Food Microbiology 50 (2015) 1e4

Contents lists available at ScienceDirect

Food Microbiology
journal homepage: www.elsevier.com/locate/fm

Inactivation of avirulent Yersinia pestis on food and food contact

surfaces by ultraviolet light and freezing*
Christopher H. Sommers*, Shiowshuh Sheen
U.S. Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, 600 East Mermaid Lane, Wyndmoor, PA 19038, USA

a r t i c l e i n f o a b s t r a c t

Article history: Yersinia pestis, the causative agent of plague, can occasionally be contracted as a naso-pharyngeal or
Received 31 July 2014 gastrointestinal illness through consumption of contaminated meat. In this study, the use of 254 nm
Received in revised form ultraviolet light (UV-C) to inactivate a multi-isolate cocktail of avirulent Y. pestis on food and food contact
16 January 2015
surfaces was investigated. When a commercial UV-C conveyor was used (5 mW/cm2/s) 0.5 J/cm2 inac-
Accepted 14 February 2015
tivated >7 log of the Y. pestis cocktail on agar plates. At 0.5 J/cm2, UV-C inactivated ca. 4 log of Y. pestis in
Available online 21 February 2015
beef, chicken, and catfish, exudates inoculated onto high density polypropylene or polyethylene, and
stainless steel coupons, and >6 log was eliminated at 1 J/cm2. Approximately 1 log was inactivated on
Keywords:
Yersinia pestis
chicken breast, beef steak, and catfish fillet surfaces at a UV-C dose of 1 J/cm2. UV-C treatment prior to
Ultraviolet freezing of the foods did not increase the inactivation of Y. pestis over freezing alone. These results
Food indicate that routine use of UV-C during food processing would provide workers and consumers some
Food contact surfaces protection against Y. pestis.
Beef Published by Elsevier Ltd.
Chicken
Catfish

1. Introduction processing intervention technologies to inactivate Y. pestis in food

products.
Yersinia species including Yersinia enterocolitica, Yersinia pseu- Recent studies have investigated the ability of Y. pestis to grow in
dotuberculosis, and Yersinia pestis are closely related species which foods, including growth at refrigeration temperature. Bhaduri and
can cause illness in humans (Paoli and Sommers, 2014). Of those 3, Phillips (2013) found that Y. pestis KIM5 did not grow in refriger-
Y. pestis is the causative agent of plague and can cause pharyngeal ated ground pork (4
C), but was able to grow at the mild refrig-
or gastrointestinal infection through the handling or consumption eration abuse temperatures of 10 and 15
C at ca. 0.05 and 0.16 log10
of meat products (Leslie et al., 2010; Albaji et al., 2005; Bin Saeed CFU/h, respectively. Y. pestis KIM5 readily grew at 20, 25 and 30
C
et al., 2005; Christie et al., 1980). The risk, and the associated at 0.26, 0.30 and 0.77 log10 CFU/h. Sommers and Cooke (2009)
morbidity and mortality, of contracting plague through the con- found that Y. pestis was capable of growth on frankfurters at
sumption or handling of foods deliberately contaminated with 10
C, while Gurtler et al. (2010) found Y. pestis capable of growth in
Y. pestis are currently unknown. Because Y. pestis is listed as a select refrigerated liquid egg. Torosian et al. (2009) found that Y. pestis
agent for both food safety and food defense (Brickner et al., 2003; was capable of growth in Brain Heart Infusion Broth at 4
C.
Khan et al., 2000; Kauffman et al., 1997; WHO, 1970) it would Other studies investigated the use of both thermal and non-
therefore be prudent to evaluate the efficacy of nonthermal food thermal intervention technologies to inactivate Y. pestis in foods.
Sommers and Niemira (2007) investigated the use of gamma ra-
diation at both refrigeration (D10 0.19 at 10
C) and frozen tem-
*
Mention of trade names or commercial products in this publication is solely for
peratures (D10 0.37 kGy at 20
C) and developed a predictive
the purpose of providing specific information and does not imply recommendation equation to describe the temperature dependent radiation inacti-
or endorsement by the U.S. Department of Agriculture. USDA is an equal oppor- vation kinetics of Y. pestis in ground meat. Sommers and Cooke
tunity provider and employer. (2009) found the gamma radiation D10 of a multi-isolate cocktail
* Corresponding author. Food Safety and Intervention Technologies Research
of Y. pestis inoculated onto phosphate buffer and frankfurters to be
Unit, Eastern Regional Research Center, USDA-ARS, 600 East Mermaid Lane,
Wydnmoor, PA 19038, USA. 0.23 and 0.31 kGy, respectively. Porto-Fett et al. (2009) determined
E-mail address: [email protected] (C.H. Sommers). the thermal D10 value of Y. pestis KIM5 suspended in 75% lean

http://dx.doi.org/10.1016/j.fm.2015.02.008
0740-0020/Published by Elsevier Ltd.
2 C.H. Sommers, S. Sheen / Food Microbiology 50 (2015) 1e4

ground beef at 48.9e60
C to range from 193 to 0.56 min, respec- then resuspended as a cocktail in 30 mL Butterfield's Phosphate
tively, with a z-value of 4.57
C. Buffer (BPB) (Applied Research Institute, Newtown, CT).
Ultraviolet light (UV-C, 254 nm), a U.S. Food and Drug Admin- One hundred mL (0.1 mL) of the Y. pestis undiluted cocktail was
istration (FDA) approved food safety intervention technology, can plated onto duplicate BHIA plates which were then allowed to dry
be used for decontamination of food and food contact surfaces (CFR, for approximately 5 min. The agar plates were then exposed to 0.5 J/
2005). UV-C exerts its bacteriocidal effect primarily through the cm2 UV-C using the UV-C conveyor as described below.
formation of DNA adducts including cyclobutane pyrimidine di- For experiments using exudates on high density polypropylene
mers and 6e4 photoproducts, either killing the bacteria or (HDPP) and polyethylene (HDPE) and stainless steel coupons (bead
rendering them unable to reproduce, and to some extent through blasted and electropolished) 0.1 mL of Y. pestis cocktail was pipetted
oxidation of proteins in the bacterial cell (Reardon and Sancar, into 0.9 mL of the sterile food exudates, mixed by vortexing for ca.
2005; Krisko and Radman, 2010). The technology has been 10 s, and 0.1 mL transferred to the chilled (4
C) food contact sur-
increasingly adopted by the health care industry for decontami- faces and exposed to UV-C as described below. Prior to experi-
nation of surfaces in medical facilities and in recent years the food mentation stainless steel coupons were sterilized by autoclaving
processing industry has expressed significant interest in using it for while HDPP and HDPE were cleaned with 70% ethanol and then
surface decontamination of foods and food contact surfaces in the subjected to 10 J/cm2 UV-C.
food processing environment (Salvage, 2014; Rutala et al., 2010; Chicken, fish, and beef pieces were inoculated on one side of the
Koutchma, 2008; Koutchma et al., 2008). UV-C is of particular ap- surface with 1.0 mL (ca. 106 CFU/ml), which was then spread on the
peal to the medical and food processing industries because it is a food surface (3  3 cm) using a sterile inoculation loop. The inoc-
green and sustainable technology which does not require the use of ulated pieces were allowed to sit in a refrigerator (4
C) for ca. 30
water or chemicals to exert its bacterial effects. min, and then passed through the conveyor to obtain the required
While a great deal of information is available on the use of ul- UV-C doses (0.25, 0.5, 1.0 and 2.0 J/cm2).
traviolet light (UV-C) to inactivate common foodborne pathogens Following exposure to UV-C, food samples were placed in sterile
(Sommers et al., 2010; Sumner et al., 1996; Stermer et al., 1987), polynylon bags (Uline, Inc., Philadelphia, PA) containing 100 ml BPB
relatively little information is available on the inactivation of and rinsed manually by massaging for ca. 30 s. The solution was
Y. pestis on foods with ultraviolet light. The purpose of this study then serially diluted in 9 ml BPB and two 0.1 mL aliquots per
was to determine the ability of Y. pestis to survive treatment with dilution were spread on BHI agar plates (BD-Difco, Sparks, MD). The
UV-C on agar plates, food contact surfaces, and foods using a plates were then incubated for 3 days at 30
C prior to enumeration
commercial UV-C conveyor. of bacterial colonies. Each experiment was conducted indepen-
dently a minimum of 3 times (n ¼ 3).
2. Materials and methods
2.4. Exposure to ultraviolet light (UV-C)
2.1. Food products
A commercial food-grade UV-C conveyor (Reyco Systems, Inc.,
Catfish fillets (2e3 oz) were obtained from the USDA-ARS Cat- Meridan, ID) was used to treat exudates inoculated with avirulent
fish Genetics Laboratory in Stoneville, Mississippi. Beef steaks (3e4 Y. pestis on surfaces of BHI agar plates, stainless steel coupons, HDPP
oz) and boneless skinless chicken breast fillets (3e4 oz) were ob- and HDPE, and the foods themselves. The UV-C intensity was
tained fresh from a local butcher. The meat, poultry, and ready-to- 5 mW/s/cm2, and 100 s of exposure provided a total UV-C dose of
eat meat products were gamma irradiated to an absorbed dose of approximately 0.5 J/cm2. All experiments were conducted in a BSL-
5 kGy at 4
C to inactivate background microflora, which was 2 laboratory.
reduced to less than to <0.1 CFU/g as determined by plating on The exudates (0.1 mL, ca. 107 CFU) were inoculated onto BHI agar
Brain Heart Infusion agar (BD-Difco, Sparks, MD). Exudate (drip/ plates, stainless steel or polypropylene coupon surfaces were run
purge) was obtained from the irradiated products by pouring through the conveyor either once, twice or four times to obtain ex-
excess liquid into sterile test tubes. Food was stored at 0e4
C until posures of 0.5 J/cm2, 1.0 J/cm2 or 2.0 J/cm2, respectively. Beef steaks,
ready for use, while exudates were stored at 20
C. chicken breast, and catfish fillets were passed through the conveyor
up to 4 times to obtain the maximum UV-C dose of 2.0 J/cm2.
UV-C intensity was monitored using a calibrated UVX Radiom-
2.2. Avirulent Yersinia pestis isolates eter (UVP Inc., Upland, CA). The temperature of the room was
approximately 15
C during the exposure to UV-C, and the food
Four avirulent Y. pestis strains (KUMA, Yokohama, KIM5, and temperature did not increase to more than 15
C at the end of the
CO99) were obtained from Dr. Susan Straley (Univ. Kentucky) and process as measured using an infrared thermometer.
Dr. Robert Brubaker (Mich. State Univ.) through Dr. George Paoli at
USDA's Eastern Regional Research Center (Wyndmoor, PA). The 2.5. Freezing
Y. pestis strains were propagated on Brain Heart Infusion Agar
(BHIA) (BD-Difco, Sparks, MD) at 30
C for approximately 3 days Catfish fillets, chicken breasts, and beef steaks were inoculated
and maintained at 0e4
C until ready for use. and treated with UV-C as described above and then frozen using
dry ice. The individual samples were then packaged in polynylon
2.3. Propagation and inoculation bags (Uline Inc., Philadelphia, PA), sealed using a Multi-Vac 300
packager to ca. 5000 Pa and stored at 20
C for ca. 30 days. To
Each Y. pestis strain was cultured independently in 30 ml BHI assess bacterial counts after freezing, the samples were thawed in a
Broth (Brain Heart Infusion, BD-Difco, Sparks, MD) in a sterile 50- refrigerator (ca. 4
C) overnight.
mL conical tube at 30
C (150 rpm) for 36 h, to a density of
approximately 108 CFU/ml, using a New Brunswick Model G24 2.6. Statistical analysis
Incubator (Edison, NJ). The cultures were then sedimented by
centrifugation (1200  g) using a Fisher Scientific Marathon Log reductions were determined according to Diehl (1995). Each
21000R Centrifuge (Needham Heights, MA). The cell pellets were experiment was conducted independently a minimum of 3 times.
 C.H. Sommers, S. Sheen / Food Microbiology 50 (2015) 1e4 3

Statistical analysis functions of MS Excel (Microsoft Corp., Red-

mond, WA) were used for routine calculations, descriptive statis-
tics, and ANOVA.

3. Results and discussion

In this study the use of UV-C light to inactivate a multi-isolate

cocktail of Y. pestis on agar plates, food contact surfaces, and
foods using a commercial UV-C conveyor was investigated. Rose
and O'Connell (2009) found that a UV-C dose of 4 mJ/cm2 inacti-
vated 4.9 and 4.4 log of Y. pestis A112 and Harbin suspended in
sterile distilled water, respectively, indicating a D10 of ca.
0.5e1.0 mJ/cm2 Paoli and Sommers (2014) obtained D10 of
0.69e1.02 mJ/cm2 for Y. pestis KIM5 or Yokohama, which either
contained or lacked the high pathogenicity island encoding several
proteins necessary or iron acquisition (PGMþ and Dpgm), on Congo
Red Agar Plates. In both of those studies low intensity lab-scale
(<100 uW/cm2) UV-C sources were used to determine D10. When
Y. pestis was inoculated onto agar plates and treated with 0.5 J/cm2
UV-C at an intensity of 5 mW/cm2/s using a UV-C conveyor >7 log Fig. 1. Inactivation of avirulent Yersinia pestis on foods by ultraviolet light (UV-C). UV-C
were inactivated. doses for beef (B), chicken (C), catfish (D)were 0.25, 0.5, 1.0 and 2.0 J/cm2. Each
experiment was conducted independently 3 times. Standard deviation is shown as
The next set of experiments investigated the ability of UV-C to
error bars.
inactivate Y. pestis suspended in exudates (drip) from chicken
breast, beef steak, and catfish fillets. Because the exudates was
relatively transparent, UV-C was able to fully penetrate (>95%) all of processes. The use of UV-C allows a waterless and chemical-free
the exudates to a depth of 2 mm. When the exudates were placed method of decontaminating food contact surfaces such as HDPE,
on bead blasted, electropolished, HDPE, and HDPP coupons, ca. 4 HDPP, and stainless steel, or more specifically the purge from the
log inactivation of Y. pestis was obtained, with no significant dif- food products which lie on the food contact surfaces. Our results
ference (p > 0.05) in inactivation for the type of food contact surface demonstrate the potential of UV-C for decontamination of purge
used or exudate used (Table 1). A >6 log reduction was obtained on and food contact surfaces. To date there has been little published
bead blasted, electropolished stainless steel, HDPE, and HDPP at a data on the use of UV-C to inactivate microorganisms suspended in
dose of 1.0 J/cm2, which was essentially a complete inactivation purge which contaminates food contact surfaces. Sommers and
(data not shown). The lower inactivation levels compared to agar Gunther demonstrated a >5 log reduction of Campylobacter jejuni
plates could be due to shadowing from particulate matter within suspended in poultry exudate on food contact surfaces by UV-C
the exudates. (Sommers and Gunther, 2013) These results are similar to UV-C
Inactivation of microorganisms on actual foods is much more inactivation obtained in similar experiments using the biothreat
difficult due to food surface topology and shadowing, which shields agent F. tularensis (Sommers et al., 2012), in which a >5 log
them from UV-C light (Gardner and Shama, 2000). In previous work reduction of F. tularensis was obtained in purge from meat, poultry,
in our laboratory we obtained ca. 0.5 log reduction of Salmonella, and catfish. It may be useful to examine the use of UV-C in com-
Staphylococcus aureus, Listeria monocytogenes, or Francisella tular- bination with natural antimicrobials for inactivation of microor-
ensis on previously frozen chicken meat, beef steak, or catfish fillets ganisms within food product exudate in the future.
(Sommers et al., 2012, 2010). In this study, refrigerated (never Because beef, chicken and fish can be frozen either before or
frozen) catfish, poultry, and beef were used to avoid potential after packaging, we wanted to determine the effect of UV-C appli-
surface topology changes due to freezing and freezer burn. In this cation prior to freezing. Previous studies with other foodborne
study ca. 0.75e1 log reductions were obtained at 1 J/cm2 UV-C pathogens have indicated a ca. 1e2 log reduction as a result of
(Fig. 1). While slightly greater inactivation was obtained at 2 J/ freezing (Rajkowski and Sommers, 2012; Sommers et al., 2011). In
cm2, there was no statistical significance as determined by ANOVA this study a ca. 1 log reduction of Y. pestis was obtained regardless of
(p > 0.05). As in our previous studies, there is no advantage to food matrix type (p > 0.05) (Table 2). As is shown in Fig. 1, a ca. 1 log
applying UV-C doses in excess of 1 J/cm2 to the meat, poultry and reduction of Y. pestis was obtained regardless of food matrix type
fish products. after 1 J/cm2 UV-C. There was no additional log reduction as a result
However, contamination of foods with microorganisms is also of UV-C exposure followed by freezing (p > 0.05) (Table 2), how-
due to transfer from food contact surfaces to the actual foods during ever, we did note that recovery of the Y. pestis required an addi-
tional day of incubation on the Brain Heart Infusion Agar.
Table 1
UV-C (0.5 J/cm2) inactivation of avirulent Y. pestis suspended in exudates on stainless
steel and plastic. Table 2
Effect of freezing on survival of avirulent Y. pestis with and without UV-C treatment.
Catfish Beef Chicken
Catfish Beef Chicken
Bead Blasted-SS 4.04 (±0.06) 3.91 (±0.11) 3.87 (±0.23)
Electropolished-SS 3.94 (±0.14) 4.03 (±0.29) 3.71 (±0.12) UV-C (1 J/cm2) 1.10 (±0.03) 1.12 (±0.06) 1.07 (±0.09)
HDPE 3.99 (±0.21) 3.78 (±0.16) 3.83 (±0.09) Freezing 1.03 (±0.02) 0.96 (±0.04) 1.07 (±0.08)
HDPP 4.01 (±0.26) 3.76 (±0.13) 3.75 (±0.19) UV-C þ Freezing 1.21 (±0.10) 1.20 (±0.05) 1.04 (±0.09)

Each experiment was conducted independently 3 times. Values are log10 reductions Each experiment was conducted independently 5 times. Values are log10 reductions
as compared to the untreated controls with the standard error of the mean in as compared to the untreated controls with the standard error of the mean in
parenthesis. There was no statistical difference in the log10 reductions as deter- parenthesis. There is no statistical difference in the log10 reductions as determined
mined by ANOVA (p > 0.05). by ANOVA (p > 0.05).
4 C.H. Sommers, S. Sheen / Food Microbiology 50 (2015) 1e4

4. Conclusions Koutchma, T., 2008. UV-C light for processing foods. IUVA News 8, 24e28.
Krisko, A., Radman, M., 2010. Protein damage and death by radiation in Escherichia
coli and Deinococcus radiodurans. PNAS (USA) 107 (32), 14373e14377.
Using a UV-C conveyor system we were able to demonstrate a Leslie, T., Whitehouse, C., Yingst, S., Baldwin, C., Kakar, F., Moflen, J., Hami, A.,
4e6 log reduction of avirulent Y. pestis suspended in beef, chicken, Mustafah, L., Omar, F., Ayazi, E., Rossi, C., Noormad, B., Ziar, N., Kakar, R., 2010.
and poultry drip (purge) on stainless steel and plastic surfaces at a Outbreak of gastroenteritis caused by Yersinia pestis in Afghanisatan. Epidemiol.
Infect. 139, 1e8.
dose of 0.5e1.0 J/cm2. A ca. 1 log reduction of avirulent Y. pestis was Paoli, G., Sommers, C., 2014. Inactivation of avirulent pgmþ and Dpgm Yersinia
obtained on boneless skinless chicken breasts, beef steaks, and pestis by ultraviolet light (UV-C). Food Microbiol. 44, 168e172.
catfish fillets. These results are consistent with other studies indi- Porto-Fett, A., Juneja, V., Tamplin, M., Luchansky, J., 2009. Validation of cooking
times and temperatures for thermal inactivation of Yersinia pestis strains KIM5
cating that use of UV-C can significantly decontaminate exudates and CDC-A1122 in irradiated ground beef. J. Food Prot. 72 (3), 564e571.
on food contact surfaces, and foods, during processing. Rajkowski, K., Sommers, C., 2012. Effect of trisodium phosphate or water dip on the
survival of Salmonella and Listeria monocytogenes inoculated catfish before and
after freezing. J. Aquat. Food Product. Technol. 1 (1), 39e47.
Acknowledgments Rose, L., O'Connell, H., 2009. UV light inactivation of bacterial biothreat agents. Appl.
Environ. Microbiol. 75, 2987e2990.
We would like to thank Robert Richardson for his technical Reardon, J., Sancar, A., 2005. Nucleotide excision repair. Prog. Nucleic Acid Res. Mol.
Biol. 79, 183e235.
assistance. Rutala, W., Gergen, M., Weber, D., 2010. Room decontamination with UV radiation.
Infect. Control Hosp. Epidemiol. 31, 1025e1029.
References Salvage, B., 2014. Strengthening Meat Safety. MeatPoultry.com: The business jour-
nal for Meat and poultry processors. Available at: http://www.meatpoultry.
com/Writers/Bryan%20Salvage/Strengthening%20meat%20safety.aspx?cck¼1
Albaji, A., Kharabsheh, S., Al-Azab, S., Al-Kayad, M., Amr, Z., Abu-Baker, M., Chu, M.,
(accessed 23.07.14.).
2005. A 12-case outbreak of pharyngeal plague following consumption of camel
Sommers, C., Gunther, N., 2013. Inactivation of Campylobacter jejuni on poultry by
meat, in north-eastern Jordan. Ann. Trop. Med. Parasitol. 99 (8), 789e793.
ultraviolet light. In: Proceedings, 2013 Midwest Poultry Forum. St. Paul Riv-
Bhaduri, S., Phillips, J., 2013. Growth of pYV bearing Yersinia pestis KIM5 in retail
erCentre, St. Paul, MN, pp. 1e4.
ground pork. Foodborne Pathog. Dis. 10 (5), 467e471.
Sommers, C., Scullen, B., Paoli, G., Bhaduri, S., 2012. Inactivation of Francisella
Bin Saeed, A., Al-Hamdan, N., Fontaine, R., 2005. Plague from eating raw camel liver.
tularensis Utah-112 on food and food contact surfaces by ultraviolet light.
Emerg. Infect. Dis. 11 (9), 1456e1457.
J. Food Process. Technol. http://dx.doi.org/10.4172/2157-7110.S11-002.
Brickner, P., Vincent, R., First, M., Nardell, E., Murray, M., Kaufman, W., 2003. The
Sommers, C., Rajkowski, K., Sheen, S., Samer, C., Bender, E., 2011. The effect of
application of ultraviolet germicidal irradiation to control transmission of
cryogenic freezing and gamma irradiation on the survival of Salmonella on
airborne disease: bioterrorism countermeasure. Public Health Rep. 118, 99e114.
frozen shrimp. J. Food Process. Technol. http://dx.doi.org/10.4172/2157-
Christie, A., Chen, H., Elberg, S., 1980. Plague in camels and goats; their role in
7110.S8eS001.
human epidemics. J. Infect. Dis. 141, 724e726.
Sommers, C., Sites, J., Musgrove, M., 2010. Ultraviolet light (254 nm) inactivation of
CFR (Code of Federal Regulations), 2005. Irradiation in the production, processing
pathogens on foods and stainless steel surfaces. J. Food Saf. 30, 70e79.
and handling of food. Code Fed. Regul. 21 (179), 450e456.
Sommers, C., Cooke, P., 2009. Inactivation of avirulent Yersinia pestis in butterfield's
Diehl, J., 1995. Safety of Irradiated Food, second ed. Marcel Dekker, New York, New
phosphate buffer and frankfurters by UVC (254 nm) and gamma radiation.
York, pp. 93e98.
J. Food Prot. 72, 755e759.
Gardner, D., Shama, G., 2000. Modeling UV-induced inactivation of microorganisms
Sommers, C., Niemira, B., 2007. Effect of temperature on the radiation resistance of
on surfaces. J. Food Prot. 63, 63e70.
Yersinia pestis suspended in raw ground pork. J. Food Saf. 27, 317e325.
Gurtler, J., Rivera, R., Zhang, H., Sommers, C., 2010. Behavior of avirulent Yersinia
Sumner, S., Wallner-Pendleton, E., Froning, G., Stetson, L., 1996. Inhibition of Sal-
pestis in liquid whole egg as affected by storage temperature, antimicrobials
monella typhimurium on agar medium and poultry skin by ultraviolet energy.
and thermal pasteurization. J. Food Saf. 30, 537e557.
J. Food Prot. 59, 319e321.
Kauffman, A., Meltzer, M., Schmid, G., 1997. The economic impact of a bioterrorist
Stermer, R., Lasater-Smith, M., Brasington, C., 1987. Ultraviolet radiation-an effective
attack:are prevention and post attack programs justifiable. Emerg. Infect. Dis. 3,
bactericide for fresh meat. J. Food Prot. 50, 108e111.
83e94.
Torosian, S., Regan, P., Doran, T., Taylor, M., Margolin, A., 2009. A refrigeration
Khan, A., Morse, S., Lillibridge, S., 2000. Public-health preparedness for biological
temperature of 4
C does not prevent static growth of Yersinia pestis in heart
terrorism in the USA. Lancet 356, 1179e1182.
infusion broth. Can. J. Microbiol. 55, 1119e1124.
Koutchma, T., Forney, L., Moraru, C., 2008. Microbial inactivation by UV light. In:
World Health Organization (WHO), 1970. Tularemia. In: Health Aspects of Chemical
Ultraviolet Light in Food Technology. Taylor & Francis CRC Press, New York, New
and Biological Weapons. World Health Organization, Geneva, SUI, pp. 105e107.
York, USA, pp. 69e99.