Salmonella enterica serovar Newport has undergone a rapid epidemic spread in dairy cattle. This provides an efficient mechanism for pathogen amplification and dissemination into the environment through manure spreading on agricultural land. The objective of this study was to determine the survival characteristics of Salmonella serovar Newport in manure and manure-amended soils where the pathogen may be amplified. A multidrug-resistant (MDR) Salmonella serovar Newport strain and a drug-susceptible (DS) strain, both bovine isolates, were inoculated into dairy manure that was incubated under constant temperature and moisture conditions alone or after being mixed with sterilized or nonsterilized soil. Salmonella serovar Newport concentrations increased by up to 400% in the first 1 to 3 days following inoculation, and a trend of steady decline followed. With manure treatment, a sharp decline in cell concentration occurred after day 35, possibly due to microbial antagonism. For all treatments, decreases in Salmonella serovar Newport concentrations over time fit a first-order kinetic model. Log reduction time was 14 to 32 days for 1 log 10 , 28 to 64 days for 2 log 10 , and 42 to 96 days for 3 log 10 declines in the organisms' populations from initially inoculated concentrations. Most-probable-number monitoring data indicated that the organisms persisted for 184, 332, and 405 days in manure, manure-amended nonsterilized soil, and manure-amended sterilized soil, respectively. The MDR strain and the DS strain had similar survival patterns.

Salmonella enteritidis colonizes the tissues of the chicken ovary and oviduct, presumably contaminating eggs and thereby contributing to human outbreaks of salmonellosis. In this study, commercial adult laying hens were given an oral inoculation of 10 8 S. enteritidis organisms. Tissues from various organs, the intestines, and the reproductive tract, including freshly laid eggs, were collected daily for up to 40 days postinoculation (p.i.). Within 2 days p.i. S. enteritidis was detected by culture in pools of the spleen, liver, heart, and gallbladder tissues, in intestinal tissues of all infected birds, and in various sections of the ovary and oviduct. Detection of organisms by immunohistochemical staining was rare for most tissues in spite of their culture-positive status, suggesting a low level of tissue colonization. However, S. enteritidis could be detected by immunohistochemical staining in oviduct tissues associated with four forming eggs, indicating the possibility of a heavier colonization in the egg during its development. In two subsequent experiments, forming eggs taken from the oviduct with their associated tissue, were found to be culture positive for S. enteritidis at a rate of 27.1 and 31.4%, while freshly laid eggs in these experiments were culture positive at the rate of 0 and 0.6%. These observations suggest that while forming eggs are significantly colonized in the reproductive tract, factors within the eggs may control the pathogen before the eggs are laid. The data show that prior to egg shell deposition, forming eggs are subject to descending infections from colonized ovarian tissue, ascending infections from colonized vaginal and cloacal tissues, and lateral infections from colonized upper oviduct tissues. The data are consistent with an ascending infection of freshly laid eggs from the cloaca, as the incidence of positive eggs in experiments 1 and 3 coincided with heavily contaminated cloacal tissues (50.7 and 80%, respectively), while no positive eggs were detected in experiment 2 when cloacal colonization was low (8.3%). The data do not support the possibility of egg invasion by bacterial translocation from the peritoneal cavity.

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Multidrug-resistant (MDR) strains of Salmonella enterica serotype Newport have been described for many years. However, the recognition of Newport strains with resistance to cephalosporin antibiotics is more recent. Plasmid-mediated CMY-2 AmpC ␤-lactamases have been identified in Salmonella in the United States, and the bla CMY-2 gene has been shown to be present in Salmonella serotype Newport. This organism is currently undergoing epidemic spread in both animals and humans in the United States, and this is to our knowledge the first description of the molecular epidemiology of this Salmonella strain in animals. Forty-two isolates were included in this study. All isolates were characterized by pulsed-field gel electrophoresis, plasmid analysis, and antibiogram. Four pulsed-field profiles with XbaI were observed. Plasmid analyses showed that although the majority of isolates harbored a single plasmid of 140 kb, this plasmid was not identical in all strains. All isolates showed the presence of the bla CMY gene by PCR. Integrons were detected in 16 of the 42 isolates; a fragment of approximately 1,000 bp, amplified with the intI-F and aadAI-R primers, confirmed the presence of the aadAI gene cassette within an integron in these 16 isolates. The potential for coselection of the bla CMY gene, if located on an MDR replicon, may not be dependent on any particular antibiotic but rather may be the result of more general antimicrobial use. If this replicon is mobile, it is to be expected that similar MDR strains of additional Salmonella serotypes will be recognized in due course.

A low-swirl burner (LSB) developed for laboratory research has been scaled to the thermal input levels of a small industrial burner. The purpose was to demonstrate its viability for commercial and industrial furnaces and boilers. The original 5.28 cm i.d. LSB using an air-jet swirler was scaled to 10.26 cm i.d. and investigated up to a firing rate of Q ‫ס‬ 586 kW. The experiments were performed in water heater and furnace simulators. Subsequently, two LSBs (5.28 and 7.68 cm i.d.) configured to accept a novel vaneswirler design were evaluated up to Q ‫ס‬ 73 kW and 280 kW, respectively. The larger vane-LSB was studied in a boiler simulator. The results show that a constant velocity criterion is valid for scaling the burner diameter to accept higher thermal inputs. However, the swirl number needed for stable operation should be scaled independently using a constant residence time criterion. NO x emissions from all the LSBs were found to be independent of thermal input and were only a function of the equivalence ratio. However, emissions of CO and unburned hydrocarbons were strongly coupled to the combustion chamber size and can be extremely high at low thermal inputs. The emissions from a large vane-LSB were very encouraging. Between 210 and 280 kW and 0.8 Ͻ Ͻ 0.9, NO x emissions of Ͻ15 ppm and CO emissions of Ͻ10 ppm were achieved. These results indicate that the LSB is a simple, low-cost, and promising environmental energy technology that can be further developed to meet future air-quality rules.

Salmonella enterica serovar Enteritidis is unable to multiply in the albumen of fresh eggs and must gain access to the yolk contents in order to multiply to a high level (>10 6 c.f.u. per ml egg contents). As human Salmonella infections resulting from the consumption of infected eggs more frequently involve serovar Enteritidis phage type (PT) 4 than other serovars or PTs, a number of isolates of various S. enterica serovars were examined for their ability to multiply to a high level in eggs over a period of 8 days storage at 20 6C. Their behaviour was compared to that of a range of defined fimbrial and flagella mutants of S. Enteritidis. Strains that did not express flagella were unable to multiply in eggs, and those deficient for curli fimbriae, including strains of S. Enteritidis PT6, displayed high-level growth in significantly fewer eggs than those able to express curli. Most S. Enteritidis strains multiplied to a high level in between 5 and 10 % of eggs during 8 days storage. One PT4 strain, though, showed high levels of growth in more than 25 % of eggs over this period, significantly higher than the other PTs or the two other isolates of PT4 tested. This ability may be important for the association of PT4 infection with the consumption of eggs. INTRODUCTIONSalmonella infection resulting from the consumption of contaminated eggs is a major public health problem in Europe and the USA and an emerging one in the Far East (Poppe, 1999). Salmonella enterica subspecies enterica serovar Enteritidis is responsible for the majority of eggassociated infections, and appears to possess factors that enable it to persist in the reproductive tissue of the hen so that it can contaminate the contents of intact eggs (Okamura et al., 2001a, b). Other serovars of S. enterica, such as Gallinarum and Pullorum, have in the past been widespread in laying flocks (Shivaprasad, 2000), without a concomitant high level of human infection.Salmonella enterica has the potential to elaborate numerous cell surface structures, including type 1 (SEF21), thin aggregative or curli (SEF17), SEF14, long polar (LPF) and plasmid-encoded (PEF) fimbriae, and flagella. Many of these have been shown to be important in the process of infection of the hen and in the colonization of the reproductive tissues. SEF14 fimbriae, which are only present in a few group D serovars (Turcotte & Woodward, 1993), mediate adherence to reproductive tissues (Ogunniyi et al., 1997) and may have a role in organ invasion and persistence (Rajashekara et al., 2000). There is some evidence that flagella are important in the subsequent invasion of internal tissues . Type 1 and curli fimbriae in Escherichia coli, analogous to SEF21 and SEF17 of S. Enteritidis, are associated with initial persistence in the avian gastrointestinal tract (La Ragione et al., 2000). There is little current evidence for a role for PEF and LPF in virulence or persistence in the chick model, or in laying hens. The role of any of these in the growth of Salmonella within the egg is not yet known. Salmonel...

Experiments were conducted in which Salmonella enteritidis Phage Type 8, Phage Type 2, and RDNC (reaction does not conform) or three isolates of Salmonella typhimurium of diverse origin were fed to adult laying hens to determine if S. enteritidis has a selective advantage over S. typhimurium, which is now rarely isolated from chicken eggs, in its capacity to invade reproductive tissues. The results revealed that S. enteritidis and S. typhimurium may be equal in their potential to colonize the tissues of the reproductive tract and eggs that are forming in the oviduct prior to oviposition. S. enteritidis, but not S. typhimurium, was isolated from egg contents after oviposition. The degree to which intestinal, hepatic, splenic, or reproductive tissues were colonized by either serotype was not seen to affect the rate of colonization of eggs forming in the oviduct or the contamination of eggs after oviposition. Virulence factors related to the difference in the association of S. enteritidis and S. typhimurium with egg-borne salmonellosis remain to be defined.

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