Editorial Type:
Article Category: Case Report
 | 
Online Publication Date: 01 Mar 2020

Mycobacteriosis in a Zoo Population of Chinese Gliding Frogs (Rhacophorus dennysi) Due to Mycobacterium marinum

MA, VetMB, DVSc, DACZM,
DVM, IPSAV, DVSc, DACZM, DECZM (Zoo Health Management),
PhD,
BSc,
PhD,
MD, FRCPC,
DVM, MSc, PhD, and
DVM, DVSc
Page Range: 14 – 20
DOI: 10.5818/19-03-186.2
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Abstract

Mycobacteriosis was implicated in the deaths of eight Chinese gliding frogs (Rhacophorus dennysi) in a zoo population over a 3 yr period. Clinical signs included nonhealing skin lesions, cloacal prolapse, hind limb weakness, weight loss, and sudden death. Abnormalities on postmortem were proliferative or ulcerative skin lesions in four of eight, pneumonia in three of eight, and gall bladder empyema in two of eight cases. All eight clinical cases had multisystemic granulomas containing acid-fast bacilli. Tissues most commonly affected were lung (seven of eight), liver (six of eight), kidney (six of eight), spleen (five of eight), and heart (five of eight). The remaining eight clinically normal frogs in the population were euthanized: eight of eight had granulomatous lesions, with acid-fast bacilli in three of eight cases. A mycobacterial species was cultured from four of the clinical cases by the Public Health Ontario Laboratory and was initially misidentified as Mycobacterium tuberculosis complex by a commercial line-probe assay (GenoType Mycobacterium CM, Hain Lifesciences, Nehren, Germany). Further diagnostic testing using 16S rRNA gene sequencing ultimately identified the mycobacterial species as Mycobacterium marinum. The correct identification of mycobacterial species is essential in epidemiological investigations at zoological facilities, and in assessing health risks to staff and to other animals in the zoo population.

Figure 1.
Figure 1.

Systemic mycobacteriosis due to Mycobacterium marinum in a Chinese gliding frog. Ulcerated skin lesions of cutaneous mycobacteriosis on the rostrum and dorsum.


Figure 2.
Figure 2.

Gall bladder empyema (arrow) and pneumonia in a Chinese gliding frog due to Mycobacterium marinum.


Figure 3.
Figure 3.

Phylogenetic tree showing the Chinese gliding frog mycobacterial isolates. The 16S rRNA consensus sequence constructs (final construct size 1417 bp) of the isolates obtained from cases 6 (lung MK411581) and 8 (lung MK411583 and ovary MK411582) were compared to each other and to valid species sequences obtained from GenBank using the UPGMA standard algorithm (BioNumerics v6.6, Applied Maths [bioMérieux], Belgium). Value at nodes and scale indicates percent similarity. All 3 Chinese gliding frog case isolates were a 100% match to Mycobacterium marinum type strains (ATCC 927 and DSM 44344) and strains M (CP000854), Europe (AB716939).


Figure 4.
Figure 4.

(A) Granulomatous pneumonia due to Mycobacterium marinum in a Chinese gliding frog (Rhacophorus dennysi). H&E. Original objective 40×. (B) Filamentous beaded acid-fast bacteria within foci of inflammation in the respiratory pillars. Ziehl–Neelsen. Original objective 100×.


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