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Effect of sunlight on the survival of Salmonella on surfaces
Abstract
To investigate the effect of simulated full-spectrum tropical sunlight on the survival of Salmonella in droplets on surfaces.
The survival on surfaces of three Zambian strains of Salmonella enterica serovars Enteritidis and Heidelberg was compared with that of a strain of S. enterica serovar Enteritidis phage type (PT) 4 with known characteristics which had been isolated from poultry in the UK. Samples were taken from surfaces every hour for 3 h and after 24 h exposure in either dark or 12 h light/12 h dark cycle conditions. Differences were analysed for significance using a one-way analysis of variance (anova). Results show that there were a significantly higher number of cells surviving on surfaces after 24 h in the dark when compared with populations exposed to a 12 h light/12 h dark cycle. Significantly more cells also survived exposure to sunlight under dirty than clean conditions.
Exposure to sunlight results in a significant decrease in numbers of Salmonella on surfaces.
Under field conditions exposure of contaminated surfaces to sunlight could be used in place of chemical methods of control as a cheaper way to reduce Salmonella contamination of surfaces.
... The intrusion of crops by wild animals, birds, reptiles, and rodents, as well as insects and nematodes, act as a vector for transferring various pathogens (Brandl, 2006). The survival and growth of microorganisms is influenced by several environmental factors and agricultural practices such as, exposure to solar UV radiation, temperature changes, humidity, and poor fertilizer regimes (Bezanson et al., 2012;Brandl, 2006;Nyeleti, Cogan, & Humphrey, 2004;Tomas-Callejas et al., 2011;Weller et al., 2017). ...
... Several studies reported that the sunlight of tropical latitudes (Davies & Evison, 1991;Nyeleti, Cogan, & Humphrey, 2004;Obiri-Danso, Paul, & Jones, 2001) and a concomitant increase in the surface temperature of produce (Tomas-Callejas et al., 2011) have an inhibitory effect against various microbial pathogens. Moreover, sunlight is found to reduce Salmonella levels in fresh water sources (Davies & Evison, 1991) and on food contact surfaces such as stainless steel (Nyeleti et al., 2004). ...
... Several studies reported that the sunlight of tropical latitudes (Davies & Evison, 1991;Nyeleti, Cogan, & Humphrey, 2004;Obiri-Danso, Paul, & Jones, 2001) and a concomitant increase in the surface temperature of produce (Tomas-Callejas et al., 2011) have an inhibitory effect against various microbial pathogens. Moreover, sunlight is found to reduce Salmonella levels in fresh water sources (Davies & Evison, 1991) and on food contact surfaces such as stainless steel (Nyeleti et al., 2004). Furthermore, microbial populations decline with decreasing nutrient availability because of failing to lower their metabolic rate to adopt the starvation condition (Fontaine, Mariotti, & Abbadie, 2003). ...
Preharvest contamination of produce with food borne pathogens has been a major food safety issue. In this study, we investigated the effect of surrounding vegetation on the survival of natural and inoculated generic Escherichia coli on watermelon rinds in an agricultural field setting. There was no significant difference (p > .05) on the populations of natural generic E. coli (1–1.46 log Most Probable Number (MPN)/sample) and coliforms (<3.99 log CFU/cm2) on watermelons harvested from low, medium, and high levels of vegetation. However, the survival rate of generic E. coli inoculated on watermelon rind discs was variable with the level of vegetation. A significant reduction in generic E. coli count was observed within 12 hr at all vegetation levels. After 108 hr, discs placed at low vegetation level had a highest die‐off reduction (3 log Colony Forming Units (CFU)/cm2) compared to medium and high vegetation levels.
Practical applications
To ensure preharvest produce safety, the Food Safety Modernization Act (FSMA) produce safety rule has suggested a time interval between last irrigation and harvest for potentially contaminating microorganisms to die‐off. However, a knowledge gap exists regarding the influence of surrounding vegetation on microbial die‐off rates on produce in the agricultural field. The findings of this study emphasize the importance of considering the surrounding vegetation while making decisions for developing preharvest risk management strategies based on microbial die‐off rate calculations.
... Harsh environmental conditions, such as direct UV radiation or lack of available nutrients, make the phyllosphere of plants an adverse environment for S. enterica [3]. In fact, S. enterica populations have been observed to naturally decline on healthy leaves [4][5][6]. ...
The human enteric bacterial pathogen Salmonella enterica causes approximately 1.35 million cases of food borne illnesses annually in the United States. Of these salmonellosis cases, almost half are derived from the consumption of fresh, raw produce. Although epiphytic S . enterica populations naturally decline in the phyllosphere, a subset of phytophagous insects have recently been identified as biological multipliers, consequently facilitating the growth of bacterial populations. We investigated whether tomato leaves with macroscopic feeding damage, caused by infestation of adult Western flower thrips ( Frankliniella occidentalis ), support higher S . enterica populations. To explore this hypothesis, we assessed S . enterica populations in response to thrips feeding by varying insect density, plant age, and the gender of the insect. As a reference control, direct leaf damage analogous to thrips feeding was also evaluated using directed, hydraulic pressure. In a supplementary set series of experiments, groups of F . occidentalis infested tomato plants were later inoculated with S . enterica to determine how prior insect infestation might influence bacterial survival and persistence. Following an infestation period, leaves visibly damaged by adult F . occidentalis supported significantly higher S . enterica populations and resulted in greater amounts of electrolyte leakage (measured as electrical conductivity) than leaves lacking visible feeding damage. Plant age did not significantly influence S . enterica populations or estimates of electrolyte leakage, independent of initial infestation. Additionally, the gender of the insect did not uniquely influence S . enterica population dynamics. Finally, applications of aggressive water bombardment resulted in more electrolyte leakage than leaves damaged by F . occidentalis , yet supported comparable S . enterica populations. Together, this study indicates that F . occidentalis feeding is one of the many potential biological mechanisms creating a more habitable environment for S . enterica .
... Specifically, the handling practices of the fresh produce may reduce the opportunities for the transfer of human pathogens to the fresh produce (54,56). In addition, the environmental conditions such as sunlight, moisture, the physical characteristics of the surface of the produce, and temperature may limit the opportunities for pathogen persistence (57)(58)(59). Of note, results indicated a variance in the range of pathogen prevalence. ...
Foodborne illness burdens individuals around the world and may be caused by consuming fresh produce contaminated with bacterial, parasite, and viral pathogens. Pathogen contamination on produce may originate at the farm and packing facility. This research aimed to determine the prevalence of human pathogens (bacteria, parasites, and viruses) on fresh produce (fruits, herbs, and vegetables) on farms and in packing facilities worldwide through a systematic review of 38 peer-reviewed articles. The median and range of the prevalence was calculated, and Kruskal–Wallis tests and logistic regression were performed to compare prevalence among pooled samples of produce groups, pathogen types, and sampling locations. Results indicated a low median percentage of fresh produce contaminated with pathogens (0%). Both viruses (p-value = 0.017) and parasites (p-value = 0.033), on fresh produce, exhibited higher prevalence than bacteria. No significant differences between fresh produce types or between farm and packing facility were observed. These results may help to better quantify produce contamination in the production environment and inform strategies to prevent future foodborne illness.
... RPCs which escape full cleaning where dirt and organic matter remain may protect any organisms which are attached. Nyeleti et al. [38] observed that Salmonella enterica appeared to survive better against ultraviolet radiation treatment on stainless steel surfaces after coating with bovine serum albumin. In the current study, treatment with 200 ppm NaClO and PAA caused an average of 2.73 and 2.62 log CFU of S. Typhimurium, respectively. ...
Salmonella serovars, one of the leading contributors to foodborne illness and are especially problematic for foods that are not cooked before consumption, such as fresh produce. The shipping containers that are used to transport and store fresh produce may play a role in cross contamination and subsequent illnesses. However, methods for quantitatively attached cells are somewhat variable. The overall goal of this study was to compare conventional plating with molecular methods for quantitating attached representative strains for Salmonella Typhimurium and Heidelberg on reusable plastic containers (RPC) coupons, respectively. We attached Salmonella enterica serovar Typhimurium ATCC 14028 and serovar Heidelberg SL486 (parent and an antibiotic resistant marker strain) to plastic coupons (2.54 cm2) derived from previously used shipping containers by growing for 72 h in tryptic soy broth. The impact of the concentration of sanitizer on log reductions between unsanitized and sanitized coupons was evaluated by exposing attached S. Typhimurium cells to 200 ppm and 200,000 ppm sodium hypochlorite (NaClO). Differences in sanitizer effectiveness between serovars were also evaluated with attached S. Typhimurium compared to attached S. Heidelberg populations after being exposed to 200 ppm peracetic acid (PAA). Treatment with NaClO caused an average of 2.73 ± 0.23 log CFU of S. Typhimurium per coupon removed with treatment at 200 ppm while 3.36 ± 0.54 log CFU were removed at 200,000 ppm. Treatment with PAA caused an average of 2.62 ± 0.15 log CFU removed for S. Typhimurium and 1.41 ± 0.17 log CFU for S. Heidelberg (parent) and 1.61 ± 0.08 log CFU (marker). Lastly, scanning electron microscopy (SEM) was used to visualize cell attachment and coupon surface topography. SEM images showed that remaining attached cell populations were visible even after sanitizer application. Conventional plating and qPCR yielded similar levels of enumerated bacterial populations indicating a high concordance between the two methods. Therefore, qPCR could be used for the rapid quantification of Salmonella attached on RPC.
... survived for up to 4 hours (Scott & Bloomfield, 1990). Survival of serovars Enteritidis, Heidelberg and Enteritidis phage type 4 on surfaces was extended to up to 24 hours under dark conditions, but when these surfaces exposed to direct sunlight, the numbers of salmonellae serovars decreased rapidly (Nyeleti et al., 2004). Thus it is not surprising that Salmonella spp. ...
The effect of variations in temperature, UV radiation, and sunlight intensity on Escherichia coli, E. coli O157:H7, Salmonella Newport, and antibiotic resistant (ABR) variants of E. coli O157:H7 and Salmonella Newport exposed to sunlight was evaluated. Bacterial strains suspended in sterile deionized water at a concentration of 8 log CFU/mL were exposed to sunlight on three different days for 180 min; control treatments were stored in the dark. The mean temperature of 30.08 and 26.57°C on day 1 and day 3, respectively, was significantly different (P < 0.05). The UV intensity was significantly different on all 3 days, and sunlight intensity significantly differed on day 3 (P < 0.05). Bacterial population decline positively correlated with temperature, sunlight, and UV intensity. Differences in bacterial population declines differed among species, ABR profile, and day of exposure (P < 0.05). On day 1 and day 2, the populations of E. coli dropped below the limit of detection (1 log CFU/mL), whereas the percentage of live cells was 67 and 6.6%, respectively. The artificial neural network model developed to predict bacterial survival under different environmental conditions suggested that Salmonella cells were more resistant than E. coli cells. The ABR strains had significantly higher numbers of viable cells after sunlight exposure (P < 0.05). Sunlight-exposed cells resuscitated in tryptic soy broth varied in maximum population density and maximum specific growth rate based on bacterial species and presence of ABR. Morphological changes such as viable but nonculturable state transition and filament formation were detected in subpopulations of sunlight-exposed bacteria. Daily fluctuations in UV and sunlight intensity can result in significant variations in bacterial decline and recovery.
HIGHLIGHTS
Salmonellosis is the second most commonly reported zoonosis in the European Union and contaminated meat from broiler chickens (Gallus gallus) is an important source of human infection. In Great Britain (GB), prevalence of Salmonella enterica in broiler flocks is low, having declined considerably since the introduction of the Salmonella National Control Programme in 2010. However, this decreasing trend has stabilised in recent years and serovars with known ability to persistently colonise hatcheries have been isolated from broiler flocks with increasing frequency, indicating that further controls on hatchery contamination are required. The broiler industry in GB has changed dramatically over the last 15 years, with greater intensification and dominance by a small number of very large companies which rely on relatively few hatcheries. An investigation of risk factors for Salmonella contamination in GB broiler hatcheries was therefore carried out so that relevant up-to-date advice on Salmonella control can be provided. Twenty-two hatcheries, representing most commercial scale GB broiler hatcheries, were visited between 2015 and 2018. Salmonella contamination was comprehensively investigated at each hatchery by collecting between 108 and 421 environmental swab samples per hatchery (6990 samples in total from all hatcheries). An in-depth questionnaire on hatchery operations was completed for each hatchery, and results were incorporated into a risk factor analysis (univariable followed by multivariable mixed effects logistic regression) to identify factors associated with Salmonella occurrence. Overall, 6.0 % (416/6990) of environmental samples were Salmonella-positive and Salmonella was isolated from 17/22 hatcheries. Ten different serovars were isolated, the most common being S. Senftenberg and S. Mbandaka which are known hatchery colonisers. Sixty-four risk factor variables were investigated. Twenty-two of these were initially retained based on univariable analyses (p ≤ 0.25) and six were ultimately left in the final multivariable model (p ≤ 0.05). Salmonella detection was positively associated with having ≥30 hatchers in regular use compared to fewer (Odds ratio [OR] 23.7, 95 % confidence interval [CI] 6.7-84.2), storing trays in process rooms (OR 28.8, CI 7.8-106.3), drying set-up trolleys in corridors (OR 15.6, CI 5.9-41.4) and having skips located in enclosed areas (OR 8.99, CI 5.89-41.35). Using a closed waste disposal system was negatively associated with Salmonella detection (OR 0.08, CI 0.04-0.18) and the odds of detecting Salmonella in hatcheries with 31-60 total workers was lower compared to hatcheries with ≤30 staff (OR 0.16, CI 0.06-0.40). Despite the complexities of hatchery enterprises, changes to a relatively small number of features may significantly reduce the occurrence of hatchery contamination.
Pre-harvest contamination of produce has been a major food safety focus. Insight into the behavior of enteric pathogens on produce in pre-harvest conditions will aid in developing pre-harvest and post-harvest risk management strategies. In this study, the attachment strength (SR) and die-off rate of E. coli on the surface of watermelon fruits and the efficacy of aqueous chlorine treatment against strongly attached E. coli population were investigated. Watermelon seedlings were transplanted into eighteen plots. Prior to harvesting, a cocktail of generic E. coli (ATCC 23716, 25922 and 11775) was inoculated on the surface of the watermelon fruits (n = 162) and the attachment strength (SR) values and the daily die-off rates were examined up to 6 days by attachment assay. After 120 h, watermelon samples were treated with aqueous chlorine (150 ppm free chlorine for 3 min). The SR value of the E. coli cells on watermelon surfaces significantly increased (P<0.05) from 0.04 to 0.99 in the first 24 h, which was primarily due to the decrease in loosely attached population, given that the population of strongly attached cells was constant. Thereafter, there was no significant change in SR values, up to 120 h. The daily die-off rate of E. coli ranged from -0.12 to 1.3 log CFU/cm2. The chlorine treatment reduced the E. coli level by 4.2 log CFU/cm2 (initial level 5.6 log CFU/cm2) and 0.62 log CFU/cm2 (initial level 1.8 log CFU/cm2), on the watermelons that had an attachment time of 30 min and 120 h respectively. Overall, our findings revealed that the population of E. coli on watermelon surfaces declined over time in an agricultural environment. Microbial contamination during pre-harvest stages may promote the formation of strongly attached cells on the produce surfaces, which could influence the efficacy of post-harvest washing and sanitation techniques.
Outbreaks associated with fresh produce have been traced to farms in several cases. Potential sources of contamination in preharvest environments have been identified, and minimizing their input is needed. In addition, understanding the fate of enteric pathogens introduced to soil and plant systems is essential to providing safe guidelines on when crops may be planted and harvested. Moisture availability and temperature are key abiotic factors affecting pathogen survival. Indigenous soil and epiphytic bacteria, however, also appear to play an important role in a pathogen's fate and thus future survival studies should routinely monitor the types and levels present. Internalization of enteric pathogens through lateral root junctions or through leaf stomata has been documented but generally requires high exposure concentrations. Plant defenses, whether basal or activated by the invading enteric pathogen, appear to inactivate internalized populations as persistence has not been observed, but this subject deserves further investigation.
The study was carried out to evaluate and compare the immune responses in mice experimentally infected with either wild-type or isogenic mutants of Salmonella enterica serovars Enteritidis (SE), Salmonella Typhimurium (ST) and Gallinarum (SG). The mutant strains were constructed by allelic replacement of some virulence-associated genes in the wild-type strains. Seven-week-old female BALB/c mice were orally or intraperitoneally inoculated by injecting bacterial suspension. To evaluate the immune responses, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot (ELISPOT) assay were conducted with serum and fecal samples. As a result, the mice group infected orally with the SE mutant strain showed the highest level of specific IgA-secreting splenocytes, compared to the other groups. The peritoneally injected groups showed the greater levels of IgG1 than the orally injected groups, which was in a good agreement with the previous studies. In addition, the mutant infected groups had the similar secretion levels of antibodies with the wild-type infected groups. These results demonstrated that the SE mutant strain elicited humoral immune response as much as wild-type, implying that it can be useful as a delivery vehicle as well as a candidate of a live attenuated vaccine.
This study was focusing on evaluating the protection of polyphosphate kinase (ppk) deleted and/or temperature-sensitive (ts) Salmonella Enteritidis (SE) as an attenuated vaccine in chickens. We constructed SEppk, SEts and SEppk::ts mutants and screened those mutants by growth capability in vitro, protection study in mice model and antibody response in chickens. Among the mutants, SEppk::ts-3 was selected because it showed higher growth capability, good protection against highly virulent SE in mice model, and good antibody response in chickens. SEppk::ts-3 also showed good protection against highly virulent SE isolate because it decreased colonization of virulent SE challenge strain in spleen, liver and cecum compared with the non-vaccinated control. The SEppk::ts-3 mutant showed cross-protection against S. Gallinarum (SG) challenge although the its cross-protection rate was a little lower than that of SG9R, a commercial vaccine against SG infection. To use for live attenuated vaccine in chickens, it should further be characterized.
Salmonella enterica subsp. enterica serotype Enteritidis is one of the major causes of gastroenteritis in humans due to consumption of poultry derivatives. Here
we report the whole-genome sequence and annotation, including the virulence plasmid, of S. Enteritidis LA5, which is a chicken isolate used by numerous laboratories in virulence studies.
An ecological model for transmission of Salmonella enterica in swine production ecosystems was developed, identifying host species, environmental reservoirs, and temporal, spatial, and functional (i.e., stage of production) dimensions. It was hypothesized that transmission was most likely within spatial and functional compartments, between hosts of the same species and abiotic compartments of the same type. Eighteen swine production systems in Illinois, USA, were sampled in four collection cycles (1998, 1999, 2000, 2003). There were 11,873 samples collected, including feces from swine and other mammals and birds, and samples from insects, pen floors, boots, feed, and water. The 460 Salmonella isolates obtained were genotyped using repetitive sequence PCR with three primers-REP, BOX, and ERIC. All isolates from 2000 and 2003 were serotyped, as well as a subsample from 1998 and 1998. Genetic relatedness was estimated from the similarity of fragmentation patterns after gel electrophoresis of PCR products. Cluster analysis identified genetically related isolates. Linking of isolates in tight clusters (similarity >or=85%) was viewed as evidence for transmission. Five farms had a sufficient number of tight clusters for hypothesis testing. The factors most differentiating isolates genetically were farm of origin and time of sampling. Isolates were also differentiated genetically by site, building, room, and pen. There was no consistent association of genotype with stage of production or host/environment reservoir. Serotype analysis confirmed that Salmonella lineages were differentiated by visit and site. Thus, Salmonella transmission was primarily over short distances, i.e., within the same pen or room, with some transmission between rooms and buildings on the same site, but with limited transmission between sites. Transmission was observed across a variety of ecological niches represented by different host species and environmental reservoirs. Genetic differences over time reflected multiple introductions into the ecosystem of different Salmonella genotypes, as well as evolutionary changes within lineages. Intervention strategies to reduce Salmonella prevalence within swine production ecosystems would be best targeted at maintaining spatial barriers to transmission, whereas intervention targeted at specific biological hosts or environmental reservoirs is less likely to be effective.
Aerosols constitute a major route of transmission for a wide range of infectious diseases in the hospital setting. The aim of this study was to determine the survival of Staphylococcus aureus on a light-activated antimicrobial coating. S. aureus suspended in phosphate-buffered saline (PBS), saliva, or horse serum was sprayed onto cellulose acetate coatings containing toluidine blue O and rose bengal and the survival of the organism on these surfaces was determined following 6 h of exposure to a 28-W domestic fluorescent lamp (light intensity = 3700 +/- 20 lux). Kills ranging from 78.9% (in horse serum) to 99.8% (in PBS) were obtained when the bacterial density on the coatings was approximately 10(5) colony-forming units/m(2). The results of this study have shown that a coating containing toluidine blue and rose bengal can achieve significant kills of S. aureus when illuminated by a domestic light source. Light-activated coatings could provide a simple, low-cost means of reducing the microbial load in hospitals and other facilities.
A study was carried out in a poultry processing plant in Lusaka, Zambia, to identify and describe bacteria found in chicken carcasses leaving the processing plant for retail outlets. The thirteen different bacteria found in the chicken carcasses included: Escherichia coli (41.7%), Staphylococcus spp. (2.49%), Pseudomonas spp. (6.71%), Klebsiella spp. (1.91%), Salmonella spp. (20.53), Citrobacter spp. (6.71%), Acinetobacter spp. (0.58%), Proteus spp. (9.02%), Flavobacterium spp. (1.15%), Streptococci spp. (1.72%), Alcaligenes spp. (0.77%), Micrococcus spp. (3.84%) and Bacillus spp. (2.88%). These results showed that the chicken carcasses entering the Zambian market are a potential source of bacteria pathogens to consumers. It is therefore suggested that much more attention should be paid to the processing plant in order to control the bacterial contamination of poultry meat.
The stability of the natural populations of fecal coliforms and fecal streptococci in raw sewage diluted 1:1,000 in seawater or phosphate-buffered water at 24 +/- 2 degrees C was markedly affected by the absence or presence of sunlight. In the absence of sunlight, these bacteria survived for days, whereas in the presence of sunlight 90% of the fecal coliforms and fecal streptococci were inactivated within 30 to 90 min and 60 to 180 min, respectively. The bactericidal effect of sunlight was shown to penetrate glass, translucent polyethylene, and at least 3.3 m of clear seawater, suggesting that the visible rather than the ultraviolet light spectrum of sunlight was primarily responsible for the observed bactericidal effect. However, these same sewage-borne bacteria were relatively resistant to the bactericidal effect of sunlight when diluted in fresh mountain stream waters. These results indicate that the presence of sunlight is a major factor controlling the survival of fecal coliforms and fecal streptococci in seawater.
To gain an understanding of the role of fimbriae and flagella in the adherence of Salmonella enterica serotype Enteritidis to inanimate surfaces, the extent of adherence of viable wild-type strains to a polystyrene microtitration plate was determined by a crystal violet staining assay. Elaboration of surface antigens by adherent bacteria was assayed by fimbriae- and flagella-specific ELISAs. Wild-type Enteritidis strains adhered well at 37 degrees C and 25 degrees C when grown in microtitration wells in Colonisation Factor Antigen broth, but not in other media tested. At 37 degrees C, adherent bacteria elaborated copious quantities of SEF14 fimbrial antigen, whereas at 25 degrees C adherent bacteria elaborated copious quantities of SEF17 fimbrial antigen. Non-fimbriate and non-flagellate knock-out mutant strains were also assessed in the adherence assay. Mutant strains unable to elaborate SEF14 and SEF17 fimbriae adhered poorly at 37 degrees C and 25 degrees C, respectively, but adherence was not abolished. Non-motile mutant strains showed reduced adherence whilst type-1, PEF and LPF fimbriae appeared not to contribute to adherence in this assay. These data indicate that SEF17 and SEF14 fimbriae mediate bacterial cell aggregation on inanimate surfaces under appropriate growth conditions.
The aim of this study was to verify the efficiency of UV irradiation in the inactivation of Salmonella spp. in treated wastewater with different levels of turbidity and exposed to increasing doses of UV irradiation. Experiments were carried out in a batch reactor and in a real scale reactor. Salmonellae obtained from clinical samples were seeded into autoclaved wastewater collected from a wastewater treatment plant (WWTP) comprising an association of a UASB reactor followed by three submerged aerated biofilters (BAF) and one tertiary filter. The results showed that salmonellae were not inactivated in effluents from the UASB reactor indicating that the presence of suspended solids was an important obstacle to UV penetration in bacteria. However, UV irradiation was efficient in inactivating Salmonella of effluents from aerated secondary and tertiary biofilm reactors.
The rate of die-off of coliform bacteria in samples of sea water exposed to daylight has been found to be very much greater than in the dark. Consequently, the possibility that sunlight has an important effect on bacterial mortality in the sea merits consideration. The paper discusses results and implications of some shore-based studies, then examines the findings of in situ experiments. Most of the work is in terms of coliform bacteria, enumerated by a membrane-filtration method using a recovery medium consisting of a lactose-peptone broth containing 3 percent Teepol 610 with phenol red as indicator and with incubation for 18 h at 37 degree C. Some work with other organisms is also discussed.
Escherichia coli and some salmonellas were exposed in seawater and freshwater to natural sunlight, visible light of comparable intensity, and light containing a similar proportion of u.v. as natural sunlight but of a much lower intensity. Direct viable bacterial counts and culturable counts on selective and non-selective media were made at intervals. The rate of decrease in numbers of culturable bacteria was significantly faster in seawater than in freshwater when exposed to natural sunlight. No significant difference was found between the rates of decrease in numbers of culturable bacteria in seawater and those in freshwater when bacteria were exposed to light with a small u.v. component of similar intensity. The effect of salinity no loss of culturability is, therefore, more significant in the presence of u.v. radiation. Direct counts by the acridine orange direct viable count method decreased much more slowly than the culturable counts in seawater but comparably with culturable counts in freshwater in natural sunlight. Direct viable counts and culturable counts decreased at a similar rate in seawater and in freshwater in visible light. This may signify the evolution of enteric bacteria towards a viable but non-culturable form in seawater when exposed to natural sunlight. The presence of humic acids significantly reduced loss of culturability but only in low salinity conditions. Salinity appears to be an important factor influencing culturability in bacteria exposed to sunlight.
Among the 23 Salmonella strains isolated from 21 diseased chicken and 2 embryonated eggs in 6 poultry farms near Lusaka City in Zambia, serovars identified were S. Gallinarum (11 strains), S. Agona (7 strains), S. Alamo (1 strain), S. Infantis (1 strain), S. Virginia (1 strain), S. Haifa (1 strain), and S. Dublin (1 strain). S Gallinarum was detected at the highest incidence and from all the poultry farms. Fourteen serovars have been reported for the chickens in Zambia so far. Three serovars (S. Alamo, S. Haifa, S. Virginia) were newly identified in this study.
To determine whether diurnal and seasonal variations in campylobacters in surface waters result from the effects of temperature and u.v. radiation, and whether natural populations of Campylobacter lari and urease-positive thermophilic campylobacters (UPTC) from birds survive better in surface waters than Camp. jejuni from sewage.
Natural populations of Camp. lari and UPTC in sea water, and Camp. jejuni in river water, were exposed to artificial sunlight (equivalent to a sunny day in June). Both populations became non-culturable within 30 min, with T90s of 15 min and 25 min, respectively. Cultures of Camp. jejuni became non-culturable within 40 min and those of Camp. coli, Camp. lari and UPTC, within 60 min. In darkness, survival was temperature-dependent. Natural populations took 12 h at 37 degrees C and 5 days at 4 degrees C to become non-culturable in sea water, and slightly less in river water. Cultures of Camp. lari and UPTCs survived for significantly longer than Camp. jejuni and Camp. coli. Loss of culturability for all isolates was most rapid at 37 degrees C and slowest at 4 degrees C. Newly isolated strains from sea water and river water behaved in an almost identical manner to NCTC strains.
Campylobacter lari and UPTCs survive for longer in surface waters than Camp. jejuni and Camp. coli, particularly in the dark. Low Campylobacter numbers in coastal waters in the summer, especially in the afternoon, are due to the combined effects of higher temperatures and higher levels of u.v. radiation.
Campylobacter lari and UPTCs from birds predominate in bathing waters in Morecambe Bay because they are better able to survive; they also originate from closer to the shore than Camp. jejuni and Camp. coli in sewage effluent, which survive poorly and die before the incoming tide reaches the shore. The predominance of Camp. jejuni in river water results from its dominance of the inputs and not from its ability to survive.
Three Salmonella enterica serovar Orion var. 15+ isolates of distinct provenance were tested for survival in various stress assays. All were less able to survive desiccation than a virulent S. Enteritidis strain, with levels of survival similar to a rpoS mutant of the S. Enteritidis strain, whereas one isolate (F3720) was significantly more acid tolerant. The S. Orion var. 15+ isolates were motile by flagellae and elaborated type-1 and curli-like fimbriae; surface organelles that are considered virulence determinants in Salmonella pathogenesis. Each adhered and invaded HEp-2 tissue culture cells with similar proficiency to the S. Enteritidis control but were significantly less virulent than S. Enteritidis in the one-day-old and seven-day-old chick model. Given an oral dose of 1 x 10(3) cfu to one-day-old chicken, S. Orion var. 15+ isolates colonised 25% of liver and spleens examined at 24 h whereas S. Enteritidis colonised 100% of organs by the same with the same dose. Given an oral dose of 1 x 10(7) cfu at seven-day old, S. Orion var. 15+ failed to colonise livers and spleens in any bird examined at 24 h whereas S. Enteritidis colonised 50% of organs by the same with the same dose. Based on the number of internal organs colonised, one of the three S. Orion var. 15+ isolates tested (strain F3720) was significantly more invasive than the other two (B1 and B7). Also, strain F3720 was shed less than either B1 or B7 supporting the concept that there may be an inverse relationship between the ability to colonise deep tissues and to persist in the gut. These data are discussed in the light that S. Orion var. 15+ is associated with sporadic outbreaks of human infection rather than epidemics.
Effects of intensity and processing time of 254 nm UV irradiation on Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Typhimurium were investigated. Intensities measured at 5.08, 10.1, 15.2, and 20.3 cm from the light source were 1.000, 500, 250, and 150 microW/cm2, respectively. Intensities of 250 or 500 microW/cm2 reduced all suspended pathogen cells in peptone water about 5 log cycles after 2 min and completely inactivated L. monocytogenes and E. coli O157:H7 after 3 min by reductions of 8.39 and 8.64 log cycles, respectively. Intensities of 250 or 500 microW/cm2 also reduced (P < or = 0.05) the tested pathogens inoculated on stainless steel (SS) chips, and E. coli O157:H7 was completely destroyed at 500 microW/cm2 for 3 min. After UV treatment for 3 min at 500 microW/cm2, all selected pathogens on chicken meat with or without skin showed reduction ranges from 0.36 to 1.28 log cycles. Results demonstrated that UV irradiation could effectively decrease pathogens in peptone water and on SS but that it was less effective on chicken meat.
The persistence of S. Enteritidis PT4 was studied on a free-range breeding chicken farm which had been depopulated following identification of the organism in breeding birds. The site was sampled periodically for 26 months after depopulation and the organism was found to persist in litter, dried faeces and feed, but not in dust within empty poultry houses, for the whole of that period. Salmonella Enteritidis PT4 was also found in soil samples after 8 months but not 13 months and in faeces from wild mice, foxes and cats but not wild birds or badgers. The organism was also found in adult and larval forms of ground beetles and centipedes. Addition of pullets to a contaminated pen or inclusion of contaminated litter, feed or beetles/larvae to feed did not result in acquisition of infection by birds.
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