ABSTRACT

We documented ectoparasites found on wild black-and-white ruffed lemurs (Varecia variegata) in the southeastern rain forests of Madagascar and describe trends in parasitism. In this study, 235 mesostigmatid mites (1 male, 87 females, 147 nymphs) identified as Liponyssella sp., in the acarine family Macronyssidae, were collected during 87% (34/39) of lemur examinations (mean number/host=7.9). The only other ectoparasite collected was the louse fly (Allobosca crassipes; 3 males, 8 females) in the dipteran family Hippoboscidae, which was collected during 26% (10/39) of lemur examinations (mean number/host=1.1). The lemur most heavily parasitized by mites was an adult female with 29 adult females and 17 nymphs, all collected from the face.

Madagascar is home to more than 100 lemur species that occupy all of the island's ecotones. Despite being the most threatened group of mammals on earth (Schwitzer et al. 2014), very little is known about the ecology of these nonhuman primates. In particular, very little is known about the ectoparasites of lemurs. Studies of lemur ectoparasites are therefore necessary to better understand their ecology before their host populations drastically decline or disappear. Ectoparasites are a significant group of the Malagasy fauna, and their evolutionary adaptations have the potential to affect the health and well being of their hosts. Because lemurs provide an ecosystem for their ectoparasites, a thorough understanding of lemur ecology should include an understanding of lemur-ectoparasite associations.

Previously, the tick Haemaphysalis lemuris and the hippoboscid louse fly Allobosca crassipes have been reported from the black-and-white ruffed lemur (Varecia variegata) in Madagascar (Junge and Louis 2002, 2005; Vaughn and McGee 2009). Ectoparasites were collected from these lemurs by one of us (A.L.B.) as part of a larger study of black-and-white ruffed lemur molecular ecology that involved live-captures of this arboreal primate (Baden et al. 2008, 2014).

Sampling was conducted from 2006 to 2008 (Tables 1, 2) in Mangevo (21°22′60″S, 47°28′0″E), a bush camp located in undisturbed, midaltitude primary rainforest in Ranomafana National Park, southeastern Madagascar. Adult and subadult black-and-white ruffed lemurs belonging to two social communities were captured by remote injection (Glander 1993). Experienced Malagasy technicians performed all captures, and a veterinarian was always present. Location at the time of capture was recorded with a handheld global positioning system unit before transporting animals back to camp where complete workups were performed.

Table 1

Thirty-nine black-and-white ruffed lemurs (Varecia variegata editorum) were sampled during three capture seasons between 2006 and 2008 in Ranomafana, Madagascar, for ectoparasites. Two ectoparasite species were found on the lemurs (Allobosca crassipes and Liponyssella sp.) with the number of parasites ranging from 0 to 46 on each individual lemur at the time of capture. NA indicates not applicable.

Thirty-nine black-and-white ruffed lemurs (Varecia variegata editorum) were sampled during three capture seasons between 2006 and 2008 in Ranomafana, Madagascar, for ectoparasites. Two ectoparasite species were found on the lemurs (Allobosca crassipes and Liponyssella sp.) with the number of parasites ranging from 0 to 46 on each individual lemur at the time of capture. NA indicates not applicable.
Thirty-nine black-and-white ruffed lemurs (Varecia variegata editorum) were sampled during three capture seasons between 2006 and 2008 in Ranomafana, Madagascar, for ectoparasites. Two ectoparasite species were found on the lemurs (Allobosca crassipes and Liponyssella sp.) with the number of parasites ranging from 0 to 46 on each individual lemur at the time of capture. NA indicates not applicable.
Table 2

Between 2006 and 2008, six black-and-white ruffed lemurs (Varecia variegata editorum), all adults, were captured during more than one season and sampled for ectoparasites in Ranomafana, Madagascar. Details of sex and intensity of ectoparasite infestation each year are provided, and years in which individuals were not captured (NC) are noted.

Between 2006 and 2008, six black-and-white ruffed lemurs (Varecia variegata editorum), all adults, were captured during more than one season and sampled for ectoparasites in Ranomafana, Madagascar. Details of sex and intensity of ectoparasite infestation each year are provided, and years in which individuals were not captured (NC) are noted.
Between 2006 and 2008, six black-and-white ruffed lemurs (Varecia variegata editorum), all adults, were captured during more than one season and sampled for ectoparasites in Ranomafana, Madagascar. Details of sex and intensity of ectoparasite infestation each year are provided, and years in which individuals were not captured (NC) are noted.

Lemurs were searched for ectoparasites using forceps and a flea comb. The time it took to examine a lemur varied, but never exceeded 15 min. Collected parasites were preserved in 90% ethanol and stored at ambient temperature until they could be brought to the US. Samples were stored in the Hunter College Primate Molecular Ecology Laboratory (New York, New York, USA) at ambient temperature until they could be sent to Auburn University and Georgia Southern University for analyses.

During lemur processing, we monitored heart rate, respiratory rate, and body temperature of the study subjects and administered a balanced electrolyte solution (Lactate Ringer Solution, Aspen Veterinary Resources LTP, Liberty, Missouri, USA) subcutaneously. Before release, captured lemurs were given AVID® microchips (Avid Identifications Systems Inc., Norco, California, USA) and fitted with unique collar-tag combinations to catalog individuals for identification. Animals recovered in breathable cloth bags (at least 3 h) and were released at the site of capture.

Protocols were approved by Stony Brook University (SBU) and Henry Doorly Zoo Institutional Animal Care and Use Committee (IACUC) standards (SBU IACUC 2005-20081449), and research permits were granted by the Tripartite Committee of the Malagasy government (Association Nationale pour la Gestion des Aires Protégées), as well as by the US Fish and Wildlife Service. No injuries or deaths resulted from sampling procedures.

Thirty-nine examinations for ectoparasites were made on 31 individual lemurs (16 males, 15 females) during three capture seasons (Table 1); six lemurs were resampled in two or three sampling years (Table 2). Two species of ectoparasites but no ticks were recorded. Eleven adult specimens of A. crassipes (Fig. 1) were collected during 26% (10/39) of lemur captures, with a mean intensity (mean per infested host) of 1.1 flies per lemur. Of the 11 parasites, three were males and the remaining eight were females. Only one lemur was parasitized by more than one fly during the same examination. The hindlimbs and groin of black-and-white ruffed lemurs were a frequent host site for A. crassipes.

Figure 1

Scanning electron microscopy imagery of a female louse fly (Allobosca crassipes) collected from a wild black-and-white ruffed lemur (Varecia variegata) in Ranomafana, Madagascar, highlights characters used to identify the species, including (A) mouthparts—ventral view, (B) mouthparts—dorsal view, (C) posterior abdomen—ventral view showing bilobed terminex, and (D) front tarsi showing short setae.

Figure 1

Scanning electron microscopy imagery of a female louse fly (Allobosca crassipes) collected from a wild black-and-white ruffed lemur (Varecia variegata) in Ranomafana, Madagascar, highlights characters used to identify the species, including (A) mouthparts—ventral view, (B) mouthparts—dorsal view, (C) posterior abdomen—ventral view showing bilobed terminex, and (D) front tarsi showing short setae.

Two hundred and thirty-five Liponyssella mites (one male, 87 females, 147 nymphs) were collected, mostly from the head of lemurs, specifically the chin, mouth, neck, and around the eyes. A total of 87% (34/39) of lemur examinations yielded mites for a mean intensity of 7.9 mites per lemur.

One of the two species of ectoparasites we recorded from black-and-white ruffed lemurs, the louse fly, has been recorded previously from this host and from other species of lemurs (Rahola et al. 2011). The other ectoparasite is a new species of mite belonging to the macronyssid genus Liponyssella and will be described separately. In contrast to Junge and Louis (2002, 2005), we did not record H. lemuris or any other species of ticks from black-and-white ruffed lemurs in this survey.

Fourteen specimens (five males, nine females) of A. crassipes were recorded from six black-and-white ruffed lemurs in Ranomafana National Park (Vaughn and McGee 2009), which represents a mean intensity of 2.3 compared with 1.1 in our study. Vaughn and McGee (2009) also collected 87 A. crassipes (49 males, 38 females) from 23 Milne-Edwards' sifakas (Propithecus edwardsi) in the same habitat, a mean intensity of 3.8, suggesting that Milne-Edwards' sifakas may be preferred over black-and-white ruffed lemurs as a host by this ectoparasitic fly. Milne-Edwards' sifakas and black-and-white ruffed lemurs are arboreal lemur species that co-occur in the eastern rainforests of Madagascar, with the former having a slightly larger body mass (5–6.5 kg; King et al. 2012) than the latter (2.6–4.1 kg; Baden et al. 2008), which may influence odorant and carbon dioxide attractants to A. crassipes adults. Allobosca crassipes has also been recorded from additional lemur species (Maa 1963, 1969; Rahola et al. 2011), which suggests relatively low host specificity by A. crassipes. However, all known hosts of this fly are relatively large lemurs (2–8 kg). Perhaps smaller lemurs do not tolerate these large ectoparasites or are more efficient groomers. Future collections of ectoparasites from lemurs should provide additional data on the host specificity of A. crassipes. One additional species of Hippoboscidae, Proparabosca alata, is known to parasitize lemurs, but it has only been recorded from Propithecus verreauxi coronatus (Theodor and Oldroyd 1965; Maa 1969; Rahola et al. 2011).

The new species of Liponyssella we recorded could be host-specific to black-and-white ruffed lemurs, because it has not previously been documented from other species. However, the only other congeneric mite species currently known from Madagascar, Liponyssella madagascariensis, has been recorded from multiple species of large lemurs (OConnor 2003; Klompen et al. 2015), suggesting this may not be the case. Unlike adult A. crassipes, which were large and easily detected, the Liponyssella mites were very small, moved quickly, and were difficult to sample. The inner inguinal region and mouth area (Fig. 2) were the most heavily parasitized regions of black-and-white ruffed lemurs by this mite, perhaps because of sparse hair, less efficient host grooming, or the presence of an abundant peripheral blood supply in those regions.

Figure 2

Liponyssella mites collected from black-and-white ruffed lemurs (Varecia variegata) in Ranomafana, Madagascar, represent a new species. These mites are shown around the mouth of a female lemur (A), and a close-up dorsal scanning electron microscopy image (B) depicts a male mite sampled as part of this study.

Figure 2

Liponyssella mites collected from black-and-white ruffed lemurs (Varecia variegata) in Ranomafana, Madagascar, represent a new species. These mites are shown around the mouth of a female lemur (A), and a close-up dorsal scanning electron microscopy image (B) depicts a male mite sampled as part of this study.

The genus Varecia is notable among lemurs for its flexible fission-fusion social dynamics (Baden et al. 2016), as well as its tendency to crèche litters of young communally (e.g., Morland 1990; Baden et al. 2013). Members of social communities exhibit variable affiliation, and inter-community interactions are limited to infrequent territorial disputes (e.g., Morland 1991; Baden 2011). Moreover, the species is known for its strict seasonal reproduction, with mating in July, and giving birth about 90–110 d later (e.g., Boskoff 1977; Rasmussen 1985). As ectoparasite levels are known to vary in accordance with the strength of social interactions (MacIntosh et al. 2012), patterns of nest sharing (Stamp et al. 2002), and hormones (Zohdy et al. 2017), future studies should consider whether and how this potentially host-specific parasite varies in presence and prevalence across these and other variables through space and time.

Allobosca crassipes is not host-specific to black-and-white ruffed lemurs, and both ectoparasites we recorded include free-living stages during their life cycles, emphasizing the importance of interactions between black-and-white ruffed lemurs and the environment for ectoparasite transmission. It is not currently known whether the louse flies and mites are vectors of any pathogens or parasites.

We thank Jean Baptiste Velontsara, Ratalata Pierre, Rajarison Emile, Solo Justin, Telo Albert, Lahitsara Jean Pierre, Razafindrakoto Georges, Leroa, Velomaro, Reychell Chadwick, Randy Junge, Felicia Knightly, and Angie Simai for their assistance and expertise during capture seasons in the field. Logistic support was provided by Madagascar Institute Conservation Environment Tropical, Centre ValBio, and Henry Doorly Zoo's Madagascar Biodiversity Partnership. Funding for fieldwork was provided to A.L.B. by the National Science Foundation (BSC-0725975), Fulbright Foundation, Primate Conservation Inc., Primate Action Fund, and Stony Brook University.

LITERATURE CITED

Baden
AL.
2011
.
Communal infant care in black-and-white ruffed lemurs (Varecia variegata)
.
PhD Dissertation
,
Department of Anthropology
,
State University of New York at Stony Brook, Stony Brook, New York
,
282
pp
.
Baden
AL,
Brenneman
RA,
Louis
EE
Jr.
2008
.
Morphometrics of wild black-and-white ruffed lemurs (Varecia variegata, Kerr 1972)
.
Am J Primatol
70
:
913
926
.
Baden
AL,
Holmes
SM,
Johnson
SE,
Engberg
SE,
Louis
EE
Jr,
Bradley
BJ.
2014
.
Species-level view of population structure and gene flow for a critically endangered primate (Varecia variegata)
.
Ecol Evol
4
:
2675
2692
.
Baden
AL,
Webster
TH,
Kamilar
JM.
2016
.
Resource seasonality and reproduction predict fission-fusion dynamics in black-and-white ruffed lemurs (Varecia variegata)
.
Am J Primatol
78
:
256
279
.
Baden
AL,
Wright
PC,
Louis
EE,
Bradley
BJ.
2013
.
Communal nesting, kinship, and maternal success in a social primate
.
Behav Ecol Sociobiol
67
:
1939
1950
.
Boskoff
KJ.
1977
.
Aspects of reproduction in ruffed lemurs (Lemur variegatus)
.
Folia Primatol
28
:
241
250
.
Glander
KE.
1993
.
Capture and marking techniques for arboreal primates
.
In
:
Estudios primatologicos en Mexico, Vol. 1
,
Estrada
A,
Rodriguez-Luna
E,
Lopez-Wilchis
R
Coates-Estrada
R,
editors
.
Universidad Veracruzana
,
Xalapa, Veracruz, Mexico
,
pp
.
299
304
.
Junge
RE,
Louis
EE.
2002
.
Medical evaluation of free-ranging primates in Betampona Reserve, Madagascar
.
Lemur News
7
:
23
25
.
Junge
RE,
Louis
EE.
2005
.
Preliminary biomedical evaluation of wild ruffed lemurs (Varecia variegata and V. rubra)
.
Am J Primatol
66
:
85
94
.
King
SJ,
Morelli
TL,
Arrigo-Nelson
S,
Ratelolahy
FJ,
Godfrey
LR,
Wyatt
J,
Tecot
SR,
Jernvall
J,
Wright
PC.
2012
Morphometrics and pattern of growth in wild sifakas (Propithecus edwardsi) at Ranomafana National Park, Madagascar
.
Am J Primatol
73
:
155
172
.
Klompen
H,
Junge
RE,
Williams
CV.
2015
.
Ectoparasites of Propithecus diadema (Primates: Indriidae) with notes on unusual attachment site selection by Haemaphysalis lemuris (Parasitiformes: Ixodidae)
.
J Med Entomol
52
:
315
319
.
Maa
TC.
1963
.
Genera and species of Hippoboscidae (Diptera): Types, synonymy, habitats and natural groupings
.
Pac Insects Monogr
6
:
1
186
.
Maa
TC.
1969
.
A revised checklist and concise host index of Hippoboscidae (Diptera)
.
Pac Insects Monogr
20
:
261
299
.
MacIntosh
AJ,
Jacobs
A,
Garcia
C,
Shimizu
K,
Mouri
K,
Huffman
MA,
Hernandez
AD.
2012
.
Monkeys in the middle: Parasite transmission through the social network of a wild primate
.
PLoS One
7
:
e51144
.
Morland
HS.
1990
.
Parental behavior and infant development in ruffed lemurs (Varecia variegata) in a northeast Madagascar rain forest
.
Am J Primatol
20
:
253
265
.
Morland
HS.
1991
.
Social organization and ecology of black and white ruffed lemurs (Varecia variegata variegata) in lowland rain forest, Nosy Mangabe, Madagascar
.
PhD Dissertation
,
Department of Anthropology
,
Yale University, New Haven, Connecticut
,
419
pp.
OConnor
BM.
2003
.
Acariformes: Parasitic and commensal mites of vertebrates
.
In
:
The natural history of Madagascar
,
Goodman
SM
Benstead
JP,
editors
.
University of Chicago Press
,
Chicago, Illinois
,
pp
.
593
602
.
Rahola
N,
Goodman
SM,
Robert
V.
2011
.
The Hippoboscidae (Insecta: Diptera) from Madagascar, with new records from the Parc National de Midongy Befotaka
.
Parasite
18
:
127
140
.
Rasmussen
DT.
1985
.
A comparative study of breeding seasonality and litter size in eleven taxa of captive lemurs (Lemur and Varecia)
.
Int J Primatol
6
:
501
517
.
Schwitzer
C,
Mittermeier
R,
Johnson
S,
Donati
G,
Irwin
M,
Peacock
H,
Ratsimbazafy
J,
Razafindramanana
J,
Louis
EE,
Chikhi
L,
et al.
2014
.
Averting lemur extinctions amid Madagascar's political crisis
.
Science
343
:
842
843
.
Stamp
RK,
Brunton
DH,
Walter
B.
2002
.
Artificial nest box use by the North Island saddleback: Effects of nest box design and mite infestations on nest site selection and reproductive success
.
N Z J Zool
29
:
285
292
.
Theodor
O,
Oldroyd
H.
1965
.
On a new genus and species of Hippoboscidae (Diptera) and its relationship to Allobosca crassipes Speiser
.
Proc R Entomol Soc Lond Ser B Taxon
34
:
91
97
.
Vaughn
SE,
McGee
EM.
2009
.
Association of Allobosca crassipes (Diptera: Hippoboscidae) with the black and white ruffed lemur (Varecia variegata variegata) and Milne-Edwards' sifaka (Propithecus edwardsi) in southeastern Madagascar
.
Pan-Pac Entomol
85
:
162
166
.
Zohdy
S,
Bisanzio
D,
Tecot
S,
Wright
PC,
Jernvall
J.
2017
.
Aggression and hormones are associated with heterogeneity in parasitism and parasite dynamics in the brown mouse lemur
.
Anim Behav
132
:
109
119
.