isolated from sick and healthy children differed only by the prevalence of adhesive cultures.     

Fermentation of mucin by bifidobacteria from rectal samples of humans and rectal and intestinal samples of animals

Bifidobacteria (246 strains in total) were isolated from rectal samples of infants and adult humans and animals, and from intestinal samples of calves. Twenty-five strains grew well on mucin: 20 from infants, two from adults, and three from goatlings. Poor or no growth on mucin was observed in 156 bifidobacterial strains of animal origin. The difference between human and animal isolates in ability to grow on mucin was significant at p < 0.001. Nine human strains with the best growth on mucin were identified as Bifidobacterium bifidum. These strains produced extracellular, membrane-bound, and intracellular mucinases with activities of 0.11, 0.53, and 0.09 μmol/min of reducing sugars per milligram of protein, respectively. Membrane-bound mucinases were active between pH 5 and 10. The optimum pH of extracellular mucinases was 6–7. Fermentation patterns in cultures grown on mucin and glucose differed. On mucin, the acetate-to-lactate ratio was higher than in cultures grown on glucose (p = 0.012). We showed that the bifidobacteria belong to the mucin-fermenting bacteria in humans, but their significance in mucin degradation in animals seems to be limited.     

Distribution of Vibrio vulnificus and Other Lactose-Fermenting Vibrios in the Marine Environment

During the summer of 1981, 3,887 sucrose-negative vibrios were isolated from seawater, sediment, plankton, and animal samples taken from 80 sites from Miami, Fla., to Portland, Maine. Of these, 4.2% were able to ferment lactose. The lactose-positive strains isolated from the various samples correlated positively with pH and turbidity of the water, vibrios in the sediment and oysters, and total bacterial counts in oysters. Negative correlations were obtained for water salinity. Numerical taxonomy was performed on 95 of the lactose-fermenting environmental isolates and 23 reference strains. Five clusters resulted, with the major cluster containing 33 of the environmental isolates and all of the Vibrio vulnificus reference strains. The 33 isolates, which produced an acid reaction in lactose broth within hours of initial inoculation, represented 20% of all lactose-fermenting vibrios studied. These isolates were nearly identical phenotypically to clinical strains of V. vulnificus studied by the Centers for Disease Control, Atlanta, Ga., and by our laboratory, and their identification was confirmed by DNA-DNA hybridization studies. V. vulnificus was isolated from all sample types and from Miami to Cape Cod, Mass., and comparison of the environmental parameters of the eight subsites yielding this species with those of all 80 subsites revealed no significant differences. The majority of the isolates were obtained from animals, with clams providing most (84%) of these. On injection into mice, 82% of the V. vulnificus isolates resulted in death. Members of the remaining four clusters contained strains which differed from V. vulnificus in such phenotypic traits as luminescence and in urease or H₂S production. None of the other reference cultures, including nine other Vibrio species, were contained in the remaining clusters, and these isolates could not be identified. Most of these were also lethal for mice. Phenotypic differences, potential pathogenicity, and geographic distribution of the five clusters were examined. It is concluded that V. vulnificus is a ubiquitous organism, both geographically and in a variety of environmental sources, although it occurs in relatively low numbers. The public health significance of this organism and of the other unidentified lactose-fermenting Vibrio species is discussed.     

Phage typing of the coagulase-positive staphylococci isolated from various species of animals

More than 200 coagulase-positive strains of animal origin have been studied by means of Staphylococcus aureus typing phages, belonging to two international sets and intended for typing staphylococci isolated from large cattle and humans, and experimental "chicken" phage A 1591. Among S. aureus strains the cultures isolated from swine, cows, chickens, and belonging to biotypes B1, C1, B2, respectively, have been mostly (in 78.5-90.0% of cases) determined by phage typing. The strains belonging to one biotype have proved to be sensitive predominantly to the same phages. In this connection further differentiation of staphylococci within individual biotypes by means of the phages used in these experiments seems to be impracticable. S. intermedius strains have been found to be completely resistant to the above phages, which confirms that S. intermedius is rightly considered to be an independent species of coagulase-positive staphylococci.     

Are zoonotic <Emphasis Type="Italic">Staphylococcus pseudintermedius</Emphasis> strains a growing threat for humans?

Staphylococcus pseudintermedius is a species often isolated from animals, as a common element of their microbiota or an agent of infection, and from people associated with an animal habitat, including owners of home pets—dogs and cats. As with many other species, adaptation of these bacteria to the human body can occur, and they become important human pathogens. 59 S. pseudintermedius strains were investigated in this study to determine the factors contributing to human body colonization: inhibition growth of human skin residents isolated from human skin (Staphylococcus epidermidis, Corynebacterium spp., Cutibacterium acnes (formerly Propionibacterium acnes)), biofilm formation, and the presence of ten genes encoding infection-promoting features (including ebpS, spsE, lukS, lukF, pvl, lip, hlgA, hlgB). The ability of human skin to be colonized and the presence of genes that promote the development of skin infections showed the significant potential of the studied strains in their adaptation to the host. However, while a comparison of the characteristics of animal strains and those isolated from human infections does not allow us to claim that we are the witnesses of the speciation of a new human pathogen, it does indicate their gradual adaptation to the human organism.     

A comprehensive evaluation of the biological properties of the causative agents of typhoid fever isolated from patients and carriers

The biological properties of 106 S. typhi cultures were studied; of these, 59 cultures were isolated from 45 chronic carriers and 47 cultures, from 23 typhoid fever patients. According to the degree of their virulence (CPD50 in the continuous cell-line culture Hep-2), the strains isolated from the patients were more virulent than those isolated from the chronic carriers. The mean value of lg CPD50 was 5.76 +/- 0.04 for the patients and 6.86 +/- 0.03 for the chronic carriers. The strains isolated from the patients showed greater variability in the degree of their virulence. The study of the plasmid spectrum showed that 9.4 +/- 5.6% of the strains contained plasmids. From the patients plasmid-containing strains were isolated more frequently than from the carriers (14.9 +/- 2.5% and 5.1 +/- 2.9%). Multiresistance to antibiotics in combination with the presence of plasmids was detected in 6 strains isolated from the typhoid patients with morbidity having the character of outbreaks.     

Antagonistic and bacteriocinogenic activity of Meningococci]

Antagonistic and bacteriocinogenic activity was studied in 169 strains of meningococci of various serological groups and with different localization in the human organism at the time of isolation. Bacteriocinogenic activity was revealed in all the meningococcus strains (by delayed antagonism method), and, in addition, antagonistic activity was found in 100 strains. The inhibitory activity was the greatest in meningococci isolated from the cerebrospinal fluid. The data obtained suggest an important inhibitory activity of meningococci, along with their resistance to the antagonistic activity of the nasopharyngeal microorganisms, in the manifestation of pathogenic properties in them.     

On the pathogenicity of nonstable Brucella L-forms and their revertants.

The pathogenicity of B. abortus 870 L-forms obtained by long-term passaging of virulent culture on media with penicillin and of revertants obtained in vitro and in vivo was studied. L-form cultures stimulated only a mild response of the reticulo-endothelial system of the animal organism, at the same time displaying a certain level of toxicity. In vitro revertants approximated to L-forms, while in vivo revertants stood closer to the initial virulent culture, as regards pathogenicity. This seems to be evidence of a potential danger of brucella L-forms for the human and animal organisms.     

Application of multilocus enzyme electrophoresis in studies of the epidemiology of Listeria monocytogenes in Denmark.

A total of 245 strains of Listeria monocytogenes were investigated. These strains were isolated from human and animal cases of listeriosis as well as from different kinds of raw and processed foods. Thirty-three electrophoretic types (ETs) were identified among the 245 strains. The strains investigated included all human clinical strains isolated in Denmark during 1989 and 1990. Seventy-three percent of the strains isolated in this period were assigned to one of only two ETs (ET 1 and ET 4). ET 1, which was found to be the most frequently occurring ET among strains isolated from human clinical cases, was also found to occur rather frequently in animal clinical cases. ET 1 was, however, found only sporadically among strains isolated from foods and food factories. The data indicate that there might be something distinctive about the physiology or ecology of the ET 1 clone which makes it more likely to bring about disease in human beings either because of high pathogenicity or because of a special ability to multiply to infectious doses in processed foods. Another type, designated ET 4, was found to be the next most frequently occurring ET, after ET 1, among human clinical isolates. This could be explained by the fact that ET 4 was found to be the most frequently occurring ET within food isolates.     

Characterization of black-pigmented Bacteroides strains isolated from animals

The aims of the study were: the isolation of strains of black-pigmented Bacteroides from the gingival sulcus of different animals, their biochemical and immunological characterization and comparison of their properties for classification within the genus. A total of 104 strains, isolated from cats, dogs, racoons and a jaguar, were characterized on the basis of fermentation of carbohydrates, metabolic end products, haemagglutination studies, enzymatic activities, catalase production and indirect immunofluorescence. No differences were observed between the strains regardless of their animal origin. The strains did not ferment carbohydrates, produce phenylacetic acid, show an array of enzyme activities or agglutinate sheep red blood cells. They were catalase-positive and so differed from the human oral strains of Bact. gingivalis. Immunofluorescence microscopy revealed that the animal strains shared at least one major antigen with Bact. gingivalis but none with Bact. asaccharolyticus. Apart from their catalase activity, the animal strains isolated were similar to those of human Bact. gingivalis strains.     

Classification and biological characteristics of coagulase-positive Staphylococci isolated from animals

A total of 165 coagulase-positive staphylococcal strains of different origin (142 S. aureus strains and 23 S. intermedius strains) were subjected to biological typing in accordance with the schemes of Hajek-Marsalek and Meyer-Witte. The former of these schemes permitted to identify 68% and the latter 18% of S. aureus strains. The cultures isolated from swine and chickens had the most uniform composition: 85-86% of the strains belonged to biotype B. 44% of the strains isolated from cows and sheep belonged to biotypes C (ecovars bovis and ovis) and A (ecovar hominis); the rest of the strains could not be identified. 96% of the strains isolated from minks were made up of S. intermedius, more than a half of them belonging to biotype E (ecovar canis). In 80% of S. aureus strains and 48% S. intermedius cultures protein A was detected. Only 9% of S. aureus strains of animal origin were found capable of producing enterotoxins (A-D). The expediency of working out a unified scheme for the biotyping of coagulase-positive staphylococci is discussed.