Coprophilous ascomycetes in Kenya: Saccobolus species from wildlife dung

Abstract

The taxonomy, occurrence and distribution of Saccobolus species was investigated from wild herbivore dung types in Kenya. Dung samples incubated in a moist chamber culture were examined for fungi over three months. Seven species, Saccobolus citrinus, S. depauperatus, S. diffusus, S. infestans, S. platensis, S. truncatus and S. versicolor were isolated from African elephant, black rhinoceros, Cape buffalo, dikdik, giraffe, hartebeest, hippopotamus, impala, waterbuck and zebra dung. Five taxa, S. citrinus, S. diffusus, S. infestans, S. platensis and S. truncatus, are new records for Kenya. The most common taxa were S. depauperatus and S. citrinus. The diversity of coprophilous Saccobolus species in wildlife dung is very high.

Full text

Mycosphere Doi 10.5943/mycosphere/3/2/2
111
Coprophilous ascomycetes in Kenya: Saccobolus species from wildlife dung
Mungai PG1, 2,3*, Chukeatirote E1,2, Njogu JG3 and Hyde KD1,2,
1Institute of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand
2School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
3Biodiversity Research and Monitoring Division, Kenya Wildlife Service, P.O. Box 40241 00100 Nairobi, Kenya
Mungai PG, Chukeatirote E, Njogu JG, Hyde KD 2012 – Coprophilous ascomycetes in Kenya:
Saccobolus species from wildlife dung. Mycosphere 3(2), 111-129, Doi 10.5943/mycosphere/3/2/2
The taxonomy, occurrence and distribution of Saccobolus species was investigated from wild
herbivore dung types in Kenya. Dung samples incubated in a moist chamber culture were examined
for fungi over three months. Seven species, Saccobolus citrinus, S. depauperatus, S. diffusus, S.
infestans, S. platensis, S. truncatus and S. versicolor were isolated from African elephant, black
rhinoceros, Cape buffalo, dikdik, giraffe, hartebeest, hippopotamus, impala, waterbuck and zebra
dung. Five taxa, S. citrinus, S. diffusus, S. infestans, S. platensis and S. truncatus, are new records
for Kenya. The most common taxa were S. depauperatus and S. citrinus. The diversity of
coprophilous Saccobolus species in wildlife dung is very high.
Key words – African elephant – diversity – moist chambers – national parks – Saccobolus citrinus
Article Information
Received 25 February 2012
Accepted 27 February 2012
Published online 17 March 2012
*Corresponding author: Paul G. Mungai – email – emu@kws.go.ke
Introduction
The genus Saccobolus Boud. (sub-
family Ascobolideae Gray, family Ascobola-
ceae Sacc.) contains over 25 species (Kirk et al.
2008). The main features of this genus include
apothecioid ascomata, clustered ascospores,
which in most cases have brown or purple-
brownish episporic pigment at maturity (van
Brummelen 1967, Kaushal & Virdi 1986,
Doveri 2004, Bell 2005), with each cluster
enclosed in a common gelatinous membrane.
The firmly clustered bundles of ascospores are
usually arranged in regular patterns (van
Brummelen 1967). Saccobolus is related to
Ascobolus Pers. from which it is differentiated
by its clustered ascospores. Another important
difference between Saccobolus and Ascobolus
is the shorter but broader asci and ascospores
of Saccobolus (van Brummelen 1967, Doveri
2004, Bell 2005). The apices of paraphyses
usually produce a coloured pigment (van
Brummelen 1967, Doveri 2004, Bell 2005).
The asci are phototropic, operculate and
protrude above the hymenial surface on
maturity. van Brummelen (1967) divided
Saccobolus into two distinct sections,
Saccobolus whose receptacles and discs are
always yellow or amber with yellow or
pigmented paraphyses and Eriobolus with
white, pale violet, violet or brown receptacles
and discs and usually colourless paraphyses.
Saccobolus comprises predominantly
cosmopolitan and coprophilous species (van
Brummelen 1967, Doveri 2004). Numerous
Saccobolus species have been recorded
globally, for instance, Europe (van Brummelen
1967), Australia and New Zealand (Bell 1983,
2005), Taiwan (Wang 2000) and South
America (Richardson 2008). In Africa, records
of Saccobolus include, S. beckii Heimerl., S.
glaber (Pers.) Lambotte, S. minimus Velen., S.
portoricensis Seaver and S. verrucisporus
Brumm from South Africa (Ebersohn & Eicker
1992) while S. citrinus Boud &

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Fig 1. Map of Kenya showing wildlife dung sampling sites

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Torrend and S. glaber were recorded from
Egypt (Abdel-Azeem et al. 2005). From Kenya
Minoura (1969) and Caretta et al. (1998)
recorded S. depauperatus (Berk. & Broome)
Rehm, S. versicolor (P. Karst.) P. Karst. and S.
violascens Boud.
The objectives of the present survey
were to 1) describe and classify Saccobolus
species found on various dung types from
Kenyan wildlife and, 2) document species
diversity and distribution of Saccobolus species
associated with different dung types.
In this study seven species of Sacco-
bolus from wildlife dung are examined and
illustrated.
Materials and Methods
For a detailed methodology refer to our
earlier work on coprophilous Ascobolus in
Kenya (Mungai et al. 2012). The sampling sites
are shown in Fig. 1.
Taxonomy of Saccobolus species from
Kenyan wildlife dung
Saccobolus citrinus Boud. & Torrend, Bull.
Soc. Mycol. Fr. 27: 131, 1911. (Figs.2A–I &
3A–F) Ascomata apothecioid, scattered or
gregarious, superficial, sessile, 180–215 μm
high, 205–315 μm diam. Receptacle outer
surface bright to lemon yellow, smooth,
without margin, subglobose, pulvinate or
lenticular at maturity. Disc convex, membra-
naceous, lemon yellow, dotted with blackish
tips of ripe protruding asci. Hypothecium and
medullary excipulum not differentiated from
ectal excipulum. Ectal excipulum thin
composed of textura globulosa, pale yellow to
yellowish grey cells, 8–20 × 8–18 μm.
Paraphyses cylindric-filiform, simple, septate,
exceeding asci, 3 μm broad, not branched,
inflated tips with abundant yellow
pigmentation. Asci 113–148 × 27–34 μm, 8-
spored, unitunicate, broadly clavate, thick-
walled, flat apex, walls turning blue in
Melzer’s reagent; stipe short, 8–10.5 × 5.5–6
μm, operculate. Ascospores 17–21 × 8–9 μm,
arranged according to van Brummelen pattern
I, ellipsoidal-fusoid, violet to brownish purple,
slightly asymetrical, with truncate or blunt
ends, verruculose, sometimes with fissures,
thick-walled; clusters elongated, 45–50.5 × 15–
20 μm, compact and firmly enclosed all round
in a narrow gelatinous envelope 2–4 μm thick.
Material examined (4 isolates) –
KENYA, Nairobi National Park, Nairobi
Province, GPS 37M0255729 9849626, altitude
1680m, impala dung, 20 August 2010, Paul
Mungai, KWSNNP020-2010; Nairobi National
Park, Nairobi Province, GPS 37M0255191
9849808, altitude 1693m, wooded grassland,
Cape buffalo dung, 20 August 2010, Paul
Mungai, KWSNNP015-2010; Nairobi National
Park, Nairobi Province, GPS 37M0257082
9850692, altitude 1668m, wooded grassland,
giraffe dung, 20 August 2010, Paul Mungai,
KWSNNP017B-2010; Nairobi National Park,
Nairobi Province, GPS 37M0254965 9850592,
altitude 1685m, wooded grassland, hippopo-
tamus dung, 20 August 2010, Paul Mungai,
KWSNNP021-2010.
Notes – Saccobolus citrinus Sect.
Saccobolus Boud. is similar to S. succineus
Brumm., (van Brummelen 1969, Doveri 2004).
However, it can be distinguished from these
species by having lemon-yellow apothecia and
notably truncate ended ascospores. In addition,
the ascospores of S. citrinus (21–24.5 × 8–9.5
µm in this examination) are narrower and more
finely warted (van Brummelen 1967, Doveri
2004, Bell 2005) while those of S. succineus
are larger. S. citrinus is quite a common species
on wildlife herbivore dung in Kenya and it is a
new record for Kenya.
Saccobolus depauperatus Berk. & Broome.,
Ann. Mag. Nat. Hist. III 15: 448, 1865. (Figs.
4A–G, 5A–C)
Ascomata apothecioid, gregarious,
superficial, sessile, 180–300 µm diam.
Receptacle globose in the early stages,
pulvinate to subglobose later, dirty white to
pale violet, with a narrow base, smooth, margin
not distinct. Disc convex, white, dotted with
pale violet tips of protruding ripe asci, outer
surface somewhat glabrous. Hymenium thick.
Hypothecium thin, of small very compacted
isodiametric cells, not well delineated.
Medullary excipulum thick, made up of small
textura globulosa cells. Ectal excipulum of
thin, textura globulosa-angularis cells 7–11 ×
5.5–10 µm, intercellular pigment not
conspicuous amongst these cells. Paraphyses

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Fig. 2 – Saccobolus citrinus (KWSNNP020-2010). A-B Ascomata on dung. C Squashed ascoma. D
Details of ectal excipulum near the margin. E Paraphyses filled with yellow pigment. F Mature asci
and ascospores, note yellow mucus (arrow). G Mature ascospore clusters, note gelatinous sheath
(arrow). H Mature ascus showing a stipe (white arrow) and an apex. I Apical portion of asci, note
closed operculum (arrow). Scale bars: A-B = 500 µm, C = 200 µm, D = 20 µm, E = 50 µm, F = 20
µm, G = 20 µm, H = 20 µm, I = 20 µm, J = 50 µm.

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Fig. 3 – Saccobolus citrinus (KWSNNP020-2010). A Ascus, note stipe (arrow). B Ascospore cluster,
note gelatinous sheath (arrow). C Free ascospore cluster and an immature ascus, note spore
arrangement (black arrow), operculum, apical ring (white arrow). D-F Features of asci, ascospores
and paraphyses, note yellow mucus (black arrow) and verruculose episporium (white arrows). Scale
bars: A-D = 20 µm, E-F = 50 µm.

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cylindric-filiform, septate, exceeding the asci,
width 2.5–4 µm, with hyaline vacuoles; tips
hyaline, not inflated. Asci 62–72 × 15.5–18.5
µm, 8-spored, unitunicate, claviform, often
curved at apex, wall turning deep blue in
Melzer’s reagent, with an operculum ca.14.5
µm, long stipitate. Ascospores 11.5–14 × 5–7
µm, single-celled, arranged according to van
Brummelen pattern II, ellipsoidal-subfusiform,
somewhat asymetrical, round or truncate at the
ends, purple-brown at maturity, smooth or
finely dotted; clusters compact, 31.5–35 ×
11.5–13.5 µm, usually surrounded by a thin
hyaline gelatinous sheath.
Material examined (10 isolates) –
KENYA, Shimba Hills National Reserve,
Coast Province, GPS S04°13’35.6”
E039°25’31.6”, altitude 373m, giraffe dung,
26 August 2009, Paul Mungai, KWSSH001-
2009; Tsavo East National Park, Coast
Province, GPS S03°21’06.4” E038°37’50.1”,
altitude 514m, riverine vegetation, Cape
buffalo dung, 23 August 2008, Paul Mungai,
KWSTE008A-2008; Aberdare Country Club
Game Sanctuary, Central Province, GPS
S00°19’28.1” E036°55’54.3”, altitude 2161m,
bushed grassland, zebra dung, 30 August 2009,
Paul Mungai, KWSACC002-2009. Nairobi
National Park, Nairobi Province, GPS
S01°21’15.1”, E036°46’54.1”, altitude 1768m,
savannah grassland, impala dung, 31 August
2009, Paul Mungai, KWSNNP008-2009;
Tsavo East National Park, Coast Province, GPS
S03°02’29.7” E038°41’35.8”, altitude 354m,
riverine vegetation, dikdik dung, 27 August
2009, Paul Mungai; KWSTE005B-2009;
Aberdare Country Club Game Sanctuary,
Central Province, GPS S00°19’28.1
E036°55’54.3”, altitude 2161m, bushed
grassland, giraffe dung, 30 August 2009, Paul
Mungai, KWSACC003-2009; Aberdare
National Park, Central Province, GPS
S00°21’35.0” E036°52’47.8”, altitude 2074m,
montane forest, African elephant dung, 30
August 2009, Paul Mungai KWSANP003-
2009; Tsavo East National Park, Coast
Province, GPS S03°21’66.6” E038°38’77.2”,
altitude 514m, riverine vegetation, dikdik dung,
23 September 2008, P. Mungai, KWSTE005A-
2008; Shimba Hills National Reserve, Coast
Province, GPS S04°14’35.4” E039°26’07.2”,
altitude 361m, hartebeest dung, 26 August
2009, Paul Mungai, KWSSH003-2009;
Kinondo Forest Reserve, Coast Province, GPS
S04°25’19.7” E039°32’60.2”, altitude 6m,
coastal tropical rain forest, dikdik dung, 19
April 2009, P. Mungai, KWSKIN004-2009;
Nairobi National Park, Nairobi Province, GPS
S01°21’25.5”, E036°47’51.6”, altitude 1748m,
savannah grassland, giraffe dung, 31 August
2009, Paul Mungai, KWSNNP009-2009.
Notes – Saccobolus depauperatus Sect
Eriobolus Sacc. is similar to S. versicolor (van
Brummelen 1967, De Sloover 2002, Doveri
2004). The two species however have some
important differences that help in their
differentiation. The latter has larger apothecia,
asci, single ascospores and ascospore clusters
(van Brummelen 1967, Doveri 2004). The
excipulum of S. depauperatus is usually of a
textura globulosa structure unlike that of S.
versicolor which is textura intricata (van
Brummelen 1967, Doveri 2004). The apices of
paraphyses and excipular cells of S.
depauperatus are also without any pigmented
intercellular mucus rendering the species to
appear whitish (van Brummelen 1967, De
Sloover 2002, Doveri 2004).
Saccobolus diffusus S.C. Kaushal & Virdi,
Willdenowia 16(1): 269, 1986. (Figs.6A–J, 7K–
R)
Ascomata apothecioid, scattered or gregarious,
superficial, sessile, 220–370 µm diam.
Receptacle at first subglobose to globose,
obconic, finally pulvinate, greenish yellow to
pale, external surface glabrous, margin entire.
Hymenium convex, dotted with black tips of
mature protruding ripe asci. Hypothecium thin,
of small textura globulosa cells. Medullary
excipulum not well delineated. Ectal excipulum
of textura angularis-globulosa cells 8–10 ×
4.5–8 µm. Paraphyses cylindric-filiform,
simple or rarely branched, septate, exceeding
asci, 2.5–4 µm broad; tips with abundant
greenish yellow pigmentation, not inflated.
Asci 114–167 × 29–39 µm, 8-spored, walls
turning blue in Melzer’s reagent, unitunicate,
broadly clavate, tapering abruptly into a short
stout and lobate stipe 13–25 × 6.5–7.5 µm;
apices subtruncate with an operculum 14.5–17
µm wide. Ascospores 21–24.5 × 8–9.5 µm,
single-celled, ellipsoidal, at first hyaline, violet
to brown, smooth or verruculose,

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Fig. 4 – Saccobolus depauperatus (KWSSH003-2009). A Ascomata on dung. B Squashed ascoma.
C Details of ectal excipulum near the margin. D Paraphyses, asci and ascospores (arrow). E-F
Details of cells of the ectal excipulum seen from outside, medullary excipulum, mature and
immature asci and ascospores. G Asci stipe (arrow). Scale bars: A = 500 µm, B = 200 µm, C = 20
µm, D = 20 µm, E-F = 50 µm, G = 20 µm.

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Fig. 5 – Saccobolus depauperatus (KWSSH003-2009). A Asci and ascospores. B Details of mature
and immature ascospores. C Mature and immature asci and ascospores. Scale bars: A = 50 µm, B =
20 µm, C = 50 µm.

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arranged according to van Brummelen pattern
I, clusters compact, 50–58 × 16–18 µm, more
pointed on one side, rather narrow gelatinous
sheath of 2–4 µm on polar region and 4–8 µm
on sides.
Material examined (1 isolate) –
KENYA, Aberdares National Park, Central
Province, GPS S00°20’23.2” E036°47’11.1”,
tropical montane forest, altitude 2075m,
waterbuck dung, 72 days incubation, 29 August
2010, Paul Mungai, KWSANP005-2010.
Notes – Saccobolus diffusus Sect.
Saccobolus is morphologically very similar to S.
citrinus but the latter has smaller apothecia,
smaller ascospore clusters and smaller
ornamented single ascospores (Kaushal & Virdi
1986, Doveri 2004). S. glaber, another similar
species, is differentiated by having lemon-
yellow apothecia, larger asci (140–275 × 25–48
µm), larger ascospore clusters (50–68 × 16–25
µm), larger individual ascospores (22–29 × 8.5–
14.5 (–16 µm) and the presence of paraphyses-
like hyphae in the upper part of the excipulum
(Kaushal & Virdi 1986, Doveri 2004). This is a
new record for Kenya.
Saccobolus infestans (Bat. & Pontual) Brumm.,
Persoonia, Suppl. 1: 204, 1967. (Figs.8A–K)
Ascomata scattered or gregarious,
superficial, sessile, white, margin not
differentiated, 150–350 μm diam. Ectal
excipulum of textura globulosa, cells 3–3.5 µm
diam. Paraphyses simple, clavate, septate,
exceeding asci, 3–4 µm broad, olive or yellow
pigmented, slightly curved, slightly inflated at
the tips. Asci 44–59 × 12–18 µm, 8-spored,
unitunicate, broadly clavate, wall blueing in
Melzer’s reagent, operculate, with a rather short
stout stipe. Ascospores 8–10 × 4.5–6 µm,
single-celled, ellipsoidal, with broadly rounded
ends, at first hyaline, finally violet to brown,
verruculose or roughened with minute dots,
arranged according to various forms of pattern
VI of van Brummelen; clusters compact,
arranged in two rows, 19–25 × 9.5–11.5 µm,
cemented together by a hyaline gelatinous
sheath. Material examined (1 isolate) –
KENYA, Nairobi National Park, Nairobi
Province, GPS 37M0257082 9850692, altitude
1668m, grassland, zebra dung, 59 days
incubation, 20 August 2010, Paul Mungai,
KWSNNP018-2010.
Notes – S. infestans is a unique species
owing to its characteristic ascospore
arrangement. This species is not likely to be
confused with other currently known
Saccobolus species (van Brummelen 1967,
Doveri 2004). S. infestans, observed only once
in a single dung type, appears to be a rare, late
sporulating coprophilous fungi species from
Kenyan wildlife. This is a new record for
Kenya.
Saccobolus platensis Gamundí & Ranalli, Nova
Hedwigia 17: 385, 1969. (Fig. 9A–F)
Ascomata apothecioid, gregarious,
superficial, sessile, 650–750 µm diam.
Receptacle pulvinate, glabrous, amber to
yellow, margin not differentiated. Disc convex
or flat. Hymenium dotted with far protruding,
almost black tips of ripe asci. Hypothecium
composed of yellow-greyish globular cells.
Medullary excipulum not well differentiated
from ectal excipulum. Ectal excipulum of
textura globulosa cells 5–8 × 5–8 µm.
Paraphyses filiform, simple, septate, in yellow
pigment, scanty, tips just slightly swollen, 3–4
µm diam. Asci 90–120 × 18–23 µm, 8-spored,
unitunicate, clavate, with a pronounced apical
ring, curved, usually with a fairly long stipe.
Ascospores 14–17 × 6–9 µm, ellipsoidal, purple
to brown at maturity, rough or with fine warts,
truncate ends, symmetrical, arranged according
to pattern I of van Brummelen; clusters
compact, 35–41 × 12–16 µm, surrounded by a
hyaline, common broad gelatinous sheath.
Material examined (3 isolates) –
KENYA, Tsavo East National Park, Coast
Province, GPS S03°21’666” E038°38’772”,
altitude 514m, savannah grassland, 18 and 57
days incubation, elephant dung, 23 September
2008, Paul Mungai, KWSTE005A-2008;
Aberdares National Park, Central Province, GPS
S00°20’23.2” E036°47’11.1”, tropical montane
forest, altitude 2075m, water-buck dung, 29
August 2010, Paul Mungai, KWSANP005-
2010. Notes – Saccobolus platensis is very
similar to S. minimus from which it is
differentiated by its smaller ascospores and
asci, irregular and rough episporium
(Richardson & Watling 1997, Doveri 2004).
The two species share a lot of similarities and

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Fig. 6 – Saccobolus diffusus (KWSANP005-2009). A Ascomata on dung. B Squash ascoma. C
Details of ectal excipulum. D Asci and ascospores among paraphyses. E Apical part of ascus. F
Gelatinous envelope around ascospore cluster. G Free mature ascospores and a dehisced ascus. H
Ascospore arrangement, paraphyses and hyaline immature spores in yellow mucus. I Ascus stipe.
J Open operculum. Scale bars: A = 500 µm, B = 200 µm, C = 20 µm, D = 50 µm, E-F = 20 µm,
G = 50 µm, H-J = 20 µm.

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Fig. 7 – Saccobolus diffusus (KWSANP005-2010). A Asci showing open operculum (black arrow),
apical portion and upper parts of paraphyses with yellow pigment (white arrow). B Ascus stipe and
paraphyses, note attachment to basal cells (arrow). C Paraphyses. D Ascospores in apical portion of
ascus, note closed operculum (arrow). E Cells of ectal excipulum. F-G Mature asci and ascospores,
note gelatinous sheath (arrow) and the yellow pigmentation on paraphyses. Scale bars: D-E = 20
µm, F-G = 50 µm.

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hence are hardly separable. This is a new
record for Kenya.
Saccobolus truncatus Vel., Monogr. Discom.
Boh. 1: 370, 1934. (Figs.10A–D, 11E–I)
Ascomata apothecioid, gregarious or
solitary, sessile, 300–400 µm diam. Receptacle
globular or lenticular, pale yellow to brownish
due to ascospore colour, smooth, inconspi-
cuous due to solitary habit and small size,
margin not differentiated. Disc convex,
yellowish, roughened by the black protruding
tips of ripe asci. Hymenium thick. Hypothecium
and medullary excipulum not well delineated
from ectal excipulum. Ectal excipulum of
hyaline thin walled textura globulosa cells.
Paraphyses cylindric-filiform, simple or bran-
ched, septate, exceeding the asci, 2–3 µm
broad; tips slightly clavate, filled with yellow
pigment. Asci 52–60.5 × 15–18 µm 8-spored,
unitunicate, wall turning blue in Melzer’s
reagent, cylindric-claviform, operculate,
gradually tapering down into a stout stalk, thick
walled, rounded to flattened apex. Ascospores
13–18 × 6–8 µm, single-celled, ellipsoidal to
fusiform, sometimes slightly assymetrical,
purple to brown at maturity, finely granulate,
thick walled, episporium peeling off with
maturity, almost smooth, truncate ends,
arranged according to pattern Ia of van
Brummelen; clusters loose, 35–44 × 11.5–15
µm characteristically contracting, shortening
and getting more rounded with age, with a
broad hyaline gelatinous sheath.
Material examined (2 isolates) –
KENYA, Tsavo East National Park, Coast
Province, GPS S03°21’666” E038°38’772”,
altitude 514m, 54 and 57 days incubation,
giraffe dung, 23 August 2008, Paul Mungai,
KWSTE002A-2008; Nairobi National Park,
Nairobi Province, wooded grassland, GPS
S01°21’15.1” E036°46’54.1”, altitude 1768m,
impala dung, 31 August 2009, Paul Mungai,
KWSNNP008-2009.
Notes – Saccobolus truncatus Sect.
Saccobolus is related to S. citrinus Boud. &
Torrend and S. minimus Velen (van
Brummelen 1967, Doveri 2004). The former is
differentiated from the latter by its smaller
dotted ascospores and paler receptacles (van
Brummelen 1967, Doveri 2004). The
ascospores of S. truncatus contract characteris-
tically at maturity (van Brummelen 1967,
Doveri 2004). This is a new record for Kenya.
Saccobolus versicolor (P. Karst.) P. Karst.,
Acta Soc. Fauna Flora Fenn. 2(6): 123, 1885.
(Figs.12A–G, 13H–L)
Ascomata apothecioid, scattered to
gregarious, superficial, sessile, 100–150 µm
diam. Receptacle at first subglobose, becoming
pulvinate or lenticular, external surface
smooth, hyaline to violaceous or whitish grey,
margin not differentiated. Disc convex, dark,
violet or almost black tips of ripe asci dotting
the hymenium, outer surface somewhat
glabrous. Hymenium rather thick. Hypothecium
thin, not well delineated. Ectal excipulum
usually poorly developed of textura-globulosa-
angularis to intricata elongated cells 9–16 × 3–
5 µm, with very pale yellowish intercellular
pigment. Paraphyses cylindric-filiform, often
branched and anastomosed, septate, exceeding
the asci, 2.5–3.5 µm broad, hyaline; tips
inflated, sometimes embedded in an
intercellular violet pigment on the upper parts.
Asci 124–146 × 30–32 µm, 8-spored,
unitunicate, broadly clavate, truncate apex,
walls turning blue in Melzer’s reagent,
operculate, short lobate stalk. Ascospores 15–
19 × 7–9 µm, single-celled, ellipsoidal-
fusiform with narrow ends, asymetrical, purple,
violaceous, becoming blackish brown with age,
smooth with minute pits, sometimes with
fissures, with a very thick epispore, episporium
of old ascospores peeling on squashing,
exposing white-creamy under-coat, arranged
according to pattern II of van Brummelen;
clusters compact, 41–46 × 15–18 µm broad,
enclosed by a granular unilateral gelatinous
envelope.
Material examined (1 isolate) –
KENYA, Nairobi National Park, Nairobi
Province, GPS 37M0255729 9849626, altitude
1680m, 18 days incubation, impala dung, 20
August 2010, Paul Mungai, KWSNNP020-
2010. Notes – The Kenyan collection of
Saccobolus versicolor Sect. Eriobolus is
notably typical with hyaline to violaceous
apothecia, asci (124–146 × 30–32 µm),
ascospores (15–18.5 × 7–9 µm), pigmented
purple-violet, becoming rough and brown with
age; clusters 41–46 × 15–18 µm (in this

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Fig. 8 – Saccobolus infestans (KWSNNP018-2009). A Details of ectal excipulum. B Mature asci
and ascospores. C Mature asci showing stipe (arrow). D Mature asci with spores among dehisced
asci. E-F, I Free mature ascospores. G-H Paraphyses, note colour (arrow). J Mature ascus with
ascospores, showing a stipe and the spore arrangement (arrows). K Ascospores, note the
arrangement (arrow). Scale bars: A = 20 µm, B = 50 µm, C-K = 20 µm.

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Fig. 9 – Saccobolus platensis (KWSTE005A-2008). A Squashed ascoma. B Details of ectal
excipulum. C Mature asci with ascospores, note open operculum (arrow). D Ascospore showing
spore arrangement and gelatinous sheath (arrow). E Ascus, note long stipe (black arrow) and closed
operculum (red arrow). F Asci and paraphyses. Scale bars: A = 200 µm, B = 20 µm, C = 50 µm, D-E
= 20 µm, F = 50 µm.

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Fig. 10 – Saccobolus truncatus (KWSTE002A-2008). A Squashed ascoma. B-D Mature
ascospores, note broad hyaline gelatinous envelope (arrow). Scale bars: A = 200 µm, B = 50 µm, C-
D = 20 µm.
examination). S. versicolor is similar to S.
caesariatus from which it is differentiated by
having hairy clusters on the external surface of
its apothecia; it is also common and
cosmopolitan in distribution while the latter is
quite uncommon (van Brummelen 1967,
Doveri 2004).
Discussion
Our specimens are typical of
Saccobolus in their morphology. The
diagnostic features of Saccobolus include
apothecioid ascomata, always clustered
ascospores which in most cases have brown or
purple-brownish episporic pigment at maturity

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and with each cluster enclosed in a common
gelatinous envelope (van Brummelen 1967,
Doveri 2004, Bell 2005). The ascospore clus-
ters are usually arranged in regular patterns
(van Brummelen 1967). The ascus walls of
Saccobolus are usually amyloid. The gelatinous
sheath on ascospores were either unilateral or
bilateral, that is surrounding the spores
completely.
The dimensions of structures such as
spores and asci are important characters when
delimiting species within the genus Sacco-
bolus. This study used morphological examina-
tions to delimit the species.
New records for Kenya are Saccobolus
citrinus, S. diffusus, S. infestans, S. platensis
and S. truncatus.
Fig. 11 – Saccobolus truncatus (KWSTE002A-2008). A-C Ascospores and asci, note open
opercula (black arrow). D Asci and paraphyses. E Asci, note open opercula (black arrow), various
stages of dehiscence and paraphyses tips (red arrows). Scale bars: A-E = 20 µm.

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Fig. 12 – Saccobolus versicolor (KWSNNP020-2010). A Squash ascoma showing asci protruding
above the hymenium (arrow). B Ascomatal wall. C-D Mature ascus and ascospores, note operculum
(arrow). E-F Free mature ascospores, note gelatinous sheath (arrow). G Paraphyses and free
ascospores(arrow). Scale bars: A = 50 µm, B-G = 20 µm.
Ecology
Twenty-four specimens of Saccobolus
sporulated on Kenyan wildlife dung. Giraffe had 5
specimens, impala 4, waterbuck 3, African
elephant 3, Cape buffalo 3, dikdik 2, zebra 2,
hippopotamus 1 and hartebeest 1. Saccobolus
depauperatus had the least substrate selectivity
and sporulated on the most kinds of dung, while
S. diffusus and S. infestans were quite rare and
were observed on a single dung type.
Interestingly, giraffe and impala dung, a browser
and a grazer, respectively, and hence totally
different feeding habits, produced the most
Saccobolus specimens.
We took note of the age of incubated dung
at which Saccobolus species sporulated.

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Fig. 13 – Saccobolus versicolor (KWSNNP020-2010). A Ascospores in the apical portions of asci, note
a portion of thicker episporium (arrow). B-C Ascospores squashed under cover slip, note peeling
episporium (arrows). D-E Mature asci with ascospores, note closed and open opercula (red and yellow
arrows). Scale bars: A-E = 20 µm.

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Saccobolus diffusus, S. infestans, S. truncatus and
S. versicolor sporulated at 40 to 70 days. S.
depauperatus, the most common species,
sporulated across the entire period of dung
incubation. This sequential sporulation indicated a
form of succession in species of Saccobolus.
Saccobolus citrinus was found only in
fresh dung from wild herbivores in Nairobi
National Park.
Our results show that there is high
coprophilous fungi species diversity in wildlife
dung. The diversity of wild animals in Kenya also
implies a corresponding high dung fungi
diversity. This virtually unexplored substrate may
hold numerous Saccobolus species currently
unknown to science with yet to be recognized
useful applications.
Acknowledgements
This work was generously supported by
Novozymes A/S of Denmark, Kenya Wildlife
Service and the Mushroom Research Foundation.
Many thanks to Dr Francesco Doveri who
meticulously guided the first author in editing and
in the taxonomy of coprophilous Saccobolus. It is
noted that without his patience and untiring
efforts this work could not have been possible.
We are also indebted to all staff and students at
the Institute of Excellence in Fungal Research,
Mae Fah Luang University, Thailand and Kenya
Wildlife Service colleagues especially Mr Moses
Otiende and Dr Edward Kariuki for their interest
and comments on this study. Ms Elsie Wambui
Maina and Asenath Akinyi are thanked for
assisting in laboratory work.
References
Abdel-Azeem AM, Kamel AK, Abo-Zied TM,
Lofty EE, Mohamed ES. 2005 – Mycobiota
survey of Wadi El-Arbae’en, Saint
Catherine Protectorate, Egypt (Unpublished
report).
Bell A. 1983 – Dung Fungi. An illustrated guide
to coprophilous fungi in New Zealand.
Victoria University Press, Wellington.
Bell A. 2005 – An illustrated guide to the
coprophilous Ascomycetes of Australia.
CBS Biodiversity Series 3.
Caretta G, Piontelli E, Savino E, Bulgheroni A.
1998 – Some coprophilous fungi from
Kenya. Mycopathologia 142, 125–134.
De Sloover JR. 2002 – On four species of
Saccobolus (Ascobolaceae, Pezizales)
rarely collected or new to Belgium.
Systematics and Geography of Plants 72,
211–224.
Doveri F. 2004 – Fungi Fimicoli Italici.
Associazione Micologica Bresadola, Trento,
Italy.
Ebersohn C, Eicker A. 1992 – Coprophilous
fungal species composition and species
diversity on various dung substrates of
African game animals. Botanical Bulletin of
Academia Sinica 33, 85–95.
Kaushal SC, Virdi SS. 1986 – Himalayan species
of Saccobolus. A coprophilous genus of
Pezizales. Willdenowia 16, 269–277.
Kirk PM, Canon PF, Stalper JA. 2008 –
Ainsworth and Bisby’s Dictionary of the
Fungi, 10th Edition. CABI Europe.
Minoura K. 1969 – Notes on some Ascomycetes
of East Africa. Transactions of the
Mycological Society of Japan 10, 41–46.
Mungai PG, Njogu JG, Chukeatirote E, Hyde KD.
2012 – Studies of coprophilous
ascomycetes in Kenya. Ascobolus species
from wildlife dung. Current Research in
Applied and Environmental Mycology 2, 1-
16.
Richardson MJ. 2008 – Records of coprophilous
fungi from the Lesser Antilles and Puerto
Rico. Caribbean Journal of Science 44,
206–214.
Richardson MJ, Watling R. 1997 – Key to fungi
on dung. Byford WJ. (ed). Bulletin of the
British Mycological Society 2, 18–46.
van Brummelen J. 1967 – A world monograph of
the genera Ascobolus and
Saccobolus(Ascomycetes, Pezizales).
Persoonia Supplement 1, 1–260.
van Brummelen J. 1969 – Studies on
Discomycetes-III. Persoonia 5, 225–231.
Wang Y-Z. 2000 – Studies on the coprophilous
Ascomycetes in Taiwan. Fungal Science 15,
43–46.