antibiotics Case Report Oral Abscess Caused by Chryseobacterium indologenes in Ball Python (Python regius); A Case Report Iradj Ashrafi Tamai 1, Babak Pakbin 2,* , Zahra Ziafati Kafi 1 and Wolfram Manuel Brück 2,* 1 Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran 1417466191, Iran; iashrafi@ut.ac.ir (I.A.T.); [email protected] (Z.Z.K.) 2 Institute for Life Technologies, University of Applied Sciences Western Switzerland Valais-Wallis, 1950 Sion 2, Switzerland * Correspondence: [email protected] (B.P.); [email protected] (W.M.B.) Abstract: Chryseobacterium indologenes is an opportunistic pathogen isolated from human infections and, rarely, from some aquatic animals. A 3-year-old male ball python (Python regius) was admitted to the veterinary clinic by a pet owner because of acute respiratory and swallowing failure. During physical examinations, oral secretions and abscesses were observed in the mouth cavity and throat of the animal. After microbiological analysis including isolation, identification, and 16s rRNA sequencing, C. indologenes was detected as the main cause of the oral abscess in this case. Phylogenetic relatedness analysis showed a close relationship between this isolate and other strains isolated from human infections. Antimicrobial susceptibility testing revealed that the isolate was multi-drug resistant. However, it was very sensitive to minocycline, ceftazidime, and tetracycline. The patient was treated by antibiotic therapy and completely recovered after two weeks. To the best of our knowledge, this is the first incidence of C. indologenes in an oral abscess in a ball python. As a result we would consider this organism as an opportunistic animal pathogen with zoonotic potentiality. Keywords: Chryseobacterium indologenes; oral abscess; ball python Citation: Tamai, I.A.; Pakbin, B.; Kafi, Z.Z.; Brück, W.M. Oral Abscess Caused by Chryseobacterium indologenes in Ball Python (Python 1. Introduction regius); A Case Report. Antibiotics Chryseobacterium species are gram-negative bacteria and have sporadically and ubiqui- 2021, 10, 686. https://doi.org/ tously been found in the environment. These bacteria have often been detected in water, 10.3390/antibiotics10060686 soil, waste, food sources, domestic animals, and aquatic environments. It was indicated Received: 4 May 2021 that Chryseobacterium species are emerging multi-drug resistant opportunistic nosocomial Accepted: 6 June 2021 pathogens and causes of different serious infections in neonates, pregnant women and Published: 8 June 2021 immunocompromised patients exposed to medical devices and environmental contami- nants [1]. Recent research indicated Chryseobacteria as the cause of some infectious disease Publisher’s Note: MDPI stays neutral outbreaks in fish and aquatic animals. C. balustinum, C. scophtalmum, C. joostei, C. piscicola with regard to jurisdictional claims in and C. arothri have been isolated from fish species such as salmon and rainbow trout [2]. published maps and institutional affil- C. indologenes have also been reported as the main infectious agent of fatal diseases in iations. newborns and other Chryseobacterium infections in human [3]. However, C. indologenes are occasionally isolated in some aquatic species such as yellow perch, Mediterranean limpets, mussels, and purple sea urchins. The isolates were mainly recovered from livers, kidneys, gills, and skin lesions. In these previous studies, all isolates were multi-resistant to a wide Copyright: © 2021 by the authors. range of antibiotics [4,5]. Investigation of genetic relatedness among the isolates showed Licensee MDPI, Basel, Switzerland. the evolutionary patterns of the pathogens [6]. Isolation of Chryseobacterium species such This article is an open access article as C. indologenes from terrestrial sources has not been reported yet. In the present study, distributed under the terms and we investigated the first case of an infected ball python (Pythonidae) with an oral abscess, conditions of the Creative Commons which, to our knowledge is the first case of Chryseobacterium infection in terrestrial animals, Attribution (CC BY) license (https:// caused by multidrug resistant C. indologenes. creativecommons.org/licenses/by/ 4.0/). Antibiotics 2021, 10, 686. https://doi.org/10.3390/antibiotics10060686 https://www.mdpi.com/journal/antibiotics AntibioticsAntibiotics2021 2021, 10, 10, 686, x FOR PEER REVIEW 22 of of 6 6 2.2. CaseCase PresentationPresentation AA three-year-old, three-year-old, 22 22 kg, kg, male male ball ball python python (Python (Python regius regius) was) submitted was submitted by a pet by owner a pet toowner the veterinary to the veterinary clinic at clinic the Faculty at the Faculty of Veterinary of Veterinary Medicine, Medicine, University University of Tehran, of Tehran,Tehran, IranTehran, due toIran respiratory due to respiratory failure, difficulty failure, swallowing,difficulty swallowing, oral secretions, oral badsecretions, breath, aggressionbad breath, andaggression anorexia. and The anorexia. snake hadThe beensnake fed had on been small fed prey, on includingsmall prey, rodents including and rodents birds, andand ◦ keptbirds, in and a terrarium. kept in a The terrarium. temperature The temperat and relativeure and humidity relative in humidity the terrarium in the were terrarium 35 C andwere 80% 35 RH.°C and Clinical 80% RH. assessments Clinical assessments of the snake of were the performedsnake were by performed the veterinarian by the ofveteri- the center.narian Respiratoryof the center. problem, Respiratory bad breath, problem, oral bad secretions, breath, andoral severalsecretions, acute and abscesses several wereacute observedabscesses in were the mouthobserved and in throatthe mouth of the and snake thro byat theof the veterinarian. snake by the Abscess veterinarian. samples Abscess were collectedsamples forwere microbiological collected for microbiological analysis and isolation analysis of theand infectious isolation agentsof the ofinfectious oral abscesses agents presentedof oral abscesses in the infected presented snake in the by infe oralcted surgery snake (Figure by oral1). surgery (Figure 1). FigureFigure 1.1.Oral Oral abscessabscess samplesample separatedseparated fromfrom the the Ball Ball Python Python mouth. mouth. Gram-negative bacilli were isolated from the abscess samples and the identities of the isolates were confirmed by biochemical tests. Antibiotic susceptibility of the isolates Antibiotics 2021, 10, 686 3 of 6 Antibiotics 2021, 10, x FOR PEER REVIEW 3 of 6 Gram-negative bacilli were isolated from the abscess samples and the identities of the isolates were confirmed by biochemical tests. Antibiotic susceptibility of the isolates was was evaluated using Kirby–Bauer disk diffusion as described by the Clinical and Labora- evaluated using Kirby–Bauer disk diffusion as described by the Clinical and Laboratory tory Standards Institute [7]. Biochemical analysis indicated that the isolate was beta-he- Standards Institute [7]. Biochemical analysis indicated that the isolate was beta-hemolytic; molytic; non-motile; indole positive; citrate negative; both methyl red and Voges–Pros- non-motile; indole positive; citrate negative; both methyl red and Voges–Proskauer’s neg- kauer’sative; glucose negative; was glucose fermented was withfermented gas production; with gas production; it was negative it wa fors negative urea and for unable urea andto grow unable on to MacConkey grow on MacConkey agar at 42 agar◦C. Antibiotic at 42 °C. Antibiotic susceptibility susceptibility testing showed testing thatshowed the thatisolate the was isolate sensitive was sensitive to minocycline, to minocycline, ceftazidime, ceftazidime, and tetracycline; and tetracycline; intermediately intermediately sensitive sensitiveto amikacin, to amikacin, ceftriaxone, ceftriaxone, ciprofloxacin, ciprofloxa enrofloxacin,cin, enrofloxacin, and levofloxacin; and levofloxacin; and resistant and re- to sistantimipenem, to imipenem, meropenem, meropenem, penicillin, penicillin, gentamycin, gentamycin, erythromycin, erythromycin, and azithromycin and azithromycin antibi- antibiotics.otics. 16s rRNA 16s rRNA gene gene sequencing sequencing was performedwas performed for definite for definite species species identification identification and andconstruction construction of the of phylogeneticthe phylogenetic tree oftree the of isolate.the isolate. The 16sThe rRNA16s rRNA gene gene of the of isolate the isolate was wasamplified amplified by conventional by conventional polymerase polymerase chain ch reactionain reaction (PCR) (PCR) using using the universalthe universal primers pri- merspA (5 pA0-AGA (5′-AGA GTT TGAGTT TGA TCC TGGTCC TGG CTC AG-3CTC 0AG-3) and′) pH and (5 pH0-AAG (5′-AAG GAG GAG GTG GTG ATC CAGATC CAG CCG CCGCA-3 0CA-3)[8].′) PCR [8]. PCR products products were were extracted, extracted, purified, purified, and and sequenced sequenced by GATC by GATC company com- pany(Cologne, (Cologne, Germany). Germany). Sequencing Sequencing of the 16sof th rRNAe 16s generRNA (NCBI, gene GenBank(NCBI, GenBank accession accession number: number:MT276587) MT276587) indicated indicated that the isolate that the was isolateChryseobacterium was Chryseobacterium indologenes indologenes. CLC software. CLC (CLC soft- wareGenomic (CLC WorkBench, Genomic WorkBench, Qiagen, Hilden, Qiagen, Germany) Hilden, was Germany) used to generatewas used the to cladogram generate the by cladogramthe neighbor-joining by the neighbor-joini clustering algorithm,ng