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Molecular identification of Cryptosporidium isolates from ill exotic pet animals in Japan including a new subtype in Cryptosporidium fayeri.

ABSTRACT: Cryptosporidium is an obligate intracellular parasite which can cause fatal diarrheal disease in exotic animals. Sugar gliders (Petaurus breviceps), hedgehogs (Atelerix albiventris), chinchillas (Chinchilla lanigera), and common leopard geckos (Eublepharis macularius) are popular exotic animals commonly sold in pet shops in Japan. We herein investigated the species and subtypes of Cryptosporidium in these animals. Cryptosporidium fayeri was detected in a sugar glider in a Japanese animal hospital. Sequence analyses of the 60-kDa glycoprotein (gp60) gene revealed that C. fayeri belonged to subtype family IVh (IVhA13G2T1), which was proposed to be a new subtype. This is the first study to report C. fayeri infection in a sugar glider. In other animals, the Cryptosporidium horse genotype, C. ubiquitum, and C. varanii were detected in two four-toed hedgehogs (A. albiventris), a chinchilla (C. lanigera), and common leopard gecko (E. macularius), respectively. The gp60 subtypes identified were VIbA13 of the horse genotype and XIId of C. ubiquitum. The present results revealed that potentially zoonotic Cryptosporidium is widespread in exotic animals in Japan.

SUBMITTER: Takaki Y

PROVIDER: S-EPMC7324920 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundThe Sanger sequencing technique has been questioned and challenged by advanced high-throughput sequencing approaches. Sanger sequencing seems to be an obsolete technology. However, there are still research problems that could be answered using the Sanger sequencing technology. Fastidious obligate anaerobic bacteria are mostly associated with abscesses in animals. These bacteria are difficult to isolate from abscesses and are frequently excluded due to the bias of conventional bacterial culturing.AimThis study demonstrated the usefulness of a broad-range polymerase chain reaction (PCR) with Sanger sequencing to identify the majority population of bacteria in abscesses from exotic pet animals.Materials and methodsThis study performed a pilot investigation of abscesses from 20 clinical cases (17 rabbits, 2 hedgehogs, and 1 sugar glider) using standard culture methods for both aerobes and anaerobes and broad-range nested PCR targeting the 16S rRNA gene followed by the Sanger sequencing technique.ResultsThe standard culture and PCR techniques detected bacteria in 9 and 17 of 20 samples, respectively. From the 17 sequencings of the 16S rRNA, 10 PCR products were found to be closely related with obligate anaerobes including Bacteroides spp., Fusobacterium spp., Prevotella spp. Phylogenetic analysis using the rpoB gene revealed that the species for the Bacteroides was thetaiotaomicron and for the Fusobacterium was varium and nucleatum. However, the amplification of the rpoB gene for the Prevotella spp. was unsuccessful. Correlations between the standard culture and PCR techniques were found in 9 (6 positive and 3 negative samples) of 20 samples. Eleven samples were discordant between the standard culture and PCR techniques which were composed of eight samples negative by culture but positive by PCR and three samples had different bacteria by the culture and PCR techniques.ConclusionAccording to this study, broad-range PCR combined with Sanger sequencing might be useful for the detection of dominant anaerobic bacteria in abscesses that were overlooked based on conventional bacterial culture.

Dataset Information

Molecular identification of Cryptosporidium isolates from ill exotic pet animals in Japan including a new subtype in Cryptosporidium fayeri.

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