Acta Scientific Microbiology (ISSN: 2581-3226)

Research Article Volume 5 Issue 8

Antimicrobial Susceptibility Profile of Bacterial Culturome of Heart Blood Samples of Big Cats Died in Zoos and Wildlife Sanctuaries in Northern India

Bhoj R Singh1*, Mathesh Karikalan2, Abhijit M Pawde2, Ravichandran Karthikeyan1, Dharmendra K Sinha1 and Himani Agri1

1Division of Epidemiology, ICAR-Indian Veterinary Research Institute, Izatnagar, India

2Centre for Wildlife, ICAR-Indian Veterinary Research Institute, Izatnagar, India

*Corresponding Author: Bhoj R Singh, Division of Epidemiology, ICAR-Indian Veterinary Research Institute, Izatnagar, India.

Received: June 23, 2022; Published: July 25, 2022


In India, little is understood precisely about the cause of death in big cats in zoos, wild and wildlife sanctuaries. The present study reports the bacterial culturome (culturable bacteria) of heart blood of leopards (5), lions (9) and tigers (26) found dead in zoos (30) or wildlife sanctuaries (10). From the samples submitted to Clinical Epidemiological study 145 bacterial (46 Gram positive and 99 Gram negative) strains (the group of isolates with a separate identity) belonging to 44 species (19 of G+ve and 25 of G-ve bacteria) were identified. From 17 (12 tiger, 1 leopard, 4 lions) heart blood samples bacteria of the single species were isolated in pure culture indicating cases of septicemia. The most common isolation as single bacteria type was of E. coli (5), followed by isolation of Alcaligenes denitrificans (2), A. feacalis (2), and one each of Acinetobacter calcoaceticus, Bacillus cereus, Paenibacillus macerans, Enterobacter (Pantoea) agglomerans, Pseudomonas aeruginosa, Staphylococcus epidermidis, S. intermedius and Streptococcus pneumoniae. Multiple drug-resistance (MDR) was detected in 73.1% and isolates belonged to 134 resistotypes. There was no significant (p, >0.05) difference in occurrence of herbal antimicrobial resistance of strains isolated from different animal of different locality. Significantly high probability (p, ≤0.04) of MDR strains and strains resistant to citral, tetracycline, nitrofurantoin, chloramphenicol and imipenem was recorded in samples from animals died in wildlife sanctuaries than those died in zoos. Of the 40 carbapenem resistant (CR) isolates identified from 16 (40.0%) heart blood samples, 21 (all Gram positive) were negative for MBL and 19 CR strains producing MBL belonged to 8 species of G-ve bacteria. The MIC of imipenem for MBL producer CR isolates ranged between 2 to 32 µg mL-1 while for those not produced MBL MIC ranged from 1.5 to 256 µg mL-1, all the carbapenem susceptible isolates had imipenem MIC between 0.001 to 1.0 µg mL-1. The study concluded multiplicity of bacteria in heart blood of big cats died in zoo and wildlife sanctuaries. Presence of multiple bacterial types in 57.5% samples suggests need for aseptic and timely collection of blood samples to understand the true etiology of fatality among big cats. Detection of MDR, ESBL and CR bacteria from 25%, 37.5% and 40% samples is alarming because of chances of spreading of AMR in the environment from animals suffering from infections with MDR strains and died in wild.

Keywords: Lion; Tiger; Leopard; Carbapenem-resistance; Herbal Drug-resistance; Blood Culture; MDR


  1. Young E. “Some Important Parasitic and Other Diseases of Lion, Panthera leo, in the Kruger National Park”. Journal of South African Veterinary Association 46 (1975): 181-183.
  2. Munson L., et al. “Diseases of Captive Cheetahs (Acinonyx jubatus jubatus) in South Africa: a 20-Year Retrospective Survey”. Journal of Zoo and Wildlife Medicine 30 (1999): 342-347.
  3. Choi JH., et al. “Morganelliasis Pneumonia in A Captive Jaguar”. Journal of wildlife Diseases 38 (2002): 199-201.
  4. Michel AL., et al. “Wildlife Tuberculosis in South African Conservation Areas: Implications and Challenges”. Veterinary Microbiology 25 (2006): 91-100.
  5. Dybas CL. “Infectious Diseases Subdue Serengeti Lions”. BioScience 59 (2009): 8-13.
  6. Carvallo FR., et al. “Necrotizing pneumonia and pleuritis associated with extraintestinal pathogenic Escherichia coli in a tiger (Panthera tigris) cub”. Journal of Veterinary Diagnosis and Investigation 22 (2010): 136-140.
  7. Green J., et al. “African Lions and Zoonotic Diseases: Implications for Commercial Lion Farms in South Africa”. Animals 10 (2020): 1692.
  8. Sharma AK., et al. “Prevalence of Pathological Conditions in Zoo/Wild Animals in India: A Retrospective Study Based on Necropsy”. 84 (2014): 937-946.
  9. Srivastava A and Chakrabarty B. “Seasonal Distribution of Deaths of Tigers (Panthera Tigris) In Indian Zoos”. Zoos' Print Journal 17 (2002): 741-743.
  10. Weinstein M P., et al. "Current Blood Culture Methods and Systems: Clinical Concepts, Technology, and Interpretation”. Clinical Infectious Diseases 23 (1996): 40-46.
  11. Carter GR. “Diagnostic Procedures in Veterinary Microbiology”. 2nd edn, Charles C Thomas Publishers: Springfield (1975).
  12. Singh BR. “Labtop for Microbiology Laboratory”. Lambert Academic Publishing: Germany (2009).
  13. Kreig NR and Holt JG. “Bergey’s Manual of Systematic Bacteriology”. Williams and Wilkins; Balitimore (1984).
  14. Clinical and Laboratory Standards Institute. “Methods for Antimicrobial Dilution and Disk Susceptibility Testing of Infrequently Isolated or Fastidious Bacteria”, M45, 3rd Clinical and Laboratory Standards Institute, Wayne, USA (2015).
  15. Clinical and Laboratory Standards Institute. “Performance Standards for Antimicrobial Disk Susceptibility Tests for Bacteria Isolated from Animals”. CLSI Supplement VET01S; Replaces VET01-S2. Clinical and Laboratory Standards Institute, Wayne, USA (2015).
  16. Singh BR., et al. “Antimicrobial Activity of Lemongrass (Cymbopogon citratus) Oil Against Microbes of Environmental, Clinical and Food Origin”. International Research Journal of Pharmacy Pharmacology 1 (2011): 228-236.
  17. Hall KK and Lyman JA. “Updated Review of Blood Culture Contamination”. Clinical Microbiology Reviews 19 (2006): 788-802.
  18. Sharma M., et al. “Molecular Analysis of Coagulase-Negative Staphylococcus Isolates from Blood Cultures: Prevalence of Genotypic Variation and Polyclonal Bacteremia”. Clinical Infectious Diseases 33 (2001): 1317-1323.
  19. Rabinowitz PM., et al. “Pet-Related Infections”. American Family Physician 76 (2007): 1314-1322.
  20. Khaki P., et al. “Glanders Outbreak at Tehran Zoo, Iran”. Iranian Journal of Microbiology 4 (2012): 3-7.
  21. Iatta R., et al. “Zoonotic and Vector-borne Pathogens in Tigers from A Wildlife Safari Park, Italy”. International Journal for Parasitology, Parasites and Wildlife 12 (2020): 1-7.
  22. Hasan MJ., et al. “Bloodstream Infection with Pandrug-Resistant Alcaligenes Faecalis Treated with Double-Dose of Tigecycline”. ID Cases 8 (2019): e00600.
  23. Huang C. “Extensively Drug-resistant Alcaligenes faecalis Infection”. BMC Infectious Diseases 20 (2020): 833.
  24. Morrison AJ and Boyce K. “Peritonitis Caused by Alcaligenes denitrificans xylosoxydans: Case Report and Review of the Literature”. Journal of Clinical Microbiology 24 (1986): 879-881.
  25. Lai CC., et al. “Recurrent Bacteremia Caused by the Acinetobacter calcoaceticus-Acinetobacter baumannii Complex”. Journal of Clinical Microbiology 50 (2012): 2982-2986,
  26. Tusgul S., et al. “Bacillus cereus Bacteraemia: Comparison Between Haematologic and Nonhaematologic Patients”. New Microbes and New Infections 15 (2016): 65-71.
  27. Noskin GA., et al. “Paenibacillus macerans Pseudobacteremia Resulting from Contaminated Blood Culture Bottles in A Neonatal Intensive Care Unit”. American Journal of Infection Control 29 (2001): 126-129.
  28. Cheng A., et al. “Bacteremia Caused by Pantoea agglomerans at A Medical Center in Taiwan, 2000-2010”. Journal of Microbiology, Immunology and Infection 46 (2013): 187-194.
  29. Kim YJ., et al. “Risk factors for Mortality in Patients with Pseudomonas aeruginosa Bacteremia; Retrospective Study of Impact of Combination Antimicrobial Therapy”. BMC Infectious Diseases 14 (2014): 161.
  30. Cheung GYC and Otto M. “Understanding the Significance of Staphylococcus epidermidis Bacteremia in Babies and Children”. Current Opinion in Infectious Diseases 23 (2010): 208-216.
  31. Iyer S and Johnson LB. “Line-Related Staphylococcus intermedius Bacteremia Misidentified as a Coagulase-Negative Staphylococcus”. Infectious Diseases in Clinical Practice 20 (2012): 433-434.
  32. Cobo F., et al. “Streptococcus pneumoniae Bacteremia: Clinical and Microbiological Epidemiology in A Health Area of Southern Spain”. Infectious Disease Reports 4 (2012): e29.
  33. Watsa M and Group WDSF. “Rigorous Wildlife Disease Surveillance”. Science 369 (2020): 145-147.
  34. Kimman T., et al. “Assessing and Controlling Health Risks from Animal Husbandry”. NJAS Wagening Journal of Life Sciences 66:2013): 7-14.
  35. Mukarati NL., et al. “Occurrence, Prevalence and Intensity of Internal Parasite Infections of African Lions (Panthera leo) in Enclosures at A Recreation Park in Zimbabwe”. Journal of Zoo and Wildlife Medicine 44 (2013): 686-693.
  36. Whitehouse-Tedd KM., et al. “Dietary Factors Associated with Faecal Consistency and Other Indicators of Gastrointestinal Health in The Captive Cheetah (Acinonyx jubatus)”. PLoS ONE 10 (2015): e0120903.
  37. Kukreti I. “92 Lions Already Dead this Year” (2020).
  38. Kumar Vijay. "Incidences of Fetal Deaths in Rescued Female Leopards (Panthera-pardus) of Shivalik Hills in Northern India”. International Journal of Biodiversity and Conservation2 (2015): 81-84.
  39. Rai S. “Uttar Pradesh: High-tension Cable Kills Leopard” (2022).
  40. Joshi R and Agarwal R. “Mortality in the Protected Leopard’s Population, Uttarakhand, North India: A Free-Ranging Wildlife Species in Threat”. International Journal of Ecosystem4 (2012): 44-53.
  41. Spickler AR., et al. “Emerging and Exotic Diseases of Animals”. Iowa State University, Ames, Iowa, USA (2010).
  42. De Alwis MCL. “Haemorrhagic Septicaemia”. Australian Centre for International Agricultural Research, Canberra, Australia (1999).
  43. Singh BR. “Emergence of Antibiotic Resistance in Captive Wildlife”. In proceedings of National Congress on Wildlife Health (NCWH), at ICAR-Indian Veterinary Research Institute, Izatnagar (6-7 January 2017).
  44. Singh BR., et al. “Bacteriological Analysis of a Lethal Outbreak of Pasteurella canis in Spotted Deer (Axis Axis) in a Zoological Park in Bareilly, India”. Journal of Bacteriology and Mycology 7 (2020): 1124.
  45. Vinodh Kumar OR., et al. “Multi-Drug Resistant (MDR), Extended Spectrum Beta-Lactamase (ESBL) and New Delhi Metallo Beta-Lactamase (NDM) Producing Escherichia Coli in Rescued Sloth Bears (Melursus Ursinus), India”. Veterinary Research Communications 45 (2021): 163-170.
  46. Ahmed AM., et al. “Zoo Animals as Reservoirs of Gram-Negative Bacteria Harboring Integrons and Antimicrobial Resistance Genes”. Applied and Environmental Microbiology 73 (2007): 6686-6690.
  47. Li Y., et al. “Prevalence of Antibiotic Resistance in Zooanimal Foods”. Proceedings of the Ninth Conference on Zoo and Wildlife Nutrition, AZA Nutrition Advisory Group, Kansas City, MO (2011).
  48. Eze EA., et al. “Congruence Between the Drug Resistance Pattern of Escherichia coli and Proteus spp. Isolated from Humans and Those from Wild Animals”. African Journal of Microbiology Research 9 (2015): 1928-1934.
  49. Jobbins SE and Alexander KA. “From Whence They Came—Antibiotic-Resistant Escherichia coli in African Wildlife”. Journal of Wildlife Diseases 51 (2015): 811-826.
  50. Vittecoq M., et al. “Antimicrobial Resistance in Wildlife”. Journal of Applied Ecology 53 (2016): 519-529.
  51. Vinodh Kumar OR., et al. “Carbapenem Resistant Escherichia coli and Pseudomonas aeruginosa in Captive Blackbucks (Antilope cervicapra) and Leopards (Panthera pardus) from India”. Veterinarski Archives 91 (2021): 73-80.
  52. Bhardwaj M., et al. “Emergence of Carbapenemase Producing Pathogens in Animals”. Pharmaceutica Analytica Acta 6 (2015): 379.
  53. Doi A., et al. “Community-acquired Pneumonia Caused by Carbapenem-Resistant Streptococcus pneumoniae: Re-Examining Its Prevention and Treatment”. International Journal of General Medicine7 (2014): 253-257.
  54. Meletis G. “Carbapenem Resistance: Overview of The Problem and Future Perspectives”. Therapeutic Advances in Infectious Disease 3 (2016): 15-21.
  55. Singh BR. “Multiple-Herbal-Antimicrobial-Resistance (MHAR) in Microbes of Animals, Birds, Fish, Food, Lizard and Water Origin”. Proceedings of International conference and 28th Annual convention of IAVMI-2014 on “Challenges and Opportunities in Animal Health at the Face of Globalization and Climate Change”. 30th October 2014 to 1st November 2014. Department of Veterinary Microbiology and Immunology, DUVASU, Mathura, India (2014): 26-29.
  56. Singh BR., et al. “Antimicrobial Resistance Profile of Enteropathogens Isolated from Diarrhea Patients: Herbal Antimicrobials, a Ray of Hope”. Annals of Pharmacology and Pharmaceutics7 (2017): 1068.
  57. Vadhana P., et al. “Emergence of Herbal Antimicrobial Drug Resistance in Clinical Bacterial Isolates”. Pharmaceutica Analltica Acta 6 (2015): 434.


Citation: Bhoj R Singh., et al. “Antimicrobial Susceptibility Profile of Bacterial Culturome of Heart Blood Samples of Big Cats Died in Zoos and Wildlife Sanctuaries in Northern India". Acta Scientific Microbiology 5.8 (2022): 21-25.


Copyright: © 2022 Bhoj R Singh., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


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