Thermal Inactivation of Avian Viral and Bacterial Pathogens in an Effluent Treatment System within a Biosafety Level 2 and 3 Enhanced Facility

Abstract

Avian influenza (AI) virus, avian paramyxovirus Type 1 (APMV-1 or NDV), reovirus, rotavirus, turkey astrovirus (TAst), avian metapneumovirus (aMPV), Marek's disease virus (MDV-1), avian parvovirus (ChPV), and Salmonella enterica serovar Enteritidis are significant biosafety level 2 (BSL-2) or biosafety level 3 enhanced (BSL-3E) pathogens of poultry that are studied in veterinary medical research laboratories worldwide. The purpose of this study was to determine the effectiveness of a moderate temperature, effluent decontamination system (EDS) to inactivate avian pathogens. First, the thermal inactivation processes for AI virus, APMV-1, reovirus, and rotaviruses were determined in phosphate-buffered saline (PBS) using in vitro assays from which thermal death rates (D_t) and changes in heat resistance (zD) of the AI virus and APMV-1 were determined at various time/temperature parameters. The PBS validation process demonstrated that 6 log₁₀ reduction was achieved following heating at 82.2°C within 30 seconds for AI virus and APMV-1, 1.8 minutes for astrovirus, while reoviruses and rotaviruses were destroyed within 3 minutes. Second, to determine whether pathogens were adequately inactivated in a moderate temperature EDS system, vials containing avian viruses (5.1-11.1 log₁₀ TCID, EID₅₀/ml, or infectious particle/ml) and bacteria (9.1 log₁₀ cfu/ml) were placed in the effluent tank and exposed to a standard cycle of 82.2°C for 6 hours. The EDS process totally inactivated enveloped and non-enveloped viruses with complete inactivation of greater than 5.1-11.1 log₁₀ TCID/ml of specific pathogens. The data from the inactivation models and the EDS validation test showed that the 6 log₁₀ reduction required for a sanitary assurance level of effluent was achieved with a significant margin of safety.

References

Alexander

D. J.

& Manvell

R. J.

(2004). Heat inactivation of Newcastle disease virus (strain Herts 33/56) in artificially infected chicken meat homogenate. Avian Pathology, 33(2), 222–225.

Alexander

D. J.

(2005). Avian influenza. Manual for diagnostic tests and vaccines for terrestrial animals (p. 2.7.12). Paris: World Organization for Animal Health.

Anonymous. (1972). Clean Water Act, 33 CFR. 301(d), 304(b), 304(g), 304(m) and 307(b). 86. United States of America.

Anonymous. (1981). Secondary Treatment Regulation. 1314(d) (1) U.S.C., 86. United States of America.

Anonymous. (1993). Standards for the Use and Disposal of Sludge. 40 CFR. 503.32 D(a)(3). United States of America.

Blümel

, Schmidt

& Willkommen

& Löwer

(2002). Inactivation of parvovirus B19 during pasteurization of human serum albumin. Transfusion, 42, 1011–1018.

Bräuniger

, Peters

, Borchers

& Kao

& Borchers

(2000). Further studies on thermal resistance of bovine parvovirus against moist and dry heat. International Journal of Hygiene and Environmental Health, 203, 71–75.

Calnek

B. W.

& Adldinger

H. K.

(1971). Some characteristics of cell-free preparations of Marek's disease virus. Avian Diseases, 15, 508–517.

Centers for Disease Control and Prevention (CDC). (2007). Viral agents. In Biosafety in microbiological and biomedical laboratories, (5th ed.). Washington, DC: U.S. Health and Human Services Publication, Government Printing Office.

10.

Chang

J. T.

, Chou

Y. C.

& Lin

M. S.

& Wang

S. R.

(2010). Inactivation strategy for pseudorabies virus in milk for production of biopharmaceuticals. Japan Journal of Veterinary Research, 58, 179–183.

11.

Cook

J. K. A.

(2000). Avian pneumovirus infections of turkeys and chickens. The Veterinary Journal, 160, 118–125.

12.

Das

, Spackman

& Pantin-Jackwood

M. J.

& Suarez

D. L.

(2009). Removal of real-time reverse transcription polymerase chain reaction (RT-PCR) inhibitors associated with cloacal swab samples and tissues for improved diagnosis of avian influenza virus by RT-PCR. Journal of Veterinary Diagnostic Investigation, 21, 771–778.

13.

Day

J. M.

& Spackman

& Pantin-Jackwood

(2007). A multiplex RT-PCR test for the differential identification of turkey astrovirus type 1, turkey astrovirus type 2, chicken astrovirus, avian nephritis virus, and avian rotavirus. Avian Diseases, 51, 681–684.

14.

Durham

P. J. K.

& Johnson

R. H.

(1985). Properties of an Australian isolate of bovine parvovirus type 1. Veterinary Microbiology, 10, 335–345.

15.

Elhafi

, Naylor

C. J.

& Savage

C. E.

& Jones

R. C.

(2004). Microwave or autoclave treatments destroy the infectivity of infectious bronchitis virus and avian pneumovirus but allow detection by reverse transcriptase-polymerase chain reaction. Avian Pathology, 33, 303–306.

16.

Emmoth

(2010). Virus inactivation—Evaluation of processes used in biowaste management. Thesis. Uppsala, Sweden: Department of Biomedical Sciences and Veterinary Public Health Swedish University of Agricultural Sciences.

17.

Estes

M. K.

, Graham

D. Y.

& Smith

E. M.

& Gerba

C. P.

(1979). Rotavirus stability and inactivation. Available at: http://vir.sgmjournals.org/cgi/content/abstract/43/2/403.

18.

Foliguet

J.-M.

& Doncoeur

(1972). Inactivation assays of enteroviruses and Salmonella in fresh and digested wastewater sludges by pasteurization. Water Research, 6, 1399–1407.

19.

Jarosinski

K. W.

, Margulis

N. G.

, Kamil

J. P.

, Spatz

S. J.

& Nair

V. K.

& Osterrieder

(2007). Horizontal transmission of Marek's disease virus requires US2, the UL13 protein kinase, and gC. Journal of Virology, 81, 10575–10587.

20.

Jin

, Zhang

& Boyd

& Tang

(2008). Thermal resistance of Salmonella enteritidis and Escherichia coli K12 in liquid egg determined by thermal-death-time disks. Journal of Food Engineering, 84, 608–614.

21.

Middleton

J. K.

, Severson

T. F.

, Chandran

, Gilli an

A. L.

& Yin

& Nibert

M. L.

(2002). Thermostability of reovirus disassembly intermediates (ISVPs) correlates with genetic, biochemical, and thermodynamic properties of major surf ace protein. Journal of Virology, 76, 1051–1061.

22.

Moce-Llivina

, Muniesa

, Pimenta-Vale

& Lucena

& Jofre

(2003). Survival of bacterial indicator species and bacteriophages after thermal treatment of sludge and sewage. Applied and Environmental Microbiology, 69, 1452–1456.

23.

Murphy

R. Y.

, Osaili

& Duncan

L. K.

& Marcy

J. A.

(2004). Thermal inactivation of Salmonella and Listeria monocytogenes in ground chicken thigh/leg meat and skin. Poultry Science, 83, 1218–1225.

24.

Nazerian

& Witter

R. L.

(1970). Cell-free transmission and in vivo replication of Marek's disease virus. Journal of Virology, 5, 388–397.

25.

Reed

L. J.

& Muench

(1938). A simple method of estimating fifty percent endpoints. The American Journal of Hygiene, 27, 493–497.

26.

Richards

G. P.

(2001). Enteric virus contamination of foods through industrial practices: A primer on intervention strategies. Journal of Industrial Microbiology and Biotechnology, 27, 117–125.

27.

Sauerbrei

& Wutzler

(2009). Testing thermal resistance of viruses. Archives of Virology, 154, 115–119.

28.

Schultz-Cherry

& King

D. J.

& Koci

M. D.

(2001). Inactivation of an astrovirus associated with Poult Enteritis Mortality Syndrome. Avian Diseases, 45, 78–82.

29.

Senne

(2008). Virus propagation in embroyonating eggs. In Dufour-Zavala

(ed.), A laboratory manual for the isolation, identification and characterization of avian pathogens. Jacksonville, FL: American Association of Avian Pathologists.

30.

Sofer

(2003). Part 3a, Plasma and plasma products (heat and solvent/detergent treatment). Virus Inactivation Series. Available at: http://biopharminternational.findpharma.com/biopharm/issue/issueDetail.jsp?id=2428.

31.

Southeast Poultry Research Laboratory (SEPRL). (2009). Biosafety/Biosecurity Guidelines, V (p. 36). Athens, GA: SEPRL.

32.

Spackman

, Pantin-Jackwood

& Day

J. M.

& Sellers

(2005). The pathogenesis of turkey origin reoviruses in turkeys and chickens. Avian Pathology, 34, 291–296.

33.

Swayne

D. E.

& Beck

J. R.

(2004). Heat inactivation of avian influenza and Newcastle disease viruses in egg products. Avian Pathology, 33, 512–518.

34.

Swayne

D. E.

& Senne

D. A.

& Beard

C. W.

(1998). Avian influenza. In Swayne

D. E.

, Glisson

J. R.

, Jackwood

M. W.

& Pearson

J. E.

& Reed

W. M.

(eds.), A laboratory manual for the isolation and identification of avian pathogens (pp. 150–155). Tallahassee, FL: American Association of Avian Pathologists.

35.

Theil

K. W.

& Reynolds

D. L.

& Saif

Y. M.

(1986). Isolation and serial propagation of turkey rotaviruses in a fetal rhesus monkey kidney (MA104) cell line. Avian Diseases, 30, 93–104.

36.

Thomas

& King

D. J.

& Swayne

D. E.

(2008). Thermal inactivation of avian influenza and Newcastle disease viruses in chicken meat. Journal of Food Protection, 71, 1214–1222.

37.

Thomas

& Swayne

D. E.

(2007). Thermal inactivation of H5N1 high pathogenicity avian influenza virus in naturally infected chicken meat. Journal of Food Protection, 70, 674–680.

38.

, Estevez

, Song

& Kapczynski

& Zsak

(2010). Generation and biological assessment of recombinant avian metapneumovirus subgroup C (aMPV-C) viruses containing different lengths of the G gene. Virus Research, 147, 182–188.

39.

Yunoki

, Tsujikawa

, Urayama

, Sasaki

, Morita

& Tanaka

(2003). Heat sensitivity of human parvovirus B19. Vox Sanguinis, 85, 67–68.

40.

Zsak

& Strother

K. O.

& Day

J. M.

(2009). Development of a polymerase chain reaction procedure for detection of chicken and turkey parvoviruses. Avian Diseases, 53, 83–88.