Comparison of commercially available kits with standard methods for detection of Salmonella strains in foods.

Six commercial kits were compared with the U.S. Food and Drug Administration (USFDA) method and the Japanese standard method for Salmonella isolation in foods. When only Salmonella serovars were tested, many of the methods performed well; however, when foods were artificially inoculated, only the USFDA method and immunomagnetic separation coupled with the xylose-lysine-brilliant green agar method (MS-XLBG) could positively detect Salmonella serovars. All seven wild-type Salmonella serovars were detected by the USFDA method, and the MS-XLBG method detected salmonellae from six samples.     

The use of a molecular hybridization method for studying the enterotoxigenic properties of Salmonella

The occurrence of the tox gene among 320 Salmonella strains of 23 serovars, differing in their origin, sensitivity to antibiotics, the presence of R-plasmids and a number of biochemical properties, has been studied by the method of DNA-DNA hybridization in situ. Essential differences in the occurrence of the tox gene have been detected both among S. typhimurium hospital strains and strains isolated in sporadic diseases, from the environment, from animals and among salmonellae belonging to different serovars. The direct correlation between the presence of the enterotoxigenicity gene and plasmids controlling resistance to antibiotics in Salmonella strains has been established. The expediency of using the method of gene probing for the study of the enterotoxigenic properties of salmonellae has been substantiated.     

Influence of environmental factors and human activity on the presence of Salmonella serovars in a marine environment

The temporal and spatial distribution of Salmonella contamination in the coastal waters of Galicia (northwestern Spain) relative to contamination events with different environmental factors (temperature, wind, hours of sunlight, rainfall, and river flow) were investigated over a 4-year period. Salmonellae were isolated from 127 of 5,384 samples of molluscs and seawater (2.4%), and no significant differences (P < 0.05) between isolates obtained in different years were observed. The incidence of salmonellae was significantly higher in water column samples (2.9%) than in those taken from the marine benthos (0.7%). Of the 127 strains of Salmonella isolated, 20 different serovars were identified. Salmonella enterica serovar Senftenberg was the predominant serovar, being represented by 54 isolates (42.5%), followed by serovar Typhimurium (19 isolates [15%]) and serovar Agona (12 isolates [9.4%]). Serovar Senftenberg was detected at specific points on the coast and could not be related to any of the environmental parameters analyzed. All serovars except Salmonella serovar Senftenberg were found principally in the southern coastal areas close to the mouths of rivers, and their incidence was associated with high southwestern wind and rainfall. Using multiple logistic regression analysis models, the prevalence of salmonellae was best explained by environmental parameters on the day prior to sampling. Understanding this relationship may be useful for the control of molluscan shellfish harvests, with wind and rainfall serving as triggers for closure.     

Dynamics of the etiological structure of salmonellosis and of the biological properties of the Salmonellae isolated in a selected territory of the Moldavian SSR 1982-1985

The etiological structure of Salmonella infections and the biological properties of salmonellae, isolated in one of the regions of the Moldavian SSR where the epidemic process of Salmonella infections reflected the regularities observed on the whole territory of this republic, were studied. Changes in the predominant serovars at the period of 1982-1985 in comparison with the preceding years were shown. Salmonellae belonging to the dominating serovars were characterized by more pronounced drug resistance in comparison with other salmonellae. Some parameters of the epidemic process of Salmonella infections were found to be related to the biological properties of the causative agents of these diseases.     

Uptake and replication of Salmonella enterica in Acanthamoeba rhysodes

The ability of salmonellae to become internalized and to survive and replicate in amoebae was evaluated by using three separate serovars of Salmonella enterica and five different isolates of axenic Acanthamoeba spp. In gentamicin protection assays, Salmonella enterica serovar Dublin was internalized more efficiently than Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium in all of the amoeba isolates tested. The bacteria appeared to be most efficiently internalized by Acanthamoeba rhysodes. Variations in bacterial growth conditions affected internalization efficiency, but this effect was not altered by inactivation of hilA, a key regulator in the expression of the invasion-associated Salmonella pathogenicity island 1. Microscopy of infected A. rhysodes revealed that S. enterica resided within vacuoles. Prolonged incubation resulted in a loss of intracellular bacteria associated with morphological changes and loss of amoebae. In part, these alterations were associated with hilA and the Salmonella virulence plasmid. The data show that Acanthamoeba spp. can differentiate between different serovars of salmonellae and that internalization is associated with cytotoxic effects mediated by defined Salmonella virulence loci.     

The importance of Shigella boydii in shigellosis morbidity in the USSR

The etiological role of S. boydii in Shigella infections registered at different territories of the USSR in 1986-1987 was analyzed. As established by this analysis, S. boydii infection occurred mainly at the territories with unsatisfactory water supply of the population; from these territories the infection spread to ther territories of the USSR. The dominating serovars causing S. boydii dysentery, as well as Shigella infections, at different territories of the USSR was shown to be identical, which was indicative of the fact that the immunological factor had no influence on the etiological structure of Shigella infections, determined by the activity of the main routes of the transfer of infection. On the whole, S. boydii serovars 2, 4 and 1 were found to prevail in the USSR.     

Serological variants of Yersinia enterocolitica circulating in different geographical areas of the Soviet Union

The serological variants of a number of Y. enterocolitica strains isolated in different regions of the USSR (1,085 strains isolated from humans and animals in the North-West of the RSFSR, 76 strains isolated from humans and animals in the Krasnodar Territory and 114 strains isolated from humans only in the area east of Lake Baikal) were determined. 25 serological variants were registered in these 3 regions of the USSR. The cultures isolated from rodents belonged mostly to serovars 06,30; 03; 05; 04,33; 019,8; 016; and from humans, to serovars 03; 05; 07,8; 016; 06,30; 09.     

Horizontal gene transfer of a ColV plasmid has resulted in a dominant avian clonal type of Salmonella enterica serovar Kentucky

Salmonella enterica continues to be a significant cause of foodborne gastrointestinal illness in humans. A wide variety of Salmonella serovars have been isolated from production birds and from retail poultry meat. Recently, though, S. enterica subsp. enterica serovar Kentucky has emerged as one of the prominent Salmonella serovars isolated from broiler chickens. Recent work suggests that its emergence apparently coincides with its acquisition of a ColV virulence plasmid. In the present study, we examined 902 Salmonella isolates belonging to 59 different serovars for the presence of this plasmid. Of the serovars examined, the ColV plasmid was found only among isolates belonging to the serovars Kentucky (72.9%), Typhimurium (15.0%) and Heidelberg (1.7%). We demonstrated that a single PFGE clonal type of S. Kentucky harbors this plasmid, and acquisition of this plasmid by S. Kentucky significantly increased its ability to colonize the chicken cecum and cause extraintestinal disease. Comparison of the completed sequences of three ColV plasmids from S. Kentucky isolated from different geographical locales, timepoints and sources revealed a nearly identical genetic structure with few single nucleotide changes or insertions/deletions. Overall, it appears that the ColV plasmid was recently acquired by a single clonal type S. Kentucky and confers to its host enhanced colonization and fitness capabilities. Thus, the potential for horizontal gene transfer of virulence and fitness factors to Salmonella from other enteric bacteria exists in poultry, representing a potential human health hazard.     

Influence of Environmental Factors and Human Activity on the Presence of Salmonella Serovars in a Marine Environment

The temporal and spatial distribution of Salmonella contamination in the coastal waters of Galicia (northwestern Spain) relative to contamination events with different environmental factors (temperature, wind, hours of sunlight, rainfall, and river flow) were investigated over a 4-year period. Salmonellae were isolated from 127 of 5,384 samples of molluscs and seawater (2.4%), and no significant differences (P < 0.05) between isolates obtained in different years were observed. The incidence of salmonellae was significantly higher in water column samples (2.9%) than in those taken from the marine benthos (0.7%). Of the 127 strains of Salmonella isolated, 20 different serovars were identified. Salmonella enterica serovar Senftenberg was the predominant serovar, being represented by 54 isolates (42.5%), followed by serovar Typhimurium (19 isolates [15%]) and serovar Agona (12 isolates [9.4%]). Serovar Senftenberg was detected at specific points on the coast and could not be related to any of the environmental parameters analyzed. All serovars except Salmonella serovar Senftenberg were found principally in the southern coastal areas close to the mouths of rivers, and their incidence was associated with high southwestern wind and rainfall. Using multiple logistic regression analysis models, the prevalence of salmonellae was best explained by environmental parameters on the day prior to sampling. Understanding this relationship may be useful for the control of molluscan shellfish harvests, with wind and rainfall serving as triggers for closure.     

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.     

Features of Salmonella serovars among food handlers in Kyushu, Japan

Salmonella were isolated from 106 (0.032%) of 331,644 fecal samples from food handlers, and from 144 of 11,478 fecal samples from symptomatic patients in Japan to determine the incidence and features of Salmonella serovars among food handlers. S. enterica subspecies enterica serovar Infantis (S. serovar Infantis) was the dominant serovar (accounting for 48.1%), followed by S. serovar Corvallis, which showed poor genetic diversity, and S. serovar Enteritidis among food handlers. The former two serovars were not dominant among symptomatic patients. The present study demonstrates the need for education on the sanitary handling of chicken eggs and chicken meat, which are possible infectious sources of these Salmonella serovars.     

Occurrence of sopE gene and its phenotypic expression among different serovars of Salmonella enterica isolated from man and animals

Salmonella pathogenesis is a complex phenomenon and a Type III secretion system plays a central role in the development of Salmonella-induced enteritis. One such Type III secretion protein is Salmonella outer protein E (SopE). Prevalence of sopE gene and its phenotypic expression (SopE protein) among different serovars of Salmonella enterica isolated from man and animals were investigated. Of 305 strains of S. enterica belonging to 11 serovars tested for the presence of sopE, 130 strains belonging to three serovars viz., Enteritidis, Gallinarum and Virchow were found to carry sopE gene irrespective of their source of isolation when tested by PCR amplification technique using its specific primers. Of these 130 strains, 112 strains were found to express SopE protein phenotypically as detected by Dot-ELISA using SopE antibody. Among the different serovars tested only serovars Gallinarum, Enteritidis and Virchow expressed SopE protein phenotypically in vitro. Role of SopE protein in pathogenesis of salmonellosis has been discussed.     

Distribution and genotypic characterization of Salmonella serovars isolated from tropical seafood of Cochin, India

Aims:  To determine the distribution of Salmonella serovars in seafood and to examine the intraserovar genetic variations in Salmonella enterica subsp. enterica serovar Rissen and Salmonella Weltevreden by polymerase chain reaction (PCR)-ribotyping and enterobacterial repetitive intergenic consensus (ERIC)-PCR methods.
Method and Results:  A total of 417 seafood samples collected over 2003–2006 from fishing harbours and fish markets of Cochin (India) was studied for presence of Salmonella serovars. Seafood samples were analysed for the presence of Salmonella by Bacteriological Analytical Manual (BAM), U.S. Food & Drug Administration (USFDA) method. The study indicated that 23·2% of the seafood samples were positive for Salmonella and a total of 241 Salmonella isolates comprising of 27 different serovars were isolated from seafood. S. Weltevreden, Salmonella Rissen, Salmonella Typhimurium and Salmonella Derby were found to be the most predominant serovars in seafood. PCR-ribotypes and ERIC-PCR profiles showed multiple genotypic profiles for S. Rissen and S. Weltevreden in seafood and the level of discrimination indices obtained was at 0·974 for S. Rissen and 0·988 for S. Weltevreden, respectively.
Conclusion:  The study highlighted the major Salmonella serovars in the seafood of Cochin (India) and molecular fingerprinting pattern revealed genetic variation among S. Rissen and S. Weltevreden.
Significance and Impact of the Study:  Widespread occurrence of Salmonella contamination in seafood and multiple clones of S. Rissen and S. Weltevreden detected in seafood, thus, indicated the diverse routes of Salmonella contamination in seafood.     

Multiyear study of sludge application to farmland: prevalence of bacterial enteric pathogens and antibody status of farm families

We describe our experience with the isolation of salmonellae from sewage sludge from four treatment plants in different geographic areas of Ohio. Over 3 years, we isolated salmonellae 50 times from 311 sludge samples. Most isolations were made after enrichment in Selenite broth (BBL Microbiology Systems, Cockeysville, Md.). The largest proportion of isolations came from the plant serving the population of Columbus, a large metropolitan area. A significantly greater number of isolations from this plant were made during the first quarter of the year. Twenty-one different serotypes were isolated, along with five untypable strains. The most frequently isolated serotype was Salmonella infantis. Five of the strains were multiply resistant to antibiotics. We also describe the prevalence of antibodies to salmonellae in members of the families residing on the farms in the study. It was found that antibodies to group C salmonellae predominated.     

Multiyear study of sludge application to farmland: prevalence of bacterial enteric pathogens and antibody status of farm families.

We describe our experience with the isolation of salmonellae from sewage sludge from four treatment plants in different geographic areas of Ohio. Over 3 years, we isolated salmonellae 50 times from 311 sludge samples. Most isolations were made after enrichment in Selenite broth (BBL Microbiology Systems, Cockeysville, Md.). The largest proportion of isolations came from the plant serving the population of Columbus, a large metropolitan area. A significantly greater number of isolations from this plant were made during the first quarter of the year. Twenty-one different serotypes were isolated, along with five untypable strains. The most frequently isolated serotype was Salmonella infantis. Five of the strains were multiply resistant to antibiotics. We also describe the prevalence of antibodies to salmonellae in members of the families residing on the farms in the study. It was found that antibodies to group C salmonellae predominated.     

Small pet aquarium frogs as a source of Salmonella.

Salmonellae were isolated from 21% of the samples of freshwater aquarium frogs tested and from 25% of the samples of aquarium water containing these frogs. The salmonellae were Salmonella arizonae, S. bovis-morbificans, S. hadar, S. saint-paul, S. typhimurium, and S. worthington. These isolations were made over a period of 9 months and from three different cities. This association of salmonellae with frogs may contribute to cases of human salmonellosis since other aquarium species have already been shown to contribute to such cases.     

Haemolysins of Salmonella, their role in pathogenesis and subtyping of Salmonella serovars

Haemolysin patterns of 175 strains of different Salmonella enterica subspecies enterica serovars isolated from different animal sources and places were determined using 11 different blood agar media made with either non-washed horse/sheep erythrocytes or with washed erythrocytes of cattle, sheep, horse, goat, rabbit, guinea pig, and human A, O and B blood groups. Study on 47 strains belonging to 10 serovars of Salmonella from buffalo meat (buffen), 42 strains of 11 serovars from goat meat (chevon): 16 strains of Salmonella enterica serovar Paratyphi B and 25 of S. enterica serovar Paratyphi B var Java from fish, meat, meat products and clinical cases; 45 isolates of S. Abortusequi from aborted mares (18), fetal contents (21), aborted donkey mares (2) and 4 reference strains, revealed that all host restricted Salmonella namely, S. enterica serovar Gallinarum, S. enterica serovar Anatum, S. enterica serovar Abortusequi and S. enterica serovar Paratyphi B could be divided into different haemolysin types based on their inability to produce haemolysis on one or more types of blood agar, while strains of all zoonotic Salmonella serovars induced haemolysis on all the 9 types of blood agar made of washed erythrocytes. None of 175 Salmonella could produce hemolytic colonies on blood agar made of non-washed horse/ sheep erythrocytes. Haemolysin type I (lysing all types of washed erythrocytes) was the commonest one among all serovars except S. Abortusequi, none of which lysed horse erythrocytes. Salmonella enterica serovar Abortusequi having hemolytic activity against sheep erythrocytes were more invasive but had lesser ability to survive in sheep mononuclear cells than non-hemolytic strains. Multiplicity of haemolysins appeared significant epidemiological tool.     

Outer membrane proteins in the characterization of salmonellae of various origins

Outer membrane proteins have been studied in 76 Salmonella strains isolated from various sources and differing in their sensitivity to antibiotics, the presence of R-plasmids and a number of enzymatic properties. The outer membranes have been isolated by the modified method of L. W. Coulton and D. T. F. Wan. The study of the isolated proteins has been carried out by means of gel and disc electrophoresis in polyacrylamide gel with sodium dodecyl sulfate according to K. Weber and M. Osborn. The composition of the proteinograms thus obtained has revealed the presence of essential differences between hospital strains and cultures isolated from animals and from the environment in sporadic infections, as well as between strains belonging to different serovars. The possibility of using the characterization of outer membrane proteins of salmonellae in epidemiological investigations is discussed.     

Recent horizontal transmission of plasmids between natural populations of Escherichia coli and Salmonella enterica.

Seventy-one natural isolates obtained from a Salmonella reference collection were examined for the presence of plasmids closely related to the Escherichia coli F plasmid. The collection consists of several serovars of the S. enterica Typhimurium complex, subspecies I, to which 99% of pathogenic salmonellae belong. Molecular genetic techniques of DNA hybridization, along with PCR and DNA sequencing, were used to examine the occurrence, distribution, and genetic diversity of F-like plasmids among Salmonella strains. The F plasmid genes examined were finO, traD, traY, and repA, which map at dispersed positions on the F plasmid of E. coli. Comparative sequence analysis of each of the four genes in Salmonella plasmids showed them to be homologous (in some cases, virtually identical) to those found in F plasmids of E. coli natural isolates. Furthermore, the frequency of F-like plasmids in Salmonella strains was approximately the same as that observed in the E. coli Reference Collection. However, in Salmonella, the distribution was confined predominately to the serovars Typhimurium and Muenchen. The unexpected finding of a shared pool of F-like plasmids between S. enterica and E. coli demonstrates the significant role of conjugation in the histories of these important bacterial species.     

Salmonellae in the African great cane rat (Thryonomys swinderianus)

Because of its large size, the African great cane rat (Thryonomys swinderianus) is valued for food and has become a popular meat in western Africa. A survey was conducted to determine the occurrence of salmonellae in cane rats. Ten strains of Salmonella sp. were isolated from eight of 25 (32%) cane rats. Salmonella ajiobo was isolated from the spleen and intestines of three cane rats; S. agama was obtained from the spleen, liver and intestines of three animals; and S. poona was isolated from the spleen and liver of two cane rats. The occurrence of salmonellae in T. swinderianus is a potential public health hazard. Humans may become exposed to infection by consumption of inadequately cooked infected cane rat meat, or by eating vegetables, sugar cane and fruits contaminated with excretions of carrier cane rats. Incidents of human salmonellosis attributable to cane rat meat have not yet been reported; however, all three serotypes isolated from the cane rats have also been isolated from stools of patients suffering from gastroenteritis in Nigeria.