Approbation of the model of acute bacterial cystitis on rabbits

DOI: 10.29296/2618723X-2021-02-04

Trofimets E.I., Toxicologist, ORCID: 0000-0003-3246-6457,
Borovkova K.Е., Head of microbiological laboratory, ORCID: 0000-0003-1571-6549,
Guschin Ya.A., Head of histology and pathomorphology department, ORCID: 0000-0002-7656-991Х,
Salmova J.V., microbiologist, ORCID: 0000-0001-9891-8634,
Nikiforova L.R., microbiologist, ORCID: 0000-0001-8710-2023,
Katelnikova A.E., PhD, Head of Specific Toxicology and Pharmacodynamics Department, ORCID: 0000-0003-3203-9869,
Kryshen K.L., PhD, Head of Toxicology and Microbiology Department, ORCID: 0000-0003-1451-7716

Institute of Pre-Сlinical Research Ltd.
3, Zavodskaya St., Build. 245, Kuzmolovsky Urban-Type Settlement,
Vsevolozhsky District, Leningrad Region, 188663, Russia

Е-mail: [email protected]


Keywords: acute cystitis Escherichia coli laboratory animals bacterial infection animal model

For citation:

Trofimets E.I., Borovkova K.Е., Guschin Ya.A., Salmova J.V. , Nikiforova L.R., Katelnikova A.E., Kryshen K.L. Approbation of the model of acute bacterial cystitis on rabbits. Laboratory Animals for Science. 2021; 2. https://doi.org/10.29296/2618723X-2021-02-04

Abstract

Urinary tract infections (UTI) are the most common diseases in outpatient and hospital practice, the most common manifestation of which is cystitis. In addition to the medical component, the social significance of this problem is extremely important. In connection with the unexpected onset, acute course, pronounced symptoms, the disease causes serious inconvenience in patients, which leads to a sharp decrease in physical and mental activity, restriction of freedom of movement. Acute cystitis (AC) is one of the most common infectious diseases that occurs mainly in young women and is manifested by dysuria, stranguria and frequent urination. In the vast majority of cases, OZ is an uncomplicated urinary tract infection caused by strains of Escherichia coli. In order to assess the effectiveness of drugs for the treatment and prevention of urinary tract diseases, as well as to study the pathogenesis of urological diseases, various models in laboratory animals are used in preclinical studies. The aim of this study was to develop a model of acute bacterial cystitis in rabbits with a visual assessment of the development of pathology based on clinical symptoms. As a test system, female rabbits of the White Giant breed were used, which are used to simulate pathologies of the genitourinary system. To simulate acute bacterial cystitis, the method of chemical damage to the mucous membrane of the urinary bladder was used, followed by the introduction of a bacterial suspension into its cavity. As damaging agents were selected: an alcoholic solution of salicylic acid and a combination of 1% protamine sulfate solution and 20% urea solution. Saline was used as a control substance. Two groups of animals were formed, which were injected with a bacterial suspension without preliminary chemical damage to the mucous membrane of the bladder. After modeling the pathology for 8 days, the animals were observed to assess the severity of the clinical manifestation of pathology and pain. During the experiment, urine samples were taken from animals three times for bacteriological and clinical analyzes, and ultrasound diagnostics of the bladder was performed. On the 8th day of the experiment, all animals were euthanized and a macroscopic assessment of the bladder tissue was performed. Histological analysis was also performed. Based on the results obtained, a model of acute bacterial cystitis was formed by chemical damage to the mucous membrane of the urinary bladder and the introduction of a suspension of E. coli. A score system for assessing the clinical manifestation of pathology and pain was developed. A pronounced manifestation of the pathological process with the preservation of clinical signs of pathology and pain, bacteriological seeding of urine throughout the experiment (8 days) was observed in animals of the group that received 0.05% alcoholic solution of salicylic acid and E. coli from rabbit feces as an inducer of pathology.

Full text avaliable in Russain only

Autors’ contributions

E.I. Trofimets – idea, literary analysis and data collection, participation in the experiment, data collection and analysis, swriting and editing of the text

M.N. Makarova – study concept and design, editing of the text, supervised the project, approved the final version of the manuscript

K.Е. Borovkova – microbiological research, analysis and recording of results

Ya.A. Guschin – conducting pathomorphological examination

J.V. Salmova – preparation of bacterial suspension, microbiological examination

L.R. Nikiforova – preparation of bacterial suspension, microbiological examination

A.E. Katelnikova – supervised the project

K.L. Kryshen – supervised the project

Conflict of interest

The authors declare no conflict of interest.

References

  1. Гяургиев Т.А. Тревожно-депрессивные расстройства у больных с хроническим рецидивирующим бактериальным циститом в стадии обострения // Вестник новых медицинских технологий. Электронное издание. 2013; 1. [Gyaurgiev T. A. Trevozhno-depressivnye rasstrojstva u bol'nyh s hronicheskim recidiviruyushchim bakterial'nym cistitom v stadii obostreniya // Vestnik novyh medicinskih tekhnologij. Elektronnoe izdanie. 2013; 1. (In Russ.)]
  2. Стручанский Л.С., Рафальский В.В. Клиническое значение и антибактериальная терапия острых циститов // Клиническая антимикробная химиотерапия. 1999; 1 (3): 84–91. [Struchanskij L.S., Rafal'skij V.V. Klinicheskoe znachenie i antibakterial'naya terapiya ostryh cistitov // Klinicheskaya antimikrobnaya himioterapiya. 1999; 1 (3): 84–91. (In Russ.)]
  3. Амдий Р.Э. Опыт применения канефрона при лечении острого неосложненного цистита у женщин // Урологические ведомости. 2016; 6 (2): 16–21. [Amdij R. E. Opyt primeneniya kanefrona pri lechenii ostrogo neoslozhnennogo cistita u zhenshchin // Urologicheskie vedomosti. 2016; 6 (2): 16-21. (In Russ.)]
  4. Desnottes J.F. et al. Experimental retrograde pyelonephritis and cystitis induced in rabbits by a group D Streptococcus sp.: serum antibody assay by a hemagglutination test // Infection and immunity. 1981; 33 (3): 647-650. PMID: 6793514
  5. Pugach J.L. Et al. Antibiotic hydrogel coated Foley catheters for prevention of urinary tract infection in a rabbit model // The Journal of urology. 1999; 162: 3 Part 1: 883-887. DOI: 10.1097/00005392-199909010-00084
  6. Niku S.D. еt al. A new method for cytodestruction of bladder epithelium using protamine sulfate and urea // The Journal of urology. 1994; 152 (3): 1025-1028. DOI: 10.1016/s0022-5347(17)32648-4
  7. Шолан Р.Ф. Результаты морфологического анализа стенки мочевого пузыря при интерстициальном цистите/синдроме болезненного мочевого пузыря в эксперименте // Вестник медицинского института «Реавиз»: реабилитация, врач и здоровье. 2019; 5 (41): 36-42. [SHolan R.F. Rezul'taty morfologicheskogo analiza stenki mochevogo puzyrya pri intersticial'nom cistite/sindrome boleznennogo mochevogo puzyrya v eksperimente // Vestnik medicinskogo instituta «Reaviz»: reabilitaciya, vrach i zdorov'e. 2019; 5 (41): 36-42. (In Russ.)]
  8. Шолан Р.Ф. Сравнительная оценка уровня фактора роста нервов в экспериментальных моделях интерстициального цистита/синдрома болезненного мочевого пузыря // Экспериментальная и клиническая урология. 2019; 3: 178-181. DOI: 10.29188/2222-8543-2019-11-3-178-181 [SHolan R. F. Sravnitel'naya ocenka urovnya faktora rosta nervov v eksperimental'nyh modelyah intersticial'nogo cistita/sindroma boleznennogo mochevogo puzyrya // Eksperimental'naya i klinicheskaya urologiya. 2019; 3: 178-181 (In Russ.)]
  9. Birder L., Andersson K.E. Animal modelling of interstitial cystitis/bladder pain syndrome // International neurourology journal. 2018; 22: Suppl 1: S3. DOI:10.5213/inj.1835062.531.
  10. Bagley R.S. The effect of experimental cystitis and iatrogenic blood contamination on the urine protein/creatinine ratio in the dog // Journal of veterinary internal medicine. 1991; 5 (2): 66-70. DOI: 10.1111/j.1939-1676.1991.tb00933.x.
  11. Senior D.F. Amoxycillin and clavulanic acid combination in the treatment of experimentally induced bacterial cystitis in cats // Research in veterinary science. 1985; 39 (1): 42-46. DOI: 10.1016/s0034-5288(18)31768-5.
  12. Choi B.H. Mast cell activation and response to tolterodine in the rat urinary bladder in a chronic model of intravesical protamine sulfate and bacterial endotoxin-induced cystitis // Molecular medicine reports. 2014; 10 (2): 670-676. DOI: 10.3892/mmr.2014.2262.
  13. Fraser M.O. A reliable, nondestructive animal model for interstitial cystitis: intravesical low-dose protamine sulfate combined with physiological concentrations of potassium chloride // Urology. 2001; 57 (6): 112. DOI: 10.1016/s0090-4295(01)01044-5.
  14. Soler R. Urine is necessary to provoke bladder inflammation in protamine sulfate induced urothelial injury // The Journal of urology. 2008; 180 (4): 1527-1531. DOI: 10.1016/j.juro.2008.06.006.
  15. Shao Y. Reduction of intercellular adhesion molecule 1 may play a role in anti-inflammatory effect of hyaluronic acid in a rat model of severe non-bacterial cystitis // World journal of urology. 2013; 31 (3): 535-540. DOI: 10.1007/s00345-012-0839-8
  16. Vysakh A. The influence of tissue antioxidant enzymes and inflammatory cascade in pathology of cystitis: an experimental rat model // Microbial pathogenesis. 2017; 113: 102-106. DOI: 10.1016/j.micpath.2017.10.035
  17. Altman D.G., Bland J.M. How to randomize // BMJ. 1999; 319: 703-704.
  18. Bland M. An Introduction to Medical Statistics (Oxford Medical Publications) 3rd edition, Oxford University Press. 2000; 422 р.
  19. Назарова А.В., Семенов Б: Применение усовершенствованной модели уретрального катетера для профилактики осложнений, возникающих после катетеризации мочевого пузыря у животных. 2019. С.73-75. [Nazarova A. V., Semenov B. S. Primenenie usovershenstvovannoj modeli uretral'nogo katetera dlya profilaktiki oslozhnenij, voznikayushchih posle kateterizacii mochevogo puzyrya u zhivotnyh. 2019. P.73-75. (In Russ.)]
  20. Директива 2010/63/EU Европейского Парламента и Совета Европейского Союза по охране животных, используемых в научных целях / пер. с англ. Под ред. М.С. Красильщиковой, И.В. Белозерцевой. СПб., 2012. 48 с.
  21. Sayers I. Approach to preventive health care and welfare in rabbits // In practice. 2010; 32 (5): 190-198. DOI: 10.1136/inp.c2228
  22. Vennen K.M., Mitchell M. A. Rabbits // Manual of Exotic Pet Practice. WB Saunders, 2009; 375-405.
  23. Cousquer G. Veterinary care of rabbits with myiasis // In practice. 2006; 28 (6): 342-349. DOI: 10.1136/inpract.28.6.342
  24. Brown C. Urolithiasis and cystotomy in the rabbit // Lab animal. 2011; 40 (3): 73-74. DOI: 10.1038/laban0311-73
  25. Rabbit Grimace Scale (RbtGS) Manual. https://www.nc3rs.org.uk/sites/default/files/documents/RbtGS%20Manual.pdf
  26. Козлов Р: Бактериологический анализ мочи: клинические рекомендации. 2014. 33 с.
  27. Пенник Д., д’Анжу М.-А. Атлас по ультразвуковой диагностике. Исследования у собак и кошек // М.-А. Д’Анжу. М.: Аквариум-Принт. 2015. 504 с.
  28. Гущин Я.А., Мужикян А.А. Влияние фиксирующих жидкостей на микроскопическую структуру органов мелких лабораторных животных // Международный вестник ветеринарии. 2014; 3: 88–95. [Gushchin YA.A., Muzhikyan A.A. Vliyanie fiksiruyushchih zhidkostej na mikroskopicheskuyu strukturu organov melkih laboratornyh zhivotnyh // Mezhdunarodnyj vestnik veterinarii. 2014; 3: 88–95. (In Russ.)]
  29. Мужикян А.А., Макарова М.Н., Гущин Я.А. Особенности гистологической обработки органов и тканей лабораторных животных // Международный вестник ветеринарии. 2014; 2: 103–109. [Muzhikyan A.A., Makarova M.N., Gushchin YA.A. Osobennosti gistologicheskoj obrabotki organov i tkanej laboratornyh zhivotnyh // Mezhdunarodnyj vestnik veterinarii. 2014; 2: 103–109.]
  30. Gallego M. Laboratory reference intervals for systolic blood pressure, rectal temperature, haematology, biochemistry and venous blood gas and electrolytes in healthy pet rabbits // Open veterinary journal. 2017; 7 (3): 203–207. DOI: 10.4314/ovj.v7i3.1
  31. Lippi I. et al. Evaluation of glomerular filtration rate estimation by means of plasma clearance of iohexol in domestic rabbits (Oryctolagus cuniculus) // American journal of veterinary research. 2019; 80: 6: 525–532. DOI: 10.2460/ajvr.80.6.525

You may be interested