Mentawai’s endemic,relictual fauna: is it evidence for Pleistocene extinctions on Sumatra?

Aim The four Mentawai islands, south‐west of Sumatra, have long been isolated from the remainder of Sundaland, resulting in a high level of endemism. We examined the distribution of 151 species of the Mentawai Islands in Sundaland and assessed various processes that may have resulted in the various biogeographical patterns. Location Southeast Asia, particularly the Mentawai Islands and nearby large landmasses (Sumatra, Java, Borneo and Peninsular Malaysia). Methods We compared the faunal composition of the Mentawai Islands for selected taxa (43 mammals, 92 reptiles and 16 amphibians) with that of the four nearby large landmasses of Sundaland using morphological comparisons and the most recent molecular phylogenetic analyses available in the literature. These comparisons yielded sister taxa, which were used to simulate species absence data for the four Sundaland landmasses under several scenarios to investigate how patterns of species absence could have arisen. Results In contrast to our expectations, several Mentawai species did not have their closest relatives on neighbouring Sumatra, but rather on the more distant Borneo, Java or Peninsular Malaysia. For mammals, the similarity between species from Mentawai and Borneo was greater than that observed between species from Mentawai and Sumatra. We conclude that the relationships represent traces of species historically distributed throughout Sundaland that became extinct in Sumatra during the Pleistocene. For reptiles and amphibians the observed pattern of species absences generally resembled the simulated pattern expected under the scenario of absence rates increasing with landmass isolation, whereas for mammals we observed more species than expected missing from Java and Sumatra, and fewer than expected from Borneo. Main conclusions The potential extinctions on Sumatra probably had two causes: changes of climate and vegetation during the Pleistocene and environmental impacts from the Toba supervolcanic eruption.     

Morphological and genetic relationships of the Crocidura monticola species complex (Soricidae: Crocidurinae) in Sundaland

Small Crocidurinae shrews (weight <8 g) from Southeast Asia have been poorly studied to date, mainly because of the difficulty to catch them and the concomitant paucity of reference specimens available in museums. Hence their systematics is still debated, and most small Crocidura shrews from Sundaland are assigned to the monticola species complex. Here, we report a study based on a survey of shrews caught with large pitfalls set on forest floors in Peninsular Malaysia. Morphometric analyses based on 14 skull measurements showed that these shrews tend to be larger with increasing altitude, but showed otherwise no consistent variation. When compared to museum specimens of the monticola species complex sampled in the Sundaland (total: 77 specimens), the Malay shrews tend also to be larger than those living on Kalimantan (Borneo) and Sumatra. All are, however, morphologically distinct from the other species, C. maxi, found in eastern Java and on the Lesser Sundas. Molecular analyses of a subset of these small shrews and based on a mitochondrial (cytochrome b) and a nuclear gene (Apolipoprotein B) suggest that samples from the central region of Peninsular Malaysia (Bukit Rengit and Ulu Gombak) genetically differ from other Malaysian populations (by about 7% K2P distance at the cyt b gene) and are more closely related to some samples from Sumatra and Borneo. These differences did not correlate with the altitudinal variation evidenced from the morphological analysis. Reference sequences from the terra typica of monticola and maxi (both species were originally described from Java) are however needed to determine if these unexpected genetic differences warrant additional taxonomic subdivision within the Sundaland.     

Phylogeographic structure in mitochondrial DNA of a South-east Asian freshwater fish, Hemibagrus nemurus (Siluroidei; Bagridae) and Pleistocene sea-level changes on the Sunda shelf

We postulated that the biogeographical history of South-east Asia contributed to extensive admixture during Pleistocene low sea levels of genetic groups of an obligate freshwater fish (the river catfish, Hemibagrus nemurus) isolated during periods of high sea levels. During Pleistocene glacial maxima, the sea level was lower than at present and the islands of the Sunda shelf (Sumatra, Borneo and Java) and the Asian mainland were connected by lowlands traversed by rivers. Restriction fragment length polymorphisms in mitochondrial DNA were documented for 140 putative H. nemurus analysed from 13 sampling sites resulting in the definition of 35 haplotypes. The high level of haplotype differentiation (mean P × 100 = 2.22, SD = 1.33) indicates that the subdivision of the ancestral H. netnurus group was extensive and probably occurred early in the Pleistocene. The occurrence of some genetically divergent groups of the H. netnurus complex occurring in sympatry in widely separated locations supports the proposition that low sea levels aided the dispersion and mingling of genetic groups. Based on both genetic and morphological evidence, the main H. nemurus line gave rise to three regional groups: (1) a morphologically distinct ‘Indochinese’ group composed of two mtDNA clades overlapping in east peninsular Malaysia; (2) a ‘Sundaic’ group composed of various lineages of differing morphology and genetic identity; (3) a genetically distinct ‘Sarawak’ group in west Borneo, similar in morphology to the ‘Sundaic’ and ‘Indochinese’ groups, but including a small, golden colour morph as a distinct dade. The morphologically similar Sundaic forms from west Java, Sumatra and west Borneo show some degree of genetic divergence, but their phylogenetic relationships are poorly resolved. The most genetically and morphologically distinct Sundaic dade, assigned to H. hoevenii, colonized the Kapuas river (west Borneo), east Sumatra and south peninsular Malaysia. Contrary to our original hypothesis and present biogeographical theory, little exchange of genetic groups has apparently occurred between the mainland and the Sunda Islands during recent glaciations.     

Population affinities of the asiatic cobra (Naja naja) species complex in south-east Asia: reliability and random resampling

The population systematics of the cobras of the genus Naja in southern Thailand, Malaysia and Indonesia are investigated, using multivariate analysis of a large number of morphological characters. These populations are found to constitute three distinct groups: a northern form, which occurs in Thailand and northern Peninsular Malaysia; an equatorial form, which occurs in southern Thailand, Peninsular Malaysia, Sumatra and Borneo; and a southern form, which occurs on Java and the Lesser Sunda Islands. The first two forms are sympatric in northern Peninsular Malaysia and southern Thailand, and therefore constitute separate species. This is of importance for the treatment of snakebite in the region. The distribution of the three forms can be related both to present ecological conditions and to Pleistocene geological and climatic events. The reliability of the results is demonstrated by the relationship between character number and congruence of patterns of geographic variation, investigated by random resampling. The pattern of geographic variation within two of the three main forms is investigated and related to current ecological conditions and Pleistocene events.     

A molecular test of Huxley's line: Cyrtandra (Gesneriaceae) in Borneo and the Philippines

A biogeographic and phylogenetic study of Cyrtandra (Gesneriaceae) in the Sundaland region (Borneo and Peninsular Malaysia) and the Philippines using nuclear ribosomal (ITS) DNA sequence data reveals a major division between the Cyrtandra floras of Sundaland and the Philippines. Palawan, the most westerly of the Philippine islands, emerges as an area of mixing between these two. The Bornean element in the Cyrtandra flora of Palawan (two species in our sample) appears to result from recent (i.e. Pleistocene) dispersal from Borneo. The remaining seven species sampled from Palawan are most closely related to those from elsewhere in the Philippines. However, the Palawan clade is sister to the other Philippine taxa, suggesting an ancient (possibly Pliocene) vicariance event. Huxley's line–a zoogeographic boundary placing Palawan and Borneo together–receives some support from this study as there is evidence of recent dispersal of Bornean flora into Palawan. However, in terms of more ancient biogeographic patterning of the region, Palawan has stronger links with the other Philippine islands.     

Mitochondrial genetic variation and population structure of the striped snakehead,Channa striata in Malaysia and Sumatra,Indonesia

We explored the genetic diversity and structure of the striped snakehead (Channa striata) across Malaysia and Sumatra (Indonesia) using the partial mtDNA CO1 gene. Twenty five populations (n = 345) were assayed and subdivided into six regions, following the physiogeographical barriers. Populations Sega (SG), Tanjung Tambutan (TR), Kajang (KJ) and Linggi (LG) are highly diversified (Hd: 0.484–0.762, π: 0.0033–0.0059) which could serve as candidates for a selective breeding programme. The only population that contributed to the total allelic richness is Takengon (CS) as it is highly differentiated from other populations and genetically variable within population. We detected two major phylogenies: 1) northwest Peninsular Malaysia and 2) all other regions of Malaysia and Sumatra, Indonesia. They are products of the physical restriction to gene flow between the two lineages by the Bintang Mountain Range. A total of 92.4% of the population pairwise comparison F_ST showed significant structuring, yet several geographically distant populations showed a close genetic relationship. The discrepancy is due to ancient population dispersal and human-mediated translocation. These major findings provide an important base study for initiating a selective breeding program. The high population genetic diversity requires independent conservation as they contain most of the total diversity in this area.     

REVISITING WALLACE'S HAUNT: COALESCENT SIMULATIONS AND COMPARATIVE NICHE MODELING REVEAL HISTORICAL MECHANISMS THAT PROMOTED AVIAN POPULATION DIVERGENCE IN THE MALAY ARCHIPELAGO

Sundaland, a biogeographic region of Southeast Asia, is a major biodiversity hotspot. However, little is known about the relative importance of Pleistocene habitat barriers and rivers in structuring populations and promoting diversification here. We sampled 16 lowland rainforest bird species primarily from peninsular Malaysia and Borneo to test the long‐standing hypothesis that animals on different Sundaic landmasses intermixed extensively when lower sea‐levels during the Last Glacial Maximum (LGM) exposed land‐bridges. This hypothesis was rejected in all but five species through coalescent simulations. Furthermore, we detected a range of phylogeographic patterns; Bornean populations are often genetically distinct from each other, despite their current habitat connectivity. Environmental niche modeling showed that the presence of unsuitable habitats between western and eastern Sundaland during the LGM coincided with deeper interpopulation genetic divergences. The location of this habitat barrier had been hypothesized previously based on other evidence. Paleo‐riverine barriers are unlikely to have produced such a pattern, but we cannot rule out that they acted with habitat changes to impede population exchanges across the Sunda shelf. The distinctiveness of northeastern Borneo populations may be underlied by a combination of factors such as rivers, LGM expansion of montane forests and other aspects of regional physiography.     

When Thailand was an island – the phylogeny and biogeography of mite harvestmen (Opiliones,Cyphophthalmi, Stylocellidae) in Southeast Asia

Aim To develop a comprehensive explanation for the biological diversity of Southeast Asia, especially in the Wallacea and Sundaland regions. This study focuses on a group of arachnids, mite harvestmen, which are thought to be an extremely old group of endemic animals that have been present in the region since most of its land supposedly formed part of the northern rim of the supercontinent Gondwana. Location Eastern Himalayas, Thai‐Malay Peninsula, Sumatra, Borneo, Java, Sulawesi, and New Guinea. Methods  Approximately 5.6 kb of sequence data were obtained from 110 South‐east Asian Cyphophthalmi specimens. Phylogenetic analyses were conducted under a variety of methods and analytical parameters, and the optimal tree was dated using calibration points derived from fossil data. Event based and paralogy‐free subtree biogeographical analyses were conducted. Results The Southeast Asian family Stylocellidae was recovered as monophyletic, arising on what is now the Thai‐Malay Peninsula and diversifying into three main clades. One clade (Meghalaya, here formally placed in Stylocellidae) expanded north as far as the eastern Himalayas, a second clade entered Borneo and later expanded back across the Sundaland Peninsula to Sumatra, and a third clade expanded out of Borneo into the entire lower part of Sundaland. Molecular dating suggested that Stylocellidae separated from other Cyphophthalmi 295 Ma and began diversifying 258 Ma, and the lineage that inhabits mostly Borneo today began diversifying between 175 and 150 Ma. Main conclusions The topology and molecular dating of our phylogenetic hypothesis suggest that Stylocellidae originated on Gondwana, arrived in Southeast Asia via the Cimmerian palaeocontinent, and subsequently diversified north, then south. Their present distribution in the Indo‐Malay Archipelago is explained largely by a diversification over the Sundaland Peninsula before western Sulawesi departed and the peninsula was extensively inundated.     

The systematics and ecology of snakeheads (Pisces: Channidae) in Peninsular Malaysia and Singapore

Seven species of snakeheads (Channidae) are known from Peninsular Malaysia and Singapore, viz. Channa bankanensis, C. gachua, C. lucius, C. marulioides, C. melasoma, C. micropeltes and C. striata. Up-to-date distribution maps of each species are presented, including new records. Their systematics is reviewed and partially revised. The taxonomic status of C. marulioides and C. melanoptera is clarified. Specimens from Peninsular Malaysia identified as C. melanoptera sensu Weber & de Beaufort, 1922, proved to be the adult form of C. marulioides s.str. The real C. melanoptera appears to be restricted to Borneo and possibly Sumatra. The life history of a blackwater species C. bankanensis is also documented, with regards to the morphological and colour-pattern changes associated with growth. An updated key to the seven species based on morphometric measurements and meristic counts is presented.     

The geography of diversification in mutualistic ants: a gene's-eye view into the Neogene history of Sundaland rain forests

We investigate the geographical and historical context of diversification in a complex of mutualistic Crematogaster ants living in Macaranga trees in the equatorial rain forests of Southeast Asia. Using mitochondrial DNA from 433 ant colonies collected from 32 locations spanning Borneo, Malaya and Sumatra, we infer branching relationships, patterns of genetic diversity and population history. We reconstruct a time frame for the ants' diversification and demographic expansions, and identify areas that might have been refugia or centres of diversification. Seventeen operational lineages are identified, most of which can be distinguished by host preference and geographical range. The ants first diversified 16-20 Ma, not long after the onset of the everwet forests in Sundaland, and achieved most of their taxonomic diversity during the Pliocene. Pleistocene demographic expansions are inferred for several of the younger lineages. Phylogenetic relationships suggest a Bornean cradle and major axis of diversification. Taxonomic diversity tends to be associated with mountain ranges; in Borneo, it is greatest in the Crocker Range of Sabah and concentrated also in other parts of the northern northwest coast. Within-lineage genetic diversity in Malaya and Sumatra tends to also coincide with mountain ranges. A series of disjunct and restricted distributions spanning northern northwest Borneo and the major mountain ranges of Malaya and Sumatra, seen in three pairs of sister lineages, further suggests that these regions were rain-forest refuges during drier climatic phases of the Pleistocene. Results are discussed in the context of the history of Sundaland's rain forests.     

Nuclear and chloroplast DNA phylogeography reveals Pleistocene divergence and subsequent secondary contact of two genetic lineages of the tropical rainforest tree species Shorea leprosula (Dipterocarpaceae) in South‐East Asia

Tropical rainforests in South‐East Asia have been affected by climatic fluctuations during past glacial eras. To examine how the accompanying changes in land areas and temperature have affected the genetic properties of rainforest trees in the region, we investigated the phylogeographic patterns of a widespread dipterocarp species, Shorea leprosula. Two types of DNA markers were used: expressed sequence tag‐based simple sequence repeats and chloroplast DNA (cpDNA) sequence variations. Both sets of markers revealed clear genetic differentiation between populations in Borneo and those in the Malay Peninsula and Sumatra (Malay/Sumatra). However, in the south‐western part of Borneo, genetic admixture of the lineages was observed in the two marker types. Coalescent simulation based on cpDNA sequence variation suggested that the two lineages arose 0.28–0.09 million years before present and that following their divergence migration from Malay/Sumatra to Borneo strongly exceeded migration in the opposite direction. We conclude that the genetic structure of S. leprosula was largely formed during the middle Pleistocene and was subsequently modified by eastward migration across the subaerially exposed Sunda Shelf.     

Phylogeny and systematics of the Pachychilidae of mainland South‐East Asia – novel insights from morphology and mitochondrial DNA (Mollusca,Caenogastropoda, Cerithioidea)

Pachychilid gastropods are a conspicuous element of the freshwater macro‐invertebrate fauna of South‐East Asia. In this region, three spatially separated groups of pachychilids can be differentiated mostly by means of their brooding strategy. The largest group comprises about 100 described species and displays an extended Sundaland distribution that ranges from north‐east India in the west to southern China in the east and Borneo in the south‐east. The most conspicuous synapomorphy shared by females is a subhaemocoelic brood pouch in which developing embryos are retained. Mitochondrial phylogenies based on partial sequences of the genes cytochrome oxidase C subunit I (COI) and 16S rRNA (16S) support the monophyly of this group of subhaemocoelic brooders. Amongst them, the two genera Brotia and Paracrostoma form a sister pair of clades both together forming the sister taxon of a third clade. This third clade itself is split into three subclades, which occupy separate ranges except for the overlap of two of them in Borneo. The subclades comprise species from (A) Thailand, Cambodia, and northern Borneo; (B) Java and south‐western Borneo; and (C) Vietnam and southern China. In contrast with their spatial structuring, all species share a widely corresponding anatomy. Characteristic key features are a pallial oviduct with a seminal receptacle and an embryonic shell with a smooth, dome‐like inflated apex. Differences in the histology of the pallial oviduct are not congruent with phylogenetic patterns as suggested by the mitochondrial trees. Given the lack of obvious distinguishing morphological characteristics, all species of the subclades A to C are assigned to a single genus, Sulcospira, with Adamietta as a junior synonym.     

Genetic Diversity of a Tropical Tree Species,Shorea leprosula Miq. (Dipterocarpaceae), in Malaysia: Implications for Conservation of Genetic Resources and Tree Improvement1

Genetic diversity and population genetic structure of Shorea leprosula was investigated using seven natural populations distributed throughout Peninsular Malaysia and one natural population from Borneo. The mean population and species level genetic diversity were exceptionally high (H_e= 0.369 ± 0.025 and 0.406 ± 0.070, respectively). Heterozygosity varied among populations, ranging from 0.326 to 0.400, with the highest values found in the populations from central Peninsular Malaysia. Correlations among ecological factors (longitude, latitude, and annual rainfall) were not significant (P > 0.05), indicating that these ecological variables were not responsible for the observed genetic differences among populations. The Bangi adult population exhibited a higher level of observed heterozygosity but lower fixation indices in comparison to its seedling population. All other seedling populations also showed positive fixation indices (f), indicating a general excess of homozygotes. This also may suggest selection against homozygotes between the seedling and adult stages. A low level of population differentiation was detected (G_ST= 0.117 with the Lambir population and GST= 0.085 without the Lambir population). Furthermore, gene flow (N_m) between populations was not significantly correlated with geographical distances for the populations within Peninsular Malaysia. Cluster analysis also did not reflect geographical proximity and gave little insight into the genetic relatedness of the populations. This may indicate that the populations sampled are part of a continuous population with fragmentation having occurred in the recent past.     

Ranging behaviour,activity, habitat use,and morphology of the Malay civet (Viverra tangalunga) on Peninsular Malaysia and comparison with studies on Borneo and Sulawesi

The Malay civet (Viverra tangalunga) is found throughout Southeast Asia, but there have been very few studies on its natural history and ecology. We present here the results from the first radio telemetry study of this species on Peninsular Malaysia and compared our findings to similar studies on Borneo and Sulawesi. From August 2004 to August 2006, we captured 11 Malay civets in Krau Wildlife Reserve and radio-tracked seven adults (four males and three females). The mean weight of males (6.6 kg) was significantly higher than females (5.8 kg). Both sexes on Peninsular Malaysia were larger than their counterparts on Borneo and Sulawesi. There was no significant difference between the mean sizes of male and female home ranges on Peninsular Malaysia; the mean home range size for both sexes was 143 ha (95% MCP), which was larger than the mean range size recorded on Borneo and Sulawesi. We found that the Malay civet is a solitary, territorial species on Peninsular Malaysia: mean range overlap was 15% for males and 0% for females, and the home range of each male overlapped one or two females. Malay civets were mainly nocturnal, with some periods of rest during the night; daytime rest sites were within dense ground cover. Lowland forest is an essential habitat for this species; although Malay civets were sometimes found in plantations, they did not venture far from forested habitat.     

Genetic structure of Melanotaenia australis at local and regional scales in the east Kimberley, Western Australia

The Kimberley region of Western Australia possesses a poorly studied freshwater fish fauna with high endemism in an aquatic landscape subject to monsoonal floods and dry season isolation. In the first population genetic study of freshwater fish in this region, the authors tested the effects of geographic barriers on genetic structure at multiple spatial scales in east Kimberley populations of the western rainbowfish, Melanotaenia australis , the most widespread and abundant species in the region. Based on allozyme comparisons, hierarchical analysis of F _ST revealed increasing genetic subdivision with spatial scale. Minimal genetic structure within creeklines demonstrated that wet season dispersal, rather than dry season isolation, determines genetic structure at small scales. At the scale of sub-catchments, a pattern of isolation by distance along creeklines was evident. Genetic subdivision between adjacent river systems was greater between rivers separated by a plateau than by lowlands. This implies greater connectivity of populations in lowland areas and may explain the greater similarity of the east Kimberly freshwater fish fauna with lowlands to the east than with the more rugged regions to the west. Similarly, greater connectivity between lowland populations may account for the on-average larger distribution of lowland Melanotaeniids.     

Systematics of the Southeast Asian mongooses (Herpestidae,Carnivora): solving the mystery of the elusive collared mongoose and Palawan mongoose

Although recent molecular studies have clarified the phylogeny of mongooses, the systematics of the Southeast Asian species was incomplete as the collared mongoose Urva semitorquata and some debatable taxa (Hose's mongoose, Palawan mongoose) were missing in the analyses. We sequenced three mitochondrial (cytochrome b, ND2, control region) and one nuclear (beta‐fibrinogen intron 7) fragments of the Southeast Asian mongooses to clarify the systematic position of the different species and populations occurring in this region. Our results showed that the collared mongoose is closely related to the crab‐eating mongoose Urva urva, these two species forming a sister‐group to the short‐tailed mongoose Urva brachyura. Despite Sumatran collared mongooses having a peculiar orange phenotype, we showed that they exhibited very little genetic divergence to individuals from Borneo. In contrast, the populations of the short‐tailed mongoose from Borneo were strongly divergent to those from Peninsular Malaysia and Sumatra, and these might represent separate species. Within the crab‐eating mongoose, we observed little geographical genetic structure. Our study suggests that Hose's mongoose is not a valid species. The Palawan mongooses did not cluster with the other populations of the short‐tailed mongoose; they were closer to the collared mongoose and should be included in this species. © 2014 The Linnean Society of London     

Hoya peninsularis (Apocynaceae,Asclepiadoideae), a new species from Peninsular Malaysia,and notes on Hoya maingayi and Gongronema wrayi

A new species of Hoya from the western part of Peninsular Malaysia is described and illustrated as H. peninsularis. It has long been confused with Hoya finlaysonii because both have leaves with prominent darker venation, but can be distinguished on the corona lobe size and corolla pubescence. In addition, Hoya maingayi, a species earlier considered ‘doubtful and excluded' is typified and illustrated for the first time, and its distribution is shown to extend to Thailand. Gongronema wrayi is recognised as an earlier name and a new synonym for Hoya omlorii, a species previously considered to occur only in Borneo and Sumatra. The name is typified. An updated checklist of Hoya of Peninsular Malaysia is provided.     

A synopsis of Cyathocalyx species (Annonaceae) in Peninsular Malaysia, Sumatra, and Borneo, with descriptions of two new species

A revised classification of the species of Cyathocalyx (Annonaceae) occurring in western Malesia (Peninsular Malaysia, Sumatra, and Borneo) is presented, with distribution maps. Fifteen species are recognized, including two that are newly described from Borneo ( C. angustipetalus and C. magnifructus ), which are illustrated. Several important nomenclatural changes are validated, including the elevation of Drepananthus carinatus var. deltoideus to specific rank as C. deltoideus , and the transfer of the name Parartabotrys hexagynus to Cyathocalyx as C. hexagynus . Other notable nomenclatural changes include the reduction of the name C. scortechinii to synonymy with C. pubescens .  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 513–532.     

Phylogenetic relationships between isolated populations of the limestone‐dwelling microsnail Gyliotrachela hungerfordiana (Gastropoda: Vertiginidae)

The vertiginid species Gyliotrachela hungerfordiana, an obligate limestone‐dwelling microsnail, is relatively widespread and is found on a large number of isolated limestone hills in Peninsular Malaysia. To elucidate the pattern of colonization of these hills, we conducted a molecular phylogenetic analysis on G. hungerfordiana subpopulations from 15 separate limestone outcrops. As outgroups, we also included five related Peninsular Malaysian Vertiginidae (Gyliotrachela frequens, Gyliotrachela transitans, Gyliotrachela salpinx, Gyliotrachela depressispira and Paraboysidia tarutao), one population each. A combined analysis of nuclear (internal transcribed spacer 1) and mitochondrial (cytochrome c oxidase 1) sequences showed that (1) G. hungerfordiana is monophyletic; (2) there is a clear geographical pattern in the phylogenetic relationships between G. hungerfordiana populations, with genetic distances increasing with geographic distance; (3) this pattern is disturbed by a few long‐distance (north‐west to south‐east and north to south) colonizations.