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Morphological Diversity of Ascobolus and Pilobolus Fungi from Wild

Herbivore Dung in Nairobi National Park, Kenya

Article · September 2015

DOI: 10.5923/j.microbiology.20150504.03

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Journal of Microbiology Research 2015, 5(4): 134-141
DOI: 10.5923/j.microbiology.20150504.03

Morphological Diversity of Ascobolus and Pilobolus

Fungi from Wild Herbivore Dung in Nairobi National
Park, Kenya
Aluoch AM1,2,*, Obonyo MA2, Okun DO3, Akinyi A4, Otiende YM1, Mungai PG4

1
Wildlife Forensics and Genetics Laboratory, Kenya Wildlife Service, Nairobi, Kenya
2
Department of Biochemistry & Molecular Biology, Egerton University, Nakuru, Kenya
3
Department of Biochemistry & Biotechnology, Kenyatta University, Nairobi, Kenya
4
Ecological Monitoring Section, Biodiversity Research and Monitoring Division, Kenya Wildlife Service, Nairobi, Kenya

Abstract The present study examined two genera of coprophilous fungi: Ascobolus and Pilobolus with the aim of
species description using their morphological diversity. Fresh dung samples from wild herbivores were collected in
different parts of Nairobi National Park in Kenya and immediately taken to the laboratory for culture by moist chamber
method. Isolates studied were obtained from dung of the following animals: white rhino, zebra, waterbuck, impala, Cape
buffalo, giraffe, Thomson’s gazelle, dikdik, hare, grant’s hartebeest, hippopotamus and eland. Five species of Ascobolus
were studied namely: Ascobolus amoenus, A. bistisii, A. calesco and A. immersus. A possible novel 4-spored Ascobolus
species was observed. Three species of Pilobolus were found: Pilobolus crystallinus var. crystallinus, P. heterosporus and
P. pullus. The most abundant species were: Ascobolus immersus and Pilobolus crystallinus var. crystallinus while the
highest diversity was observed in waterbuck dung samples with a total of five different species.
Keywords Ascomycetes, Coprophilous, Wildlife, Zygomycetes

the class Zygomycetes and can be identified through their

1. Introduction characteristic sporangiophores that have a swollen
extension referred to as collumelae and a sporangium that
Coprophilous fungi are saprobic thus an important part of host the spores at the top [6]. They are observed within two
the wildlife ecosystem since they aid in recycling nutrients or three days of incubation of dung at room temperature
in animal dung [1]. In addition, they are thought to with alternate periods of natural light and darkness [7].
influence digestive efficiency of animals as well as being Pilobolus are naturally obligate coprophilous species and
part of nutrition for certain arthropods living in herbivore only grow on dung materials [8]. They are attached to the
dung [2]. dung by a swollen trophocyst which is semi immersed in
Diversity studies on coprophilous fungi are essential the dung [9]. These trophocysts are normally ovoid to
since they give an indication of the state of the environment globose while the rhizoidal extensions are long and
as they reveal the extent to which environmental stressors cylindrical [9]. Pilobolus have straight unbranched
contribute to degradation of the environment [3]. High sporangiophores which grow towards light [9]. The
fungi species diversity demonstrates an undisturbed sporangiophores have orange pigments at the base and near
ecosystem that is suitable for the flourishing of wildlife [4]. the subsporangial vesicles. The sporangia are hemispherical
According to Tibuhwa et al. (2011), macro fungi diversity in shape with resistant walls and contain the spores which
decreases in areas where agriculture is practiced as opposed are spherical or ellipsoid depending on species [9].
to protected wildlife areas with little external environmental The genus Ascobolus belongs to the class Ascomycetes
stress factors. Species richness is important as an index of which consists of a group of phototropic fungi which
the community structure which is useful for conservation release their ascospores towards light. In general, members
[5]. of this genus take longer than Pilobolus to fruit (and is
The members of fungi in the genus Pilobolus belong to observed after about seven to twenty days following
incubation) compared to those of Pilobolus [7], [10], [11].
* Corresponding author:
[email protected] (Aluoch AM)
The fruit body of Ascobolus has a disk and a receptacle
Published online at http://journal.sapub.org/microbiology which vary in shape from: perithecoid, pyriform, cylindrical,
Copyright © 2015 Scientific & Academic Publishing. All Rights Reserved cupulate, scutullate, discoid and lenticular to pulvinate [12].
 Journal of Microbiology Research 2015, 5(4): 134-141 135

They have characteristic thick walled spores that are glass Petri dishes lined with Whatman cellulose filter paper.
contained in asci. The ascospores are normally single celled, Those that were dry were moistened with distilled water. The
eight within each asci, colored and can be smooth or rough Petri dishes were positioned near a window to permit natural
[13]. light to induce sporulation (van Brummelen, 1967). The set
Previous studies on the genus Ascobolus undertaken on up was monitored daily for sporulation under a
wildlife dung in Kenya indicate that there is high species stereomicroscope (LEICA Microsystems®). Upon
richness [14]. However, there are no specific studies on sporulation, fruiting bodies were picked using a sharp sterile
Pilobolus species in Kenya. Pilobolus is especially needle and placed on microscope slides with a drop of water
important due to its role in transmission of bronchitis in and covered in a cover slip then examined under a compound
animals as a vector for lungworms [15]. Therefore, it is microscope. The morphological features such as shape, size,
necessary to establish which species are present in the park colour, height and width were observed and measured for
and their abundance. each fruiting body using LAS Suite software®. In order to
This study was focused on species diversity of obtain an average spore length 10 spores were measured
coprophilous Ascobolus and Pilobolus from wild herbivores from each fruiting body. Apart from measurements, the
with the aim of identifying the different species present in features were also observed and unique characteristics
herbivore dung. The results of this study will provide recorded for the morphological descriptions. The observed
reference for future biodiversity studies and ecological morphological features were used to identify the species
monitoring of the Nairobi National Park ecosystem. following keys developed by van Brummelen (1967) and
Viriato (2008).
In order to get the diversity index for the fungi observed in
2. Materials & Methods the dung samples, Shannon-weinner index and Simpson
diversity index were calculated [16]. Chao’s species richness
Wildlife herbivore dung was collected from Nairobi estimation was used to calculate the expected highest species
National Park in Kenya. The park is situated approximately 7 abundance [5], [17].
km from Nairobi city centre with a savannah ecosystem
comprising of scattered acacia and open grass plains. The
park covers an area of 117km2 with central coordinates 3. Results
1°22′24″S, 36°51′32″E. Wild herbivore dung samples were
randomly collected from different locations of the park. 3.1. Isolates and Morphology
During collection, all sites were marked using GPS and used Ascobolus amoenus Oudem., Hedwigia 21: no. 11 (1882)
to generate a map of sample collection as shown (Fig. 1). (Fig. 2A-F).
Fresh dung samples were collected and placed in paper Ascomata apothecioid, solitary or gregarious,
envelopes and labelled. The following information was semi-immersed, 500–600 μm high, 600–700 μm diam.
recorded for each sample: vegetation type in the surrounding, Receptacle at first closed then irregularly opening at the top,
and animal species voiding the dung. cupulate, brown. Disc flat to convex, greenish yellow,
numerous tips of ripe asci protruding and dotting the
hymenium. Hypothecium thin discontinuous and composed
of isodiametric and oblong cells 4–6 μm. Medullary
excipulum of textura globulosa thin cells. Ectal excipulum of
textura globulosa-angularis brownish cells 10–20 × 6–10 μm.
Paraphyses filiform, hyaline, tips rarely inflated, embedded
in greenish-yellow mucus 2–4 μm broad. Asci 180–245 ×
50–60 μm, 8-spored, narrowly clavate, rounded and curved
with walls that characteristically stain blue in Melzer’s
reagent. Ascospores 30–40 × 21–22 μm, biseriate, ellipsoidal,
violet, and finally brownish, with thin gelatinous sheath.
Material examined
KENYA, Nairobi County, incubated for seven to fourteen
days, Nairobi National Park, GPS S1°35’45.11”
Figure 1. Map of Nairobi National Park showing sample collection points
E36°85’70.55”, altitude 1647 m, waterbuck, 23 July 2013, P.
Mungai, KWSNNP005-2013; Nairobi National Park,
Upon collection, the herbivore dung was transported to the Nairobi County, GPS S1°35’91.55” E36°84’48.32”, altitude
KWS laboratory. Once in the laboratory, the dung samples 1649m, impala, 23 July 2013, P. Mungai,
were incubated in moist chamber under natural conditions of KWSNNP007-2013; Nairobi National Park, Nairobi County,
light and temperature for up to twenty one days. This was GPS S1°35’42.28” E36°85’68.92”, altitude 1650m,
achieved by placing the dung in labelled 100mm by 15mm hartebeest, 26 August 2013, P. Mungai, KWSNNP020-2013;
136 Aluoch AM et al.: Morphological Diversity of Ascobolus and Pilobolus Fungi
from Wild Herbivore Dung in Nairobi National Park, Kenya

Nairobi National Park, Nairobi County, GPS S1°34’77.08” Ascospores 53–58 × 30–33 μm, irregularly biseriate,
E36°85’17.54”, altitude 1648m, Thomson’s gazelle, 26 ellipsoidal, rounded at the ends, purple, irregularly
August 2013, P. Mungai, KWSNNP012-2013. distributed at the end of the ascus surrounded by thin
Notes gelatinous sheath.
Ascobolus amoenus sect Dasyobolus [18] is closely Material examined
similar to A. Elegans [19] but it can be differentiated by its KENYA, Nairobi National Park, Nairobi County,
smaller ascospores. This collection differs from that Specimens, dung incubate for 7-9 days. GPS S 1°35’43.02”
described by Oudemans (1882) in the size of the asci. The E 36°85’68.79”, altitude 1657 m, white rhino, 23 July 2013,
latter has asci with smaller diameter of about 35–40 μm. The P. Mungai, KWSNNP001-2013; Nairobi National Park,
ascospores were observed to have double walls. Nairobi County, GPS S 1°35’75.45” E 36°84’63.72”,
altitude 1649 m, zebra, 23 July 2013, P. Mungai,
KWSNNP006-2013; Nairobi National Park, Nairobi County,
GPS S 1°35’43.02” E 36°85’68.79”, altitude 1657 m,
waterbuck, 23 July 2013, P. Mungai, KWSNNP003-2013;
Nairobi National Park, Nairobi County, GPS S1°34’77.08”
E36°85’17.54”, altitude 1648 m, Thomson’s gazelle, 26
August 2013, P. Mungai, KWSNNP012-2013; Nairobi
National Park, Nairobi County GPS S1°85’07.59”
E37°02’58.18”, altitude 1658 m, dikdik, 10 January 2014, A.
Aluoch, KWSNNP024-2014.
Notes
Ascobolus bistisii Sect. Dasyobolus [20] is similar to
Ascobolus immersus in many ways morphologically.
However, this species has regularly ellipsoid ascospores
while those of A. immersus [21] are subcylindrical with
rounded ends. This species is quite common in Kenya
wildlife dung as observed in this study.

Figure 2. Ascobolus amoenus (KWSNNP005-2013). A. Ascomata on

substrate (arrow). B. Specimen squashed in glycerol. C. 8-spored ascus. D.
Asci. E. Ascomatal wall. F. asci and paraphyses. Scale bars: A = 2000 μm, B
= 500 μm, C = 20 μm, D = 50 μm, E = 50 μm, F = 50 μm

Ascobolus bistisii Gamundí & Ranalli, Nova Hedwigia 10:

347 (1966) [1965] (Fig. 3A-F)
Ascomata cleistothecioid early stages, hymenium exposed
later on, gregarious, superficial or semi-immersed, 600–700
μm high 400–500 μm diameter. Receptacle, subglobose,
brown, dotted with few protruding, finger-like asci, barrel
shaped, widest at equatorial part, with a hardly differentiated
margin. Disc convex, light yellow to brown. Hypothecium
very thin of isodiametric cells. Medullary excipulum of
textura angularis cells 5–10 × 6–20 μm. Ectal excipulum of
textura angularis 15–20 × 7–8 μm. Paraphyses
cylindric-filiform, with tips not inflated, embedded in clear
Figure 3. Ascobolus bistisii (KWSNNP001-2013). A. Ascoma squashed
mucus, numerous septae, 3–4 μm. Asci 400–500 × 100–105 in water. B. Ascospores at the tip of asci. C. Ascomatal wall. D-E.
μm, 8-spored, broadly clavate-cylindrical, operculate with Paraphyses. F. Ascospores. Scale bars: A = 200 μm, B = 50μm, C = 20μm,
dome-shaped apex 30–40 μm wide, wall weakly amyloid. D = 20μm, E = 20μm, F = 20μm
 Journal of Microbiology Research 2015, 5(4): 134-141 137

Ascobolus calesco A.E. Bell & Mahoney, Fungal Planet, Ascobolus immersus Pers., Neues Mag. Bot. 1: 115 (1794)
no. 11-21: 21: [2] (2007). (Fig. 4A-F) (Fig. 5A-F).
Ascomata apothecioid, scattered or gregarious, Ascomata clestothecoid at first, gregarious or scattered,
semi-immersed 800 μm high, 700 μm diam. Receptacle deep immersed or superficial, sessile, 700–1000 μm high,
yellow to yellowish-brown, subglobose, barrel shaped, 600–800 μm diam. Receptacle deep yellow to
margin not differentiated. Disc globular flat ripe asci yellowish-brown or greenish-brown, subglobose, margin not
protruding above the hymenium. Hypothecium not well differentiated. Disc flat or convex without margin, shiny, a
differentiated from Medullary excipulum. Ectal excipulum few ripe asci protruding above the hymenium. Hypothecium
of textura angularis brown cells, 14–21 × 7–11 μm. very thin, of isodiametric cells. Medullary excipulum thin, of
Paraphyses filiform, hyaline, simple or sparingly branched at textura globulosa or angularis hyaline cells. Ectal excipulum
the base, septate, exceeding asci, 2–4 μm broad, tips not of somewhat horizontally elongated textura angularis
swollen, embedded in greenish-yellow mucus. Asci 600 × yellowish-brown thick walled cells, 22–43 × 11–17μm.
100 μm, 8-spored, unitunicate Ascospores 48–57 × 27–33 Paraphyses filiform, simple or sparingly branched at the base,
μm, biseriate, single-celled, ellipsoidal, purple. Gelatinous septate, exceeding asci 2–3.5 μm broad, embedded in
sheath hyaline, surrounding each ascospore. greenish-yellow mucus. Asci broadly clavate 460–675 ×
Material examined 95–115 μm, 8-spored, unitunicate broadly clavate to
sacciform rounded above. Ascospores 55–60 × 30–35 μm,
KENYA, Nairobi National Park, Nairobi County, one
biseriate, single-celled, subcylindrical, ends markedly
specimen, dung incubated for seven to fourteen days, GPS S
rounded, violet becoming purple at maturity. Gelatinous
1°35’42.28” E 36°85’68.92”, altitude 1650 m, hartebeest,
sheath on each spore, hyaline, broader on sides and narrow
26th August 2013, P. Mungai, KWSNNP020-2013.
on polar region.
Notes Material examined
The Kenya Ascobolus calesco Sect. Dasyobolus described KENYA, Nairobi National Park, Nairobi County, seven
here agrees with the description of A calesco as described by specimens, dung incubated for 7-15 days, GPS S 1°34’27.03”
Bell and Mahoney (2007) [22]. E 36°82’22.23”, altitude 1657 m, buffalo, 23 July 2013 P.
Mungai, KWSNNP010-2013; Nairobi National Park,
Nairobi County, GPS S 1°35’43.02” E 36°85’68.79”,
altitude 1657 m, white rhino, 23 July 2013, P. Mungai,
KWSNNP001-2013; Nairobi National Park, Nairobi County,
GPS S 1°35’45.11” E 36°85’70.55”, altitude 1647 m, zebra,
23 July 2013, P. Mungai, KWSNNP002-2013; Nairobi
National Park, Nairobi County, GPS S 1°35’43.02” E
36°85’68.79”, altitude 1657 m, waterbuck, 23 July 2013, P.
Mungai, KWSNNP003-2013; Nairobi National Park,
Nairobi County, GPS S 1°35’42.28” E 36°85’68.92”,
altitude 1650 m, hartebeest, 26 August 2013, P. Mungai,
KWSNNP020-2013; Nairobi National Park, Nairobi County,
GPS S1°34’77.08” E36.85’17.54”, altitude 1648 m,
Thomson’s gazelle, 26 August 2013, P. Mungai,
KWSNNP012-2013; Nairobi National Park, Nairobi County
GPS S1°85’07.59” E37°02’58.18”, altitude 1658 m, white
rhino, 10 January 2014, A. Aluoch, KWSNNP031-2014;
Nairobi National Park, Nairobi County GPS S1°85’07.59”
E37°02’58.18”, altitude 1658 m, hare, 10 January 2014, A.
Aluoch, KWSNNP026-2014; Nairobi National Park, Nairobi
County GPS S1°85’07.59” E37°02’58.18”, altitude 1658 m,
eland, 10 January 2014, A. Aluoch, KWSNNP037-2014;
Nairobi National Park, Nairobi County GPS S1°84’64.98”
E37°02’65.83”, altitude 1619 m, zebra, 14 January 2014, A.
Aluoch, KWSNNP039-2014.
Notes
Ascobolus immersus Sect. Dasyobolus [21] is quite
Figure 4. Ascobolus calesco (KWSNNP020-2013). A. Ascomata on
substrate. B. Ascoma squashed in lactic acid. C. Ascomatal wall. D-E
common in dung from Kenya wildlife and domestic
Ascospores. F Paraphyses. Scale bars: A = 1000 μm, B = 500 μm, C = 20 μm, herbivore dung. It is easily distinguished from other
D = 20 μm, E = 20 μm, F = 20μm members of the Ascobolus genus by its characteristically
138 Aluoch AM et al.: Morphological Diversity of Ascobolus and Pilobolus Fungi
from Wild Herbivore Dung in Nairobi National Park, Kenya

large ascospores. Each spore is surrounded by a gelatinous

sheath. They can grow immersed on soft surfaces while
growing on the surface of more dense surfaces.

Figure 6. 4-spored Ascobolus sp. (KWSNNP020-2013). A. Ascomata on

Figure 5. Ascobolus immersus (KWSNNP001-2013). A. Ascomata on substrate. B. Ascoma squashed in water. C. 4-spored asci. D. Paraphyses. E.
substrate (arrows). B. Ascoma squashed in water. C. 8-spored mature asci. Ascomatal wall. F-G. Ascospores. Scale bars: A = 1000 μm, B = 500 μm, C
D. Open operculum (arrow). E. Paraphyses. F. Mature ascopores surrounded = 50 μm, D = 20 μm, E = 20 μm, F = 20μm, G = 20μm
by gelatinous sheath. G. Ascomatal wall. Scale bars: A=1000μm, B= 500μm,
C=200μm, D=50μm, E=20μm, F=20μm, G=20μm Pilobolus crystallinus (F. H. Wigg.) Tode var. crystallinus,
Schrift. Berl. Gesell. Naturf. Freunde 5: 47 (1784) (Fig. 7
Ascobolus sp (KWSNNP020-2013 hartebeest dung) (Fig.
A-F)
6A-G)
Trophocyst subglobose, 370×360 µm with rhizoidal
Ascomata apothecioid, scattered, immersed, 380 μm high,
extension up to 980 µm, yellowish pigmentation.
540 μm diam. Receptacle deep brown, subglobose, margin
Sporangiophore long-cylindrical, unbranched, phototrophic,
not differentiated. Disc flat ripe asci protruding above the
4mm×100 µm. Sporangial wall black, hemispherical to
hymenium. Hypothecium thin with globose cells. Medullary
ovoid 480×250 µm. Columella smooth walled.
excipulum of yellowish brown cells of various thickness.
Subsporangial vesicle smooth walled a little orange
Ectal excipulum of textura angularis yellowish-brown cells,
pigmentation elliptical 700×530 µm. Spores yellow, grainy
10–25 × 10–12 μm. Paraphyses filiform, tips not swollen,
content, smooth wall, ellipsoid 8×5 µm.
embedded in greenish-yellow mucus. Asci 365–400 × 60–65
μm, 4-spored sacciform, rounded above. Ascospores 35–45 Material examined
× 20–25 μm, uniseriate, single-celled, subcylindrical, ends KENYA, Nairobi National Park, Nairobi County,
markedly rounded, purple Gelatinous sheath absent. incubated for four to seven days, GPS S 1°35’45.11” E
Material examined 36°85’50.89”, altitude 1646 m, zebra, 23 July 2013, P.
Mungai, KWSNNP004-2013; Nairobi National Park,
KENYA, Nairobi National Park, Nairobi County,
Nairobi County, incubated for three to seven days, GPS S
hartebeest 26th August 2013 dung incubated for seven days,
1°34’50.02” E 36°84’82.34”, altitude 1649 m, buffalo, 23
GPS S 1°35 E 36°86, altitude 1650 m, P. Mungai,
July 2013, P. Mungai, KWSNNP009-2013; Nairobi National
KWSNNP020-2013.
Park, Nairobi County GPS S1°85’07.59” E37°02’58.18”,
Notes altitude 1658 m, Grant’s gazelle, 10 January 2014, A.
This species differs from those described earlier due to the Aluoch, KWSNNP027-2014; Nairobi National Park, Nairobi
fact that it contained four ascospores in each ascus. The County GPS S1°84’64.98” E37°02’65.83”, altitude 1619 m,
ascospores are seen clustered on one end of the ascus. zebra, 14 January 2014, A. Aluoch, KWSNNP032-2014.
 Journal of Microbiology Research 2015, 5(4): 134-141 139

Notes Park, Nairobi County GPS S1°85’07.59” E37°02’58.18”,

Our specimen closely agrees with the Pilobolus altitude 1658 m, giraffe, 10 January 2014, A. Aluoch,
crystallinus var. crystallinus described in the original KWSNNP034-2014.
diagnosis by F. H. Wigg. Tode Schrift. Berl. Gesell. Naturf. Notes
Freunde 5: 47 (1784) [23]. Some specimens had spore sizes The isolated material agrees with the original diagnosis
differing slightly from those described earlier. Pilobolus [24]. However, the rhizoidal extensions for some of the
crystallinus var. crystallinus differs from Pilobolus identified species were longer than the 300 µm of the
crystallinus var. kleinii by having pale yellow spores while described species.
those of the latter are bright yellow.

Figure 8. Pilobolus pullus (KWSNNP009-2013). A. Pilobolus on

Figure 7. Pilobolus crystallinus var. crystallinus (KWSNNP004-2013). A. substrate. B. Pilobolus squashed in glycerol. C. Columella and
Pilobolus on substrate. B. Sporangium and Columella. C. Trophocyst and sporangiophore. D. Sporangium. E-F. Zygospores. Scale bars: A = 2000 µm,
rhizoidal extension. D. Zygospores. E. Trophocyst. F. Zygospores. Scale B = 500 µm, C = 200 µm, D =50 µm, E = 50 µm, F = 20 µm
bars: A = 2000 µm, B = 500 µm, C = 200 µm, D = 50 µm, E = 50 µm, F = 50
µm Pilobolus heterosporus Palla, Öst. bot. Z. 50: 349 (1900)
(Fig. 9A-F)
Pilobolus pullus Massee, Bull. Misc. Inf., Kew: 160 (1901)
Trophocysts ovoid to globose, short ellipsoid
(Fig. 8A-F)
200.27×280.46 µm with rhizoidal extensions little
Trophocysts ovoid to globose, hyaline 180 µm diameter.
pigmentation. Sporangiophores long cylindrical, 1.5mm ×
Sporangiophore long-cylindrical, 720×90 μm. Sporangia
69.23 µm. Sporangia black, hemispherical to ovoid
black, hemispherical 270×140 μm. Columella with smooth
169.7×212.29 µm, resistant wall. Columella conical, little
walls. Subsporangial vesicle with smooth wall hyaline, little
pigmented. Subsporangial vesicles ovoid and ellipsoid,
pigmentation, ovoid, 370×200 μm. Zygospores 9×5 μm
412.31×344.65µm. Zygospores 5.6-10.08×5.15-6.1 µm
yellow, homogenous content, subcylindrical.
yellowish grainy content, ovoid, cylindrical.
Material examined
Material examined
KENYA, Nairobi National Park, Nairobi County,
KENYA, Nairobi National Park, Nairobi County,
incubated for three to seven days, GPS S 1°34’50.02” E
incubated for three to seven days, GPS S1°85’07.59”
36°84’82.34”, altitude 1649 m, buffalo, 23 July 2013, P.
E37°02’58.18”, altitude 1658 m, dikdik, 10 January 2014 A.
Mungai, KWSNNP009-2013; Nairobi National Park,
Aluoch, KWSNNP024-2014.
Nairobi County, incubated for four to seven days, GPS S
1°35’45.11” E 36°85’50.89”, altitude 1646 m, zebra, 23 July Notes
2013, P. Mungai, KWSNNP004-2013; Nairobi National This species shows similarities to those described by
140 Aluoch AM et al.: Morphological Diversity of Ascobolus and Pilobolus Fungi
from Wild Herbivore Dung in Nairobi National Park, Kenya

Viriato (2007). The described species is differentiated from Table 1. Host animals and fungal species described in each
the others due to the different shaped irregular zygospores Number Number of
with grainy content. Animal of dung Pilobolus Ascobolus species
piles identified
White
7 - 2 2
Rhino
Zebra 9 2 2 4
Water
3 1 3 4
buck
Impala 7 1 1 2
Buffalo 9 1 1 2
Giraffe 8 1 1 2
Thomson's
2 1 3 4
gazelle
Hartebeest 9 1 4 5
Dikdik 2 2 1 3
Hare 2 - 1 1
Grant’s
3 1 2 3
gazelle
Hippopota
1 - 2 2
mus
Eland 3 2 1 3

4. Discussion
According to the findings of the current study, five species
from the genus Ascobolus were identified namely: A.
Figure 9. Fig. 9. Pilobolus heterosporus (KWSNNP021-2013). A.
Pilobolus on substrate. B. Pilobolus squashed in glycerol. C. Columella and
amoenus, A. bistisii¸ A. calesco, A. immersus and fifth
sporangiophore. D. Rhizoidal extension. E-F. Zygospores. Scale bars: A = putatively new species (a species with 4-spored asci) that has
2000 µm, B = 500 µm, C = 200 µm, D =50 µm, E = 50 µm, F = 20 µm never been described to the best of our knowledge. The most
abundant species from the animal dung samples was A.
3.2. Diversity Index immersus (28.9%). In addition, there were three Pilobolus
There were a total of 8 species reported from 13 different species identified, that is: P. crystallinus var. crystallinus, P.
animal hosts (table 1). Three from the genus Pilobolus and 5 heterosporus and P. pullus. Pilobolus crystallinus var
from the genus Ascobolus. The highest recorded species was crystallinus (25.2%) was the most abundant species in this
Ascobolus immersus followed by Pilobolus crystallinus. genus.
These were abundant in a large variety of the animal dung Fungi from the genus Pilobolus were observed to grow
samples collected. However, since the number of samples and die out within the first week of incubation in a moist
collected for each animal species was not equal, the number chamber. However, for some samples, there was still new
of coprophilous fungi species found in each dung sample growth of Pilobolus even after ten days of incubation. This
was used to calculate diversity index. There was no growth could be attributed to the fact that some of the spores
of any of the two genera of interest in this study in two rhino, dispersed within the plates were falling on the dung and
two impala, three buffalo, and six giraffe dung samples. The growing again. This may mean that Pilobolus genus do not
maximum fungi species of interest in this study was require passage in the animal gut or treatment to sporulate.
observed in waterbuck dung which had 3 Ascobolus species The observed diversity and expected diversity indicate
and 2 Pilobolus species. that there is high species richness of coprophilous fungi in
Across the 68 dung samples collected, there were 8 Nairobi National Park. This is likely to indicate that even
different coprophilous fungal species recorded. Since most though the park is located near the capital city, surrounded
of them were rare, Chao’s formula to obtain the estimated by upcoming industrial areas as well as increasing human
richness. This gave an estimated richness as 15 different settlements, the park ecosystem itself is relatively well
species per dung sample compared to the observed richness preserved. This is comparable to the work of Tibuwa et al.
of 5 species. (2011) who studied macro fungi diversity in Serengeti-Mara
 Journal of Microbiology Research 2015, 5(4): 134-141 141

ecosystem and reported high diversity in woodland and [9] A. Viriato, “Pilobolus species found on herbivore dung from
grassland areas as compared to areas with human settlements. the Sao Paulo Zoological Park, Brazil,” Acta Bot. Brasilica,
vol. 22, pp. 614–620, 2008.
The theory that there is little interference in the ecosystem is
also supported by the fact that there was no outright [10] O. Piasai, L. Manoch, and others, “Coprophilous ascomycetes
dominant species that are adapted to altered environment from Phu Luang Wildlife Sanctuary and Khao Yai National
conditions since the difference between the abundance of the Park in Thailand,” Kasetsart J. (National Sci., vol. 43, pp.
34–40, 2009.
observed species was not very significant [3], [25].
Future studies on morphological diversity of these [11] P. Mungai, K. D. Hyde, L. Cai, J. Njogu, and K. Chukeatirote,
coprophilous fungi genera and comparison with the results of “Coprophilous ascomycetes of northern Thailand,” Curr. Res.
this study will aid in monitoring the changes in the park Environ. Appl. Mycol., vol. 1, pp. 135–159, 2011.
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ACKNOWLEDGEMENTS [13] A. Bell, An illustrated guide to the coprophilous Ascomycetes
of Australia, Fungal Bio. Utrecht, The Netherlands: CBS
This work was made possible by the generous support of Biodiversity Series 3, 2005.
the Kenya Wildlife Service technical and scientific staff.
[14] P. G. Mungai, J. G. Njogu, E. Chukeatirote, and K. D. Hyde,
“Studies of coprophilous ascomycetes in Kenya. Ascobolus
species from wildlife dung,” Curr. Res. Appl. Environ. Mycol.,
vol. 2, no. 1, pp. 1–16, 2012.
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