Implications of experimental technique for analysis and interpretation of data from animal experiments: outliers and increased variability resulting from failure of intraperitoneal injection procedures

Das, Rose Gaines; North, Delia

doi:10.1258/002367707781282802

Cited by 59 publications

(38 citation statements)

References 16 publications

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“…Variability in the response of mice to i.p. injections has been documented (13,14). To reduce such variability to a minimum, all mice were handled and injected by the same experienced investigator (S.H.…”

Section: Mouse Strainsmentioning

confidence: 99%

Genetic Linkages for Thyroxine Released in Response to Thyrotropin Stimulation in Three Sets of Recombinant Inbred Mice Provide Evidence for Shared and Novel Genes Controlling Thyroid Function

Hamidi

Aliesky²,

Williams³

et al. 2013

Thyroid

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Genetic susceptibility for thyroid sensitivity to TSH stimulation was distinct for three families of inbred mouse lines. These observations parallel the human situation with multiple genetic loci contributing to the same trait and different loci associated with the same trait in different ethnic groups. Of the genetic loci highlighted in mice, three overlap with, or are located up or downstream, of human TSH-controlling genes. Other studies show that human disease genes can be identified through cross-species gene mapping of evolutionary conserved processes. Consequently, our findings suggest that novel thyroid function genes may yet be revealed in humans.

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Section: Mouse Strainsmentioning

confidence: 99%

Genetic Linkages for Thyroxine Released in Response to Thyrotropin Stimulation in Three Sets of Recombinant Inbred Mice Provide Evidence for Shared and Novel Genes Controlling Thyroid Function

Hamidi

Aliesky²,

Williams³

et al. 2013

Thyroid

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“…18 F-FDG has been administered via the intraperitoneal method, and after 60 min of uptake there is little difference in the image biodistribution of the 18 F-FDG between intraperitoneal and intravenous methods (6,7). However, there can be poor intraperitoneal injections 10%-20% of the time (8,9) if the injection is made erroneously, such as directly into the bowel (9). Because these poor intraperitoneal injections are not always immediately recognized, they may lead to outlying results, which may distort or invalidate statistical analysis (9).…”

mentioning

confidence: 99%

“…However, there can be poor intraperitoneal injections 10%-20% of the time (8,9) if the injection is made erroneously, such as directly into the bowel (9). Because these poor intraperitoneal injections are not always immediately recognized, they may lead to outlying results, which may distort or invalidate statistical analysis (9). The main limitation of the intraperitoneal method is that it is dependent on the pharmacokinetics of the radiotracer; thus, many radiotracers other than 18 F-FDG cannot be used with this method.…”

mentioning

confidence: 99%

“…Injection errors are often not recognized and can lead to misadministration of the radiotracer into the interstitium, which can negatively affect the outcome of the preclinical research (9,15). The frequency and degree of errors, even with experienced staff, are often underestimated (15) and can be a limiting factor in obtaining useful data from mouse small-animal PET.…”

mentioning

confidence: 99%

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Evaluation of Mouse Tail-Vein Injections Both Qualitatively and Quantitatively on Small-Animal PET Tail Scans

Vines¹,

Green²,

Kudo³

et al. 2011

Journal of Nuclear Medicine Technology

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Quantitative small-animal PET of mice requires successful delivery of radiotracers into the venous system. Intravenous injection of radiotracers via lateral tail veins is the most commonly used method of administration and can be technically challenging. Evaluation of the quality of an intravenous injection is necessary to determine whether small-animal PET is quantitatively accurate. The purpose of this study was to evaluate and compare the quality of 50 consecutive intravenous injections into mouse tail veins using both quantitative and qualitative methods. Methods: During 18 F-FDG intravenous injection, qualitative assessment of the injection was performed and classified according to specific criteria as good, intermediate, or poor. Small-animal PET scans of the body and tail were acquired, and tail injection sites were quantitatively assessed in terms of percentage injected dose per gram and classified as low, medium, or high uptake of 18 F-FDG. Qualitative and quantitative methods were compared. To assess baseline amounts of 18 F-FDG in the tail without a tail injection, 3 additional mice were injected by the intraperitoneal method, imaged, and quantitatively assessed in the same manner. The in vivo imaging data were validated on 7 additional mice by sacrificing them after scans, removing their tails, rescanning the tails, and then measuring the tail radioactivity ex vivo in a g-counter and correlating it with the in vivo amount. Results: Validation of in vivo imaging to ex vivo data yielded an excellent correlation, with an r 2 value of 0.95. Comparison of qualitative and quantitative methods yielded 45 matching results (42 good and low, 2 intermediate and medium, and 1 poor and high). There were 5 cases of mismatching results (1 false-negative and 4 false-positive) between qualitative and quantitative methods. Low-uptake tail injections were comparable to the intraperitoneal injection values. Using qualitative methods, accuracy was true 90% (45/50) of the time. The overall rate of successful intravenous injections was 92% (46/50) using quantitative methods. Conclusion: Qualitative assessment is all that is necessary if the intravenous injection is classified as good. In intermediate, poor, or uncertain classifications, a scan of the tail should be performed for quantitative assessment.

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“…Leucopenia was induced 96 h before induction of pneumonia in anaesthetised animals (60 mg/kg sodium thiopental intraperitoneally) by infusing a bolus dose of 30 mg/kg cyclophosphamide (Asta-Medica, Mérignac, France) through the penile vein [7]. Immunocompetent animals received saline alone.…”

Section: Methodsmentioning

confidence: 99%

Animal Model Methodology: Immunocompetent or Leucopenic Rats, Which Is the Best? Results from a Model of Experimental Pneumonia due to Derepressed Cephalosporinase-Producing <b><i>Enterobacter cloacae</i></b>

Jacolot¹,

Judel

Chaumais

et al. 2012

Chemotherapy

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Background: The aim of this study was to compare the bactericidal activity of cefepime plus amikacin against experimental pneumonia induced by a stably derepressed cephalosporinase-producing Enterobacter cloacae strain in immunocompetent and leucopenic rats. Methods: Sixty Wistar rats were used. Leucopenia was induced in half of them by a single intravenous administration of 30 mg/kg cyclophosphamide, while the remaining rats received the same volume of saline. All rats were infected 96 h later by tracheal instillation of 8 log₁₀ colony-forming units of E. cloacae. Twelve rats (6 immunocompetent and 6 leucopenic) were sacrificed 6 h later to assess the initial bacterial burden to the lungs. Then, the remaining 48 rats received a combination of 60 mg/kg cefepime twice a day and 25 mg/kg amikacin once a day given intraperitoneally or the same volume of saline. Six rats per group (leucopenic or not, treated or not) were sacrificed 12 and 30 h after therapy started. Results: Spontaneous bacterial clearance with time was observed only in immunocompetent rats. Compared to untreated animals, antibiotic administration induced a decrease in lung bacterial titres in immunocompetent and leucopenic rats. The difference was statistically significant only in leucopenic rats. Conclusions: The use of leucopenic rats reduced spontaneous bacterial clearance in the lungs and increased the bactericidal effect of the antibiotic combination and ultimately the confidence in the reliability of the results.

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Implications of experimental technique for analysis and interpretation of data from animal experiments: outliers and increased variability resulting from failure of intraperitoneal injection procedures

Cited by 59 publications

References 16 publications

Genetic Linkages for Thyroxine Released in Response to Thyrotropin Stimulation in Three Sets of Recombinant Inbred Mice Provide Evidence for Shared and Novel Genes Controlling Thyroid Function

Genetic Linkages for Thyroxine Released in Response to Thyrotropin Stimulation in Three Sets of Recombinant Inbred Mice Provide Evidence for Shared and Novel Genes Controlling Thyroid Function

Evaluation of Mouse Tail-Vein Injections Both Qualitatively and Quantitatively on Small-Animal PET Tail Scans

Animal Model Methodology: Immunocompetent or Leucopenic Rats, Which Is the Best? Results from a Model of Experimental Pneumonia due to Derepressed Cephalosporinase-Producing <b><i>Enterobacter cloacae</i></b>

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