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Research Article |

Insight of Molecular Prevalence on Antibiotic Sensitive H. pylori Biotypes from Apparently Healthy and Clinical Illness Felines and Sheep

Helicobacter pylori is gram-negative bacteria may transmit through human food chain causing serious health problems in humans worldwide upon misusing antibiotic therapy for animals. Aim of the study represent the incidence of variants biotypes H. pylori susceptible to antibiotic in apparently healthy or clinical illness felines and sheep through amplification of 16srRNA. Four stomachs of 3 apparently healthy and diarrheal feline and two stool of constipated and pan-leukopenia felines, in addition to five gastric sheep from 2 normal and 3 congested plus one milk, selected from 52 felines and 83 sheep, respectively based on traditional cultivation and biochemical differentiation in total twelve H. pylori isolates that confirmed by amplification of 16srRNA, not being recognized by v3-v4 primer as nitrate gram negative bacteria. H. pylori isolates were grouped upon urease and nitrate reduction reaction in total percent 50% for each of weak and strong urease biotypes, including 33.3% & 66.6% for (+ve) or (-ve) nitrate reductive biotypes, respectively. Sensitivity of these biotypes was determined against fourteen antibiotic discs by antimicrobial susceptibility test to find highest sensitivity non-producing nitrate biotypes from felines is (87.5%), lesser than sheep (100%) but almost positive nitrate reductive isolates is less susceptible in percent 25%. Moderate sensitivity of weak urease biotypes represents 50% against amikacin, opposite to strong urease isolates (33.3%). Highest sensitivity strong urease biotypes show 83.3% against clarithromycin and levofloxacin, compared to weak urease biotypes 50 & 66.6%, respectively. Bio-typing H. pylori is preferable for programming eradication in molecular surveying normal or clinical illness animals.

Antimicrobial Susceptibility Test (AST), H. pylori, Nitrate, Urease, 16srRNA, Sheep and Felines

APA Style

Enany, M. E. S., Fadel, H. M., Abdelsadek, U. H. A., Ahmed, M. M. M., Kholief, M. E. A. G. (2024). Insight of Molecular Prevalence on Antibiotic Sensitive H. pylori Biotypes from Apparently Healthy and Clinical Illness Felines and Sheep. Animal and Veterinary Sciences, 12(1), 37-47. https://doi.org/10.11648/avs.20241201.15

ACS Style

Enany, M. E. S.; Fadel, H. M.; Abdelsadek, U. H. A.; Ahmed, M. M. M.; Kholief, M. E. A. G. Insight of Molecular Prevalence on Antibiotic Sensitive H. pylori Biotypes from Apparently Healthy and Clinical Illness Felines and Sheep. Anim. Vet. Sci. 2024, 12(1), 37-47. doi: 10.11648/avs.20241201.15

AMA Style

Enany MES, Fadel HM, Abdelsadek UHA, Ahmed MMM, Kholief MEAG. Insight of Molecular Prevalence on Antibiotic Sensitive H. pylori Biotypes from Apparently Healthy and Clinical Illness Felines and Sheep. Anim Vet Sci. 2024;12(1):37-47. doi: 10.11648/avs.20241201.15

Copyright © 2024 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

1. Sharma S, Prasad K, Chamoli D, Ayyagari A. Antimicrobial susceptibility pattern & biotyping of Helicobacter pylori isolates from patients with peptic ulcer diseases. The Indian Journal of Medical Research. 1995; 102: 261-6. PMID: 8557318.
2. Mowat C, Williams C, Gillen D, Hossack M, Gilmour D, Carswell A, et al. Omeprazole, Helicobacter pylori status, and alterations in the intragastric milieu facilitating bacterial N-nitrosation. Gastroenterology. 2000; 119(2): 339-47. https://doi.org/10.1053/gast.2000.9367.
3. Schulz C, Schütte K, Mayerle J, Malfertheiner P. The role of the gastric bacterial microbiome in gastric cancer: Helicobacter pylori and beyond. Therapeutic advances in gastroenterology. 2019; 12: https://doi.org/10.1177/1756284819894062.
4. Mladenova-Hristova I, Grekova O, Patel A. Zoonotic potential of Helicobacter spp. Journal of Microbiology, Immunology and Infection. 2017; 50 (3): 265-9. https://doi.org/10.1016/j.jmii.2016.11.003.
5. Lee A, Fox JG, Otto G, Murphy J. A small animal model of human Helicobacter pylori active chronic gastritis. Gastroenterology. 1990; 99(5): 1315-23. https://doi.org/10.1016/0016-5085 (90)91156-Z.
6. Mobley HL. Urease. Helicobacter pylori: physiology and genetics. Amer Soc Microbiol, Washington DC. 2001: 177-91. PMID: 21290711 Bookshelf ID: NBK2408.
7. Eriksson L, Valtonen M, editors. Seasonal changes in renal urea concentration in the reindeer (Rangifer tarandus L.). Annales Zoologici Fennici; JSTOR. 1974: http://www.jstor.org/stable/23731738.
8. Megraud F. H pylori antibiotic resistance: prevalence, importance, and advances in testing. Gut. 2004; 53(9): 1374-84. https://doi.org/10.1136/gut.2003.022111.
9. Justino MC, Ecobichon C, Fernandes AF, Boneca IG, Saraiva LM. Helicobacter pylori has an unprecedented nitric oxide detoxifying system. Antioxidants & Redox Signaling. 2012; 17(9): 1190-200. DOI: 10.1089/ars.2011.4304.
10. Shiotani A, Yanaoka K, Iguchi M, Saika A, Itoh H, Nishioka S. Helicobacter pylori infection reduces intraluminal nitric oxide in humans. Journal of gastroenterology. 1999; 34: 668-74. https://doi.org/10.1007/s005350050317.
11. Li Q, Yu H. The role of non-H.pylori bacteria in the development of gastric cancer. American Journal of Cancer Research. 2020; 10(8): 2271. PMID: 32905382; PMCID: PMC7471357.
12. Knorr J, Ricci V, Hatakeyama M, Backert S. Classification of Helicobacter pylori virulence factors: Is CagA a toxin or not? Trends in microbiology. 2019; 27(9): 731-8. https://doi.org/10.1016/j.tim.
13. Burkitt MD, Duckworth CA, Williams JM, Pritchard DM. Helicobacter pylori-induced gastric pathology: insights from in vivo and ex vivo models. Disease models & mechanisms. 2017; 10(2): 89-104. https://doi.org/10.1242/dmm.027649.
14. Al Sulami A, Al Kiat H, Bakker L, Hunoon H. Primary isolation and detection of Helicobacter pylori from dyspeptic patients: a simple, rapid method. EMHJ-Eastern Mediterranean Health Journal, 14(2), 268-276, 2008. 2008. https://iris.who.int/handle/10665/117434.
15. Patel SK, Pratap CB, Verma AK, Jain AK, Dixit VK, Nath G. Pseudomonas fluorescens-like bacteria from the stomach: a microbiological and molecular study. World Journal of Gastroenterology: WJG. 2013; 19(7): 1056. doi: 10.3748/wjg. v 19.i7.1056.
16. Heimesaat MM, Fischer A, Plickert R, Wiedemann T, Loddenkemper C, Göbel UB, et al. Helicobacter pylori induced gastric immunopathology is associated with distinct microbiota changes in the large intestines of long-term infected Mongolian gerbils. PloS one. 2014; 9(6): e100362. https://doi.org/10.1371/journal.pone.0100362.
17. Thyagarajan S, Ray P, Das BK, Ayyagari A, Khan AA, Dharmalingam S, et al. Geographical difference in antimicrobial resistance pattern of Helicobacter pylori clinical isolates from Indian patients: Multicentric study. Journal of gastroenterology and hepatology. 2003; 18(12): 1373-8. https://doi.org/10.1046/j.1440-1746.2003.03174.x.
18. Mishra K, Srivastava S, Garg A, Ayyagari A. Antibiotic susceptibility of Helicobacter pylori clinical isolates: comparative evaluation of disk-diffusion and E-test methods. Current microbiology. 2006; 53: 329-34. https://doi.org/10.1007/s00284-006-0143-1.
19. Raubenheimer K, Bondonno C, Blekkenhorst L, Wagner K-H, Peake JM, Neubauer O. Effects of dietary nitrate on inflammation and immune function, and implications for cardiovascular health. Nutrition reviews. 2019; 77(8): 584-99. https://doi.org/10.1093/nutrit/nuz025.
20. Graham DY, Miftahussurur M. Helicobacter pylori urease for diagnosis of Helicobacter pylori infection: A mini review. Journal of advanced research. 2018; 13: 51-7. https://doi.org/10.1016/j.jare.2018.01.006.
21. Dunn BE, Phadnis SH. Structure, function and localization of Helicobacter pylori urease. The Yale journal of biology and medicine. 1998; 71(2): 63. PMID: 10378351; PMCID: PMC2578883.
22. Loeffler IK. Euthanasia in veterinary field projects. Field Manual for Small Animal Medicine. 2018: 289-306. https://doi.org/10.1002/9781119380528.ch12.
23. Han S, Flamm R, Hachem C, Kim H, Clarridge J, Evans D, et al. Transport and storage of Helicobacter pylori from gastric mucosal biopsies and clinical isolates. European Journal of Clinical Microbiology and Infectious Diseases. 1995; 14: 349-52. https://doi.org/10.1007/BF02116531
24. Sabbagh P, Mohammadnia-Afrouzi M, Javanian M, Babazadeh A, Koppolu V, Vasigala VR, et al. Diagnostic methods for Helicobacter pylori infection: ideals, options, and limitations. European Journal of Clinical Microbiology & Infectious Diseases. 2019; 38: 55-66. https://doi.org/10.1007/s10096-018-3414-4.
25. Omar M, Aboelazm AA, El-Glil A, Reem R. Impact of Helicobacter pylori Eradication on Absolute Telomere Length in Gastric Mucosa. Afro-Egyptian Journal of Infectious and Endemic Diseases. 2016; 6(3): 121-7. DOI: 10.21608/AEJI.2016.9954.
26. Bertino E, Giribaldi M, Baro C, Giancotti V, Pazzi M, Peila C, et al. Effect of prolonged refrigeration on the lipid profile, lipase activity, and oxidative status of human milk. Journal of pediatric gastroenterology and nutrition. 2013; 56(4): 390-6. DOI: 10.1097/MPG.0b013e31827af155.
27. Howe R. JJS Snell, DFJ Brown & C. Roberts, Eds. Quality Assurance: Principles and Practice in the Microbiology Laboratory: Public Health Laboratory Service, UK, Journal of Antimicrobial Chemotherapy. 2000; 46(5): 865- ISBN 0-091144-452.
28. Agharid- Manssour, M. H. and Ahmed, Y. A. Polymerase chain reaction as diagnostic method for Helicobacter pylori in comparison with other conventional methods. Zag Vet J. (2008) 36(1): 85-91.
29. Harper CG, Xu S, Rogers AB, Feng Y, Shen Z, Taylor NS, et al. Isolation and characterization of novel Helicobacter spp. from the gastric mucosa of harp seals Phoca groenlandica. Diseases of aquatic organisms. 2003; 57(1-2): 1-9. doi: 10.3354/dao057001.
30. Park C, Smibert R, Blaser M, Vanderzant C, Stern N. Campylobacter in:" Compendium of methodes for the Microbiological examination of food". Speck. M (ed) American public Health Association, Washington, DC; 1984. Print ISSN: 0090-0036 | Electronic ISSN: 1541-0048.
31. Collee JG, Miles R, Watt B. Tests for identification of bacteria. Mackie and McCartney practical medical microbiology. 1996; 14: 131-49.
32. Chattopadhyay S, Patra R, Ramamurthy T, Chowdhury A, Santra A, Dhali G, et al. Multiplex PCR assay for rapid detection and genotyping of Helicobacter pylori directly from biopsy specimens. Journal of clinical microbiology. 2004; 42(6): 2821-4. https://doi.org/10.1128/jcm.
33. Hoshina S, Kahn SM, Jian W, Green PH, Neu HC, Chin N, et al. Direct detection and amplification of Helicobacter pylori ribosomal 16S gene segments from gastric endoscopic biopsies. Diagnostic microbiology and infectious disease. 1990; 13(6): 473-9. https://doi.org/10.1016/0732-8893 (90)90079-B.
34. Tiwari SK, Khan AA, Manoj G, Ahmed S, Abid Z, Habeeb A, et al. A simple multiplex PCR assay for diagnosing virulent Helicobacter pylori infection in human gastric biopsy specimens from subjects with gastric carcinoma and other gastro‐duodenal diseases. Journal of applied microbiology. 2007; 103 (6): 2353-60. https://doi.org/10.1111/j.1365-2672.2007.03478.x.
35. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: Cold spring harbor laboratory press; 1989. ISBN: 978-1-936113-42-2.
36. Wu X, Zhang H, Chen J, Shang S, Wei Q, Yan J, et al. Comparison of the fecal microbiota of holes high-throughput Illumina sequencing of the V3–V4 region of the 16S rRNA gene. Applied microbiology and biotechnology. 2016; 100: 3577-86. https://doi.org/10.1007/s00253-015-7257
37. Standards Laboratory Clinical for Committee National susceptibility antimicrobial for standards performance "NCCLS" (2001): testing. Supplement M 100-S111. Villanova, PA, USA.
38. Chen D, Cunningham SA, Cole NC, Kohner PC, Mandrekar JN, Patel R. Phenotypic and molecular antimicrobial susceptibility of Helicobacter pylori. Antimicrobial agents and chemotherapy. 2017; 61(4): 10.1128/aac. 02530-16. https://doi.org/10.1128/aac.02530-16.
39. Singh S, Yadav AS, Singh SM, Bharti P. Prevalence of Salmonella in chicken eggs collected from poultry farms and marketing channels and their antimicrobial resistance. Food Research International. 2010; 43(8): 2027-30. https://doi.org/10.1016/j.foodres.2010.06.001.
40. Carp I. IBM SPSS statistics for windows, version 22.0. Armonk, NY: IBM Corp. 2013.
41. Simoes LC, Simões M. Biofilms in drinking water: problems and solutions. Rsc Advances. 2013; 3(8): 2520-33. https://doi.org/10.1039/C2RA22243D.
42. Keikha M, Karbalaei M. Probiotics as the live microscopic fighters against Helicobacter pylori gastric infections. BMC gastroenterology. 2021; 21(1): 1-18. https://doi.org/10.1186/s12876-021-01977-1.
43. Shariati A, Arshadi M, Khosrojerdi MA, Abedinzadeh M, Ganjalishahi M, Maleki A, Heidary M, Khoshnood S. The resistance mechanisms of bacteria against ciprofloxacin and new approaches for enhancing the efficacy of this antibiotic. Front Public Health. 2022; 10: 1025633. doi: 10.3389/fpubh.2022.1025633. PMID: 36620240; PMCID: PMC9815622.
44. Mohammed HH, Abbas SH, Hayallah AM, Abuo-Rahma GE, Mostafa YA. Novel urea linked ciprofloxacin-chalcone hybrids having antiproliferative topoisomerases I/II inhibitory activities and caspases-mediated apoptosis. Bioorganic Chemistry. 2021; 106: 104422. DOI: 10.1016/j.bioorg.2020.104422.
45. Sizar O, Rahman S, Sundareshan V. Amikacin. In: StatPearls [Internet]. Treasure Island (FL): Stat Pearls [Updated 2023 Jul 17]. Publishing; Available from: https://www.ncbi.nlm.nih.gov/books/NBK430908.
46. Domnin PA, Parfenov VA, Kononikhin AS, Petrov SV, Shevlyagina NV, Arkhipova AY, et al. Combined impact of magnetic force and spaceflight conditions on Escherichia coli physiology. International Journal of Molecular Sciences. 2022; 23(3): 1837. https://doi.org/10.3390/ijms23031837.
47. Joosten M, Lindén S, Rossi M, Tay ACY, Skoog E, Padra M, et al. Divergence between the highly virulent zoonotic pathogen Helicobacter heilmannii and its closest relative, the low-virulence “Helicobacter ailurogastricus” sp. nov. Infection and Immunity. 2016; 84(1): 293-306. DOI: https://doi.org/10.1128/iai.01300-15.
48. Gang JG, Yun SK, Choi KM, Lim WJ, Park JK, Hwang SY. Significance of urease distribution across Helicobacter pylori membrane. Journal of microbiology and biotechnology. 2001; 11(2): 317-25. doi: 10.1128/iai.66.11.5060-5066.1998.
49. Zeng Q, An S. Identifying the Biogeographic Patterns of Rare and Abundant Bacterial Communities Using Different Primer Sets on the Loess Plateau. Microorganisms. 2021. 9; 9(1): 139. doi: 10.3390/microorganisms 9010139. PMID: 33435426; PMCID: PMC7827256.
50. Yu J, Wang Y, Xiao Y, Li X, Xu X, Zhao H, Wu L, Li J. Effects of chronic nitrate exposure on the intestinal morphology, immune status, barrier function, and microbiota of juvenile turbot (Scophthalmus maximus). Ecotoxicology and Environmental Safety. 2021; 207: 111287. https://doi.org/10.1016/j.ecoenv.2020.111287.
51. Sarraseca A, Milne E, Metcalf MJ, Lobley GE. Urea recycling in sheep: effects of intake. British Journal of Nutrition. 1998 Jan; 79(1): 79-88. PMID: 9505805 DOI: 10.1079/bjn19980011.
52. Pich OQ, Merrell DS. The ferric uptake regulator of Helicobacter pylori: a critical player in the battle for iron and colonization of the stomach. Future Microbiol. 2013 Jun; 8(6): 725-38. doi: 10.2217/fmb.13.43. PMID: 23701330; PMCID: PMC3852439.
53. Shutter MC, Akhondi H. Tetracycline. In: StatPearls [Internet]. Treasure Island (FL): StatPearls [Updated 2023 Jun 5]. Publishing; Available from: https://www.ncbi.nlm.nih.gov/books/NBK549905.
54. Amoxicillin, PubChem Compound Summary for CID 33613, PubChem [Internet]. Bethesda (MD): National Library of Medicine (US), National Center for Biotechnology Information; 2004. Available from: https://pubchem.ncbi.nlm.nih.gov/compound/Amoxicillin.
55. Marily C. Roberts, Tetracycline resistance determinants: mechanisms of action, regulation of expression, genetic mobility, and distribution, FEMS Microbiology Reviews, October 1996, P: 1–24, Vol: 19, Issue 1, https://doi.org/10.1111/j.1574-6976.1996.tb00251.x.
56. Bhattacharjee B, Das A, Das G, Ramesh A. Urea-Based Ligand as an Efflux Pump Inhibitor: Warhead to Counter Ciprofloxacin Resistance and Inhibit Collagen Adhesion by MRSA. ACS Applied Bio Materials. 2022 Mar 28; 5(4): 1710-20. PMID: 35344332. DOI: 10.1021/acsabm.2c00092.
57. Leitsch D. A review on metronidazole: an old warhorse in antimicrobial chemotherapy. Parasitology. 2019 Aug; 146(9): 1167-78. PMID: 29166971 DOI: 10.1017/S0031182017002025.
58. Raghu B, SARMA GR, Venkatesan P. Effect of anti-tuberculosis drugs on the iron-sequestration mechanisms of mycobacteria. Indian Journal of Pathology and Microbiology. 1995 Jul 1; 38(3): 287-92. PMID: 8819661.
59. McKinney JD, Lee JY, O'Neill RE, Goldfarb A. Overexpression and purification of a biologically active rifampicin-resistant beta subunit of Escherichia coli RNA polymerase. Gene. 1987; 58(1): 13-8. doi: 10.1016/0378-1119(87)90024-2. PMID: 3319782.
60. Takemori N, Ooi HK, Imai G, Hoshino K, Saio M. Possible mechanisms of action of clarithromycin and its clinical application as a repurposing drug for treating multiple myeloma. Ecancermedicalscience. 2020 Aug 18; 14: 1088. doi: 10.3332/ecancer.2020.1088. PMID: 33014130; PMCID: PMC7498274.
61. Levofloxacin, PubChem Compound Summary for CID 149096, PubChem [Internet]. Bethesda (MD): National Library of Medicine (US), National Center for Biotechnology Information; 2004. Available from: https://pubchem.ncbi.nlm.nih.gov/compound/Levofloxacin.
62. Jacoby GA. Mechanisms of resistance to quinolones. Clinical infectious diseases. 2005 Jul 15; 41(Supplement_2): S120-6. https://doi.org/10.1086/428052.