Isolation of Bifidobacteria from feces of chimpanzees in the wild

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CommunicationIntestinal bacteria in wild animals have not been thoroughly studied due to technical problems such as poor laboratory equipment in fi eld conditions.The lack of electricity, clean water, and availability of CO 2 gas makes work with anaerobic cultures diffi cult.In the case of wild apes in Africa, even buildings are often lacking.However, these constraints can be overcome by the use of molecular-based techniques.Indeed, bacteriological surveys based on the 16S rRNA gene can be conducted in all research fi elds of wild apes if fresh feces are available.In a previous study, fresh feces were collected aseptically in ethanol to fi x bacteria for DNA preservation.After transportation to our laboratory in Kyoto, we successfully analyzed the bacterial 16S rRNA gene in the feces of chimpanzees in the wild (Fujita and Kageyama, 2007;Uenishi et al., 2007;Ushida, 2008).However, 16S rDNA-basis phylogenetic analyses of bacteria provide poor information on their functions in the intestine.Live bacterial cultures are essential to estimate their function in the physiology, particularly the nutrition, of the host (Gibson and Macfar-lane, 1995).In this report, we show the successful isolation of bifi dobacteria from the feces of wild chimpanzees living in the tropical forest of Bossou, Guinea (Matsuzawa, 2006), though the identifi cation of the isolates remains incomplete.In this experiment, all equipment, anaerobic media in particular, was prepared in Kyoto and transported to Bossou.TOS-propionate agar plates (Yakult, Tokyo) and LBS agar plates (Becton Dickinson and Company, Cockeysville, MD, USA) were prepared according to the manufacturerʼs instructions.Plates were placed in plastic bags (Techbarrier ® , Mitsubishi Chemicals, Tokyo) with Anaeropack ® (Mitsubishi Gas Chemicals, Tokyo).Plastic bags were closed by heat-sealing.TOS-propionate stubs and LBS stubs were also prepared under O 2 -free CO 2 using Hungate-type tubes (Bellco, Vineland, NJ, USA).Transportation to the research fi eld took 1 week after preparation.Fresh feces of 10 chimpanzees were successfully collected at the end of the rainy season of 2007 and rapidly transported (within ca.30 min of defecation) to a house in Bossou Village as indicated in a previous study (Ushida et al., 2006).If chimpanzees were located at a distant location from the village, sampling was abandoned.Collected feces were mixed with 9 volumes of a phosphate buffer (pH 6.5, 0.05 M) within a sterile 50-ml