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Determination of close genetic relatedness of the major Salmonella enteritidis phage types by pulsed-field gel electrophoresis and DNA sequence analysis of several Salmonella virulence genes.

Salmonella enteritidis (SE) is an important cause of egg-associated outbreaks in both Europe and the United States. Phage typing has become an important epidemiologic tool in identifying the source of outbreaks. Limitations of phage typing have become apparent with wholesale egg distributors that have multiple suppliers in an area where a particular phage type is endemic. Several different molecular typing methods were evaluated for their discriminatory power to identify genetic differences among different SE phage types isolated in Europe and the United States. Pulsed-field gel electrophoresis (PFGE) identified a single DNA pattern among the different SE phage types. Comparison of the nucleotide sequence for several Salmonella virulence genes failed to identify a single nucleotide change in the gene sequences from most SE isolates, regardless of phage type. On the basis of these results, the different SE phage types appear to be genetically related or clonal. However, with primers 1283 and Opa4, it was possible to differentiate not only SE isolates from different geographic locations but those within a specific geographic locale as well by random amplified polymorphic DNA polymerase chain reaction. Any chance for discerning genetic differences among isolates will need to rely on molecular techniques other than PFGE.

Animals↗

Occurrence of Salmonella typhi and Salmonella paratyphi in Jordan.

In order to justify the surveillance control system and hygiene policy in Jordan, this study evaluated the occurrence of diarrhoea during the period 1988-2000, focusing on cases caused by Salmonella typhi and Salmonella paratyphi. From January 1988 to December 2000, the number of notified diarrhoeal cases by the Ministry of Health in Jordan was 1,399,563 million. Other groups of patients confined to the Governorate of Amman was diagnosed at Al-Battikhi Medical Laboratories. One-way ANOVA and Least Significant Difference (LSD) were carried out for statistical analysis. The number of reported diarrhea cases was 1,399,563, 53.0% were males, and 47.0% were females, among them, 80.3% were < 20 years and 19.7%, were > 20 years. Out of 245,255 patients tested for S. typhi and S. pararyphi, positive stool culture were 1992 (0.6%). Out of these, 960 (48.2%) were males and 1,032 (51.8%) were females (P = 0.028). The highest incidence rate (10.8) was observed in the year 1993, while the lowest incidence rate (0.9) was found in year 2000. A significant difference (P < 0.001) was found between the number of S. typhi and S. Paratyphi cases and year. The seasonal variation was also found to be significant (P < 0.0001), with the summer period showing the highest incident rate. A significant difference (P < 0.001) was observed between number of typhoid and paratyphoid cases and districts. A significance difference between number of typhoid and paratyphoid cases with age and sex. The group most affected was school age and adolescence. The demographic situation plays an important role in reporting typhoid and paratyphoid cases, where there might be an urgent indication for a better surveillance control system on water resources and disposal systems. S. typhi and S. paratyphi antibiotics resistance pattern showed they were resistant to tetracycline (56.0%, 58.0%), ampicillin (45.0%, 48.0%), trimethoprim (43.0%, 47.0%), cephtazidime (12.0%, 13.5%) chloramphenicol (6.8%, 7.2%), gentamycin (3.0%, 4.0%) neomycin (2.1. 1.8%), calvulanic acid (augmentin (1.4%, 2.2%) and norofloxacin (0.92%, 1.1%). Susceptibility to amikacin, ciprofloxacin, cetfriaxone, ofloxacine, imepenim, cefixime and cefotaxime was 100.0%. The increase in percentage of antibiotic resistant strain might indicate a need for a further prescribing policy for treatment.

Adolescent↗

Long-term post-Salmonella reactive arthritis due to Salmonella Blockley.

We describe the case of a patient who became ill with Salmonella Blockley food poisoning while working in Cyprus in August 1994. As his diarrhoea resolved he began to suffer from lower limb joint pains which were diagnosed as acute salmonella reactive arthritis. His condition deteriorated, then improved somewhat over a period of 2 years, but he continued to suffer symptoms over 5 years after infection. This case predates other reported cases of S. Blockley infection in Cyprus by 4 years. S. Blockley is associated with chickens, and the chicken meal is the probable source of his infection. This case is of interest since it demonstrates the emergence of the serovar outside South East Asia where it is common, and shows that information on the incidence and duration of reactive arthritis caused by serovars other than S. Enteritidis and S. Typhimurium is limited.

Adult↗

Detection and characterization of variant Salmonella genomic island 1s from Salmonella Derby isolates.

The purpose of this study is to investigate the distribution and structure of Salmonella genomic island 1 (SGI1) among Salmonella enterica serovar Derby isolates from swine and their rearing environment. Three variants of SGI1s, specifically SGI1-A, C, and I, were identified by PCR mapping. The results of macro-restriction analysis and DNA sequencing of SGI1 flanking regions revealed that there are at least two genomic lineages of Derby strains bearing SGI1s.

Animals↗

Antigen sharing of Salmonella typhi non-flagellar proteins with other salmonellae and some shigellae and Escherichia coli.

Cathodal moving protein components were identified in agarose gel electrophoresis of the Veronal buffer extract of a non-motile strain of S. typhi (8393, Colindale). Rabbit antiserum was raised against these cationic proteins; it had both agglutinating and precipitating activity. A total of 80 salmonella strains belonging to serogroups A, B, C1, C2, D, E1 and E2 including 26 S. typhi and 10 S. paratyphi A were tested against this antiserum in a slide agglutination test; all strains were agglutinated. Among 94 other bacterial strains tested, the antiserum agglutinated all 16 strains of Shigella flexneri, 2 of 5 Shigella sonnei, 5 of 34 E. coli and 1 of 8 Citrobacter species. These results show that there is antigenic sharing of the non-flagellar proteins of S. typhi with many other salmonellae as well as with some shigellae and E. coli.

Agglutination Tests↗

Study on the pH of ogi, Nigerian fermented weaning food, and its effect on enteropathogenic Escherichia coli, Salmonella typhi and Salmonella paratyphi.

The pH values of both cooked and uncooked ogi samples were determined and the survival of nalidixic acid-resistant enteropathogenic Escherichia coli OB 26 34/20, Salmonella typhi and Salmonella paratyphi A when seeded into cooked ogi were studied. The pH values of ogi ranged from pH 3.0 to 3.9. Cooked ogi had a slightly lower pH value than uncooked ogi. Our survival experiments showed that the inoculated enteric pathogens were considerably inhibited in cooked ogi during storage for 24 h. The antibacterial effect of cooked ogi was more pronounced on enteropathogenic E. coli with a log10 5 decline in bacterial count within 6 h.

Diarrhea, Infantile↗

The effect of oral and parenteral typhoid vaccination on the rate of infection with Salmonella typhi and Salmonella paratyphi A among foreigners in Nepal.

We studied the incidence of enteric fever among travelers and foreign residents who attended an expatriate clinic in Kathmandu, Nepal, from February 1987 to June 1988. There were 42 cases of enteric fever; 20 were caused by Salmonella typhi and 22 by Salmonella paratyphi A. Among 18 unvaccinated foreigners who had enteric fever, S typhi was isolated from 67%, and S paratyphi A from 33%, a ratio similar to the local Nepalese population. Among 22 vaccinated foreigners, S typhi was isolated from 35%, compared with 65% with S paratyphi A. Nine percent of tourists had received the oral Ty21A typhoid vaccine. However, among seven vaccinated tourists who became infected with S typhi, four (57%) had received the oral vaccine. Typhoid vaccine efficacy for tourists was calculated and showed an overall protective rate of 90% against enteric fever in general, 95% protection against S typhi, and 72% to 75% protection against S paratyphi A. We conclude that typhoid vaccine should be recommended to all travelers to the Indian subcontinent, and since S paratyphi A is the predominant cause of enteric fever among vaccinated travelers, consideration should be given to an effective vaccine against S paratyphi A when that becomes available.

Administration, Oral↗

[A trial with attenuated live Salmonella typhimurium vaccine for protection against an experimental Salmonella infection in pigeons].

A live, attenuated Salmonella typhimurium vaccine, available on the market, was administered to six pigeons. The strain of vaccine could not be identified in the droppings of pigeons up to ten days after administration. The pigeons as well as controls were experimentally infected with a Salmonella typhimurium var. copenhagen pigeon strain four weeks after vaccination. Differences in excretion of the var. copenhagen strain and in the appearance of diarrhoea and polyuria between the vaccinated and non-vaccinated pigeons were not detected.

Animals↗

Enterotoxin activity of a Salmonella typhimurium of equine origin in vivo in rabbits and the effect of Salmonella culture lysates and cholera toxin on equine colonic mucosa in vitro.

To determine whether a strain of Salmonella typhimurium (UCD 1755) of equine origin had enterotoxin activity, 2 ml of a cell-free culture lysate of strain UCD 1755 and approximately 10(9) viable strain UCD 1755 organisms were inoculated into ligated small intestinal segments of rabbits. Intestinal segments inoculated with viable strain UCD 1755 organisms and those inoculated with a cell-free culture lysate of strain UCD 1755 had significant (P less than 0.05) accumulations of fluid 10 hours after inoculation when compared with ligated intestinal segments either inoculated with sterile brain-heart infusion broth or left empty. There was not a statistically significant difference between fluid accumulation of intestinal segments inoculated with viable strain UCD 1755 and that of segments inoculated with a cell-free culture lysate of strain UCD 1755. The responses of equine colonic mucosa to culture filtrates of 2 strains of salmonella typhimurium (UCD 1755 and SL 1027) and purified cholera toxin were studied in vitro. Isolated smaples of colonic mucosa were incubated for 4 hours at 37 C in Krebs-Henseleit bicarbonate buffer (KHB) alone, KHB plus culture lysate of strain UCD 1755, KHB plus culture lysate of strain SL 1027, and KHB plus 1 microgram of cholera toxin/ml. Cyclic adenosine monophosphate (cAMP) content of each sample was determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Studies on the pathogenesis of Salmonella typhimurium and Salmonella choleraesuis var kunzendorf infection in weanling pigs.

Twenty-six 4-week-old pigs were randomly allotted to 4 groups: group 1--orally inoculated with Salmonella typhimurium; group 2--orally dosed with S choleraesuis; and groups 3 and 4, with surgically constructed intestinal loops--loops inoculated with either S typhimurium or S choleraesuis. One pig each from groups 1 and 2 was killed at 8, 12, 24, 48, 72, 96, and 120 hours after inoculation. One pig each from groups 3 and 4 was killed at 2, 4, 6, 8, 12, and 24 hours after intestinal loop inoculation. Inoculation of S typhimurium resulted in acute enterocolitis of variable severity, whereas inoculation of S choleraesuis resulted initially in septicemia followed by formation of large necrotic and ulcerative lesions in the colonic mucosa. The most consistent systemic lesion of S choleraesuis infection was interstitial pneumonia and multifocal hepatic necrosis. Salmonella typhimurium and S choleraesuis were ultrastructurally within enterocytes of ligated ileal loops. Intracellular bacteria were morphologically intact, occurred free in the cytoplasm and membrane bound, and caused no detectable cytotoxic effect to the cell. Both S typhimurium and S choleraesuis penetrated the intestinal mucosa and were isolated from mesenteric lymph nodes at 2 hours after inoculation.

Animals↗

Immunogenic complexes obtained from Salmonella typhi-murium and Salmonella typhi Ty2 by the bacterial acetone powder method.

An immunogenic complex was isolated from Salmonella typhi-murium and another one from Salmonella typhi Ty2. Both were prepared by the bacterial acetone powder method which eliminated the cell wall, the DNA almost completely and the membrane phospholipids. The complexes were denominated "New Vaccines". The S. typhi-murium new vaccine induced, even at doses of 0.5 microgram dry weight per mouse, a high degree of protection against the challenge of the virulent microorganism. By immunoelectrophoresis, 21 antigen-antibody systems could be detected, two of them corresponding to O antigens. The S. typhi Ty2 new vaccine induced better protection than the standard vaccine (heat-phenol inactivated typhoid vaccine) when both vaccines were compared in the relative potency test. Moreover, the new vaccine had very low toxicity when inoculated in humans at doses of 1 microgram dry weight, able to elicite a high antibody titre (1/1,790 mean of 10 sera) in 75% of the tested population, estimated by the complement fixation test. In contrast, the standard vaccine induced a low antibody titre (1/222, mean of 5 sera) in 50% of the humans inoculated with 1 X 10(8) bacterial cells. The new vaccine did not induce undesirable effects whereas the standard vaccine induced an important inflammatory process in 100% of the cases, with intense local pain in 67% after 24 h post-first inoculation as well as other less severe symptoms.

Acetone↗

Salmonella agona isolated from fish meal and a Salmonella strain isolated from shrimps in Lebanon.

Salmonella agona was isolated from an imported fish meal specimen and a Salmonella strain not listed in the Kauffmann-White Scheme was isolated from a shrimp specimen obtained from the Mediterranean coast of Lebanon. The biochemical and serological properties of the S. agona fish meal strain was identical to a standard strain. The serotype of the shrimp strain was O13,22 Vi. With the exception of hydrogen sulfide production the biochemical properties of the shrimp strain were similar to S. agona.

Agglutination Tests↗

[Epidemiological studies on salmonella in a certain area ("Walcheren project"). II. Presence of salmonella in the mesenteric lymph nodes and rectal contents of normal pigs slaughtered in Walcheren (author's transl].

During a period of twelve months, all pigs slaughtered on the former Isle of Walcheren were examined for the presence of Salmonella in the mesenteric lymph nodes and rectal contents. Salmonella was detected in 22.3% of 7,756 animals studied. Fifty different serotypes were isolated, two of these (S. menhaden and S. mikawasima) being isolated for the first time in the Netherlands. Data on the farms was collected, a differentiation being made between the type of farm (breeding, breeding and fattening, and fattening farms), the type of feed (pellets, pellets plus meal and meal) and the number of fattening pigs on each farm. The results show that there is a slight difference in the proportion of pigs responding positively to pellet rather than meal feeding, viz. 20.9% against 23.7%. A striking feature consists in the fact that this difference is much larger in favour of pellet feeding on the smaller farms (up to one hundred fattening pigs), viz. 12.9% against 21.0%. Another finding was that the average number of serotypes isolated on farms on which pellets were fed was smaller than that on farms on which meal was fed.

Animal Feed↗

[Dynamics of agglutination in birds artificially infected with Salmonella gallinarum-pullorum and Salmonella typhimurium and treated with sulfaguanidine and furazolidone].

Canadian Leghorn chickens were used to study the effect of sulfaguanidine and furazolidon on the dynamics of blood agglutinin formation. The birds were artificially infected with Salmonella gallinarum-pullorum and Salmonella typhimurium. At various intervals following infection they were treated with sulfaguanidine 1% tablets and 0.04% furazolidon. The level of agglutinins was determined by the methods of Huddleson (whole blood agglutination reaction), the fast serum agglutination, the method after Wright, and the indirect hemagglutination reaction. It was found that the agglutinin level in birds treated in the course of ten days with the chemotherapeutics mentioned prior to their infection did not change. The use of these chemotherapeutics parallel to the infection of the test birds and ten days later inhibited agglutinin formation. This was more pronouncedly expressed in birds treated with furazolidon. The delayed use of these therapeutic means did not inhibit substantially the level of blood agglutinins.

Agglutinins↗

Tumor necrosis factor-alpha production in swine after oral or respiratory challenge exposure with live Salmonella typhimurium or Salmonella choleraesuis.

A series of experiments was conducted to document tumor necrosis factor-alpha (TNF) activity in serum of swine after inoculation with Salmonella spp endotoxin and after oral or respiratory tract challenge exposure with live Salmonella spp. For experiment 1, a potentially lethal dose of S typhimurium endotoxin (25 micrograms/kg of body weight) was administered IV, and serum TNF activity was measured. High TNF (approx 700 IU/ml) activity at 1 to 2 hours after administration of the inoculum was associated with death, whereas lower TNF (approx 30 IU/ml) activity was associated with a general prolonged state of shock. For experiment 2, pigs were administered a nonlethal dose (5 micrograms/kg, IV) of either S typhimurium or S choleraesuis endotoxin. Difference in the ability to induce porcine serum TNF activity was not observed between strains. During experiment 3, pigs were inoculated with 10(4) colony-forming units of S typhimurium chi 4232 either orally by gelatin capsule (GC) or by intranasal (IN) instillation. A late serum TNF response (17 IU/ml) was measured at 6 weeks after IN inoculation. A serum TNF response was not detected in GC-inoculated pigs. All tissues and feces were test-negative for S typhimurium prior to the 6-week TNF response. Serum TNF activity may be related to clearance of S typhimurium after respiratory tract exposure, but it is not important to or indicative of clearance of orally presented S typhimurium in swine. During experiment 4, pigs were inoculated with 10(6) colony-forming units of S typhimurium chi 4232 similarly as for experiment 3.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Intranasal↗

Fate of Salmonella enteritidis and Salmonella typhimurium during the fermentation of siljo.

The behaviour of Salmonella enteritidis and S. typhimurium in control gruel and fermenting siljo was evaluated. S. enteritidis and S. typhimurium reached maximum levels of 10(8) cfu/ml and 10(6) cfu/ml, respectively within 48 hours in the control gruel. In fermenting "siljo", pH fell to < 5.0 within 48 hours and titratable acidity increased markedly. This resulted in the complete inhibition of S. enteritidis in 48 hours and S. typhimurium in 72 hours. Acid production by the fermenting lactic acid bacteria and components of mustard powder could have inhibited the test organisms. Since siljo is usually ready for consumption after 72 hours of fermentation, contaminating salmonellae would be eliminated before the product is consumed.

Colony Count, Microbial↗

Evaluation of oral, subcutaneous, and nasal administration of Salmonella enteritidis-immune lymphokines on the potentiation of a protective heterophilic inflammatory response to Salmonella enteritidis in day-old chickens.

We have previously reported that the prophylactic administration of factor(s) from T-cell supernatants derived from Salmonella enteritidis-immune chickens (ILK) have a favorable effect in controlling or eliminating salmonellosis in neonatal poultry. Experimentally, we have used the intraperitoneal injection as the standard method of administering ILK to neonatal poultry. However, this method is neither easy, practical, nor economical for the poultry industry. In the present study, we evaluated the effectiveness of oral (p.o.), intranasal (i.n.), and subcutaneous (s.c.) administration of ILK for ease of delivery, induction of protective resistance against Salmonella enteritidis (Se) organ invasion, and the ability to activate peripheral blood heterophils in day-old chickens. In the first experiments, delivery of ILK p.o., i.n., and s.c. significantly (P < 0.01) increased the resistance of day-old chickens to Se organ invasion. The level of protection was equivalent to that induced by the i.p. route. Administration of a comparable protein control (bovine serum albumin, BSA) by the 3 routes induced no protective effect against Se organ invasion. Likewise, a significant increase was found in the number of circulating heterophils within 4 h of administration of the ILK by all routes. In the 2nd experiment, the function of the heterophils from ILK-treated birds was compared with that of the control cells in adherence, chemotaxis, and phagocytosis assays. The heterophils from birds given ILK i.p., s.c., p.o., or i.n. had significantly (P < 0.01) increased functional activities when compared to the activities of the heterophils from the control birds. These studies indicate that the delivery of ILK either orally or parenterally, routes which can be used by the poultry industry, can confer protection to chickens against a localized enteric Se organ invasion by potentiating the systemic heterophilic innate response.

Administration, Intranasal↗

Criteria for the standardization of Salmonella mutagenicity tests: results of a collaborative study. II. Studies to investigate the effect of bacterial liquid culture preparation conditions on Salmonella mutagenicity test results.

The influence of various parameters and growth conditions in the "overnight culture" of Salmonella typhimurium strains on mutagenicity test results was investigated. A number of factors were first suspected to be of some importance for the quantitative outcome of the mutagenicity test. None of them, however, was found to influence the results to such a marked extent as to be a major source of variability. Only the brand of nutrient broth used for the propagation of the bacteria proved finally to have a certain effect on the number of (spontaneous and induced) revertant colonies, although no precise and quantitative statements can be made with regard to a possible standardization of this experimental segment in the Salmonella mutagenicity test. The occurrence of such unpredictable but noticeable influences is, however, evidence for the importance of an intralaboratory optimization and standardization of all parts of the test procedure.

Animals↗