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Article ; Online: Pathogen Prevalence Estimates and Diagnostic Methodology Trends in Laboratory Mice and Rats from 2003 to 2020.

Albers, Theresa M / Henderson, Kenneth S / Mulder, Guy B / Shek, William R

Journal of the American Association for Laboratory Animal Science : JAALAS

2023 Volume 62, Issue 3, Page(s) 229–242

Abstract: Rodents used in biomedical research are maintained as specific pathogen-free (SPF) by employing biosecurity measures that eliminate and exclude adventitious infectious agents known to confound research. The efficacy of these practices is assessed by ... ...

Abstract	Rodents used in biomedical research are maintained as specific pathogen-free (SPF) by employing biosecurity measures that eliminate and exclude adventitious infectious agents known to confound research. The efficacy of these practices is assessed by routine laboratory testing referred to as health monitoring (HM). This study summarizes the results of HM performed at Charles River Research Animal Diagnostic Services (CR-RADS) on samples submitted by external (non-Charles River) clients between 2003 and 2020. Summarizing this vast amount of data has been made practicable by the recent introduction of end-user business intelligence tools to Excel. HM summaries include the number of samples tested and the percent positive by diagnostic methodology, including direct examination for parasites, cultural isolation and identification for bacteria, serology for antibodies to viruses and fastidious microorganisms, and polymerase chain reaction (PCR) assays for pathogen-specific genomic sequences. Consistent with comparable studies, the percentages of pathogen-positive samples by diagnostic methodology and year interval are referred to as period prevalence estimates (%P
MeSH term(s)	Rats ; Animals ; Mice ; Prevalence ; Polymerase Chain Reaction ; Bacteria ; Helicobacter ; Pasteurellaceae ; Housing, Animal
Language	English
Publishing date	2023-05-01
Publishing country	United States
Document type	Journal Article
ISSN	2769-6677
ISSN (online)	2769-6677
DOI	10.30802/AALAS-JAALAS-22-000097
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Role of housing modalities on management and surveillance strategies for adventitious agents of rodents.

Shek, William R

ILAR journal

2007 Volume 49, Issue 3, Page(s) 316–325

Abstract: Specific pathogen-free (SPF) rodents for modern biomedical research need to be free of pathogens and other infectious agents that may not produce disease but nevertheless cause research interference. To meet this need, rodents have been rederived to ... ...

Abstract	Specific pathogen-free (SPF) rodents for modern biomedical research need to be free of pathogens and other infectious agents that may not produce disease but nevertheless cause research interference. To meet this need, rodents have been rederived to eliminate adventitious agents and then housed in room- to cage-level barrier systems to exclude microbial contaminants. Because barriers can and do fail, routine health monitoring (HM) is necessary to verify the SPF status of colonies. Testing without strict adherence to biosecurity practices, however, can lead to the inadvertent transfer of unrecognized, inapparent agents among institutions and colonies. Microisolation caging systems have become popular for housing SPF rodents because they are versatile and provide a highly effective cage-level barrier to the entry and spread of adventitious agents. But when a microisolation-caged colony is contaminated, the cage-level barrier impedes the spread of infection and so the prevalence of infection is often low, which increases the chance of missing a contamination and complicates the corroboration of unexpected positive findings. The expanding production of genetically engineered mutant (GEM) rodent strains at research institutions, where biosecurity practices vary and the risk of microbial contamination can be high, underscores the importance of accurate HM results in mitigating the risk of the introduction and spread of microbial contaminants with the exchange of mutant rodent strains among investigators and institutions.
MeSH term(s)	Animals ; Animals, Laboratory ; Environment, Controlled ; Housing, Animal/standards ; Infection Control/methods ; Mice ; Rats ; Rodent Diseases/prevention & control
Language	English
Publishing date	2007-12-06
Publishing country	England
Document type	Journal Article
ZDB-ID	2192062-X
ISSN	1930-6180 ; 1084-2020
ISSN (online)	1930-6180
ISSN	1084-2020
DOI	10.1093/ilar.49.3.316
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Article ; Online: Serendipitous Discovery of a Novel Murine Astrovirus Contaminating a Murine Helper T-cell Line and Incapable of Infecting Highly Immunodeficient Mice.

Ricart Arbona, Rodolfo J / Kelly, Sean / Wang, Chuanwu / Dhawan, Rajeev K / Henderson, Kenneth S / Shek, William R / Williams, Simon H / Altan, Eda / Delwart, Eric / Wolf, Felix / Lipman, Neil S

Comparative medicine

2020 Volume 70, Issue 4, Page(s) 359–369

Abstract: The unexpected seroconversion of sentinel mice in our facility to murine T lymphotrophic virus (MTLV) positivity led to our identification of a novel murine astrovirus that we designated murine astrovirus 2 (MuAstV-2). During our investigation, MuAstV-2 ... ...

Abstract	The unexpected seroconversion of sentinel mice in our facility to murine T lymphotrophic virus (MTLV) positivity led to our identification of a novel murine astrovirus that we designated murine astrovirus 2 (MuAstV-2). During our investigation, MuAstV-2 was found to be a contaminant of the T helper cell line (D10. G4.1) that was used to generate the MTLV antigen that we included in the multiplex fluorometric immunoassay (MFIA) that we used for sentinel screening. We eventually determined that cross-reactivity with the astrovirus generated a positive result in the MTLV assay. A confirmatory immunofluorometric assay (IFA) using the same MTLV-infected cell line yielded a similar result. However, the use of antigen prepared from MTLV-infected neonatal mouse thymus did not reproduce a positive result, leading us to suspect that the seroreactivity we had observed was not due to infection with MTLV. A mouse antibody production test showed that mice inoculated with naïve D10. G4.1 cells and their contact sentinels tested positive for MTLV using cell-line generated antigen, but tested negative in assays using MTLV antigen produced in mice. Metagenomic analysis was subsequently used to identify MuAstV-2 in feces from 2 sentinel mice that had recently seroconverted to MTLV. Two closely related astrovirus sequences (99.6% capsid identity) were obtained and shared 95% capsid amino acid identity with the MuAstV-2 virus sequenced from the D10. G4.1 cell line. These viruses are highly divergent from previously identified murine astroviruses, displaying <30% capsid identity, yet were closely related to murine astrovirus 2 (85% capsid identity), which had recently been isolated from feral mice in New York City. A MuAstV-2 specific PCR assay was developed and used to eradicate MuAstV-2 from the infected colony using a test and cull strategy. The newly identified MuAstV2 readily transmits to immunocompetent mouse strains by fecal-oral exposure, but fails to infect NOD-
MeSH term(s)	Animals ; Astroviridae ; Astroviridae Infections/metabolism ; Astroviridae Infections/virology ; Cell Line ; Feces/virology ; Genome, Viral ; Immunocompetence/genetics ; Mice/virology ; T-Lymphocytes, Helper-Inducer/immunology
Language	English
Publishing date	2020-07-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2006425-1
ISSN	2769-819X ; 0023-6764 ; 1532-0820
ISSN (online)	2769-819X
ISSN	0023-6764 ; 1532-0820
DOI	10.30802/AALAS-CM-19-000106
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Article ; Online: Outcomes After Aortopulmonary Window for Hypoplastic Pulmonary Arteries and Dual-Supply Collaterals.

Bauser-Heaton, Holly / Ma, Michael / McElhinney, Doff B / Goodyer, William R / Zhang, Yulin / Chan, Frandics P / Asija, Ritu / Shek, Jennifer / Wise-Faberowski, Lisa / Hanley, Frank L

The Annals of thoracic surgery

2019 Volume 108, Issue 3, Page(s) 820–827

Abstract: Background: Our institutional approach to tetralogy of Fallot with major aortopulmonary collateral arteries (MAPCAs) emphasizes early unifocalization and complete repair (CR). In the small subset of patients with dual-supply MAPCAs and confluent but ... ...

Abstract	Background: Our institutional approach to tetralogy of Fallot with major aortopulmonary collateral arteries (MAPCAs) emphasizes early unifocalization and complete repair (CR). In the small subset of patients with dual-supply MAPCAs and confluent but hypoplastic central pulmonary arteries (PAs), our surgical approach is early creation of an aortopulmonary window (APW) to promote PA growth. Factors associated with successful progression to CR and mid-term outcomes have not been assessed. Methods: Clinical data were reviewed. PA diameters were measured offline from angiograms prior to APW and on follow-up catheterization >1 month after APW but prior to any additional surgical interventions. Results: From November 2001 to March 2018, 352 patients with tetralogy of Fallot/MAPCAs underwent initial surgery at our center, 40 of whom had a simple APW with or without ligation of MAPCAs as the first procedure (median age, 1.4 months). All PA diameters increased significantly on follow-up angiography. Ultimately, 35 patients underwent CR after APW. Nine of these patients (26%) underwent intermediate palliative operation between 5 and 39 months (median, 8 months) after APW. There were no early deaths. The cumulative incidence of CR was 65% 1 year post-APW and 87% at 3 years. Repaired patients were followed for a median of 4.2 years after repair; the median PA:aortic pressure ratio was 0.39 (range, 0.22 to 0.74). Conclusions: Most patients with tetralogy of Fallot/MAPCAs and hypoplastic but normally arborizing PAs and dual-supply MAPCAs are able to undergo CR with low right ventricular pressure after APW early in life. Long-term outcomes were good, with acceptable PA pressures in most patients.
MeSH term(s)	Abnormalities, Multiple/diagnostic imaging ; Abnormalities, Multiple/surgery ; Aortopulmonary Septal Defect/diagnostic imaging ; Aortopulmonary Septal Defect/surgery ; Cardiac Surgical Procedures/methods ; Cohort Studies ; Collateral Circulation/physiology ; Computed Tomography Angiography/methods ; Databases, Factual ; Female ; Follow-Up Studies ; Hospitals, Pediatric ; Humans ; Imaging, Three-Dimensional ; Infant, Newborn ; Male ; Pulmonary Atresia/diagnostic imaging ; Pulmonary Atresia/mortality ; Pulmonary Atresia/surgery ; Retrospective Studies ; Risk Assessment ; Survival Analysis ; Tetralogy of Fallot/diagnostic imaging ; Tetralogy of Fallot/mortality ; Tetralogy of Fallot/surgery ; Time Factors ; Treatment Outcome
Language	English
Publishing date	2019-04-10
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	211007-6
ISSN	1552-6259 ; 0003-4975
ISSN (online)	1552-6259
ISSN	0003-4975
DOI	10.1016/j.athoracsur.2019.03.022
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Article ; Online: Chapter 11 Microbiological Quality Control for Laboratory Rodents and Lagomorphs

Shek, William R. / Smith, Abigail L. / Pritchett-Corning, Kathleen R.

Laboratory Animal Medicine

Abstract: Mice (Mus musculus), rats (Rattus norvegicus), other rodent species, and domestic rabbits (Oryctolagus cuniculus) have been used in research for over 100 years. During the first half of the 20th century, microbiological quality control of lab animals was ...

Abstract	Mice (Mus musculus), rats (Rattus norvegicus), other rodent species, and domestic rabbits (Oryctolagus cuniculus) have been used in research for over 100 years. During the first half of the 20th century, microbiological quality control of lab animals was at best rudimentary as colonies were conventionally housed and little or no diagnostic testing was done. Hence, animal studies were often curtailed and confounded by infectious disease (Mobraaten and Sharp, 1999; Morse, 2007; Weisbroth, 1999). By the 1950s, it became apparent to veterinarians in the nascent field of comparative medicine that disease-free animals suitable for research could not be produced by standard veterinary disease control measures (e.g., improved sanitation and nutrition, antimicrobial treatments) in conventional facilities. Henry Foster, the veterinarian who founded Charles River Breeding Laboratories in 1948 and a pioneer in the large-scale production of laboratory rodents, stated in a seminar presented at the 30th anniversary of AALAS, “After a variety of frustrating health-related problems, it was decided that a major change in the company’s philosophy was required and an entirely different approach was essential”. Consequently, he and others developed innovative biosecurity systems to eliminate and exclude pathogens (Allen, 1999). In 1958, Foster reported on the Cesarean-originated barrier-sustained (COBS) process for the large-scale production of specific pathogen-free (SPF) laboratory rodents (Foster, 1958). To eliminate horizontally transmitted pathogens, a hysterectomy was performed on a near-term dam from a contaminated or conventionally housed colony. The gravid uterus was pulled through a disinfectant solution into a sterile flexible film isolator where the pups were removed from the uterus and suckled on axenic (i.e., germ-free) foster dams. After being mated to expand their number and associated with a cocktail of nonpathogenic bacteria to normalize their physiology and prime their immune system, rederived rodents were transferred to so-called barrier rooms for large-scale production. The room-level barrier to adventitious infection entailed disinfection of the room, equipment, and supplies, limiting access to trained and properly gowned personnel, and the application of new technologies such as high-efficiency particulate air-filtration of incoming air (Dubos and Schaedler, 1960; Foster, 1980; Schaedler and Orcutt, 1983; Trexler and Orcutt, 1999). The axenic and associated rodents mentioned in the COBS process are collectively classified as gnotobiotic to indicate that they have a completely known microflora. By contrast, barrier-reared rodent colonies are not gnotobiotic because they are housed in uncovered cages and thus acquire a complex microflora from the environment, supplies, personnel, and other sources. Instead, they are described as SPF to indicate that according to laboratory testing, they are free from infection with a defined list of infectious agents, commonly known as an ‘exclusion’ list.
Keywords	covid19
Publisher	Elsevier; PMC
Document type	Article ; Online
DOI	10.1016/b978-0-12-409527-4.00011-0
Database	COVID19

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Article ; Online: Microbiological Quality Control for Laboratory Rodents and Lagomorphs

Shek, William R. / Smith, Abigail L. / Pritchett-Corning, Kathleen R.

Laboratory Animal Medicine

Abstract	Mice (Mus musculus), rats (Rattus norvegicus), other rodent species, and domestic rabbits (Oryctolagus cuniculus) have been used in research for over 100 years. During the first half of the 20th century, microbiological quality control of lab animals was at best rudimentary as colonies were conventionally housed and little or no diagnostic testing was done. Hence, animal studies were often curtailed and confounded by infectious disease (Mobraaten and Sharp, 1999; Morse, 2007; Weisbroth, 1999). By the 1950s, it became apparent to veterinarians in the nascent field of comparative medicine that disease-free animals suitable for research could not be produced by standard veterinary disease control measures (e.g., improved sanitation and nutrition, antimicrobial treatments) in conventional facilities. Henry Foster, the veterinarian who founded Charles River Breeding Laboratories in 1948 and a pioneer in the large-scale production of laboratory rodents, stated in a seminar presented at the 30th anniversary of AALAS, “After a variety of frustrating health-related problems, it was decided that a major change in the company’s philosophy was required and an entirely different approach was essential”. Consequently, he and others developed innovative biosecurity systems to eliminate and exclude pathogens (Allen, 1999). In 1958, Foster reported on the Cesarean-originated barrier-sustained (COBS) process for the large-scale production of specific pathogen-free (SPF) laboratory rodents (Foster, 1958). To eliminate horizontally transmitted pathogens, a hysterectomy was performed on a near-term dam from a contaminated or conventionally housed colony. The gravid uterus was pulled through a disinfectant solution into a sterile flexible film isolator where the pups were removed from the uterus and suckled on axenic (i.e., germ-free) foster dams. After being mated to expand their number and associated with a cocktail of nonpathogenic bacteria to normalize their physiology and prime their immune system, rederived rodents were transferred to so-called barrier rooms for large-scale production. The room-level barrier to adventitious infection entailed disinfection of the room, equipment, and supplies, limiting access to trained and properly gowned personnel, and the application of new technologies such as high-efficiency particulate air-filtration of incoming air (Dubos and Schaedler, 1960; Foster, 1980; Schaedler and Orcutt, 1983; Trexler and Orcutt, 1999). The axenic and associated rodents mentioned in the COBS process are collectively classified as gnotobiotic to indicate that they have a completely known microflora. By contrast, barrier-reared rodent colonies are not gnotobiotic because they are housed in uncovered cages and thus acquire a complex microflora from the environment, supplies, personnel, and other sources. Instead, they are described as SPF to indicate that according to laboratory testing, they are free from infection with a defined list of infectious agents, commonly known as an ‘exclusion’ list.
Keywords	covid19
Publisher	Elsevier; PMC
Document type	Article ; Online
DOI	10.1016/b978-0-12-409527-4.00011-0
Database	COVID19

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Article ; Online: Microbiological Quality Control for Laboratory Rodents and Lagomorphs

Shek, William R. / Gaertner, Diane J.

Laboratory Animal Medicine

Keywords	covid19
Publisher	PMC
Document type	Article ; Online
DOI	10.1016/b978-012263951-7/50013-2
Database	COVID19

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Article ; Online: Multiplexed fluorometric immunoassay testing methodology and troubleshooting.

Wunderlich, Michelle L / Dodge, Megan E / Dhawan, Rajeev K / Shek, William R

Journal of visualized experiments : JoVE

2011 , Issue 58

Abstract: To ensure the quality of animal models used in biomedical research we have developed a number of diagnostic testing strategies and methods to determine if animals have been exposed to adventitious infectious agents (viruses, mycoplasma, and other ... ...

Abstract	To ensure the quality of animal models used in biomedical research we have developed a number of diagnostic testing strategies and methods to determine if animals have been exposed to adventitious infectious agents (viruses, mycoplasma, and other fastidious microorganisms). Infections of immunocompetent animals are generally transient, yet serum antibody responses to infection often can be detected within days to weeks and persist throughout the life of the host. Serology is the primary diagnostic methodology by which laboratory animals are monitored. Historically the indirect enzyme-linked immunosorbent assay (ELISA) has been the main screening method for serosurveillance. The ELISA is performed as a singleplex, in which one microbial antigen-antibody reaction is measured per well. In comparison the MFIA is performed as a multiplexed assay. Since the microspheres come in 100 distinct color sets, as many as 100 different assays can be performed simultaneously in a single microplate well. This innovation decreases the amount of serum, reagents and disposables required for routine testing while increasing the amount of information obtained from a single test well. In addition, we are able to incorporate multiple internal control beads to verify sample and system suitability and thereby assure the accuracy of results. These include tissue control and IgG anti-test serum species immunoglobulin (αIg) coated bead sets to evaluate sample suitability. As in the ELISA and IFA, the tissue control detects non-specific binding of serum immunoglobulin. The αIg control (Serum control) confirms that serum has been added and contains a sufficient immunoglobulin concentration while the IgG control bead (System Suitability control), coated with serum species immunoglobulin, demonstrates that the labeled reagents and Luminex reader are functioning properly.
MeSH term(s)	Animals ; Fluorometry/methods ; Immunoassay/methods
Keywords	covid19
Language	English
Publishing date	2011-12-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/3715
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Article: A comparison of mouse parvovirus 1 infection in BALB/c and C57BL/6 mice: susceptibility, replication, shedding, and seroconversion.

Henderson, Kenneth S / Pritchett-Corning, Kathleen R / Perkins, Cheryl L / Banu, Laila A / Jennings, Steven M / Francis, Brian C / Shek, William R

Comparative medicine

2015 Volume 65, Issue 1, Page(s) 5–14

Abstract: This study characterized the effects of challenge with a field isolate of mouse parvovirus 1 (MPV1e) in C57BL/6NCrl (B6) and BALB/cAnNCrl (C) mice. We found that C mice were more susceptible to MPV1e infection than were B6 mice; ID50 were 50 to 100 times ...

Abstract	This study characterized the effects of challenge with a field isolate of mouse parvovirus 1 (MPV1e) in C57BL/6NCrl (B6) and BALB/cAnNCrl (C) mice. We found that C mice were more susceptible to MPV1e infection than were B6 mice; ID50 were 50 to 100 times higher after gavage and 10-fold higher after intraperitoneal injection in B6 as compared with C mice. To evaluate the host strain effect on the pathogenesis of MPV1e, B6 and C mice were inoculated by gavage. Feces and tissues, including mesenteric lymph nodes (MLN), ileum, spleen and blood, were collected for analysis by quantitative PCR (qPCR) to assess infection and fecal shedding and by RT-qPCR to evaluate replication. Peak levels of MPV1e shedding, infection, and replication were on average 3.4, 4.3, and 6.2 times higher, respectively, in C than in B6 mice. Peaks occurred between 3 and 10 d after inoculation in C mice but between 5 and 14 d in B6 mice. Multiplexed fluorometric immunoassays detected seroconversion in 2 of 3 C mice at 7 d after inoculation and in all 3 B6 mice at 10 d. By 56 d after inoculation, viral replication was no longer detectable, and fecal shedding was very low; infection persisted in ileum, spleen, and MLN, with levels higher in C than B6 mice and highest in MLN. Therefore, the lower susceptibility of B6 mice, as compared with C mice, to MPV1e infection was associated with lower levels of infection, replication, and shedding and delayed seroconversion.
MeSH term(s)	Animals ; Disease Susceptibility/virology ; Feces/virology ; Fluorometry ; Immunoassay ; Mice ; Mice, Inbred BALB C/virology ; Mice, Inbred C57BL/virology ; Parvoviridae Infections/blood ; Parvoviridae Infections/physiopathology ; Reverse Transcriptase Polymerase Chain Reaction ; Seroconversion/physiology ; Species Specificity ; Time Factors ; Virus Replication/physiology ; Virus Shedding/physiology
Language	English
Publishing date	2015-02
Publishing country	United States
Document type	Comparative Study ; Journal Article
ZDB-ID	2006425-1
ISSN	1532-0820 ; 0023-6764
ISSN	1532-0820 ; 0023-6764
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Large-scale rodent production methods make vendor barrier rooms unlikely to have persistent low-prevalence parvoviral infections.

Shek, William R / Pritchett, Kathleen R / Clifford, Charles B / White, William J

Contemporary topics in laboratory animal science

2005 Volume 44, Issue 4, Page(s) 37–42

Abstract: A recent article in Contemporary Topics in Laboratory Animal Science by Pullium and colleagues expressed the opinion that because no other source could be found for a parvoviral contamination detected in sentinel mice prior to deployment, the infection ... ...

Abstract	A recent article in Contemporary Topics in Laboratory Animal Science by Pullium and colleagues expressed the opinion that because no other source could be found for a parvoviral contamination detected in sentinel mice prior to deployment, the infection apparently came from the unspecified vendor, even though no antibodies were ever detected in mice within 3 weeks of arrival. As this opinion may be shared by others and expresses some of the deep frustration in trying to detect the source of parvoviral infection in facilities using cage-level bioexclusion housing, Charles River Laboratories (CRL) feels it important to contribute to scientific dialogue by claiming to be the unnamed vendor in the Pullium article and discussing why a parvoviral contamination in a CRL barrier room would be detected rapidly. We show that viral infections in CRL barrier rooms rapidly reach high prevalence and that such contaminations historically have been detected quickly, and we describe why we feel enhancements in current monitoring methods provide for even more rapid detection of parvoviruses. Furthermore, we present substantial evidence that the barrier rooms that served as the source of the customer-suspect sentinel mice remain free of all parvoviruses, in light of monitoring of hundreds of mice by all available techniques. Therefore, although an initial list of all possible sources of contamination prudently should include vendors, the evidence is overwhelming that this vendor was not the source of the parvoviral contamination discussed in the Pullium paper.
MeSH term(s)	Animals ; Animals, Laboratory ; Contract Services ; Disease Outbreaks/prevention & control ; Disease Outbreaks/veterinary ; Housing, Animal ; Mice ; Parvoviridae Infections/diagnosis ; Parvoviridae Infections/prevention & control ; Parvoviridae Infections/veterinary ; Rodent Diseases/diagnosis ; Rodent Diseases/prevention & control ; Rodent Diseases/virology ; Sentinel Surveillance/veterinary ; Serologic Tests/veterinary
Language	English
Publishing date	2005-07
Publishing country	United States
Document type	Comment ; Comparative Study ; Journal Article
ISSN	1060-0558
ISSN	1060-0558
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