J Mol Biol, 2004 Jul 16, 340(4), 753 - 66 Stepwise dissection of the Hin-catalyzed recombination reaction from synapsis to resolution; Sanders ER et al.; The Hin DNA invertase promotes a site-specific DNA recombination reaction in the Salmonella chromosome . The native Hin reaction exhibits overwhelming selectivity for promoting inversions between appropriately oriented recombination sites and requires the Fis regulatory protein, a recombinational enhancer, and a supercoiled DNA substrate . Here, we report a robust recombination reaction employing oligonucleotide substrates and a hyperactive mutant form of Hin . Synaptic complex intermediates purified by gel electrophoresis were found to contain four Hin protomers bound to two recombination sites . Each Hin protomer is associated covalently with a cleaved DNA end . The cleaved complexes can be ligated into both parental and recombinant orientations at equivalent frequencies, provided the core residues can base-pair, and are readily disassembled into separated DNA fragments bound by Hin dimers . Kinetic analyses reveal that synapsis occurs rapidly, followed by comparatively slow Hin-catalyzed DNA cleavage . Subsequent steps of the reaction, including DNA exchange and ligation, are fast . Thus, post-synaptic step(s) required for DNA cleavage limit the overall rate of the recombination reaction. J Food Prot, 2004 Jun, 67(6), 1257 - 60 Heat treatments to enhance the safety of mung bean seeds; Hu H et al.; Salmonella enterica serovars and Escherichia coli O157:H7 have been associated with contaminated seed sprout outbreaks . The majority of these outbreaks have been traced to sprout seeds contaminated with low levels of pathogens . E . coli O157:H7 strains can grow an average of 2.3 log CFU/g over 2 days during seed germination, and Salmonella can achieve an average growth of 3.7 log CFU/g . Therefore, it is important to find an effective method to reduce possible pathogenic bacterial populations on the seeds prior to sprouting . Our objective was to assess the effectiveness of various dry heat treatments on reducing E . coli O157:H7 and Salmonella populations on mung beans intended for sprout production and to determine the effect of these treatments on seed germination . Mung beans were inoculated with five-strain cocktails of E . coli O157:H7 and of Salmonella serovars harboring the green fluorescent protein gene and then air dried overnight . Heat treatments were performed by incubating the seeds at 55 degrees C for various periods of time . Heat-treated seeds were then assessed for the efficacy of the heat treatment and the effects of heat treatment on germination rates . After inoculation and drying, 6 log CFU/g E . coli O157:H7 and 4 log CFU/g Salmonella were detected on the seeds . Following heat treatment, pathogenic bacterial populations on the seeds were below detectable levels (<1 log CFU/g), but the germination rate of the seed was not affected . Thus, the risk of contamination and the presence of pathogens in the finished sprouts were greatly reduced via the seed heat treatment process. J Food Prot, 2004 Jun, 67(6), 1201 - 8 Simulation model for enumeration of Salmonella on chicken as a function of PCR detection time score and sample size: implications for risk assessment; Oscar TP; A data gap commonly identified in risk assessments is the lack of quantitative information on the contamination of food with pathogens . A simulation model that predicts the incidence and distribution of Salmonella contamination on chicken as a function of PCR detection time score and sample size was developed with data from challenge studies with preenrichment samples that were composed of 25 g of chicken and 225 ml of buffered peptone water inoculated with 10(0.7) to 10(6) Salmonella and incubated at 37 degrees C . At 0, 2, 4, 6, 8, 10, 12, and 24 h of incubation, subsamples were collected and tested for Salmonella by PCR, and a PCR detection time score based on the widths of the bands in the electrophoresis gel was obtained for each preenrichment sample . Standard curves relating PCR detection time score to initial density of Salmonella inoculated were developed for sterile and nonsterile preenrichment samples . Presence of other microorganisms in the preenrichment sample decreased the PCR detection time score at low (<10(2) per 25 g) but not at high (>10(2) per 25 g) initial densities of Salmonella and resulted in a nonlinear standard curve rather than the linear standard curve obtained for sterile samples . The predicted incidence and distribution of Salmonella contamination on chicken increased in a nonlinear manner as sample size increased from 25 to 500 g . The new method reduced the time and cost of Salmonella enumeration by eliminating the selective enrichment, selective plating, and confirmation steps of the traditional most-probable-number method . Results are useful for risk assessment because they consider the uncertainty of the standard curve predictions and because they provide distributions of Salmonella contamination for different size samples of chicken that can be directly used in risk assessment. J Food Prot, 2004 Jun, 67(6), 1177 - 83 Reduction of Salmonella by two commercial egg white pasteurization methods; Robertson WR et al.; The effect of pH, processing temperatures, and preheating steps in two commercial egg white pasteurization procedures (Armour and Standard Brands methods) were evaluated using a five-strain cocktail of Salmonella . We devised a benchtop pasteurization system that would more closely resemble the two commercial processes than could the traditional capillary tube method . The pasteurization methods both require hydrogen peroxide to be metered into the egg white stream between a required initial preheat step and the main heating regimen . Both processes were evaluated at three pH levels (pH 8.2, 8.6, 9.0), at four temperatures (51.7 degrees C/125 degrees F, 53.1 degrees C/127.5 degrees F, 54.4 degrees C/130 degrees F, 55.8 degrees C/132.5 degrees F), and over four residence times to allow calculation of D-values at each temperature . When compared at the minimum allowable time and temperatures for each process, our results showed at least a 1-log greater log reduction (P < 0.05) for the Standard Brands method than the Armour method in 10 of 12 of the pH and temperature combinations tested . Almost all runs at any given temperature showed more reduction at pH 9.0 than at pH 8.2 except for the Standard Brands method at 54.4 degrees C and 55.8 degrees C, which showed the most consistent reduction across all three pH levels tested . Analysis of the preheat portion of the two methods showed that there was no contribution (P > 0.05) toward Salmonella reduction when compared with the identical process without the preheating step . We generally observed a greater reduction of Salmonella with egg white at pH 9.0 that is typical of older, off-line processing than with low pH egg white (i.e., 8.2) that is typical of modern in-line processing facilities . This difference was as much as 3.5 log cycles depending on the processing conditions . The data has been used to make recommendations for minimum processing conditions for hydrogen peroxide-based egg white pasteurization. J Food Prot, 2004 Jun, 67(6), 1157 - 62 Radiosensitization of Escherichia coli and Salmonella Typhi in ground beef; Chiasson F et al.; The radiosensitization of two pathogenic bacteria, Escherichia coli and Salmonella Typhi, was evaluated in the presence of thyme and its principal essential oil constituents (carvacrol and thymol) in ground beef . Ground beef was inoculated with E . coli or Salmonella Typhi (10(5) CFU/g), and each compound was added separately at various concentrations (0 to 3.5%, wt/wt) . The antimicrobial potential of carvacrol, thymol, and thyme was evaluated in unirradiated meat by determining the MIC in percentage (wt/wt) after 24 h of storage at 4 +/- 1 degree C . Results showed a MIC of 0.88 +/- 0.12%, 1.14 +/- 0.05%, and 2.33 +/- 0.32% for E . coli in the presence of carvacrol, thymol, and thyme, respectively . MICs of 1.15 +/- 0.02%, 1.60 +/- 0.01%, and 2.75 +/- 0.17% were observed for Salmonella Typhi in the presence of the same compounds, respectively . The best antimicrobial compound (i.e., carvacrol) was selected and added to the sterilized ground beef along with ascorbic acid (0.5%, wt/wt) and tetrasodium pyrophosphate (0.1%, wt/wt) . Meat samples (10 g) were packed in air and then irradiated in a 60Co irradiator at doses of 0 to 0.7 kGy for the determination of E . coli radiation D10 and 0 to 2.25 kGy for the determination of Salmonella Typhi radiation D10 . Addition of carvacrol increased the relative sensitivity of both bacteria 2.2 times . The radiation D10 was reduced from 0.126 +/- 0.0039 to 0.057 +/- 0.0015 kGy for E . coli and from 0.519 +/- 0.0308 to 0.235 +/- 0.0158 kGy for Salmonella Typhi . The addition of tetrasodium pyrophosphate did not affect significantly (P > 0.05) the radiosensitization of either bacterium . However, the presence of ascorbic acid in the media reduced significantly (P < or = 0.05) the radiosensitivity of both bacteria . An additive effect of carvacrol addition and packaging under modified atmosphere conditions (60% O2-30% CO2-10% N2) was also observed on bacterial radiosensitization at 4 degrees C . Compared with the control packed under air, modified atmosphere packaging conditions in the presence of carvacrol and tetrasodium pyrophosphate improved the relative sensitivity of E . coli by 2.7 times and Salmonella Typhi by 9.9 times. J Food Prot, 2004 Jun, 67(6), 1111 - 5 Frozen chicken nuggets and strips--a newly identified risk factor for Salmonella Heidelberg infection in British Columbia, Canada; MacDougall L et al.; Salmonella enterica var . Heidelberg was isolated from an unusual food source during routine case follow-up, prompting a case control investigation of frozen chicken nuggets and strips . Most frozen nuggets and strips are raw; however, par-frying lends a cooked appearance . As such, suitable food preparation precautions might not be undertaken by consumers . Cases were confirmed in the laboratory between 1 January and 1 April 2003 . Controls were generated through forward-digit dialing and individually matched by age category . Telephone interviews were conducted, and limited sampling of unopened product was performed . Eighteen matched pairs were interviewed . The odds of infection were 11 times higher in individuals who had consumed frozen processed chicken nuggets and strips (95% confidence interval, 1.42 < odds ratio < 85.20) . One-third of cases and controls considered frozen nuggets and strips to be precooked, and one quarter used the microwave, an ill-advised cooking method . Consumer misconceptions contributed to the risk of infection . Clear labels identifying nuggets and strips as raw poultry are needed. J Food Prot, 2004 Jun, 67(6), 1092 - 103 Survival and recovery of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on lettuce and parsley as affected by method of inoculation, time between inoculation and analysis, and treatment with chlorinated water; Lang MM et al.; The effects of method for applying inoculum and of drying time after inoculation on survival and recovery of foodborne pathogens on iceberg lettuce and parsley were studied . Five-strain mixtures of Escherichia coli O157:H7, Salmonella, or Listeria monocytogenes were applied to lettuce and parsley by dip, spot, or spray inoculation methods . Inocula were dried for 2 h at 22 degrees C or for 2 h at 22 degrees C and then 22 h at 4 degrees C before being treated with water (control) or chlorine (200 microg/ml) . Significantly higher populations (CFU per lettuce or parsley sample) of E . coli O157:H7 and Salmonella (alpha = 0.05) were recovered from dip-inoculated produce than from spot- or spray-inoculated produce . This difference was attributed to larger numbers of cells adhering to lettuce and parsley subjected to dip inoculation . Populations of E . coli O157:H7 and Salmonella recovered from lettuce inoculated by spot and spray methods were not significantly different, but populations recovered from spot-inoculated parsley were significantly higher than those recovered from spray-inoculated parsley, even though the number of cells applied was the same . Significantly different numbers of L . monocytogenes were recovered from inoculated lettuce (dip > spray > spot); populations recovered from dip-inoculated parsley were significantly higher than those recovered from spot- or spray-inoculated parsley, which were not significantly different from each other . Populations of pathogens recovered from lettuce and parsley after drying inoculum for 2 h at 22 degrees C were significantly higher than or equal to populations recovered after drying for 2 h at 22 degrees C and then for 22 h at 4 degrees C . Significant differences (water > chlorine) were observed in populations of all pathogens recovered from treated lettuce and parsley, regardless of inoculation method and drying time . It is recommended that spot inoculation with a drying time of 2 h at 22 degrees C followed by 22 h at 4 degrees C be used to determine the efficacy of chlorine and other sanitizers in killing foodborne pathogens on lettuce and parsley. Biometals, 2004 Jun, 17(3), 337 - 42 Recombinant human lactoferrin treatment for global health issues: iron deficiency and acute diarrhea; Bethell DR et al.; Iron deficiency and diarrhea are two of the most significant issues for global health . Iron deficiency anemia is the most common nutritional deficiency in the world, affecting nearly 25% of the world population (UNICEF/WHO 1999) . The prevalence of iron deficiency in developing countries is illustrated by comparison with other deficiencies: iron deficiency affects 3.5 billion people, while vitamin A and iodine deficiency affect 0.3 billion people and 0.8 billion people, respectively . The prevalence is highest among young children and women of childbearing age (particularly pregnant women) . It is estimated that national productivity levels could be raised as much as 20% by correcting iron deficiency in developing countries . Recombinant human lactoferrin (rhLF), expressed and extracted from rice seed, is being evaluated by Ventria Bioscience for use as a dietary supplement to treat iron deficiency and/or iron deficiency anemia . Diarrhea is also a major world health issue . Sixty percent of children who die under age five die of pneumonia, diarrhea or measles . World Health Organization oral rehydration solution (WHO-ORS) is one of the major medical advances in the past 50 years, saving the lives of 1 to 2 million children annually . Many studies have demonstrated similar efficacy of rice-based ORS . There are studies documenting the reduced frequency of diarrhea in breast-fed children and this health improvement is attributed to the antimicrobial action of the human milk proteins lactoferrin and lysozyme . In vitro data document the growth inhibition of the diarrheal associated organisms: rotavirus, ETEC, cholera, salmonella, and shigella by human lactoferrin (hLF) and human lysozyme . Using Ventria's ExpressTec system, we have expressed human lactoferrin and human lysozyme in rice . In a rice-based ORS formulation, these proteins have the potential to provide not only the benefits of reduced stool volume and improved weight gain, but also shorten the course of diarrheal episodes via antimicrobial activity against the causative agent. Prev Vet Med, 2004 Jun 10, 64(1), 1 - 14 Estimation of infection prevalence from correlated binomial samples; Condon J et al.; Infection prevalence in a population often is estimated from grouped binary data expressed as proportions . The groups can be families, herds, flocks, farms, etc . The observed number of cases generally is assumed to have a Binomial distribution and the estimate of prevalence is then the sample proportion of cases . However, the individual binary observations might not be independent--leading to overdispersion . The goal of this paper was to demonstrate random-effects models for the estimation of infection prevalence from data which are correlated and in particular, to illustrate a nonparametric random-effects model for this purpose . The nonparametric approach is a relatively recent addition to the random-effects class of models and does not appear to have been discussed previously in the veterinary epidemiology literature . The assumptions for a logistic-regression model with a nonparametric random effect were outlined . In a demonstration of the method on data relating to Salmonella infection in Irish pig herds, the nonparametric method resulted in the classification of herds into a small number of distinct prevalence groups (i.e . low, medium and high prevalence) and also estimated the relative frequency of each prevalence category in the population . We compared the estimates from a logistic model with a nonparametric distribution for the random effects with four alternative models: a logistic-regression model with no random effects, a marginal model using a generalised estimating equation (GEE) and two methods of fitting a Normally distributed random effect (the GLIMMIX macro and the NLMIXED procedure both in SAS) . Parameter estimates from random-effects models are not readily interpretable in terms of prevalences . Therefore, we outlined two methods for calculating population-averaged estimates of prevalence from random-effects models: one using numerical integration and the other using Monte Carlo simulation. Public Health Rep, 2004 Jul-Aug, 119(4), 435 - 42 A review of outbreaks of waterborne disease associated with ships: evidence for risk management; Rooney RM et al.; OBJECTIVE: The organization of water supply to and on ships differs considerably from that of water supply on land . Risks of contamination can arise from source water at the port or during loading, storage, or distribution on the ship . The purpose of this article is to review documented outbreaks of waterborne diseases associated with passenger, cargo, fishing, and naval ships to identify contributing factors so that similar outbreaks can be prevented in the future . METHODS: The authors reviewed 21 reported outbreaks of waterborne diseases associated with ships . For each outbreak, data on pathogens/toxins, type of ship, factors contributing to outbreaks, mortality and morbidity, and remedial action are presented . RESULTS: The findings of this review show that the majority of reported outbreaks were associated with passenger ships and that more than 6,400 people were affected . Waterborne outbreaks due to Enterotoxigenic Escherichia coli, noroviruses, Salmonella spp, Shigella sp, Cryptosporidium sp, and Giardia lamblia occurred on ships . Enterotoxigenic E . coli was the pathogen most frequently associated with outbreaks . One outbreak of chemical water poisoning also occurred on a ship . Risk factors included contaminated port water, inadequate treatment, improper loading techniques, poor design and maintenance of storage tanks, ingress of contamination during repair and maintenance, cross-connections, back siphonage, and insufficient residual disinfectant . CONCLUSIONS: Waterborne disease outbreaks on ships can be prevented . The factors contributing to outbreaks emphasize the need for hygienic handling of water along the supply chain from source to consumption . A comprehensive approach to water safety on ships is essential . This may be achieved by the adoption of Water Safety Plans that cover design, construction, operation, and routine inspection and maintenance. Przegl Epidemiol, 2004, 58(1), 213 - 8 {Typhoid fever in a child--a case report}; Kuchar E et al.; We report a case of typhoid fever in an 8 years old boy . The child was initially admitted to a local hospital where pneumonia, myocarditis with heart failure, pyelonephritis, liver and pancreatic failure as well as cholelithiasis were suspected . Zinaceff and Amikin were administered and after 8 days the child was referred to the cardiology department of a regional reference hospital due to heart failure symptoms . There the diagnosis of sepsis was established, and the antibiotics changed to Pipril and Amikin . The child however did not improve and after two days he was transferred to an intensive care unit . The previous anti-microbial therapy was continued for another 7 days until the results of stool culture revealing Salmonella sp . were available . Subsequently the boy was admitted to our clinic . Based on the clinical course, Widal test and isolating of the Salmonella typhi from the stool samples typhoid fever was diagnosed. Przegl Epidemiol, 2004, 58(1), 85 - 101 {Foodborne infections and intoxications in Poland in 2002}; Przybylska A; A total of 26,734 bacterial foodborne infections and intoxications were registered in 2002 (incidence 69.9/100,000 population) . S . Enteritidis was found in 94.8% of collective outbreaks (in Poland 4 sick people and more) and 90.4% cases in outbreaks caused by Salmonella of animal's source . The main vehicle of foodborne and waterborne outbreaks in 2002 was food prepared from various raw materials (3 and more) of animal's source (35.6% cases in outbreaks) and the next--from eggs (20.9% cases in outbreaks) . Private homes prevailed (47.1% outbreaks, 66.2% outbreaks caused by Salmonella) among the places of the ready made food production . Ten epidemics with 100 and more cases each, were registered . Six deaths were noted in outbreaks in 2002 (2--in the result of salmonellosis of the animal's source, 1--in the result of staphylococcal infection and 3--after chemical poisoning). Przegl Epidemiol, 2004, 58(1), 67 - 76 {Salmonellosis in Poland in 2002}; Gonera E et al.; Total number of salmonellosis cases slightly increased . In 2002, 20,688 cases were reported (19,881 in previous year), incidence rate = 54.1 per 100,000 population . 68% of patients were hospitalized, but percentage of hospitalized cases with extraintestinal manifestations was much higher--above 91% . The seasonal peak was observed as in previous year in July and August . Since 1995 (when Salmonella strains were isolated in 91% patients) decreasing trend in confirmation of clinical diagnosis is observed . The most frequent isolated type remained Salmonella Enteritidis--above 86% of cases . Only four other serotypes (Typhimurium, Hadar, Virchow and Infantis) were detected in all of 16 voivodeships of Poland . The age, sex, urban/rural distribution of salmonellosis cases remains stable; the highest incidence was registered among children aged 2 (474.0/100,000) . Extraintestinal manifestations of salmonellosis (septicaemia, meningitis, pneumonia peritonitis and other) were observed in 113 patients (the highest number since 1994). J Vasc Surg, 2004 Jul, 40(1), 30 - 5 Infected aortic aneurysms: clinical outcome and risk factor analysis; Hsu RB et al.; PURPOSE: Infected aortic aneurysms are difficult to treat, and are associated with significant mortality . Hospital survival is poor in patients with severe aortic infection, Salmonella species infection, Staphylococcus aureus infection, aneurysm rupture, and suprarenal aneurysm location . We reviewed the clinical outcome in 46 patients with primary infected aortic aneurysms and identified clinical variables associated with prognosis . METHODS: Data were collected by means of retrospective chart review . Univariate and multivariate logistic regression models were used for risk factor analysis . RESULTS: Between August 1995 and March 2003, 48 patients with primary infected aortic aneurysms were treated at our hospitals . Two patients with negative culture results were excluded . Of the remaining 46 patients, 35 patients had aortic aneurysms infected with Salmonella species and 11 patients had aortic aneurysms infected with microorganisms other than Salmonella species . There were 20 suprarenal infections and 26 infrarenal infections . Surgical debridement and in situ graft replacement were performed in 35 patients, with an early mortality rate of 11% . The incidence of late prosthetic graft infection was 10% . The 90-day mortality rate in patients operated on was 0% for elective operation and 36% for nonelective operation (P =.006, Fisher exact test) . Independent predictors of aneurysm-related death were advanced age, non-Salmonella infection, and no operation . CONCLUSION: With timely surgical intervention and prolonged antibiotic treatment, in situ graft replacement provides an excellent outcome in patients with primary infected aortic aneurysms and elective operation . Mortality is still high in patients undergoing urgent operation . Advanced age, non-Salmonella infection, and no operation are major determinants of mortality. J Med Assoc Thai, 2004 Apr, 87(4), 400 - 4 Percutaneous ultrasound-guided fine needle aspiration of abdominal lymphadenopathy in AIDS patients; Veerapand P et al.; OBJECTIVE: To analyse the causes of abdominal lymphadenopathy in AIDS patients and evaluate the performance success rate, diagnostic yielding rate and safety of ultrasound (US)-guided fine needle aspiration (FNA) . MATERIAL AND METHOD: Medical records of HIV seropositive with abdominal lymphadenopathy who underwent percutaenous US-guided FNA from August 1997 to January 2002 were retrospectively reviewed . Records of clinical and ultrasonographic findings, numbers of FNA performed, numbers of obtained specimen, laboratory results and postoperative problems were analysed . Performance success is defined as obtaining a specimen while diagnostic yielding is performance success with laboratory result . RESULTS: FNA was done 72 times in 71 patients obtaining specimens in 68 cases (performance success rate = 94.4%) with positive laboratory result in 63 cases (diagnostic yielding rate = 87.5%) . The culture gave results in 56 cases; M . tuberculosis (TB) = 36, TB with other organisms = 5, Mycobacterium avium complex = 2, Nontuberculous mycobacterium = 10, C . neoformans = 1, Salmonella with positive fungal stain = 1, E . coli with P . mirabilis = 1 . Five cases had positive AFB without culture specimen and one revealed positive AFB stain but negative culture . Lymphoma was found in only one case . No postoperative hemorrhage, peritonitis or perforated bowel in patient or needle stick injury to medical staff was reported . CONCLUSION: Abdominal lymphadenopathy at Bamrasnaradura Hospital is mostly caused by TB and the second most common is nontuberculous mycobacterium . Percutaneous FNA under US guidance could yield definite diagnosis with a high performance success rate, high diagnostic yielding rate and was safe. J Chemother, 2004 Apr, 16(2), 128 - 33 Effects of acetyl salicylate and ibuprofen on fluoroquinolone MICs on Salmonella enterica serovar typhimurium in vitro; Coban AY et al.; In this study, the effects of acetylsalicylate and ibuprofen at 2, 4 and 8 mM concentration were investigated on ofloxacin, ciprofloxacin, levofloxacin and pefloxacin minimum inhibitory concentrations (MICs) for 14 Salmonella enterica serovar typhimurium clinical isolates, one standard strain (SZH KUEN 557), SH7616 (acr mutant), SH5014 (parent strain of acr mutant) and PP120 (soxRS mutant) strains . All isolates were susceptible to the 4 fluoroquinolones . In the presence of 2, 4 and 8 mM acetylsalicylate and ibuprofen, 2- to 8-fold increases were observed in fluoroquinolone MICs . This rise was higher, especially in the presence of acetylsalicylate . In spite of this rise, none of the MICs were in the range of resistance limits in vitro . Except for a 2-fold increase in levofloxacin MICs, we did not observe any difference in MICs of ofloxacin, ciprofloxacin, and pefloxacin in the presence of 2, 4 and 8 mM acetylsalicylate and ibuprofen for SH7616 and PP120 strains . According to the in vitro results of this study, it can be suggested that use of acetylsalicylate or ibuprofen together with clinical treatment of bacteria, especially bacteria which show intermediate resistance, will cause resistance . However, since clinical data are insufficient, further studies are needed. Rozhl Chir, 2004 May, 83(5), 209 - 16 {Bacterial aortitis}; Sebesta P et al.; A cohort of 14 patients with bacterial destruction of various segments of the aortic wall is presented . The Salmonella enteritidis strain was predominantly responsible . Most patients had typical history of symptomatic trias of sepsis, abdominal and/or back pain and positive blood cultures . CT scan showed pseudoaneurysm within the thoracic, subphrenic or subrenal aorta as well as acute hemorrhage in three patients . One of these was excluded from invasive treatment due to hopeless prognosis . In one patient primary aortoduodenal phistula was responsible for GI bleeding . Five patients were operated and prosthetic replacement of subrenal or iuxtarenal aortic portion together with aortorenal bypass in a couple of cases was performed . In eight patients stentgrafts (SG) of various types were deployed completed with femorofemoral crossover bypass when necessary . All patients were subject to long-standing antibiotic therapy . Two patients expired following SG insertion, all operated patients survived . Average follow-up has been 1 year (1-22 months) so far . A groin abscess was later drained in one patient . Neither CT nor isotope scanning showed persistent or recurrent infectious or hemorrhagic foci in any survivors whatsoever . The authors review and consider the doubtful indication of aortic SG deployment into the septic terrain in selected cases . Midterm results might justify its use in overly debilitated patients otherwise not eligible for radical operation due to its prohibitive risk. Virus Genes, 2004 Aug, 29(1), 87 - 98 Homology between two different Salmonella phages: Salmonella enterica serovar Typhimurium phage P22 and Salmonella enterica serovar Anatum var . 15 + phageepsilon34; Salgado CJ et al.; A distinguishing feature of many microorganisms, belonging to the Gram negative group of bacteria, is the presence of the lipopolysaccharide on their cell surface . Salmonella is a prominent member of this group of bacteria . Many Salmonella phages use the LPS as the initial receptor in the infection process and they can distinguish subtle changes in the LPS molecules . The phage protein that is responsible for recognition of these cells is the tail or tailspike protein (TSP) . Those TSPs, which use LPS as a receptor, are prokaryotic LPS-binding proteins . As an initial step in using phage TSPs as model systems for a detailed molecular genetic analysis of protein-LPS interactions, a comparison of two phages and their TSPs from two different Salmonella bacterial viruses (phages), Salmonella enterica serovar Typhimurium phage P22 and Salmonella enterica serovar Anatum var . 15 + phage epsilon34, is being carried out . This present study shows significant viral protein homology between many viral structural proteins from these two phages including their TSPs . Significantly this report suggests a general structural motif for part of the TSP of phages and suggests that a more detailed comparative analysis of these TSPs is warranted. J Biol Chem, 2004 Aug 27, 279(35), 36470 - 80 Epub 2004 Jun 23. Investigation of the structural requirements in the lipopolysaccharide core acceptor for ligation of O antigens in the genus Salmonella: WaaL "ligase" is not the sole determinant of acceptor specificity; Kaniuk NA et al.; The ligation of O antigen polysaccharide to lipid A-core oligosaccharide is a late step in the formation of the complex glycolipid known as lipopolysaccharide . Although the process has been localized to the periplasmic face of the inner membrane, details of the ligation mechanism have not been resolved . To date, there is only one gene product (WaaL, often referred to as "ligase") known to be required . There exists a requirement for a specific lipid A-core oligosaccharide acceptor structure for ligation activity, and it has been proposed that the WaaL protein imparts this acceptor specificity . Here the structural requirements in the core oligosaccharide acceptor for O antigen ligation are investigated in prototype serovars of Salmonella enterica . Complementation experiments in mutants with defined core oligosaccharide structure indicate that the specificity of the ligation reaction for a particular core oligosaccharide structure is not dependent on the WaaL protein alone . The data provide the first indication of a more complicated recognition process involving additional cellular components. Antimicrob Agents Chemother, 2004 Jul, 48(7), 2712 - 5 Incidence of the recently described sulfonamide resistance gene sul3 among German Salmonella enterica strains isolated from livestock and food; Guerra B et al.; The sul3 gene recently described in Escherichia coli was found in 22 of 512 (4.3%) German Salmonella isolates from different regions and sources and of different serotypes, antimicrobial resistance phenotypes, and genomic groups . This is the first report on the prevalence of sul3 among Salmonella strains, and the findings support the strong potential of this determinant to spread within bacterial populations. Antimicrob Agents Chemother, 2004 Jul, 48(7), 2510 - 7 Salmonella genomic island 1 multidrug resistance gene clusters in Salmonella enterica serovar Agona isolated in Belgium in 1992 to 2002; Doublet B et al.; Salmonella genomic island 1 (SGI1) harbors a multidrug resistance (MDR) gene cluster which is a complex class 1 integron . Variant SGI1 MDR gene clusters conferring different MDR profiles have also been identified in several Salmonella enterica serovars and classified as SGI1-A to -F . A retrospective study was undertaken to characterize MDR regions from serovar Agona strains harboring SGI1 isolated from poultry in Belgium between 1992 and 2002 . A total of 171 serovar Agona strains, displaying resistance to at least one antibiotic, were studied for the presence of SGI1 . SGI1 was detected in 94 serovar Agona strains . The most prevalent variant was SGI1-A (85%), which harbors within the SGI1 complex class 1 integron a common region (CR1) containing orf513, a putative transposase gene, adjacent to the dfrA10 trimethoprim resistance gene . A new variant SGI1 named SGI1-G was identified in two strains . It consisted of the pse-1 gene cassette, as in SGI1-B, but with additional insertion of the orf513/dfrA10 region structure . Seven strains displaying the typical SGI1 MDR profile (Ap Cm Ff Sm Sp Su Tc) showed genetic variation at the 3' end of SGI1 . These strains harbored the insertion of the CR1 containing orf513 as in SGI1-A, -D, and -G . However, downstream the right end of CR1, they presented different 7.4- to 8.5-kb deletions of the SGI1 3' end that extended to the chromosomal genes yieE and yieF . These results suggest a possible role of CR1 in deletion formation, as has been reported for some insertion sequences . Pulsed-field gel electrophoresis analysis showed that all the serovar Agona SGI1-carrying strains belonged to a single clone . Thus, SGI1 is largely encountered in serovar Agona strains isolated from poultry in Belgium, the most prevalent variant being SGI1-A . SGI1 MDR region undergoes recombinational events resulting in a diversity of MDR gene clusters. Antimicrob Agents Chemother, 2004 Jul, 48(7), 2364 - 9 Antibiotic resistance conferred by a class I integron and SXT constin in Vibrio cholerae O1 strains isolated in Laos; Iwanaga M et al.; Changes in the drug susceptibility pattern were observed in Vibrio cholerae O1 isolated in the Lao People's Democratic Republic during 1993 to 2000 . In this study, 50 V . cholerae O1 strains were selected during this period for studying the presence of class I integron and SXT constin . Twenty-four streptomycin-resistant strains out of 26 isolated before 1997 contained a class I integron harboring the aadA1 gene cassette . Twenty-four strains isolated after 1997 contained an SXT constin (a large conjugative element) . Twenty of the strains were resistant to chloramphenicol, tetracycline, streptomycin, and trimethoprim-sulfamethoxazole, while four strains were susceptible to the antibiotic tested . The resistance genes included in the SXT constins were floR, tetA, strAB, and sulII, which encode resistance to chloramphenicol, tetracycline, streptomycin, and sulfamethoxazole, respectively . The antibiotic resistance gene cluster was found to be deleted in the four susceptible strains . SXT(LAOS) did not contain dfrA1 or dfr18, which confer resistance to trimethoprim in SXT(ET) and SXT(MO10), respectively . A hot spot region of SXT(LAOS) was sequenced, and we identified two novel open reading frames showing homology to sO24 (exonuclease) and sO23 (helicase) of the genomic island associated with the multidrug resistance region of Salmonella enterica serovar Typhimurium DT104 . Analysis of SXT(LAOS) showed that there is a continuous flux of genes among V . cholerae SXT constins which should be carefully monitored. Antimicrob Agents Chemother, 2004 Jul, 48(7), 2355 - 63 Single-nucleotide polymorphism mutation spectra and resistance to quinolones in Salmonella enterica serovar Enteritidis with a mutator phenotype; Levy DD et al.; Resistance to quinolone antibiotics has been associated with single-nucleotide polymorphisms (SNPs) in the quinolone resistance-determining region (QRDR) of gyrA . Mutations in the gyrA gene were compared by using mutant populations derived from wild-type Salmonella enterica serovar Enteritidis and its isogenic mutS::Tn10 mutator counterpart . Spontaneous mutants arising during nonselective growth were isolated by selection with either nalidixic acid, enrofloxacin, or ciprofloxacin . QRDR SNPs were identified in approximately 70% (512 of 695) of the isolates via colony hybridization with radiolabeled oligonucleotide probes . Notably, transition base substitution SNPs in the QRDR were dramatically increased in mutants derived from the mutS strain . Some, but not all, antibiotic-resistant mutants lacking QRDR SNPs were resistant to tetracycline and chloramphenicol, consistent with alterations in nonspecific efflux pumps or other membrane transport mechanisms . Changing the selection conditions shifted the mutation spectrum . Selection with ciprofloxacin was least likely to yield a mutant harboring either a QRDR SNP or chloramphenicol resistance . Selection with enrofloxacin was more likely to yield mutants containing Ser83-->Phe mutations, whereas selection with ciprofloxacin or nalidixic acid favored recovery of Asp87-->Gly mutants . Fluoroquinolone-resistant Salmonella strains isolated from veterinary or clinical settings frequently display a mutational spectrum with a preponderance of transition SNPs in the QRDR, the pattern found in vitro among mutS mutator mutants reported here . Both the preponderance of transition mutations and the varied mutation spectra reported for veterinary and clinical isolates suggest that bacterial mutators defective in methyl-directed mismatch repair may play a role in the emergence of quinolone and fluoroquinolone resistance in feral settings. Clin Microbiol Infect, 2004 Jul, 10(7), 595 - 7 Rhabdomyolysis induced by Salmonella enterica serovar Typhi bacteraemia; Fisk DT et al.; Rhabdomyolysis has been reported infrequently with salmonella infection . Since 1964, there have been at least 22 reports associated with gastroenteritis or bacteraemia . Twenty cases have been associated with non-typhoidal strains of Salmonella, with single reports of Salmonella enterica serovars Paratyphi and Typhi . A second case of typhoid fever associated with rhabdomyolysis was recently diagnosed in Ann Arbor, USA in a traveller returning from an endemic area . Prompt diagnosis and treatment resulted in a good outcome . Salmonella infection should be considered by clinicians as a possibility in the differential diagnosis of rhabdomyolysis. Lett Appl Microbiol, 2004, 38(4), 265 - 70 Cloth-based hybridization array system for the detection of multiple antibiotic resistance genes in Salmonella enterica subsp . enterica serotype Typhimurium DT104; Gauthier M et al.; AIMS: A simple DNA macroarray system was developed for detection of antibiotic resistance and other marker genes associated with the multidrug-resistant food pathogen Salmonella enterica subsp . enterica serotype Typhimurium DT104 . METHODS AND RESULTS: A multiplex polymerase chain reaction (PCR) incorporating digoxigenin-dUTP was used to simultaneously amplify seven marker sequences, with subsequent rapid detection of the amplicons by hybridization with an array of probes immobilized on polyester cloth and immunoenzymatic assay of the bound label . This system provided sensitive detection of the different genetic markers in the S . Typhimurium DT104 genome, giving positive reactions with as few as 10 CFU, and the hybridizations were highly specific, with no reactions of amplicons with heterologous probes on the array . CONCLUSIONS: This cloth-based hybridization array system (CHAS) provides a simple, cost-effective tool for monitoring S . Typhimurium DT104 in foods and their production environment . SIGNIFICANCE AND IMPACT OF THE STUDY: The CHAS is a simple and cost-effective tool for the simultaneous detection of amplicons generated in a multiplex PCR, and the concept is broadly applicable to the detection and characterization of food pathogens. Infect Immun, 2004 Jul, 72(7), 4297 - 301 Oral immunization with an rfaH mutant elicits protection against salmonellosis in mice; Nagy G et al.; Loss of the transcriptional antiterminator RfaH results in virulence attenuation (>10(4)-fold increase in 50% lethal dose) of the archetypal Salmonella enterica serovar Typhimurium strain SL1344 by both orogastric and intraperitoneal routes of infection in BALB/c mice . Oral immunization with the mutant efficiently protects mice against a subsequent oral infection with the wild-type strain . Interestingly, in vitro immunoreactivity is not confined to strain SL1344; rather, it is directed also towards other serovars of S . enterica and even Salmonella bongori strains. Infect Immun, 2004 Jul, 72(7), 4138 - 50 Flagella and chemotaxis are required for efficient induction of Salmonella enterica serovar Typhimurium colitis in streptomycin-pretreated mice; Stecher B et al.; Salmonella enterica subspecies 1 serovar Typhimurium is a common cause of gastrointestinal infections . The host's innate immune system and a complex set of Salmonella virulence factors are thought to contribute to enteric disease . The serovar Typhimurium virulence factors have been studied extensively by using tissue culture assays, and bovine infection models have been used to verify the role of these factors in enterocolitis . Streptomycin-pretreated mice provide an alternative animal model to study enteric salmonellosis . In this model, the Salmonella pathogenicity island 1 type III secretion system has a key virulence function . Nothing is known about the role of other virulence factors . We investigated the role of flagella in murine serovar Typhimurium colitis . A nonflagellated serovar Typhimurium mutant (fliGHI) efficiently colonized the intestine but caused little colitis during the early phase of infection (10 and 24 h postinfection) . In competition assays with differentially labeled strains, the fliGHI mutant had a reduced capacity to get near the intestinal epithelium, as determined by fluorescence microscopy . A flagellated but nonchemotactic cheY mutant had the same virulence defects as the fliGHI mutant for causing colitis . In competitive infections, both mutants colonized the intestine of streptomycin-pretreated mice by day 1 postinfection but were outcompeted by the wild-type strain by day 3 postinfection . Together, these data demonstrate that flagella are required for efficient colonization and induction of colitis in streptomycin-pretreated mice . This effect is mostly attributable to chemotaxis . Recognition of flagellar subunits (i.e., flagellin) by innate immune receptors (i.e., Toll-like receptor 5) may be less important. Infect Immun, 2004 Jul, 72(7), 3987 - 4003 The two murein lipoproteins of Salmonella enterica serovar Typhimurium contribute to the virulence of the organism; Sha J et al.; Septic shock due to Salmonella and other gram-negative enteric pathogens is a leading cause of death worldwide . The role of lipopolysaccharide in sepsis is well studied; however, the contribution of other bacterial outer membrane components, such as Braun (murein) lipoprotein (Lpp), is not well defined . The genome of Salmonella enterica serovar Typhimurium harbors two copies of the lipoprotein (lpp) gene . We constructed a serovar Typhimurium strain with deletions in both copies of the lpp gene (lpp1 and lpp2) by marker exchange mutagenesis . The integrity of the cell membrane and the secretion of the effector proteins through the type III secretion system were not affected in the lpp double-knockout mutant . Subsequently, the virulence potential of this mutant was examined in a cell culture system using T84 intestinal epithelial and RAW264.7 macrophage cell lines and a mouse model of salmonellosis . The lpp double-knockout mutant was defective in invading and inducing cytotoxic effects in T84 and RAW264.7 cells, although binding of the mutant to the host cell was not affected when compared to the wild-type (WT) serovar Typhimurium . The motility of the mutant was impaired, despite the finding that the number of flagella was similar in the lpp double knockout mutant and the WT serovar Typhimurium . Deletion in the lpp genes did not affect the intracellular survival and replication of Salmonella in macrophages and T84 cells . Induction of the proinflammatory cytokines tumor necrosis factor alpha and interleukin-8 (IL-8) was significantly reduced in macrophages and T84 cells infected with the lpp double-knockout mutant . The levels of IL-8 remained unaffected in T84 cells when infected with either live or heat-killed WT and lpp mutant, indicating that invasion was not required for IL-8 production and that Toll-like receptor 2 signaling might be affected in the Lpp double-knockout mutant . These effects of the Lpp protein could be restored by complementation of the isogenic mutant . The lpp double-knockout mutant was avirulent in mice, and animals infected with this mutant were protected from a lethal challenge dose of WT serovar Typhimurium . The severe combined immunodeficient mice, on the other hand, were susceptible to infection by the lpp double-knockout mutant . The serovar Typhimurium mutants from which only one of the lpp (lpp1 or lpp2) genes was deleted were also avirulent in mice . Taken together, our data indicated that Lpp specifically contributed to the virulence of the organism. Infect Immun, 2004 Jul, 72(7), 3803 - 11 Oral tolerance in T cells is accompanied by induction of effector function in lymphoid organs after systemic immunization; Parameswaran N et al.; The physiological ramifications of oral tolerance remain poorly understood . We report here that mice fed ovalbumin (OVA) exhibit oral tolerance to subsequent systemic immunization with OVA in adjuvant, and yet they clear systemic infection with a recombinant OVA-expressing strain of Salmonella enterica serovar Typhimurium better than unfed mice do . Mice fed a sonicated extract of S . enterica serovar Typhimurium are also protected against systemic bacterial challenge, and the protection is Th1 mediated, as feeding enhances clearance in interleukin-4-null (IL-4(-/-)) and IL-10(-/-) mice but not in gamma interferon-null (IFN-gamma(-/-)) mice . When T-cell priming in vivo is tracked temporally in T-cell receptor-transgenic mice fed a single low dose of OVA, CD4 T-cell activation and expansion are restricted largely to mucosal lymphoid organs . However, T cells from spleens and peripheral lymph nodes of fed mice proliferate and secrete IFN-gamma when restimulated with OVA in vitro, indicating the presence of primed T cells in systemic tissues following oral exposure to antigen . Nonetheless, oral tolerance can be observed in the fed mice as reduced recall responses following subsequent systemic immunization with OVA in adjuvant . Soluble OVA administered systemically has similar effects in vivo, and the "tolerance" seen in both cases can be partially reversed if the initial priming is made more immunogenic . Together, the results indicate that antigen exposure under poor adjuvantic conditions, whether oral or systemic, may lead to T-cell commitment to effector rather than proliferative capabilities, necessitating a reassessment of therapeutic modalities for induction of oral tolerance in allergic or autoimmune states. FEMS Microbiol Lett, 2004 Jul 1, 236(1), 103 - 7 Inhibition of Salmonella enterica serovars by microcin J25; Vincent PA et al.; Escherichia coli microcin J25 (MccJ25) is a 2107-Da peptide antibiotic whose uptake into E . coli is mediated by the outer-membrane receptor FhuA and the inner membrane proteins TonB, ExbB, ExbD, and SbmA . A survey of the sensitivity of several Salmonella enterica serovars showed that the antibiotic was highly active against some serovars, while S . Typhimurium, S . Derby, and some S . Enteritidis strains were completely resistant . Resistant strains became hypersensitive to MccJ25 when given the fhuA gene of E . coli, indicating that insensitivity is due to the inability of the FhuA protein to mediate penetration of MccJ25 . Whereas in E . coli MccJ25 targets RNA polymerase, in S . Typhimurium it inhibits not only RNA synthesis but also cell respiration . Fluorescence viability staining showed that S . Typhimurium cells exposed to MccJ25 remain viable but are unable to form colonies. FEMS Microbiol Lett, 2004 Jul 1, 236(1), 65 - 72 The steady-state orgA specific mRNA levels in Salmonella enterica serovar Typhimurium are repressed by oxygen during logarithmic growth phase but not early-stationary phase; Russell DA et al.; The orgA gene from Salmonella enterica serovar Typhimurium is involved in promoting cellular invasion of the pathogen . Its exact role in virulence is still unclear mainly due to difficulties in understanding its complex regulation . In this study a novel competitive RT-PCR (cRT-PCR) system was developed to measure the steady-state orgA specific mRNA levels in cells under various growth parameters . Previous studies have been inconsistent regarding oxygen regulation of orgA . Using our system we found that oxygen repressed the copy levels 3.5-fold in cells grown only to logarithmic phase . Oxygen repression was not observed in cells grown to early-stationary phase, a parameter that has previously been demonstrated to be the most invasive stage of growth . The importance of NaCl in orgA gene regulation is also illustrated . Significant increases in copy numbers were observed after growth in high NaCl conditions . Measuring the steady-state mRNA levels using cRT-PCR provides an accurate insight into prokaryotic gene regulation prior to translation. Tohoku J Exp Med, 2004 Jun, 203(2), 129 - 32 Cerebrospinal fluid cytokines in Salmonella urbana encephalopathy; Minami K et al.; We present a case report of encephalopathy associated with Salmonella urbana infection in a child . A 5-year-old boy was admitted to our clinic with convulsions and coma . Cerebrospinal fluid (CSF) interleukin-6 (IL-6) and IL-8 were elevated at onset and were decreased within normal limit on the fifth day . Residual neurological deficits included severe mental deficits and spastic tetraplegia . High levels of CSF proinflammatory cytokines might be related to central nervous system (CNS) disease activity . Although encephalopathy is a rare complication of non-typhi Salmonella infection, it should be borne in mind as an occasionally serious and potentially lethal disease. Wei Sheng Yan Jiu, 2004 May, 33(3), 284 - 7 {Toxicological effects of copper on human intestinal Caco-2 cells}; Liu Z et al.; OBJECTIVE: To study the toxicological effects of copper on human intestinal Caco-2 cells . METHODS: The effects of copper on human intestinal Caco-2 cells were evaluated by MTT conversion, intra-cellular production of reactive oxygen species (ROS), cloning efficiency and changes of P-glycoprotein (P-gp) activities in Caco-2 cells . In addition, Caco-2 cells and Salmonella enteritidis served as the models to examine the effect of copper on the host-parasite interaction . RESULTS: Copper induced a dose-dependent decrease of cell viability measured by MTT assay; by using the fluorescent probe 2, 7-dichlorofluorescin diacetate, a significant increase of ROS production in Cu-treated cells was detected; the significant time- and dose-dependent decrease of P-gp activities in Cu-treated cells was demonstrated by increased accumulation of rhodamine 123 in cells . Copper treatment also increased the efficiency of the bacterial invasion, but decreased the number of bacteria surviving in the intracellular environment . CONCLUSION: Copper induced a variety of toxicological endpoints which might be caused by oxidative damage, but the effect of copper on cell-bacteria interaction needs to be further investigated. J Appl Toxicol, 2004 May-Jun, 24(3), 177 - 81 Determination of the rat tissue partitioning of endotoxin in vitro for physiologically-based pharmacokinetic (PBPK) modeling; Ross IA et al.; The biosynthetically double-labeled lipopolysaccharide (LPS), containing (3)H-labeled on the fatty acyl-chains and (14)C-labeled on the glucosamine of Salmonella enterica serotype typhimurium, was isolated from bacteria grown in proteose peptone-beef extract (PPBE) medium in the presence of labeled precursors; 133 micro Ci/ml of {2-(3)H} acetate sodium salt and 0.167 micro Ci/ml of N-acetyl{D-1-(14)C}glucosamine . The LPS was extracted from the bacteria with 90% phenol/chloroform/petroleum ether, purified and stored in 0.1% (v/v) triethylamine/10 mM Tris HCl at -70 degrees C . Tissue slices and portions of the meninges were prepared and incubated in artificial cerebrospinal fluid (CSF) or Krebs phosphate buffer (Krebs) containing 150 ng/ml LPS with {(3)H} LPS (0.004 micro Ci/ml, sp . act . 28 micro Ci/mg LPS) . The tissues were incubated under 95% oxygen/5% carbon dioxide at 37 degrees C with constant agitation until steady-state uptake was reached (60 min) . At the end of the incubation period, tissues were processed for radioactivity measurement . The rat tissue partitioning of LPS in artificial CSF for brain and Krebs for other organs was measured by using the ratio of tissue to medium at the steady state in vitro . The following results were obtained from the study: Heart, 0.15; liver, 0.19; spleen, 0.12; kidney, 0.18; stomach, 0.17; small intestine, 0.18; brain stem, 0.10; cerebellum, 0.11; meninges, 0.77; hippocampus, 0.12; hypothalamus, 0.12; frontal cortex, 0.09 and caudate nucleus, 0.10 . This information, along with plasma or blood/buffer partition coefficients, is a requisite for constructing a physiologically-based pharmacokinetic (PBPK) model of endotoxins for quantitative risk assessment . Chemotherapy, 2004 Jun, 50(2), 88 - 91 Antimicrobial susceptibility of 128 Salmonella enterica serovar typhi and paratyphi A isolates from northern India; Safdar A et al.; Most systemic Salmonella enterica serovar typhi and paratyphi A infections diagnosed in the United States (up to 70%) are acquired during travel to regions of high endemicity . Increasing resistance to agents commonly used for the treatment of such infections (including multidrug resistant isolates) is being reported from several areas of the world (Southeast Asia, Africa, Latin America) . Since regional differences in susceptibility patterns may exist, we sought to determine the frequency of antimicrobial resistance among blood and stool isolates (n = 128) from patients in Northwestern India . Salmonella enterica serotype typhi (n = 101) isolates from 14 patients were susceptible to all agents tested . Among 55 isolates with single drug resistance, 44 (81%) were resistant to chloramphenicol . Multidrug resistant (>/=3 drugs) Salmonella enterica was more common in pediatric patients (10 of 30) compared to adults (10 of 71 patients; p = 0.05) . All isolates (S . enterica serovar typhi and serovar paratyphi A) were susceptible to ciprofloxacin and ceftriaxone . Travelers to Northwestern India may still receive trimethoprim-sulfamethoxazole, or ciprofloxacin for effective chemoprophylaxis if indicated . Ceftriaxone and ciprofloxacin remain favorable choices for treatment of patients with enteric fever in this region . Am J Trop Med Hyg, 2004 Jun, 70(6), 670 - 5 The etiology of febrile illness in adults presenting to Patan hospital in Kathmandu, Nepal; Murdoch DR et al.; In Nepal, many infections remain poorly characterized, partly due to limited diagnostic facilities . We studied consecutive febrile adults presenting to a general hospital in Kathmandu, Nepal . Of the 876 patients enrolled, enteric fever and pneumonia were the most common clinical diagnoses . Putative pathogens were identified in 323 (37%) patients, the most common being Salmonella enterica serotype Typhi and S . enterica serotype Paratyphi A (117), Rickettsia typhi (97), Streptococcus pneumoniae (53), Leptospira spp . (36), and Orientia tsutsugamushi (28) . Approximately half of the Salmonella isolates were resistant to nalidixic acid . No clinical predictors were identified to reliably distinguish between the different infections . These findings confirm the heavy burden of enteric fever and pneumonia in Kathmandu, and highlight the importance of murine typhus, scrub typhus, and leptospirosis . Given the lack of reliable clinical predictors, the development of cheap and accurate diagnostic tests are likely to be of great clinical utility in this setting. Vet Res, 2004 May-Jun, 35(3), 291 - 8 Humoral immune response in hens naturally infected with Salmonella Enteritidis against outer membrane proteins and other surface structural antigens; Ochoa-Reparaz J et al.; A simple procedure for obtaining surface exposed antigens of Salmonella Enteritidis is described . A heat treatment of whole bacteria in saline solution induced the release of small membrane vesicles containing outer membrane components as well as surface appendage components, such as fimbriae and flagellin . The characterization of the structural components of this extract, called HE, was established by SDS-PAGE and immunoblotting using polyclonal and monoclonal specific antibodies . Five major groups of proteins were identified: flagellin, porins, OmpA, SEF21 and SEF14 fimbriae . The immunogenicity of these proteins was studied by immunoblotting with serum samples from naturally infected hens . Flagellin, porins, OmpA, SEF14 and SEF21 fimbriae were immunogenic in the S . Enteritidis infected hens (frequency of reactants: 47.3, 97.3, 64.7, 50.0 and 60.8%, respectively); porins also reacted with sera from non infected hens (66.7%) . The immunogenicity of these antigens in infected birds provide promise that they may serve as components of an effective subcellular vaccine for poultry salmonellosis . J Biol Chem, 2004 Sep 10, 279(37), 38618 - 25 Epub 2004 Jun 18. Transcriptional control of the antimicrobial peptide resistance ugtL gene by the Salmonella PhoP and SlyA regulatory proteins; Shi Y et al.; The PhoP/PhoQ two-component system is a master regulator that governs the ability of Salmonella to cause a lethal infection in mice, the adaptation to low Mg(2+) environments, and resistance to a variety of antimicrobial peptides . We have recently established that the PhoP-activated ugtL gene is required for resistance to the antimicrobial peptides magainin 2 and polymyxin B . Here we report that ugtL transcription requires not only the PhoP protein but also the virulence regulatory protein SlyA . The PhoP protein footprinted two regions of the ugtL promoter, mutation of either one of which was sufficient to abolish ugtL transcription . Although the SlyA protein is a transcriptional activator of the ugtL gene, it footprinted the ugtL promoter at a region located downstream of the transcription start site . The PhoP protein footprinted the slyA promoter, indicating that it controls slyA transcription directly . The slyA mutant was hypersensitive to magainin 2 and polymyxin B, suggesting that the virulence attenuation exhibited by slyA mutants may be caused by hypersensitivity to antimicrobial peptides . We propose that the PhoP and SlyA proteins control ugtL transcription using a feed-forward loop design. Vet Immunol Immunopathol, 2004 Aug, 100(3-4), 151 - 64 Age at primary infection with Salmonella enterica serovar Typhimurium in the chicken influences persistence of infection and subsequent immunity to re-challenge; Beal RK et al.; Salmonella enterica remains one of the most important food-borne pathogens of humans and is often acquired through consumption of infected poultry meat or eggs . Control of Salmonella infections in chicken is therefore an important public health issue . Infection with S . enterica serovar Typhimurium results in a persistent enteric infection without clinical disease in chickens of more than 3 days of age, and represents a source for contamination of carcass at slaughter and entry into the human food chain . Data presented indicate a profound effect of age at initial exposure on the persistence of infection and a lesser effect on the development of effective immunity to re-challenge . The percentage of birds positive for Salmonella was high until 8-9 weeks of age, regardless of the age at which the birds were infected (1, 3 or 6 weeks) . The birds infected at 3 and 6 weeks of age produced a more rapid and higher antibody response (IgY and IgA) than those infected at 1 week of age, but in all cases infection persisted for a considerable period despite the presence of high antibody levels . Following a re-challenge infection with S . Typhimurium, all three previously-infected groups had fewer bacteria in the gut, spleen and liver compared with age-matched birds receiving a parallel primary infection . However, the birds primary infected at 3 and 6 weeks of age cleared infection more rapidly than those infected at a younger age . Interestingly older-primed birds had higher specific T lymphocyte proliferative responses and specific circulating levels of IgY antibody at time of re-challenge . Although birds initially infected at 1 week of age and those that were previously uninfected produced a stronger antibody response following re-challenge, they were slower to clear Salmonella from the gut than the older-primed groups which expressed a stronger T lymphocyte response . The data presented indicate that clearance of Salmonella from the gut is age-dependent and we propose that this relates to the increased competence of the enteric T cell response . The findings that Salmonella persists beyond 8-9 weeks, irrespective of age at exposure, has implications for the broiler sector and indicates the need to remain Salmonella free throughout the rearing period . Moreover, the re-challenge data demonstrates that infection at a young age is less effective in producing protective immunity than in older chickens . This feature of the development of protective immunity needs to be considered when developing vaccines for the broiler sector of the poultry industry. Cytokine, 2004 Jul 7, 27(1), 15 - 24 Interleukin-8 production by THP-1 cells stimulated by Salmonella enterica serovar Typhimurium porins is mediated by AP-1, NF-kappaB and MAPK pathways; Vitiello M et al.; Interleukin-8 (IL-8) is released in response to inflammatory stimuli, such as bacterial products . Either porins or lipopolysaccharide (LPS) stimulated THP-1 cells to release IL-8 after 24 h . We have previously reported that stimulation of monocytic cells with Salmonella enterica serovar Typhimurium porins led to the activation of mitogen-activated protein kinase cascades and of protein tyrosine kinases (PTKs) . In this report, we demonstrate, using two potent and selective inhibitors of MEK activation by Raf-1 (PD-098059) and p38 (SB-203580), that both ERK1/2 and p38 pathways play a key role in the production of IL-8 by porins and LPS . Porin-stimulated expression of activating protein 1 (AP-1) and correlated IL-8 release is also inhibited by PD-098059 or SB-203580 indicating that the Raf-1/MEK1-MEK2/MAPK cascade is required for their activation . Also PTKs modulate the pathway that control IL-8 gene expression, in fact its expression is abolished by tyrphostin . By using N-acetyl-leucinyl-leucinyl-norleucinal-H (ALLN) an inhibitor of nuclear factor-kappaB (NF-kappaB) activity, we also observed IL-8 release modulation . Our results elucidate some of the molecular mechanisms by which AP-1 and NF-kappaB regulate IL-8 release and open new strategies for the design of specific molecules that will modulate IL-8 effects in various infectious diseases. Emerg Infect Dis, 2004 Jun, 10(6), 985 - 7 Salmonella-based rodenticides and public health; Painter JA et al.; Several countries still permit strains of Salmonella enterica serotype Enteritidis, a leading cause of gastrointestinal illness in humans, to be used in rat baits . To assess the human health risk associated with such rat bait, we first reviewed historic data on health hazards associated with Ratin, a rodenticide that was used in Europe until the early 1960s . Ratin caused outbreaks of human illness, including several deaths . We then compared S . Enteritidis isolated from a current commercial product, Biorat, with S . Enteritidis from Ratin and found that the strains were both phage type 6a . Based on the similarity of the strains, currently available Salmonella-based rodenticides likely are as great a threat to public health as past strains were . Health officials should be aware that the continued use of Salmonella-based rodenticides is a risk to public health and should take appropriate measures to prevent use in their jurisdictions. Poult Sci, 2004 Jun, 83(6), 997 - 1002 Ovarian cell-mediated immune response to Salmonella enteritidis infection in laying hens (Gallus domesticus); Barua A et al.; The aim of this study was to examine the response of cell-mediated ovarian immunity against Salmonella infection in hens . Laying hens were injected intraperitoneally with PBS (control) or Salmonella enteritidis (SE) . Ovarian stroma containing stromal follicles, small white follicles (SWF), and third largest (F3) and the largest (F1) follicles were collected 12 or 24 h after inoculation and fixed in periodate-lysine-paraformaldehyde . Frozen sections were stained first for CD3+, CD4+, or CD8+ T cells and then for SE by a double immunostaining method . Immunoreaction products for SE were detected in the ovarian stroma, theca of stromal follicles, SWF, F3, and F1 at 12 and 24 h after inoculation . Immunopositive T-cell subsets were localized in the stroma and theca of follicles in birds inoculated with or without SE . The populations of CD3+, CD4+, and CD8+ T cells were significantly greater in the stroma and the theca of follicles 12 h after SE inoculation than in those of control birds (P < 0.01) . Their frequencies were further increased in those tissues 24 h after inoculation (P < 0.01) . Injection of SE did not cause significant differences in the CD4+:CD8+ T-cell ratio as both subsets increased proportionately . The current results indicate that the population of T-cell subsets increases in the ovarian stroma and the follicular tissues in response to SE invasion within 12 h of inoculation . Thus, cell-mediated immune response against SE, their products, or both may be induced in the hen ovary. Clin Infect Dis, 2004 Jul 1, 39(1), 61 - 7 Epub 2004 Jun 14. A clinical, microbiological, and pathological study of intestinal perforation associated with typhoid fever; Nguyen QC et al.; One of the most serious complications of typhoid fever is intestinal perforation . Of 27 patients admitted to a provincial hospital in the Mekong Delta region of Vietnam who had gastrointestinal perforation secondary to suspected typhoid fever, 67% were male, with a median age of 23 years and a median duration of illness of 10 days . Salmonella enterica subspecies enterica serotype Typhi (S . Typhi) was isolated from 11 (41%) of 27 patients; of 27 patients, only 4 (15%) had positive cultures from gut biopsies . S . Typhi DNA was detected by polymerase chain reaction for all perforation biopsy samples . Detailed histological examination of the gastrointestinal mucosa at the site of perforation in all cases showed a combination of discrete acute and chronic inflammation . Acute inflammation at the serosal surface indicated additional tissue damage after perforation . Immunohistochemical results showed that the predominant infiltrating cell types at the site of perforation were CD68+ leukocytes (macrophages) or CD3+ leukocytes (T lymphocytes). Science, 2004 Jun 18, 304(5678), 1805 - 7 Salmonella modulates vesicular traffic by altering phosphoinositide metabolism; Hernandez LD et al.; Salmonella enterica, the cause of food poisoning and typhoid fever, induces actin cytoskeleton rearrangements and membrane ruffling to gain access into nonphagocytic cells, where it can replicate and avoid innate immune defenses . Here, we found that SopB, a phosphoinositide phosphatase that is delivered into host cells by a type III secretion system, was essential for the establishment of Salmonella's intracellular replicative niche . SopB mediated the formation of spacious phagosomes following bacterial entry and was responsible for maintaining high levels of phosphatidylinositol-three-phosphate {PtdIns(3)P} in the membrane of the bacteria-containing vacuoles . Absence of SopB caused a significant defect in the maturation of the Salmonella-containing vacuole and impaired bacterial intracellular growth. J Bacteriol, 2004 Jul, 186(13), 4124 - 33 The PmrA-regulated pmrC gene mediates phosphoethanolamine modification of lipid A and polymyxin resistance in Salmonella enterica; Lee H et al.; The PmrA/PmrB regulatory system of Salmonella enterica controls the modification of lipid A with aminoarabinose and phosphoethanolamine . The aminoarabinose modification is required for resistance to the antibiotic polymyxin B, as mutations of the PmrA-activated pbg operon or ugd gene result in strains that lack aminoarabinose in their lipid A molecules and are more susceptible to polymyxin B . Additional PmrA-regulated genes appear to participate in polymyxin B resistance, as pbgP and ugd mutants are not as sensitive to polymyxin B as a pmrA mutant . Moreover, the role that the phosphoethanolamine modification of lipid A plays in the resistance to polymyxin B has remained unknown . Here we address both of these questions by establishing that the PmrA-activated pmrC gene encodes an inner membrane protein that is required for the incorporation of phosphoethanolamine into lipid A and for polymyxin B resistance . The PmrC protein consists of an N-terminal region with five transmembrane domains followed by a large periplasmic region harboring the putative enzymatic domain . A pbgP pmrC double mutant resembled a pmrA mutant both in its lipid A profile and in its susceptibility to polymyxin B, indicating that the PmrA-dependent modification of lipid A with aminoarabinose and phosphoethanolamine is responsible for PmrA-regulated polymyxin B resistance. J Bacteriol, 2004 Jul, 186(13), 4056 - 66 Regulation of the Ysa type III secretion system of Yersinia enterocolitica by YsaE/SycB and YsrS/YsrR; Walker KA et al.; Yersinia enterocolitica biovar 1B contains two type III secretion systems (TTSSs), the plasmid-encoded Ysc-Yop system and the chromosomally encoded Ysa-Ysp system . Proteins secreted from the Ysa TTSS (Ysps) have only been detected in vitro when cells are cultured at 26 degrees C in a high-NaCl medium . However, the exact role of the Ysa TTSS is unclear . Thus, investigations into the regulation of this system may help elucidate the role of the Ysps during the life cycle of Y . enterocolitica . Here we present evidence that the AraC-like regulator YsaE acts together with the chaperone SycB to regulate transcription of the sycByspBCDA operon, a phenomenon similar to that seen in the closely related Salmonella SPI-1 and Shigella flexneri Mxi-Spa-Ipa TTSSs . Deletion of either sycB or ysaE results in a twofold reduction in the activity of a sycB-lacZ fusion compared to the wild type . In a reconstituted Escherichia coli system, transcription of sycB was activated sixfold only when both YsaE and SycB were present, demonstrating that they are necessary for activation . ysrR and ysrS are located near the ysa genes and encode a putative two-component regulatory system . Mutations in either gene indicated that both YsrR and YsrS were required for secretion of Ysps . In addition, transcription from sycB-lacZ and ysaE-lacZ fusions was decreased 6.5- and 25-fold, respectively, in the ysrS mutant compared to the wild type . Furthermore, in the absence of NaCl, the activity of ysaE-lacZ was reduced 25-fold in the wild-type and DeltaysrS strains, indicating that YsrS is probably required for the salt-dependent expression of the ysa locus . These results suggest that the putative two-component system YsrRS may be a key element in the regulatory cascade for the Ysa TTSS. J Drug Target, 2003, 11(8-10), 481 - 8 Attenuated salmonella and Shigella as carriers for DNA vaccines; Xu F et al.; The discovery that genes can be functionally transferred from bacteria to mammalian cells has suggested the possible use of bacterial vectors as gene delivery vehicles for vaccines . Attenuated invasive human intestinal bacteria, such as Salmonella and Shigella, have been used as plasmid DNA vaccine carriers and their potency has been evaluated in several animal models . This delivery system allows the administration of DNA vaccines together with associated bacterial immunostimulators directly to professional antigen presenting cells via human mucosal surfaces . Various strategies have been taken to improve the use of this delivery system to achieve robust immune responses at both mucosal and systemic sites of the immunized animals. J Drug Target, 2003, 11(8-10), 471 - 9 The use of live attenuated bacteria as a delivery system for heterologous antigens; Garmory HS et al.; Live attenuated mutants of several pathogenic bacteria have been exploited as potential vaccine vectors for heterologous antigen delivery by the mucosal route . Such live vectors offer the advantage of potential delivery in a single oral, intranasal or inhalational dose, stimulating both systemic and mucosal immune responses . Over the years, a range of strategies have been developed to allow controlled and stable delivery of antigens and improved immunogenicity where required . Most of these approaches have been evaluated in Salmonella vaccine vectors and, as a result, several live attenuated recombinant Salmonella vaccines are now in human clinical trials . In this review, these strategies and their use in the development of a delivery system for the Yersinia pestis V antigen are described. Rev Esp Quimioter, 2004 Mar, 17(1), 37 - 43 {Change in resistance to quinolones and betalactams in different serogroups of Salmonella during the last decade in a Madrid hospital}; Guerri Santos ML et al.; The prevalence of antibiotic resistance was studied in 3230 strains of Salmonella enterica isolated in the Hospital Universitario La Paz in Madrid, Spain, from 1991 to 2001 . Betalactam antibiotic resistance has been notorious in serogroup B4; the highest prevalence of ampicillin resistance (84%) was reached in 2000 and that of amoxicillin-clavulanic acid (45%) in 1996 . Resistance to cephalosporins has been controlled, although in 2000 cefazolin resistance reached 37% in serogroup C2-8 . An increase in first generation quinolone resistance was detected in every serogroup, especially in D9 and C2-8, which showed an increase from 6% and 15% in 1991, respectively, to 40% and 85% in 2001 . Although important resistance to ciprofloxacin has not yet been detected, the activity of fluoroquinolones against Salmonella must be closely monitored. Rev Esp Quimioter, 2004 Mar, 17(1), 29 - 36 {Antimicrobial drug resistance in non-typhi Salmonellae in Castilla y Leon}; Delgado Ronda M et al.; We studied the antibiotic susceptibility of 309 Salmonella isolates obtained from three hospitals serving the provinces of Salamanca, Avila and Zamora in the region of Castilla y Leon (mid-west Spain) . The susceptibility to 18 antibiotics was studied using the agar dilution method, according to NCCLS guidelines, and the most common multiresistance phenotypes were determined for each province . We observed clear susceptibility differences between the two main serotypes found, S . enteritidis and S . typhimurium . Seventy percent of S . typhimurium were resistant to amoxicillin . In 44% of these isolates, amoxicillin resistance was associated with resistance to streptomycin, sulfonamides, tetracyclines and chloramphenicol . S . enteritidis was susceptible to most antibiotics tested; amoxicillin resistance was observed in 23.3%, and nalidixic acid resistance in 49.6% . Resistance to nalidixic acid was higher in S . enteritidis than in any other serotypes . According to NCCLS breakpoints, no strain was resistant to fluoroquinolones . However, according to MENSURA criteria, 9% of S . typhimurium isolates were resistant to ciprofloxacin . Resistance to cotrimazole and gentamicin was less than 10% for all the serotypes tested . The results indicate that S . typhimurium showed greater resistance and a high multidrug resistance rate . Conversely, S . enteritidis showed high resistance only to amoxicillin and nalidixic acid, though in most cases there was no correlation between this resistance and reduced susceptibility to fluoroquinolones. Eur J Cardiothorac Surg, 2004 Jul, 26(1), 225 - 7 Endovascular stent-graft for thoracic aorta aneurysm caused by Salmonella; Nikos K et al.; We describe the placement of an endovascular stent-graft in a patient with mycotic aneurysm of the descending thoracic aorta caused by Salmonella . Endovascular grafting combined with antibiotic therapy in thoracic mycotic aneurysms might represent an alternative to conventional surgery in patients with high operative risk. Eur J Cardiothorac Surg, 2004 Jul, 26(1), 221 - 4 Endovascular repair for multiple Salmonella mycotic aneurysms of the thoracic aorta presenting with Cardiovocal syndrome; Ting AC et al.; Salmonella mycotic thoracic aortic aneurysm is a rare but life-threatening condition . We report a 59-year-old man with two Salmonella mycotic thoracic aortic aneurysms, presented with fever and chills associated with hoarseness due to left vocal cord palsy (Cardiovocal syndrome) . Successful endovascular repair was performed using two Talent thoracic stent-graft devices deployed separately to cover the two mycotic aneurysms . Subsequent computed tomography at 12 months after the operation confirmed exclusion of the two pseudoaneurysms with no endoleak . With potent antibiotics and careful surveillance program, endovascular repair is a possible alternative to conventional open surgery in the management of mycotic thoracic aortic aneurysms, especially in high-risk patients. Emerg Infect Dis, 2004 May, 10(5), 795 - 801 Multidrug-resistant strains of Salmonella enterica Typhimurium, United States, 1997-1998; Rabatsky-Ehr T et al.; To evaluate multidrug-resistant strains of Salmonella enterica serotype Typhimurium, including definitive type 104 (DT104) in the United States, we reviewed data from the National Antimicrobial Resistance Monitoring System (NARMS) . In 1997 to 1998, 703 (25%) of 2,767 serotyped Salmonella isolates received at NARMS were S . Typhimurium; antimicrobial susceptibility testing and phage typing were completed for 697 . Fifty-eight percent (402) were resistant to > or = 1 antimicrobial agent . Three multidrug-resistant (> or = 5 drugs) strains accounted for (74%) 296 of all resistant isolates . Ceftriaxone resistance was present in 8 (3%), and nalidixic acid resistance in 4 (1%), of these multidrug-resistant strains . By phage typing, 259 (37%) of S . Typhimurium isolates were DT104, 209 (30%) were of undefined type and 103 (15%) were untypable . Fifty percent (202) of resistant (> or = 1 drug) isolates were DT104 . Multidrug-resistant S . Typhimurium isolates, particularly DT104, account for a substantial proportion of S . Typhimurium isolates; ceftriaxone resistance is exhibited by some of these strains. Nucleic Acids Res . 2004 Jun 15;32(10):e82. The recognition and modification sites for the bacterial type I restriction systems KpnAI, StySEAI, StySENI and StySGI; Kasarjian JK et al.; Using an in vivo plasmid transformation method, we have determined the DNA sequences recognized by the KpnAI, StySEAI, StySENI and StySGI R-M systems from Klebsiella oxytoca strain M5a1, Salmonella eastbourne, Salmonella enteritidis and Salmonella gelsenkirchen, respectively . These type I restriction-modification systems were originally identified using traditional phage assay, and described here is the plasmid transformation test and computer program used to determine their DNA recognition sequences . For this test, we constructed two sets of plasmids, pL and pE, that contain phage lambda and Escherichia coli K-12 chromosomal DNA fragments, respectively . Further, using the methylation sensitivities of various known type II restriction enzymes, we identified the target adenines for methylation (listed in bold italics below as A or T in case of the complementary strand) . The recognition sequence and methylation sites are GAA(6N)TGCC (KpnAI), ACA(6N)TYCA (StySEAI), CGA(6N)TACC (StySENI) and TAAC(7N)RTCG (StySGI) . These DNA recognition sequences all have a typical type I bipartite pattern and represent three novel specificities and one isoschizomer (StySENI) . For confirmation, oligonucleotides containing each of the predicted sequences were synthesized, cloned into plasmid pMECA and transformed into each strain, resulting in a large reduction in efficiency of transformation (EOT). J Biotechnol, 2004 Jul 1, 111(1), 1 - 8 Surface plasmon resonance immunosensor using self-assembled protein G for the detection of Salmonella paratyphi; Oh BK et al.; A surface plasmon resonance (SPR) based immunosensor using self-assembled protein G was developed for the detection of Salmonella paratyphi . In order to endow a solid substrate binding affinity to protein G, the free amine (-NH2) of protein G was substituted into thiol (-SH) using 2-iminothiolane . Thus, self-assembled protein G was fabricated on gold (Au) substrate . The formation of protein G layer on Au surface, and the binding of antibody and antigen in series were confirmed by SPR spectroscopy . The surface morphology analysis of the protein G layer on Au surface was performed by atomic force microscope (AFM) . Consequently, an immunosensor based on SPR for the detection of S . paratyphi using self-assembled protein G was developed with a detection range of 10(2)-10(7) CFU/ml . The current fabrication technique of a SPR immunosensor for the detection of S . paratyphi could be applied to construct other immnosensors or protein chips. Microb Pathog, 2004 Jul, 37(1), 11 - 9 Lipid raft microdomains mediate class A scavenger receptor-dependent infection of Brucella abortus; Kim S et al.; Brucella abortus is a facultative intracellular bacterium that can survive inside macrophages . Intracellular replication of B . abortus requires the VirB complex, which is highly similar to the conjugative DNA transfer system . In this study, we showed that a class A scavenger receptor (SR-A) of macrophages is required to internalize B . abortus and contributes to the establishment of bacterial infection in mice . Macrophages from SR-A-deficient mice inhibited internalization and intracellular replication of both wild type strain and the virB4 mutant, and that bacterial proliferation was inhibited in SR-A-deficient mice . Adding lipopolysaccharide from B . abortus and Salmonella enterica serovar Typhimurium, but not from Escherichia coli, to macrophages inhibited bacterial internalization . VirB-dependent bacterial internalization induced localization of SR-A into detergent-resistant membrane lipid rafts . These results indicate that B . abortus internalizes into macrophages by using SR-A as a receptor and that the VirB type IV secretion system of B . abortus regulates signal transduction dependent on SR-A to form replicative phagosomes, and which is mediated by lipid rafts. Mol Ther, 2004 Jun, 9(6), 895 - 901 Retroviral-mediated gene transfer restores IL-12 and IL-23 signaling pathways in T cells from IL-12 receptor beta1-deficient patients; Bosticardo M et al.; Genetic deficiency of human IL-12 receptor beta1 chain (IL-12Rbeta1) results in increased vulnerability to weakly pathogenic strains of Mycobacteria and Salmonella . This phenotype results from the combined lack of IL-12 and IL-23 signaling as both cytokine receptors share IL-12Rbeta1 . Such infections can be treated by administration of antibiotics and IFN-gamma; however, patients can succumb to infections despite these treatments . Reversion of patients' susceptibility by corrective gene transfer could prevent the infectious episodes, thus providing a beneficial alternative . We therefore evaluated the feasibility of retroviral-mediated gene correction of T cells obtained from patients carrying "null" mutations of IL-12Rbeta1 . Transduction of the IL-12Rbeta1 cDNA restored the expression of IL-12Rbeta1 and resulted in the reconstitution of a functional IL-12 signaling pathway, as demonstrated by STAT4 phosphorylation and IFN-gamma production . IFN-gamma production in response to IL-23 was also corrected after gene transfer . These results indicate that the biological defects of T cells from patients carrying IL-12Rbeta1 deficiency can be corrected by gene transfer and form the basis for further development of gene therapy for this disease. Vaccine, 2004 Jun 30, 22(20), 2524 - 32 A Salmonella enterica serovar Typhi vaccine expressing Yersinia pestis F1 antigen on its surface provides protection against plague in mice; Morton M et al.; A recombinant strain of attenuated Salmonella enterica serovar Typhi surface-expressing Yersinia pestis F1 antigen was generated by transforming strain BRD1116 (aroA aroC htrA) with plasmid pAH34L encoding the Y . pestis caf operon . BRD1116/pAH34L was stable in vitro and in vivo . An immunisation regimen of two intranasal doses of {Formula: see text} cfu of BRD1116/pAH34L given intranasally to mice 7 days apart induced the strongest immune response compared to other regimens and protected 13 out of 20 mice from lethal challenge with Y . pestis . Intranasal immunisation of mice constitutes a model for oral immunisation with Salmonella vaccines in humans . Thus, the results demonstrate that attenuated strains of S . enterica serovar Typhi which express Y . pestis F1 antigen may be developed to provide an oral vaccine against plague suitable for use in humans. J Zoo Wildl Med, 2004 Mar, 35(1), 60 - 4 Survey for Haemoproteus spp., Trichomonas gallinae, Chlamydophila psittaci, and Salmonella spp . in Galapagos Islands columbiformes; Padilla LR et al.; Endemic free-ranging Galapagos doves (Zenaida galapagoensis) and introduced rock doves (Columba livia) were surveyed in several islands of the Galapagos archipelago to establish sample prevalence of hemoparasites, Trichomonas gallinae, Chlamydophila psittaci, and Salmonella species . A Haemoproteus sp., the only hemoparasite identified, was found in 89% of the Galapagos doves sampled but not in the rock doves . Trichomonas gallinae was detected by polymerase chain reaction in 44% of rock doves from San Cristobal but in none of the Galapagos doves . Chlamydophila psittaci was detected from cloacal swabs in 6% of the Galapagos doves but in none of the rock doves sampled . All positive cases of C . psittaci occurred on Espanola, where the crude sample prevalence was 24% . A polymerase chain reaction-based Salmonella test failed to show evidence of this organism from any birds sampled. MMWR Morb Mortal Wkly Rep, 2004 Jun 11, 53(22), 484 - 7 Outbreak of Salmonella serotype Enteritidis infections associated with raw almonds--United States and Canada, 2003-2004; Centers for Disease Control and Prevention (CDC); On May 12, 2004, the Oregon State Public Health Laboratory identified a cluster of five patients infected with Salmonella enterica serotype Enteritidis (SE) isolates that were matched by using two-enzyme pulsed-field gel electrophoresis (PFGE) . The five patients were from four Oregon counties; their onsets of illness occurred during February-April 2004 . A subsequent investigation, still ongoing, has identified a total of 29 patients in 12 states and Canada with matching SE isolates, since at least September 2003 . Seven patients have been hospitalized; no one has died . Raw almonds distributed throughout the United States and internationally have been implicated as the source of the SE infections . As of May 21, approximately 13 million pounds of raw almonds had been recalled by the producer. Epidemiol Infect, 2004 Jun, 132(3), 443 - 53 The effect of temperature on food poisoning: a time-series analysis of salmonellosis in ten European countries; Kovats RS et al.; We investigated the relationship between environmental temperature and reported Salmonella infections in 10 European populations . Poisson regression adapted for time-series data was used to estimate the percentage change in the number of cases associated with a 1 degree C increase in average temperature above an identified threshold value . We found, on average, a linear association between temperature and the number of reported cases of salmonellosis above a threshold of 6 degrees C . The relationships were very similar in The Netherlands, England and Wales, Switzerland, Spain and the Czech Republic . The greatest effect was apparent for temperature 1 week before the onset of illness . The strongest associations were observed in adults in the 15-64 years age group and infection with Salmonella Enteritidis (a serotype of Salmonella) . Our findings indicate that higher temperatures around the time of consumption are important and reinforce the need for further education on food-handling behaviour. Zh Mikrobiol Epidemiol Immunobiol, 2004 Mar-Apr, (2), 85 - 7 {Adhesiveness and antibiotic resistance of the causative agents of salmonella infection in Gomel' region}; Tapal'skii DV et al.; The adhesive capacity, resistance to antibiotics and biological properties of Salmonella strains of different serogroups, circulating in the Gomel region, were under study . Resistance to antibiotics and changes in biological properties were accompanied by an increased adhesiveness of these strains . A high degree of adhesiveness was noted in S . hadar, a new for Belarus strains. Tohoku J Exp Med, 2004 May, 203(1), 47 - 52 Phenoxazine compounds produced by the reactions with bovine hemoglobin show antimicrobial activity against non-tuberculosis mycobacteria; Shimizu S et al.; We studied the anti-microbial effects of phenoxazines produced by the reaction of o-aminophenol or its derivatives with bovine hemoglobin, on seven species of mycobacteria such as Mycobacterium tuberculosis, Mycobacterium marinum, Mycobacterium intracellulare, Mycobacterium scrofulaceum, Mycobacterium fortuitum, Mycobacterium kansasii and Mycobacterium smegmatis and some bacteria such as Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, Listeria monocytogeneses . These phenoxazines, including 2-amino-4, 4alpha-dihydro-4alpha, 7-dimethyl-3H-phenoxzine-3-one (Phx-1), 3-amino-1, 4alpha-dihydro-4alpha, 8-dimethyl-2H-phenoxazine-2-one (Phx-2), and 2-aminophenoxazine-3-one (Phx-3), prevented the proliferation of four non-tuberculosis mycobacteria including M . scrofulaceum, M . kansasii, M . marinum, and M . intracellulare dose-dependently, though the inhibitory effects of these phenoxazines differed according to the species of mycobacteria . However these phenoxazines failed to prevent the proliferation of M . tuberculosis, M . fortuitum, and M . smegmatis, and the concerned bacteria other than mycobacteria . The present results may contribute to development of novel antibiotics against non-tuberculolsis mycobacteria. Epidemiol Mikrobiol Imunol, 2004, 53(2), 70 - 3 {Occurrence of phage types of non-typhoid Salmonella serovar in the Slovak Republic 2000-2003}; Majtanova L; Salmonellae of non-typhoidal serovars are the most important pathogens involved in foodborne diseases in humans all over the world . The incidence rates of two major Salmonella serovars, i.e . S . enterica serovar Enteritidis (S . Enteritidis) and S . enterica serovar Typhimurium (S . Typhimurium), in the Slovak Republic in 2000-2003 are given . Over the period studied, 829 S . Enteritidis strains and 258 S . Typhimurium strains isolated from patients with salmonellosis were investigated in the National Reference Centre for Salmonella Phage Typing . The S . Enteritidis strains were differentiated into 16 phage types, with phage type 8 being dominant since found in 73.6%, 53.8%, 62.8% and 45.6% of strains in 2000, 2001, 2002 and 2003, respectively . The following most frequent phage types were 4 and 13a . New phage types, i.e . 15, 5, 25 and 14b, were identified from salmonellosis outbreaks in 2003 . The S . Typhimurium strains were also differentiated into 16 phage types with phage type DT104 strains being prevalent and showing an increase from 7.4% in 2000 to 44.6% in 2003; 54.2% of them were resistant and of R type ACSSut . The second most frequent phage type in 2000-2001 was 2b in 2003 DT41 . The frequency of the other phage types was not epidemiologically significant. J Med Microbiol, 2004 Jul, 53(Pt 7), 705 - 9 Protection in a mouse peritonitis model mediated by iron-regulated outer-membrane protein of Salmonella typhi coupled to its Vi antigen; Chibber S et al.; Vi polysaccharide and iron-regulated outer-membrane proteins (IROMPs) were extracted and purified from the standard strain of Salmonella typhi, Ty2 . These were then conjugated by chemical coupling using the carbodimide method . Vi-IROMP conjugate was tested for its ability to protect against colonization by S . typhi in different organs . Mice immunized with 2.5 microg Vi-IROMP conjugate showed the most protection, as the least bacterial colonization of spleen, liver and Peyer's patches was observed . Peritoneal macrophages obtained from conjugate-treated mice phagocytosed bacteria efficiently . Circulating antibodies and the delayed type hypersensitivity response elucidated by mouse foot-pad swelling was significantly higher in conjugate-treated animals . These results clearly demonstrate that an IROMP and polysaccharide conjugate of S . typhi prepared from the same strain has the potential to protect animals against challenge. J Clin Microbiol, 2004 Jun, 42(6), 2581 - 6 Multiplex PCR for distinguishing the most common phase-1 flagellar antigens of Salmonella spp; Herrera-Leon S et al.; Most Salmonella serotypes alternatively express either phase-1 or phase-2 flagellar antigens, encoded by the fliC and fljB genes, respectively . Flagellar phase reversal for the identification of both flagellar antigens is not necessary at the genetic level . Variable internal regions of the fliC genes encoding the H:i, H:r, H:l,v, H:e,h, H:z(10), H:b, and H:d antigens have been sequenced; and the specific sites for each antigen in selected Salmonella serotypes have been determined . These results, together with flagellar G-complex variable internal sequences obtained by the Foodborne and Diarrheal Diseases Branch at the Centers for Disease Control and Prevention in Atlanta, GA, have been used to design a multiplex PCR to identify the G-complex antigens as well as the H:i, H:r, H:l,v, H:e,h, Hz(10), H:b, and H:d first-phase antigens . These antigens are part of the most common Salmonella serotypes possessing first-phase flagellar antigens . Salmonella enterica serotype Enteritidis is identified by adding a specific primer pair published previously . This multiplex PCR includes 13 primers . A total of 161 Salmonella strains associated with 72 different serotypes were tested . Each strain generated one first-phase-specific antigen fragment ranging from 100 to 500 bp; Salmonella serotype Enteritidis, however, generated two amplicons of 500 bp that corresponded to the G complex and a 333-bp serotype-specific amplicon, respectively . Twenty-three strains representing 19 serotypes with flagellar genes different from those targeted in this work did not generate any fragments . The method is quick, specific, and reproducible and is independent of the phase expressed by the bacteria when they are tested. J Clin Microbiol, 2004 Jun, 42(6), 2432 - 7 SHV-12-like extended-spectrum-beta-lactamase-producing strains of Salmonella enterica serotypes Babelsberg and Enteritidis isolated in France among infants adopted from Mali; Weill FX et al.; From December 2002 to June 2003, 14 cultures of Salmonella enterica serotype Babelsberg and 6 cultures of serotype Enteritidis, isolated in France from internationally adopted children, were identified at the French National Reference Center for Salmonella . All serotype Babelsberg isolates were related, as determined by pulsed-field gel electrophoresis, and all serotype Enteritidis strains displayed the same phage type . All serotype Enteritidis and seven serotype Babelsberg isolates produced an SHV-12-like extended-spectrum beta-lactamase as determined by sequencing of PCR products and by isoelectrofocusing . Some serotype Enteritidis isolates exhibited additional antimicrobial resistance (aminoglycosides, tetracycline, chloramphenicol, sulfonamides, and trimethoprim) . Our investigation indicated that these Salmonella isolates were certainly acquired in the same orphanage in Bamako, Mali, before the children were adopted by French families . An inappropriate use of ceftriaxone was probably the cause of the emergence of such strains . There is an urgent need to determine the origin of the contamination and to introduce adequate antibiotic protocols into this orphanage to prevent further transmission and dissemination . Screening for infections and follow-up, adapted to the origin of the internationally adopted children, should be recommended. Appl Environ Microbiol, 2004 Jun, 70(6), 3706 - 14 Uptake and replication of Salmonella enterica in Acanthamoeba rhysodes; Tezcan-Merdol D et al.; 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. Appl Environ Microbiol, 2004 Jun, 70(6), 3618 - 23 Establishment of a real-time PCR-based approach for accurate quantification of bacterial RNA targets in water, using Salmonella as a model organism; Fey A et al.; Quantitative PCR (Q-PCR) is a fast and efficient tool to quantify target genes . In eukaryotic cells, quantitative reverse transcription-PCR (Q-RT-PCR) is also used to quantify gene expression, with stably expressed housekeeping genes as standards . In bacteria, such stable expression of housekeeping genes does not occur, and the use of DNA standards leads to a broad underestimation . Therefore, an accurate quantification of RNA is feasible only by using appropriate RNA standards . We established and validated a Q-PCR method which enables the quantification of not only the number of copies of target genes (i.e., the number of bacterial cells) but also the number of RNA copies . The genes coding for InvA and the 16S rRNA of Salmonella enterica serovar Typhimurium were selected for the evaluation of the method . As DNA standards, amplified fragments of the target genes were used, whereas the same DNA standards were transcribed in vitro for the development of appropriate RNA standards . Salmonella cultures and environmental water samples inoculated with bacteria were then employed for the final testing . Both experimental approaches led to a sensitive, accurate, and reproducible quantification of the selected target genes and RNA molecules by Q-PCR and Q-RT-PCR . It is the first time that RNA standards have been successfully used for a precise quantification of the number of RNA molecules in prokaryotes . This demonstrates the potential of this approach for determining the presence and metabolic activity of pathogenic bacteria in environmental samples. Appl Environ Microbiol, 2004 Jun, 70(6), 3582 - 7 Medium-chain fatty acids decrease colonization and invasion through hilA suppression shortly after infection of chickens with Salmonella enterica serovar Enteritidis; Van Immerseel F et al.; The most common source of Salmonella infections in humans is food of poultry origin . Salmonella enterica serovar Enteritidis has a particular affinity for the contamination of the egg supply . In this study, the medium-chain fatty acids (MCFA), caproic, caprylic, and capric acid, were evaluated for the control of Salmonella serovar Enteritidis in chickens . All MCFA were growth inhibiting at low concentrations in vitro, with caproic acid being the most potent . Contact of Salmonella serovar Enteritidis with low concentrations of MCFA decreased invasion in the intestinal epithelial cell line T84 . By using transcriptional fusions between the promoter of the regulatory gene of the Salmonella pathogenicity island I, hilA, and luxCDABE genes, it was shown that all MCFA decreased the expression of hilA, a key regulator related to the invasive capacity of Salmonella . The addition of caproic acid (3 g/kg of feed) to the feed of chicks led to a significant decrease in the level of colonization of ceca and internal organs by Salmonella serovar Enteritidis at 3 days after infection of 5-day-old chicks . These results suggest that MCFA have a synergistic ability to suppress the expression of the genes required for invasion and to reduce the numbers of bacteria in vivo . Thus, MCFA are potentially useful products for reducing the level of colonization of chicks and could ultimately aid in the reduction of the number of contaminated eggs in the food supply. Appl Environ Microbiol, 2004 Jun, 70(6), 3485 - 92 Effects of physical properties of feed on microbial ecology and survival of Salmonella enterica serovar Typhimurium in the pig gastrointestinal tract; Mikkelsen LL et al.; A two-by-two factorial experiment with pigs was conducted to study the effect of feed grinding (fine and coarse) and feed processing (pelleted and nonpelleted) on physicochemical properties, microbial populations, and survival of Salmonella enterica serovar Typhimurium DT12 in the gastrointestinal tracts of pigs . Results demonstrated a strong effect of diet on parameters measured in the stomachs of the pigs, whereas the effect was less in the other parts of the gastrointestinal tract . Pigs fed the coarse nonpelleted (C-NP) diet showed more solid gastric content with higher dry matter content than pigs fed the fine nonpelleted (F-NP), coarse pelleted (C-P), or fine pelleted (F-P) diet . Pigs fed the C-NP diet also showed significantly increased number of anaerobic bacteria (P < 0.05), increased concentrations of organic acids, and reduced pH in the stomach . In addition, pigs fed the C-NP diet showed increased in vitro death rate of S . enterica serovar Typhimurium DT12 in content from the stomach (P < 0.001) . Pigs fed the C-NP diet had a significantly higher concentration of undissociated lactic acid in gastric content than pigs fed the other diets (P < 0.001) . A strong correlation between the concentration of undissociated lactic acid and the death rate of S . enterica serovar Typhimurium DT12 was found . In the distal small intestine, cecum, and midcolon, significantly lower numbers of coliform bacteria were observed in pigs fed the coarse diets than in pigs fed the fine diets (P < 0.01) . Pigs fed the C-NP diet showed the lowest number of coliform bacteria in these segments of the gastrointestinal tract . Pigs fed the coarse diets showed increased concentration of butyric acid in the cecum (P < 0.05) and colon (P < 0.10) compared with pigs fed the fine diets . It was concluded that feeding a coarsely ground meal feed to pigs changes the physicochemical and microbial properties of content in the stomach, which decreases the survival of Salmonella during passage through the stomach . In this way the stomach acts as a barrier preventing harmful bacteria from entering and proliferating in the lower part of the gastrointestinal tract. Biochem Biophys Res Commun, 2004 Jul 2, 319(3), 1001 - 9 Cloning of IP15, a pancreatitis-induced gene whose expression inhibits cell growth; Ropolo A et al.; We describe the cloning and expression of the mouse gene interferon-inducible-protein 15 (IP15), whose activation is related to the acute phase of experimental pancreatitis . Analysis of its structure indicates that it encodes a putative transmembrane protein of 137 amino acids . This gene contains a predicted IFN-stimulable-response element . In vivo studies showed that IP15 is strongly activated in pancreas early during caerulein-induced pancreatitis . In situ hybridization of IP15 mRNA showed that its expression is restricted to acinar cells . IP15 was also induced in pancreas under systemic-lipopolysaccharide treatment and in intestine under Salmonella infection . In vitro studies using NIH3T3 fibroblasts showed that IP15 is induced by IFN-alpha . Growth rate was significantly lower in cells transfected with pcDNA4/IP15 plasmid . In addition, cells expressing IP15 showed less capacity to develop colonies after antibiotic selection . In conclusion, we identified a new interferon-inducible gene that is activated early in pancreas with pancreatitis and whose expression inhibits cell growth. Vet Immunol Immunopathol, 2004 Jul, 100(1-2), 81 - 97 Effects of furazolidone pretreatment of Salmonella enteritidis PT4 at sub- and suprainhibitory concentrations on phagocytosis and intracellular survival in chicken macrophages; Chadfield MS et al.; The antimicrobial effect of the nitrofuran derivative furazolidone at sub- and suprainhibitory concentrations on Salmonella enteritidis PT4 and the influence with regard to interaction with avian macrophages was investigated in this study . Phagocytosis of furazolidone-sensitive (FzS) S . enteritidis with chicken macrophages in the presence of furazolidone at concentrations of 1/8, 1/4, 1/2 and 8x MIC resulted in an increase in the rate of phagocytic killing of approximately 3-, 6-, 6.5- and 9-fold, respectively, with 1/2 and 8x MIC concentrations producing statistically significant (P<0.05) increases in phagocytosis . Treatment of the FzS Salmonella with furazolidone at concentrations of 4x and 10x MIC, for 15 min prior to phagocytosis, also significantly (P<0.005) increased phagocytic uptake when compared with untreated bacteria . The rate of phagocytosis monitored over 90 min was highest between 30 and 60 min with the furazolidone pretreated salmonella, compared with the delayed rate of the control between 60 and 90 min . Exposure of FzS and FzR strains with suprainhibitory concentrations of furazolidone at 4x, 8x and 10x MIC for 30 min prior to phagocytosis demonstrated an increase in bacterial killing . Exposure of strains to sub-inhibitory concentrations of furazolidone led to an increase in chemiluminescence during phagocytosis with macrophages, suggesting an increase in oxidative metabolism in the macrophages as a result of an increase in activation and phagocytosis . Pretreatment of the strains with suprainhibitory concentrations of furazolidone for 30 min prior to phagocytosis demonstrated a similar increase in oxidative metabolism in the macrophages . Measurement of the amount of 14C-furazolidone associated with chicken