Origin and evolution of the sulawesi macaques 1. Electrophoretic analysis of hemoglobins

The monkeys on the island of Sulawesi (Celebes), Indonesia, comprise seven species ofMacaca, that isM. maura, M. tonkeana, M. hecki, M. nigrescens, M. nigra, M. ochreata, andM. brunnescens. Hemoglobins from 248 individuals of these seven species were analyzed by isoelectric focusing electrophoresis (IEFE) and by starch gel electrophoresis in the presence of urea (USGE). Eighteen phenotypes consisting of eight molecular types were identified by IEFE analysis. The speciestonkeana inhabiting the central part of the island revealed 11 phenotypes, while peripheral species such asnigrescens andbrunnescens carried only 3 and 2 phenotypes, respectively. On USGE, three α chains and three β chains were identified and named α1, α2, and α6, and β1, β3, and β5, respectively. The α1 chain has the same mobility as the α chains of other macaques, while the α2 chain is less positively charged than α1, and α6 is the least positive among these α chains. The α2 chain is widely distributed in the Sulawesi macaques as the major component. Four species,ochreata, tonkeana, maura, andnigrescens, carried the α1 and α6 chains as minor components. The electrophoretic mobility of β1 was the same as that of other macaques, while β3 and β5 were more positively charged and less positively charged than β1, respectively. All of the Sulawesi species had β3 in high or low gene frequencies and inmaura, tonkeana, andbrunnescens, this type was most abundant. β5 chain existed in the species of the northern peninsula, as the major type. The subordinate type was β3 innigra andnigrescens and β1 inhecki. On the other hand, β1 was most frequently observed inochreata.     

Evidence of gene flow between Sulawesi macaques

A pilot field study was conducted in Sulawesi (Indonesia) to assess the status of macaque populations on the island. Wild and captive animals were sampled, mainly in border areas between presumed different species. The five species investigated were Macaca maurus, M. tonkeana, M. hecki, M. nigrescens, and M. nigra, for which morphological and gene frequency data suggested the presence of hybridization zones. Some individuals within these zones showed intermediate or mosaic morphology between parental forms. These individuals also had intermediate gene frequencies for most of the polymorphic systems investigated. Karyotypes were identical in all species, and no cytogenetic barrier to hybridization existed between species. A review of the recent literature also provided evidence for hybridization between Sulawesi macaques. Clinical frequencies in both morphological and biomolecular traits perhaps can be best explained by the operation of gene flow between the various forms of macaques on the island. However, additional data are necessary before current classification schemes are revised. The unique opportunity and need of further study of Sulawesi macaques for a range of evolutionary questions is emphasized.     

Species differences of male loud calls and their perception in sulawesi macaques

Playback experiments were conducted to investigate interspecific discrimination of male loud calls in Sulawesi macaques. Loud calls of four macaque species living in Sulawesi (Macaca tonkeana, M. maurus, M. hecki, andM. nigrescens) and a control stimulus (an 8-sec frequency modulated sound) were played back to semi-free-ranging Tonkean macaques (M. tonkeana). A preliminary acoustic analysis indicated that the calls of these four species differ in some spectral and temporal features. In the playback experiments, Tonkean macaques responded in a similar manner to conspecific calls and calls of two other species,M. maurus andM. hecki. In contrast, animals responded more weakly to the call ofM. nigrescens and the control stimulus. Males responded more strongly than females to all stimuli, while females appeared to be more discriminating for species differences than males. Analyses on the acoustic features of loud calls suggested that high frequency, wide frequency range, and repetition of sound units at a high rate elicit quick responses from animals.     

Morphological studies on the Sulawesi macaques I: Phyletic analysis of body color

The body color of Sulawesi macaques was measured quantitatively and compared among the different monkeys. As a result, divergence models for extant Sulawesi macaques, withtonkeana as the starting point and fading as the sole direction of color change, were inferred as follows: (1) fading slightly on the upper half of the body—nigra, fading more on the proximal part of the body—nigrescens; (2) fading over the whole body—maura; (3) fading greatly on the legs—hecki; and (4) fading on the distal part of the body—ochreata, fading more over the whole body, including the proximal part of the body—brunnescens. The color changed progressively in the order of (1) through (4). The divergence model, excluding the position ofhecki (3), supports the speciation model ofFooden (1969). If the proto-Sulawesi macaques had a body color pattern similar to the livingnemestrina, darkening would have been necessary for the evolution of the Sulawesi macaques after their immigration, and it may have been acquired as an adaptation to the ground (forest floor) living nature of the Sulawesi macaques, together with influences deriving from the insularity and/or from the absence of predators.     

Hybridization and population genetics of two macaque species in Sulawesi, Indonesia

This study investigates hybridization and population genetics of two species of macaque monkey in Sulawesi, Indonesia, using molecular markers from mitochondrial, autosomal, and Y-chromosome DNA. Hybridization is the interbreeding of individuals from different parental taxa that are distinguishable by one or more heritable characteristics. Because hybridization can affect population structure of the parental taxa, it is an important consideration for conservation management. On the Indonesian island of Sulawesi an explosive diversification of macaques has occurred; seven of 19 species in the genus Macaca live on this island. The contact zone of the subjects of this study, M. maura and M. tonkeana, is located at the base of the southwestern peninsula of Sulawesi. Land conversion in Sulawesi is occurring at an alarming pace; currently two species of Sulawesi macaque, one of which is M. maura, are classified as endangered species. Results of this study indicate that hybridization among M. maura and M. tonkeana has led to different distributions of molecular variation in mitochondrial DNA and nuclear DNA in the contact zone; mitochondrial DNA shows a sharp transition from M. maura to M. tonkeana haplotypes, but nuclear DNA from the parental taxa is homogenized in a narrow hybrid zone. Similarly, within M. maura divergent mitochondrial DNA haplotypes are geographically structured but population subdivision in the nuclear genome is low or absent. In M. tonkeana, mitochondrial DNA haplotypes are geographically structured and a high level of nuclear DNA population subdivision is present in this species. These results are largely consistent with a macaque behavioral paradigm of female philopatry and obligate male dispersal, suggest that introgression between M. maura and M. tonkeana is restricted to the hybrid zone, and delineate one conservation management unit in M. maura and at least two in M. tonkeana.     

Origin and evolution of the sulawesi macaques 2. complete amino acid sequences of seven β chains of three molecular types

Seven β chains were identified as the typical molecular types carried by the seven species of Sulawesi macaques based on isoelectric focusing and urea starch gel electrophoresis. These β chains include the β3 chains ofmaura, tonkeana, nigra, andbrunnescens, β1 chains ofhecki andochreata and β5 chain ofnigra. The results of chromatography on cation-exchange and reversed phase columns and the amino acid compositions of the tryptic peptides suggested substitutions at the 9th and 13th amino acids from the N-terminal. Sequence analyses of these seven β chains from the N-terminal to the 18th amino acid and those of purified tryptic peptides from βT3 to βT15 by Edman degradation revealed the following facts: (1) the amino acid sequences of the β3 chains carried by the four species coincided with each other and as did those of the β1 chains of the two named species; and (2) the 9th and 13th amino acids were Lys and Thr in β3, Asn and Asn in β1, and Asp and Thr in the β5 chain, respectively. These three β chains are related with each other by at least two-base changes. The evolution of the β chains of the Sulawesi macaques was inferred to be as follows. (1) The β3 chain might have been dominant β chain in the past among Sulawesi macaques, since peripheral species separately carried this chain; (2) the β1 and β5 chains might have derived from a “missing link” because of more than two-base substitutions between β3 and β1 and between β3 and β5; (3) eight other macaque species, including the lion-tailed macaque (M. silenus), bear Asn and Thr at these two positions, while the Barbary macaque (M. sylvanus) has Thr and Thr; and (4) thus, if the parsimonious rule is followed, the type with Asn-Thr is the most plausible “missing link,” since only the Asn-Thr type can combine these five β chains by minimum one-base change. Two genetic events are postulated in the evolutionary process of the Sulawesi β chains: the first Lys-Thr type (β3) was distributed over the whole island, and next Asn-Thr, the common type in other macaques, produced Asn-Asn (β1) and Asp-Thr (β5).     

Hematological values and parasite fauna in free-rangingMacaca hecki and theM. hecki/M. tonkeana hybrid group of Sulawesi Island, Indonesia

Hematological values and parasitological fauna of free-rangingMacaca hecki and the hybrid group betweenM. hecki/M. tonkeana of Sulawesi Island, Indonesia, were investigated. The hematological values, especially the red cell number (RBC), were lower than those of other macaque species, indicating that Sulawesi macaques are slightly anemic. Several parasites including Plasmodium sp., trombiculid mites, andTrichuris trichiura were identified. Although infection by Plasmodium was observed with considerable frequency, no clear relationship between its infection and the occurrence of anemia was found. Trombiculid mites and eggs ofAnatrichosoma sp. were detected in foci of the ears of most monkeys. The infection with a trombiculid mite is the first recorded occurrence in free-ranging wild Sulawesi macaques. Gastrointestinal parasites were identified from their eggs in fecal samples, where five species of nematoda and one trematoda species were found.     

Morphological variation within a macaque hybrid zone

A hybrid zone exists between Macaca tonkeana and Macaca hecki (Primates: Cercopithecidae), centered along the Tawaeli-Toboli road in the narrow isthmus that connects North and Central Sulawesi, Indonesia. The current study demonstrates morphological substructure from north to south across the hybrid zone. Macaques to the northwest of the Tawaeli-Toboli road more closely resemble M. hecki, and macaques to the southeast of the road resemble M. tonkeana. While morphology shifted for both males and females over a distance of 1,500-2,000 m, adult males were significantly more M. tonkeana-like across the morphological gradient. This suggests that in the study area, males of M. tonkeana-like morphology are dispersing into hybrid groups at the expense of M. hecki-like males. A permutation analysis of diagnostic character states indicated that associations existed among several morphological traits. This could be due to the operation of one or several nonexclusive evolutionary processes, including recent secondary contact, pleiotropic effects, physical linkage of loci, natural selection against hybrids, the influx of parental types, or assortative mating. Continued environmental perturbation associated with the Tawaeli-Toboli road is likely to be a significant factor in the future of the M. tonkeana/M. hecki hybrid interaction.     

Macaca nigra on Bacan Island,Indonesia: Its morphology,distribution, and present habitat

We studied the monkeys inhabiting Bacan Island on the Maluku Archipelago, Indonesia. We compared them morphologically with Sulawesi black macaques (Macaca nigra).We also assessed their distribution on the island and on neighboring islands and the influence of human activity on them. We found that the Bacan black macaques are essentially similar to Sulawesi macaques; the variations in each locality are comparable. The monkeys probably inhabit the whole island or, at least, the lower forest thereon. Human activities — cultivation, logging, catching and killing monkeys — have not much affected their population. Thus they constitute a good stock of Macaca nigra,though the main population in Sulawesi is endangered.     

Origin of the Sulawesi macaques (Cercopithecidae: Macaco) as suggested by mitochondrial DNA phylogeny

One of the sharpest biogeographical transitions in the world occurs between the Indonesian islands of Borneo and Sulawesi; this transition is demarcated by Wallace's line. Macaque monkeys represent an interesting anomaly to faunal distributions in this region as they occur on both sides of Wallace's line, with Macacafascicularis, M. nemestrina and other species to the west and seven Sulawesi species to the east. We have investigated macaque evolution and dispersal in the Sunda region and Sulawesi using phylogenetic analysis of mitochondrial DNA sequences. Female philopatry of macaques, which causes sharp geographic clustering of maternally inherited mitochondrial DNA haplotypes, makes mitochondrial phytogenies particularly useful for investigating ancient patterns of dispersal. Results of this study suggest the following: (1) M. fascicularis is not a sister taxon to any species of Sulawesi macaque; (2) haplotypes of some M. nemestrina have a sister relationship to northern and central Sulawesi macaques, while haplotypes of other M. nemestrina have a sister relationship to soudiern Sulawesi macaques; (3) Sulawesi was probably colonized by macaques twice, once to the base of the northern peninsula now occupied by M. hecki and once to the southwestern peninsula now occupied by M, mama; and (4) within north/central and southern Sulawesi, patterns of dispersal are largely consistent with contemporary and past geography of the island, with the exception of a geographically discontinuous relationship between M. nigra and a portion of M. tonkeana from a region in northwest central Sulawesi.     

Identification of Entamoeba polecki with Unique 18S rRNA Gene Sequences from Celebes Crested Macaques and Pigs in Tangkoko Nature Reserve,North Sulawesi,Indonesia

Unique species of macaques are distributed across Sulawesi Island, Indonesia, and the details of Entamoeba infections in these macaques are unknown. A total of 77 stool samples from Celebes crested macaques (Macaca nigra) and 14 stool samples from pigs were collected in Tangkoko Nature Reserve, North Sulawesi, and the prevalence of Entamoeba infection was examined by PCR. Entamoeba polecki was detected in 97% of the macaques and all of the pigs, but no other Entamoeba species were found. The nucleotide sequence of the 18S rRNA gene in E. polecki from M. nigra was unique and showed highest similarity with E. polecki subtype (ST) 4. This is the first case of identification of E. polecki ST4 from wild nonhuman primates. The sequence of the 18S rRNA gene in E. polecki from pigs was also unique and showed highest similarity with E. polecki ST1. These results suggest that the diversity of the 18S rRNA gene in E. polecki is associated with differences in host species and geographic localization, and that there has been no transmission of E. polecki between macaques and pigs in the study area.     

External characteristics and associated developmental changes in two species of Sulawesi macaques,Macaca tonkeana andM. hecki,with special reference to hybrids and the borderland between the species

The degree of intergradation between two species of Sulawesi macaques,Macaca tonkeana andM. hecki, was studied by examining the diagnostic external characteristics of more than 100 monkeys kept as pets by natives. Two possible hybrid monkeys were found and both originated from the borderland between the two species, located in the most proximal region of the northern peninsula of Sulawesi. The previously postulated wide area of integradation between the two species at the possible contact zone was, however, not recognized, and typical examples oftonkeana orhecki were found to be present on the two sides of a narrow “hybrid” zone which was defined by direct observations. Furthermore, despite considerable individual variations, we were able to allocate most monkeys to one or other of the species. Each of ten external characteristics of the members of both species more or less encompassed the individual variations, but may undergo changes with the development of the monkeys. The mechanisms of reproduction of hybrid monkeys and the maintenance of differences between the species are discussed.     

A novel GH43 α-l-arabinofuranosidase from Humicola insolens: mode of action and synergy with GH51 α-l-arabinofuranosidases on wheat arabinoxylan

A novel α-l-arabinofuranosidase (α-AraF) belonging to glycoside hydrolase (GH) family 43 was cloned from Humicola insolens and expressed in Aspergillus oryzae. ¹H-NMR analysis revealed that the novel GH43 enzyme selectively hydrolysed (1→3)-α-l-arabinofuranosyl residues of doubly substituted xylopyranosyl residues in arabinoxylan and in arabinoxylan-derived oligosaccharides. The optimal activity of the cloned enzyme was at pH 6.7 and 53 °C. Two other novel α-l-arabinofuranosidases (α-AraFs), both belonging to GH family 51, were cloned from H. insolens and from the white-rot basidiomycete Meripilus giganteus. Both GH51 enzymes catalysed removal of (1→2) and (1→3)-α-l-arabinofuranosyl residues from singly substituted xylopyranosyls in arabinoxylan; the highest arabinose yields were obtained with the M. giganteus enzyme. Combinations (50:50) of the GH43 α-AraF from H. insolens and the GH51 α-AraFs from either M. giganteus or H. insolens resulted in a synergistic increase in arabinose release from water-soluble wheat arabinoxylan in extended reactions at pH 6 and 40 °C. This synergistic interaction between GH43 and GH51 α-AraFs was also evident when a GH43 α-AraF from a Bifidobacterium sp. was supplemented in combination with either of the GH51 enzymes. The synergistic effect is presumed to be a result of the GH51 α-AraFs being able to catalyse the removal of single-sitting (1→2)–α-l-arabinofuranosyls that resulted after the GH43 enzyme had catalysed the removal of (1→3)–α-l-arabinofuranosyl residues on doubly substituted xylopyranosyls in the wheat arabinoxylan.     

Genetic characterization of long-tailed macaques (Macaca fascicularis) on Tabuan Island Indonesia

Protein and mitochondrial DNA variations (D-loop region PCR-RFLP) were analyzed for 7 serum and 40 clot samples collected from long-tailed macaques (Macaca fascicularis) living on Tabuan Island, Indonesia. Protein polymorphisms were examined electrophoretically for 5 and 12 kinds of protein in serum and erythrocytes, respectively. Each of the protein loci tested showed a monomorphic pattern. Polymorphisms were detected in the analysis of the D-loop-containing region of mtDNA (PCR-RFLP) using 32 restriction endonucleases. Two haplotypes, differing 1.03% in sequence divergence were observed, and both were previously undetected in other local populations. Based on genetic features and differences in pelage color as outlined inFooden's (1995) morphological analysis, the present results suggest that long-tailed macaques on Tabuan Island are a unique population. From the genetic analyses performed here, Tabuan monkeys are considered to be the same species group as those populations of Sumatra and Java (Fooden, 1995).     

The Human–Macaque Interface: Conservation Implications of Current and Future Overlap and Conflict in Lore Lindu National Park, Sulawesi, Indonesia

Here, I examine overlapping resource use of forest and cultivated resources by villagers and tonkean macaques (Macaca tonkeana) in Lore Lindu National Park, Sulawesi, Indonesia. An integrative research design was employed, using methods from primatology and cultural anthropology, in conjunction with quantitative measurements of cacao crop loss. The results indicate that the current patterns of overlapping resource use may not be severely affecting the tonkean macaques or villagers in Lindu. The research does, however, point to possible future difficulty if cacao crop raiding by macaques increases, and as changing socioeconomic trends and loss of traditional folklore result in further modification of macaque habitat. Successful strategies to ameliorate human–macaque conflict may require efforts aimed at the adoption of alternative buffer zone crops that use shade-management systems, the deliberate protection of important macaque food species, and increasing local tolerance to crop raiding by exploring the role of macaques in forest regeneration.     

Flexibility in Diet and Activity Patterns of <Emphasis Type="Italic">Macaca tonkeana</Emphasis> in Response to Anthropogenic Habitat Alteration

I examined how Sulawesi Tonkean macaques (Macaca tonkeana) respond in terms of their diet and activity patterns to anthropogenic habitat alteration in Lore Lindu National Park, Sulawesi, Indonesia. Anthropogenic habitat alteration comprised clearing of forest for agriculture and small-scale forest product collection. I quantified the diet and activity of 2 groups (CH and Anca), occupying habitats with different levels of alteration, via scan sampling. Tree abundance, key food specific density, and fruit production were greater in the minimally altered habitat (CH), substantiating the characterization of the group’s habitat as higher quality. For the group in the heavily altered habitat (Anca), alternative foods, e.g., insects, fungus, young and mature leaves, shoots, and stems, accounted for a significantly greater proportion of the diet. Dietary diversity is significantly lower in the Anca group, with 52% of their diet being palm fruits from Arenga pinnata. The activity patterns of the Anca group—more time foraging, less time moving, and more time resting than the CH group—reflect the lower resource availability in their habitat and their reliance on more alternative food items, coupled with their extremely small group size (6–9 individuals). The group may be at the optimal size in which foraging efficiency is maximized for the habitat, a response, in conjunction with dietary and behavioral flexibility, to alteration of their habitat. The results are contextualized with respect to the conservation value of human-modified landscapes.     

Intergroup encounters in wild moor macaques (<Emphasis Type="Italic">Macaca maurus</Emphasis>)

We studied intergroup encounters among moor macaques at the Karaenta Nature Reserve, South Sulawesi, Indonesia. Group B has been observed on the basis of individual identification since 1988. We analyzed 85 encounters between members of Group B and members of neighboring groups from September 1990 to November 1998. The average frequency of intergroup encounters was 0.035/hour. Neither the presence of females in estrus nor rainfall had an effect on encounter frequency. Behaviors of moor macaques during intergroup encounters differed from those of Japanese macaques. In moor macaques, no intergroup interactions with body contact were observed during encounters, and females never directed aggression toward members of different groups. The present study did not confirm the prediction of the model ofvan Schaik (1989). Extension of the existing models is required to explain the difference in female dominance styles among macaques by socioecological factors.     

Prevalence of enteric parasites in pet macaques in Sulawesi, Indonesia

On the Indonesian island of Sulawesi, nonhuman primate pets come into frequent contact with humans, presenting the possibility of zoonotic and anthropozoonotic disease transmission. We collected fecal samples from 88 pet macaques representing six of the seven macaque species currently recognized as endemic to Sulawesi (Macaca nigra, M. nigrescens, M. hecki, M. tonkeana, M. maura, and M. ochreata) as well as two non-endemic species (M. fascicularis and M. nemestrina) in order to determine the prevalence of intestinal parasitic infection in this population. Seven taxa of intestinal protozoa (Blastocystis hominis, Iodamoeba bütschlii, Entamoeba coli, Entamoeba hartmanni, Chilomastrix mesnili, Endolimax nana, and Retortamonas intestinalis) and three taxa of nematodes (hookworm, Trichuris spp., and Ascaris spp.) were detected. The overall parasitization rate was 59.1%. Commensal organisms predominated in this population. Parasitization was not statistically correlated with macaque age group, sex, species, or location, or with the owner's level of education. These findings are discussed in the context of primate pet ownership practices in Sulawesi.     

Contributions to the taxonomy of the genera Pachysternum and Cyrtonion (Coleoptera, Hydrophilidae, Sphaeridiinae)

Pachysternum loxodonta sp. n. from the Republic of the Congo and Pachysternum sulawesicum sp.n. from Sulawesi Island, Indonesia are described. The generic status of Pachysternum sculpticolle (Régimbart, 1907) is revised, the species is transferred to the genus Cyrtonion and its differences from C. ghanense Hansen, 1989 are discussed. Taxonomy of Pachysternum is discussed, dividing the genus prelusively into three species groups, one comprising all known Oriental and Eastern Palearctic species, and two comprising the Afrotropical species. Species with unclear positions within the genus Pachysternum or with questionable generic status are left as “Species incertae sedis”.