Supplementary information
1. DNA larvae extraction protocol
This protocole is adapted from QIAamp DNA Mini kit Tissue instructions to be applied for
DNA extractions from larvae stored in Ethanol.
1- Homogenize flask and sample 10ml in a 15ml Falcon Tube.
Centrifuge 10 min at 3000T/min (14G).
Remove the supernatant.
2- Add 1.5 ml of PBS and transfer in a 1.5 ml Eppendorf.
Centrifuge10 min at 3000T/min.
Remove the supernatant.
Repeat step 2.
3- Add 100µl buffer ATL and freeze samples at -20°C.
4- Mix samples with a Pellet-Mixer.
5- Add 20 µl of Proteinase K, mix by vortexing and incubate at 56°C all the night. Do not
centrifuge.
6- Add 200 µl Buffer AL, mix by vortexing, and incubate at 70°C for 30 min.
7- Add 200 µl of ethanol (96-100%), mix slowly.
8- Carefully apply the mixture from step 7 to the QIAamp Mini spin column (in a 2ml
collection tube) without wetting the rim. Centrifuge 1 min at 10000 rpm. Place the QIAamp
Mini-spin column in a clean 2 ml collection tube and discard the tube containing the filtrate.
9- Carefully open the QIAamp Mini spin column and add 500 µl buffer AW1 without wetting
the rim. Close the cap, and centrifuge at 10 000 rpm for 1 min. Place the QIAamp Mini-spin
column in a clean 2 ml collection tube and discard the collection tube containing the filtrate.
10- Carefully open the QIAamp Mini-spin column and add 500 µl buffer AW2 without
wetting the rim. Close the cap, and centrifuge at 10 000 rpm for 1 min. Centrifuge again at
full speed for 1 min.
11- Place the QIAamp Mini spin column in a clean 1.5 ml eppendorf and discard the old
collection tube with the filtrate. Add 100 µl TE/Tween 0,05%. Incubate at room temperature
for 1 min and Centrifuge at 10 000 rpm for 1 min.
12- Repeat Step 11.
2. DNA dried feces extraction protocol
Only modifications on the Stool Pathogen Detection protocol from QIAamp DNA Stool kit
for DNA extraction from dried feces are indicated.
1- Weigh 100mg dried stool in a 2 ml micro-centrifuge tube and add 300 µl PBS 1x. Incubate
at room temperature for 10 min.
3- Incubate at 70°C all the night.
8’- Repeat step 8 one time.
9- Pipet 25 µl proteinase K into a new 1.5 ml microcentrifuge tybe.
11- Add 500 µl Buffer AL and vortex for 15 s.
12- Incubate at 70°C for 30 minutes.
13- Add 700 µl of ethanol (96-100% and mix by vortexing.
18- Transfer the QIAamp spin column into a new, labeled 1.5 ml microcentrifuge tube.
Carefully open the QIAamp spin column and pipet 100 µl TE/Tween 0,05% directly onto the
QIAamp membrane. Close the cap and incubate at room temperature for 1 min, then
centrifuge at full speed for 1 min to elute DNA.
3. Precisions on helminths identification in bonobo samples and pathogenicity
From the egg size and shape of the helminths observed in bonobos, we can suggest more
precise identification. The egg size and shape of the Strongyloides sp. observed herein is
consistent with S. fuelleborni (Hasegawa et al. 1983; Dupain et al. 2009). According to egg
size and host-specificity, Capillaria was probably C. brochieri, a species described in bonobos
by Justine (1988). Enterobius was likely to be E. anthropopitheci (Sandosham 1950, Hugot
1993, Hasegawa et al. 2005). One of the bonobo samples had a very high CPL for Enterobius
(2200 egg/g). Coprophagy behavior, observed in bonobos in another study area (Sakamaki,
2010) but also in bonobos of Manzano (Narat, et al. in prep), probably increases re-infection
and may account for this surprising result. The Toxocara sp. found in bonobos of Manzano
(and never described before in this species) is probably a spurious infection according to its
life cycle, caused for example by consumption of food contaminated by canid feces (dogs are
used by local people for traditional net hunting in the forest during the entire day). According
to their shape and size, the Troglodytella sp. detected in bonobos is compatible with being T.
abrassarti (described in chimpanzees). Occurring at high prevalence in African great apes,
this ciliate is probably a symbiont known to facilitate fiber fermentation (Pomajbíková et al.
2010). Among the strongylids found in bonobos, molecular data confirmed the presence of O.
stephanostomum, Necator sp. and a Nematodirus-like species in bonobos. The latter might be
N. weinbergii, which was identified by Railliet and Henry in 1909, and is the only species of
this genus that has been described in a primate (chimpanzee) (Neveu-Lemaire, 1917). O.
stephanostomum was found in several primate species in Uganda (chimpanzees and several
monkey species) (Krief et al. 2008; 2010; Ghai et al. 2014).