Automatic generation of alloreactivity-reduced donor lymphocytes and hematopoietic stem cells from the same mobilized apheresis product.

INTRODUCTION: In vitro or in vivo depletion of alloreactive T cells can facilitate haplo-identical hematopoietic stem cell transplantation (HSCT). Very satisfactory transplant outcomes were thus reported for TCRαß/CD19-depleted hematopoietic stem/progenitor cell (HSPC) grafts. The current semi-automatic manufacturing process on the CliniMACS Plus, although robust, still requires a significant amount of manual labor to be completed. Towards advancing and further facilitating large scale cell processing, a new TCRαß/CD19 depletion module combined with the previously described CD45RA depletion module (to serve as allo-reactivity attenuated donor lymphocyte infusion) was established on the CliniMACS Prodigy. METHODS: We evaluated six apheresis products from G-CSF-mobilized volunteer donors which were split automatically by the Prodigy, one portion each depleted of CD45RA+ or of TCRαß+ and CD19+ cells. We investigated critical quality attributes for both products. Products were assessed for recovery of HSPCs and mature subsets, as well as depletion efficiency of targeted cells using flow cytometry. Effects of apheresis and product age post 48 h storage at 2-6 °C as well as freeze-thawing on product viability and recovery of WBC and HPSCs were assessed by flow cytometry. RESULTS: Ten sequential automatic processes were completed with minimal hands-on time beyond tubing set installation. Depletion efficiency of CD45RA+ resp. TCRαß+ and CD19+ cells was equivalent to previous reports, achieving mean depletions of 4 log of targeted cells for both products. HSPC products retained TCRγδ+ and NK cells. 48 h storage of apheresis product was associated with the expected modest loss of HSPCs, but depletions remained efficient. Depleted products were stable until at least 72 h after apheresis with stem cell viabilities > 90%. Freeze-thawing resulted in loss of NK cells; post-thaw recovery of viable CD45+ and HSPCs was > 70% and in line with expectation. CONCLUSION: The closed, GMP-compatible process generates two separate medicinal products from the same mobilized apheresis product. The CD45RA-depleted products contained functional memory T cells, whereas the TCRαß/CD19-depleted products included HSPCs, TCRγδ+ and NK cells. Both products are predicted to be effectively depleted of GVH-reactivity while providing immunological surveillance, in support of haplo-identical HSCT.

Risk factors for antimicrobial resistance among fecal Escherichia coli from residents on forty-three swine farms.

Fecal Escherichia coli (n = 555) were isolated from 115 residents on 43 farrow-to-finish swine farms to determine the prevalence of antimicrobial resistance and associated risk factors. Susceptibility to 21 antimicrobials was determined and the overall prevalence of antimicrobial resistance was 25.8%. Pair-wise difference in prevalences of resistance to individual antimicrobials was significant between isolates from residents on farms that fed medicated swine rations compared to those that did not (p = 0.013). Cross-resistance among antimicrobials of same class and multidrug-resistance were observed. Logistic regression models revealed the following risk factors positively associated with antimicrobial resistance: use of antimicrobials in pigs on farms; number of hours per week that farmers spent in their pig barns; handling of sick pigs; and intake of antimicrobials by farm residents. This study indicates that occupational exposure of farmers to resistant bacteria and use of antimicrobials in pig farming may constitute a source of resistance in humans, although the human health impacts of such resistance is unknown. The consumption of antimicrobials by farmers appeared to constitute a significant risk for resistance development. Fecal E. coli from farm residents may act as a reservoir of resistance genes for animal and/or human pathogens.

The characterization of Salmonella serovars isolated from apparently healthy slaughtered pigs at Addis Ababa abattoir, Ethiopia.

We looked for Salmonella in all 278 apparently healthy pigs slaughtered between September 2004 and May 2005 at the only pig-slaughtering slaughterhouse in Addis Ababa, Ethiopia. We used standard methods and tested caecal contents, mesenteric lymph nodes, and carcass swabs from each pig (missing only one carcass swab). Of the 278 pigs, 120 (43%) were positive; of the 833 samples 173 (21%) were positive. Thirty-three percent of the isolates were multi-resistant (including 46/48 isolates of S. Hadar, but none of the 39 isolates of S. Eastbourne or of the 37 of S. Saintpaul). Resistance to streptomycin (32.4%), tetracycline (31.8%) and nitrofurantoin (27.2%) was relatively high. The most common pattern of MDR observed was to nitrofurantoin, streptomycin and tetracycline (Resistance type NitStrTet). Our results indicate that salmonellae are prevalent in pigs slaughtered at Addis Ababa abattoir and a large proportion of the isolates were multi-drug resistant.

Impact of experimentally induced serotonin deficiency by tryptophan depletion on sleep EEG in healthy subjects.

The tryptophan depletion test is a research strategy to investigate the functional consequences of decreasing the brain serotonin metabolism. Because serotonin is involved in sleep regulation and the regulation of affective states, we studied the acute polysomnographic effects of tryptophan depletion and expected to induce similar changes of sleep EEG as observed in depressed patients. A total of 12 healthy subjects (mean age 34 +/- 3 years) had eight polysomnograms, divided in two blocks of 4 consecutive nights. After one adaptation and 1 baseline night, subjects received a low-protein diet on day 3 and 4 until midday. On day 4 at 18.00 h, they drank an amino acid mixture either devoid of tryptophan or containing 2.3 g of tryptophan (placebo control) in randomized and double-blind order, resulting in an 85% decrease (tryptophan depletion) and a 144% increase (placebo control) of serum tryptophan at 22.00 h. After tryptophan depletion but not placebo, significant effects on sleep EEG were observed in terms of decreased non-rapid eye movement (non-REM) stage 2, increase of wake %, and of rapid eye movement (REM) density compared with baseline. REM latency was not altered, however the first and second REM period interval were significantly shorter after tryptophan depletion. This study underlines the impact of the serotonergic system on sleep maintenance and on REM sleep.

Diversity in antimicrobial resistance and other characteristics among Salmonella typhimurium DT104 isolates.

Multiresistant Salmonella enterica subspecies enterica serovar Typhimurium definitive type 104 (S. Typhimurium DT104 or DT104) bacteria are important pathogens in animals and humans. DT104 isolates are often called pentaresistant strains that spread clonally. The purpose of this study was to determine phenotypic, genotypic, and epidemiologic characteristics of 175 S. Typhimurium DT104 strains isolated from food-producing animals in Canada. More than 90% of the isolates were resistant to ampicillin (Amp), chloramphenicol (Chl), florfenicol (Flo), sulfisoxazole (Sul), and tetracycline (Tet), 53% of the isolates were additionally resistant to spectinomycin (Spc) and streptomycin (Str), and 28% to kanamycin (Kan) and neomycin (Neo). Sixty-one percent of the strains harbored a single 60-MDa plasmid, 21% contained 60- and 2.0-MDa plasmids, and 4% had 60, 4.6- and 2.0-MDa plasmids. Resistance to Kan and Neo was encoded by the aminoglycoside aphA-1 gene on 2.0-MDa plasmids, whereas resistance to trimethoprim (Tmp) and Sul was encoded by the dhfrIb gene on 4.6-MDa plasmids. Polymerase chain reactions (PCR) showed the presence of integrons with the ant (3")-Ia aminoglycoside adenyltransferase and the bla(PSE-1) beta-lactamase gene cassettes, and the presence of the flost gene in all but one strain resistant to Spc and Str, Amp, and Chl and Flo, respectively. DT104 isolates from cattle at six feedlots represented a separate clone; they were sensitive to Str and Spc and lacked the ant (3")-Ia gene. Pulsed-field gel electrophoresis (PFGE) using Bln I, Spe I, and Xba I resulted in 15, 12, and 8 PFGE patterns, respectively. In summary, we observed considerable diversity in phenotypic, genotypic, and epidemiological characteristics among the DT104 isolates.

Trends in antimicrobial resistance of Salmonella isolated from animals, foods of animal origin, and the environment of animal production in Canada, 1994-1997.

The purpose of our study was to determine the occurrence, magnitude, trends, and relationships regarding antibiotic resistance of Salmonella isolated from animals, animal food products, and the environment of animals. We examined 621 strains of 67 different serovars isolated in 1994, 721 strains of 75 different serovars isolated in 1995, 1,219 strains of 83 different serovars isolated in 1996, and 1,336 Salmonella strains of 92 different serovars isolated in 1997, for resistance to 17 antibiotics at one to three different concentrations with the agar dilution method. The overall resistance magnitude regressed from 9.2% in 1994 to 8.1% in 1997. Resistance to streptomycin (30.4% of 3,897 isolates), tetracycline (27.3%), and sulfisoxazole (23.7%) was highest. Resistance to streptomycin, tetracycline, kanamycin, and gentamicin declined during the 4-year period. Notable increases in resistance to ampicillin, chloramphenicol, and neomycin occurred during the 1994-1997 years. None of the isolates was resistant to amikacin. None of the isolates was resistant to ciprofloxacin at 1, 2, and 4 microg/ml. Salmonella bredeney isolates from turkeys showed a decreased sensitivity to ciprofloxacin and were resistant at the low level of 0.125 microg/ml, but none of these isolates was resistant at 1 microg/ml. Resistance to nalidixic acid correlated significantly with decreased sensitivity to ciprofloxacin; 122 of 127 (96%) isolates resistant to nalidixic acid at 32 microg/ml were resistant to ciprofloxacin at 0.125 microg/ml but sensitive at 1 microg/ml. Resistance to S. typhimurium to each of the seven antibiotics ampicillin, chloramphenicol, kanamycin, neomycin, streptomycin, sulfisoxazole, and tetracycline increased persistently during each of the years 1994-1997, but none of the S. typhimurium isolates showed decreased sensitivity to ciprofloxacin. Clinical isolates of Salmonella were twice as frequently resistant to the antimicrobials in the test panel than isolates obtained during surveys. Salmonella isolates from turkeys were more frequently resistant than isolates from pigs, cattle, and chickens.

Construction of mini-Tn10luxABcam/Ptac-ATS and its use for developing a bacteriophage that transduces bioluminescence to Escherichia coli O157:H7.

Mini-Tn10luxABcam/Ptac-ATS was constructed in order to develop a luciferase-transducing bacteriophage for detecting Escherichia coli O157:H7. The transposon was designed to deliver a 3.6-kb insertion that confers n-decanal-dependent bioluminescence and resistance to chloramphenicol and was constructed using mini-Tn10cam/Ptac-ATS in the plasmid pNK2884 and luxAB from Vibrio harveyi. PhiV10, a temperate bacteriophage infecting common phage types of Escherichia coli O157:H7, was mutagenized as a prophage in E. coli O157:H7 strain R508. PhiV10::luxABcamA1-23 was rescued from the strain by propagating it on a strain lacking the bacteriophage and the vector containing the transposon. The bacteriophage transduced n-decanal-dependent bioluminescence to E. coli O157:H7 strain R508 that was measurable approximately 1 h post infection.

Salmonella enteritidis in Canada.

During the last few years the prevalence of Salmonella enteritidis in people in Canada has increased slightly from 9 to 12% of all Salmonella isolates. Nation-wide surveys showed a low prevalence of S. enteritidis in poultry flocks: S. enteritidis was isolated from environmental samples of 2.7% of layer flocks and of 3% of broiler flocks. The overall prevalence of S. enteritidis-contaminated eggs from two flocks with infected hens was less than 0.06%. With the exception of S. enteritidis phage type (PT) 4, which has not been isolated from poultry in Canada, the phage types that occur most commonly in poultry (PT 8, 13 and 13a) are also the most prevalent in people. Phage typing was the most effective method for subdividing S. enteritidis strains. Eighty-three percent of the S. enteritidis isolates from poultry and its environment were sensitive to antimicrobial agents. The reasons why strains of the same phage type differ in virulence need to be elucidated.

Drug resistance, plasmids, biotypes and susceptibility to bacteriophages of Salmonella isolated from poultry in Canada.

Salmonella isolates from 295 layer and 294 broiler flocks in Canada were examined to determine resistance to antimicrobial agents, plasmid profiles, biochemical properties, and susceptibility to polyvalent bacteriophages. Except for the high number of strains resistant to spectinomycin (97.8%), the frequency of drug resistance of Salmonella isolates from layer flocks was low. None of 457 isolates from layer flocks was resistant to amikacin or ciprofloxacin, and less than 2% of the strains were resistant to cephalothin, chloramphenicol, cotrimoxazole, gentamicin, kanamycin, neomycin, nitrofurantoin, and/or polymyxin B. About 3% of the strains were resistant to ampicillin, carbenicillin and/or tetracycline, whereas 8% of the strains were resistant to sulfisoxazole. Salmonella anatum var. O15+ and S. typhimurium var. copenhagen strains were resistant to multiple antimicrobial agents. None of 1159 Salmonella strains from broiler flocks was resistant to amikacin, cephalothin, ciprofloxacin or polymyxin B, less than 1% of the strains were resistant to chloramphenicol, 2% were resistant to ampicillin, carbenicillin and/or chloramphenicol; 5-7% were resistant to the aminoglycosides gentamicin, kanamycin and/or neomycin; 6% were resistant to nitrofurantoin; 10% to tetracycline; 14% to sulfisoxazole; and 99% to spectinomycin. A high percentage of S. binza, S. anatum var. O15+, S. schwarzengrund and S. heidelberg strains were resistant to antimicrobial agents. Some of the single or multiple resistances were encoded by conjugative plasmids or by plasmids that were thermosensitive for transfer. Eight percent of S. heidelberg strains did not produce hydrogen sulfide. Ninety-seven percent of the Salmonella strains were susceptible to the lytic effect of polyvalent bacteriophages.

Tagging and elimination of plasmids in Salmonella of avian origin.

This study compared the effectiveness of a number of procedures designed to label and eliminate plasmids that may play a role in virulence in Salmonella. Twenty strains of Salmonella of 9 serovars were subjected to 3 methods for labelling plasmids with transposons. Strains containing labelled and unlabelled plasmids were exposed to physical and chemical curing agents. Plasmids in 9 of 20 strains of Salmonella were tagged by conjugation with a donor Escherichia coli containing a temperature-sensitive RP4 plasmid that carried the Tn1 transposon. Plasmids in 2 of 5 strains of Salmonella were labelled by conjugation with a donor E. coli that contained a F' tslac::Tn5 plasmid. Transduction of Salmonella with a P22 bacteriophage that carried a temperature-sensitive Tn10 transposon resulted in chromosomal insertion of Tn10 in 2 of 10 strains. Use of chemical curing agents resulted in curing of plasmids in only 6 of 17 strains. Two strains were cured by ethidium bromide, two by a combination of ethidium bromide and novobiocin, two by a combination of imipramine and methylene blue, and none by acridine orange, novobiocin, sodium dodecyl sulfate or rifampicin. In contrast, plasmids in 14 of 17 Salmonella strains were eliminated by incubation at 45.5 degrees C.

Characterization of Salmonella isolates from beef cattle, broiler chickens and human sources on Prince Edward Island.

Non-typhoid Salmonella serovars remain a potential threat to human health, and beef cattle and broiler chickens are possible sources of these organisms on Prince Edward Island (PEI). In this study, the ceca of beef cattle belonging to fasted and non-fasted groups, and broiler chickens were examined for Salmonella at the time of slaughter. The characteristics of the isolates, including antimicrobial resistance patterns and virulence genes, were studied along with the isolates obtained from cases of human salmonellosis on PEI during the study period (1996-97). The prevalence of Salmonella in beef cattle was 4.6% (11/240). The rate was significantly higher in fasted cattle (7.46%), than in non-fasted cattle (0.94%). The prevalence rate in chickens was 32.5% (39/120). In beef cattle, Salmonella typhimurium phage type (PT) or definitive type (DT) 104 which was resistant to ampicillin, chloramphenicol, streptomycin, sulfisoxazole and tetracycline, was the most predominant type (64%). In chickens, S. heidelberg, with resistance to gentamicin, streptomycin and sulfisoxazole, predominated. Of 26 isolates from humans, the most common serovar was S. typhimurium, including a multidrug-resistant strain of DT104. Examination by PCR revealed presence of the virulence gene invA in all serovars, and the spvC gene in all S. typhimurium isolates, of both beef cattle and human origin. Among the other serovars the latter gene was found in 7 human isolates, but in none of the chicken or beef isolates. All but 3 of the spvC-positive isolates possessed a 90 kilobasepair (kbp) plasmid suggesting that the 3 isolates had the spvC gene on their chromosome. These findings were confirmed by plasmid DNA isolation using 3 different protocols and by sequence analysis of the spvC-PCR product.

Antibiotic resistance of Salmonella isolated from hog, beef, and chicken carcass samples from provincially inspected abattoirs in Ontario.

The emergence of antimicrobial-resistant Salmonella organisms, especially Salmonella Typhimurium DT104, has been reported in many countries, including the United States and Canada. The purposes of this study were to determine the antimicrobial resistance patterns of Salmonella isolated from hog, beef, and chicken carcasses from provincially inspected abattoirs in Ontario and to determine the agreement between the agar dilution method and the microbroth dilution method for measurement of antimicrobial resistance of the isolates. Antimicrobial resistance of Salmonella isolates from hogs (n = 71), beef (n = 24), and chicken (n = 295) to amikacin, ampicillin, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, streptomycin, sulfamethoxazole, and tetracycline was determined using the two methods. None of the 390 isolates were resistant to ciprofloxacin at levels of 0.125 microg/ml. All chicken and hog isolates were sensitive to amikacin, whereas all beef isolates were sensitive to both amikacin and gentamicin. Multiple antimicrobial resistance (resistance to more than one antimicrobial) was found in 29% of bovine isolates and 42% of porcine isolates using both methods for testing and in 42% by the agar dilution and 33% by the microbroth dilution methods in the chicken isolates. Overall, there was good agreement between the two test methods for resistance to most of the antimicrobials, with disagreement found in the results in 1.3% of the isolates for ampicillin and sulfamethoxazole, 8.2% for streptomycin, 5.6% for cephalothin, and 1.0% of the isolates for tetracycline. The lack of agreement between the two test methods was found mostly among the chicken isolates.

Classification of grossly detectable abnormalities and conditions seen at postmortem in Canadian poultry abattoirs according to a hazard identification decision tree.

This study was designed to review all grossly detectable abnormalities and conditions (GDACs) encountered in poultry in Canadian abattoirs to determine which have potential to cause adverse health effects for the consumer. Review of the literature and consultation with scientists in the field of microbiology, epidemiology, poultry pathology, chemistry, and meat inspection served to generate an inventory of GDACs, and a decision tree containing algorithms was developed to identify GDACs potentially representing a health hazard to consumers. Through the use of the decision tree, GDACs were classified into different categories with regard to the risk they represent to humans. A number of GDACs were identified as being of potential concern from a food safety perspective, namely Erysipelas, fowl cholera, Campylobacteriosis, clostridial diseases, hepatitis/enteritis associated with Helicobacter, Listeriosis, Salmonella infections (nontyphoid infections, Salmonella arizonae, pullorum disease, and fowl typhoid), Staphylococcosis, and Toxoplasmosis. Further characterization--i.e., hazard characterization, exposure assessment, and risk characterization--is required to quantify or better characterize the probability that products derived from affected carcasses may affect the consumer as well as the resulting consequences. Risk assessment is a dynamic process. Results presented in this paper are based on available information and expert opinion. As new information is obtained, the inventory of GDACs and their classification may be modified.

Relation of plasmids to virulence and other properties of salmonellae from avian sources.

A collection of 185 isolates of 34 serovars of Salmonella from avian sources was examined for plasmids, drug resistance, biochemical properties, serum resistance, and virulence. No serovars other than S. enteritidis, S. typhimurium, and S. heidelberg showed evidence of serovar-associated plasmids. All S. enteritidis isolates carried a single plasmid of 36 Mdal and were resistant to guinea pig serum; one strain that was tested was virulent. Of 27 isolates of S. typhimurium, 11 possessed a 60-Mdal plasmid and 17 harbored a 2.3-Mdal plasmid. Among isolates of S. heidelberg, 21 of 24 carried a 2.2-Mdal plasmid. The only biochemical property that varied was fermentation of inositol, which tended to be related to serovar. Of 172 isolates, 54 were resistant to at least one drug. Multiple drug resistance was usually associated with R plasmids, and transmissible plasmids that encoded resistance to chloramphenicol and gentamicin were demonstrated. Of 117 isolates tested, 43 were resistant to guinea pig serum. Resistance appeared to be a characteristic of isolates rather than serovar and could not be related to plasmids. Twenty-five isolates highly resistant to guinea pig serum were all susceptible to the bactericidal action of chicken serum. In tests for virulence using intraperitoneally (i.p.) and orally inoculated Balb/c mice and day-old chicks, only i.p.-inoculated chicks proved useful in demonstrating large differences among isolates: LD50's ranged from 10(0) to 10(8).

Effect of chlorination of drinking water on experimental salmonella infection in poultry.

The type of drinker used for poults influenced the level of free available chlorine (FAC) in chlorinated water as well as the total plate count, fecal coliform count, and number of salmonellae in chlorinated and non-chlorinated drinking water. Nipple drinkers maintained higher levels of FAC in drinking water than Swish-cups, Swish-cups maintained higher levels than MarkIII, and MarkIII maintained higher levels than trough drinkers. The level of FAC retained in the water in trough drinkers was insufficient to exert a bactericidal effect against coliforms and salmonellae. Chlorination of drinking water and the resulting diminished number or absence of salmonellae in the drinking water did not lower the number of salmonellae per gram of cecal contents in challenged or unchallenged but exposed poults. The number of salmonellae per gram of cecal contents decreased significantly (P less than 0.01) in poults between 14 and 21 days of age, irrespective of whether or not the poults drank chlorinated water.

The tryptophan depletion test. Impact on sleep in healthy subjects and patients with obsessive-compulsive disorder.

UNLABELLED: The tryptophan depletion test is a research tool to study the functional consequences of decreasing the brain serotonin metabolism. Since serotonin is involved in sleep regulation and assumed to be of high importance in the etiology of psychiatric disorders, the acute polysomnographic effects of tryptophan depletion were studied in healthy subjects and patients with obsessive compulsive disorder (OCD). According to the reciprocal interaction model of non-REM and REM-sleep regulation we expected that tryptophan depletion in healthy controls should provoke alterations of sleep similar to depression, whereas we assumed that these effects would be more pronounced in patients with OCD. METHODS: 12 healthy subjects with a mean age of 34 years and 12 patients suffering from OCD with a mean age of 30 years had 4 polysomnographic investigations. After 1 adaption and 1 baseline night subjects received a low protein diet on day 3 and 4 until midday. On day 4 at 18.00 h subjects ingested an aminoacid mixture devoid of tryptophan. This procedure resulted in a decrease of 85% in healthy subjects and 80% in OCD patients at 22.00 h. RESULTS: The tryptophan depletion led to more pronounced disturbances of sleep continuity in OCD patients than in healthy subjects in terms of an increase of wake time and a decrease of total sleep time. In both groups a decrease of sleep stage 2 could be observed. Healthy subjects showed significant alterations of phasic REM parameters as REM density and total number of rapid eye movements, what was not the case for OCD patients. CONCLUSIONS: Our results indicate the important role of the serotonergic system for sleep maintainance and the phasic aspects of REM sleep. Furthermore our data demonstrate that the tryptophan depletion test is a useful tool to evaluate the hypothesis of a serotonergic involvement in sleep regulation and the etiology of psychiatric disorders.

Treatment of obsessive compulsive disorder in the elderly: a review and case example.

Obsessive compulsive disorder (OCD) is a prevalent form of psychopathology in the elderly, yet limited evaluation of the disorder in this age group has occurred. We review the literature and describe a case of OCD effectively treated in an 80-year-old man. Case study reports suggest that elderly persons are responsive to selective serotonin uptake inhibitors, although medication selection and dosage may need to be adjusted as a result of the medical conditions sometimes present in the elderly. Elderly persons appear able to benefit from exposure and response prevention, although behavioral intervention has not been frequently used. We describe here the first case report where exposure and response prevention procedures were successfully used and this intervention was not confounded with psychopharmacologic treatment.

Salmonella contamination of hatching and table eggs: a comparison.

This study determined and compared Salmonella contamination rates of pools of surplus, early and culled hatching eggs from layer and broiler breeder flocks, and of pools of early and regular table eggs from layer flocks. Each pool contained 6 eggs. Five methods were used for the isolation of Salmonella. Nine of 126 pools of culled layer hatching eggs, 2 of 126 pools of surplus layer hatching eggs, and one of 126 pools of early layer hatching eggs were contaminated with Salmonella. All 126 pools of broiler breeder surplus, and early and culled hatching eggs tested negative for Salmonella. All 168 pools of regular table eggs tested negative for Salmonella, whilst one of 84 pools of early table eggs contained Salmonella agona. The pools of culled layer hatching eggs and surplus layer hatching eggs that contained S. typhimurium were derived from the same breeder operation. Similarly, the pools of culled and early layer hatching eggs that contained S. heidelberg were derived from one breeder operation. Pools of culled hatching eggs were more frequently contaminated with Salmonella than other hatching or table eggs. Pools containing eggs that were both cracked and dirty were more frequently contaminated with Salmonella than all other pools of eggs. The overall Salmonella contamination rate of the table eggs was 0.07 to 0.4%. Critical control points (macroscopic classification of the eggs as cracked and dirty) were validated microbiologically.

Virulence of Salmonella enteritidis phagetypes 4, 8 and 13 and other Salmonella spp. for day-old chicks, hens and mice.

Virulence of three Canadian poultry strains of Salmonella enteritidis, namely phagetypes (PT) 4, 8 and 13, and one Salmonella heidelberg strain was assessed in orally and intraperitoneally inoculated one-day old chickens and compared to the virulence of a human S. enteritidis PT 4 strain from the United Kingdom (UK). The two PT 4 strains were also compared in orally inoculated adult laying hens. In addition, orally inoculated Balb/c mice were used to evaluate virulence of the above strains and two strains of Salmonella typhimurium containing different plasmids. In orally inoculated one-day old chickens, the UK S. enteritidis PT 4 strain was more virulent than the Canadian PT 4 strain. The UK PT 4 strain was also more virulent and invasive in adult laying hens than the Canadian PT 4 strain. The S. enteritidis PT 8 strain and one S. typhimurium strain isolated from a chicken hatchery were the most virulent for orally inoculated Balb/c mice. This strain of S. typhimurium contained the 60 megadalton plasmid associated with virulence for Balb/c mice which was not present in the S. typhimurium strain isolated from a pig with septicemic disease.

Characterization of Salmonella enteritidis strains.

A study was conducted to characterize 318 Salmonella enteritidis strains that were mainly isolated from poultry and their environment in Canada. Biotype, phagetype (PT), plasmid profile (PP), hybridization with a plasmid-derived virulence sequence probe, antibiotic resistance, outer membrane proteins (OMPs), and lipopolysaccharide (LPS) profiles were determined. Relationships of these properties to one another, and their diagnostic and pathogenic significance were assessed. Biotyping indicated that failure to ferment rhamnose was sometimes useful as a marker for epidemiologically related strains. Phagetyping was the most effective method for subdividing S. enteritidis; it distinguished 12 PTs. Phagetype 13 was occasionally associated with septicemia and mortality in chickens. The strains belonged to 15 PPs. A 36 megadalton (MDa) plasmid was found in 97% of the strains. Only the 36 MDa plasmid hybridized with the probe. Seventeen percent of the strains were drug resistant; all strains were sensitive to ciprofloxacin. Thirty-five of 36 strains possessed the same OMP profile, and 36 of 41 strains contained smooth LPS.