Isolation and identification of avirulent strains of Bacillus anthracis from environmental samples in Central Java, Indonesia

Research Article

International Journal of One Health, 7(2): 204-211

https://doi.org/10.14202/IJOH.2021.204-211

Ully Apriliana^1,2, Hendra Wibawa¹, Endang Ruhiat¹, Tri Untari³, and Soedarmanto Indarjulianto³

1. Disease Investigation Center Wates, Jalan Raya Jogja-Wates Km 27 Po Box 18, Wates, Yogyakarta, 55602, Indonesia.
2. Department of Internal Medicine, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Jl. Fauna 2, Karangmalang, Yogyakarta, 55281, Indonesia.
3. Department of Internal Medicine , Faculty of Veterinary Medicine, Universitas Gadjah Mada, Jl. Fauna 2, Karangmalang, Yogyakarta, 55281, Indonesia.

Background and Aim: Anthrax is a non-contagious infectious disease caused by Bacillus anthracis. The bacteria form spores that are resistant to extreme conditions and can contaminate the environment for decades. This study aimed to detect and characterize B. anthracis found in endemic areas of anthrax in Yogyakarta and Central Java province, Indonesia.

Materials and Methods: Soil samples were collected from Gunungkidul regency, Yogyakarta province (n=315) and Boyolali regency, Central Java province (n=100). Additional soil samples (n=10) and straw samples (n=5) were obtained from Pati regency, Central Java province. The isolation and identification of B. anthracis were performed using conventional methods: Morphology of bacteria colony in solid media, Gram staining, capsule staining, spores staining, and motility test. Isolates were further identified using polymerase chain reaction (PCR) against Ba813, lef (pXO1), and capC (pXO2) gene. An avirulent vaccine strain of B. anthracis (strain 34F2) was used as a control.

Results: Only four samples grew on blood agar with a ground-glass appearance, white-gray colony (Gunungkidul and avirulent strain) or yellowish (Boyolali and Pati). All were Gram-positive, presented chains, square-ended rods, spores, and were then identified as B. anthracis. Boyolali, Pati, and avirulent strain isolates had slightly different characteristics, including the growth of non-mucoid in the bicarbonate agar medium, and their uncapsulated form. The PCR showed two Gunungkidul isolates which amplified three genes, including Ba813, lef, and capC. Contrarily, the other isolates did not amplify the capC gene.

Conclusion: Gunungkidul isolates were identified as virulent strains of B. anthracis while Boyolali and Pati isolates were proposed as avirulent strains. This is the first report of isolation and identification of avirulent strains of B. anthracis in Central Java, Indonesia. Keywords: avirulent Bacillus anthracis, Boyolali, identification, pati, polymerase chain reaction multiplex.

Keywords: avirulent Bacillus anthracis, Boyolali, identification, pati, polymerase chain reaction multiplex.

How to cite this article: Apriliana U, Wibawa H, Ruhiat E, Untari T, Indarjulianto S (2021) Isolation and identification of avirulent strains of Bacillus anthracis from environmental samples in Central Java, Indonesia, Int J One Health, 7(2): 204-211.

Received: 29-04-2021 Accepted: 23-08-2021 Published online: 13-10-2021

Corresponding author: Soedarmanto Indarjulianto E-mail: indarjulianto@ugm.ac.id

DOI: 10.14202/IJOH.2021.204-211

Copyright: Apriliana, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.