Y‐chromosome and mtDNA variation confirms independent domestications and directional hybridization in South American camelids

Investigations of genetic diversity and domestication in South American camelids (SAC) have relied on autosomal microsatellite and maternally‐inherited mitochondrial data. We present the first integrated analysis of domestic and wild SAC combining male and female sex‐specific markers (male specific Y‐chromosome and female‐specific mtDNA sequence variation) to assess: (i) hypotheses about the origin of domestic camelids, (ii) directionality of introgression among domestic and/or wild taxa as evidence of hybridization and (iii) currently recognized subspecies patterns. Three male‐specific Y‐chromosome markers and control region sequences of mitochondrial DNA are studied here. Although no sequence variation was found in SRY and ZFY, there were seven variable sites in DBY generating five haplotypes on the Y‐chromosome. The haplotype network showed clear separation between haplogroups of guanaco–llama and vicuña–alpaca, indicating two genetically distinct patrilineages with near absence of shared haplotypes between guanacos and vicuñas. Although we document some examples of directional hybridization, the patterns strongly support the hypothesis that llama (Lama glama) is derived from guanaco (Lama guanicoe) and the alpaca (Vicugna pacos) from vicuña (Vicugna vicugna). Within male guanacos we identified a haplogroup formed by three haplotypes with different geographical distributions, the northernmost of which (Peru and northern Chile) was also observed in llamas, supporting the commonly held hypothesis that llamas were domesticated from the northernmost populations of guanacos (L. g. cacilensis). Southern guanacos shared the other two haplotypes. A second haplogroup, consisting of two haplotypes, was mostly present in vicuñas and alpacas. However, Y‐chromosome variation did not distinguish the two subspecies of vicuñas.     

Camelid Ig V genes reveal significant human homology not seen in therapeutic target genes,providing for a powerful therapeutic antibody platform

Camelid immunoglobulin variable (IGV) regions were found homologous to their human counterparts; however, the germline V repertoires of camelid heavy and light chains are still incomplete and their therapeutic potential is only beginning to be appreciated. We therefore leveraged the publicly available HTG and WGS databases of Lama pacos and Camelus ferus to retrieve the germline repertoire of V genes using human IGV genes as reference. In addition, we amplified IGKV and IGLV genes to uncover the V germline repertoire of Lama glama and sequenced BAC clones covering part of the Lama pacos IGK and IGL loci. Our in silico analysis showed that camelid counterparts of all human IGKV and IGLV families and most IGHV families could be identified, based on canonical structure and sequence homology. Interestingly, this sequence homology seemed largely restricted to the Ig V genes and was far less apparent in other genes: 6 therapeutically relevant target genes differed significantly from their human orthologs. This contributed to efficient immunization of llamas with the human proteins CD70, MET, interleukin (IL)-1β and IL-6, resulting in large panels of functional antibodies. The in silico predicted human-homologous canonical folds of camelid-derived antibodies were confirmed by X-ray crystallography solving the structure of 2 selected camelid anti-CD70 and anti-MET antibodies. These antibodies showed identical fold combinations as found in the corresponding human germline V families, yielding binding site structures closely similar to those occurring in human antibodies. In conclusion, our results indicate that active immunization of camelids can be a powerful therapeutic antibody platform.     

Prevalence of Eimeria spp. (Apicomplexa: Eimeriidae) in Oregon Llamas1

Eimeria alpacae, E. punoensis, E. lamae, and E. macusaniensis were identified in fecal samples from 189 llama (Lama glama (L.)) adults and 50 llama crias (animals less than one year of age of any species in the genus Lama) from central and western Oregon. In both adults and crias, E. alpacae was the most common species found. The least common was E. macusaniensis, which was found in only two adults. Overall prevalence and numbers of animals with mixed infections was approximately twice as high in crias as in adults.     

Speciation and genetic divergence of three species of charr from ancient Lake El'gygytgyn (Chukotka) and their phylogenetic relationships with other representatives of the genus Salvelinus

The diversity of phenotypically different and often reproductively isolated lacustrine forms of charrs of the genus Salvelinus represents a substantial problem for taxonomists and evolutionary biologists. Based on the analysis of variability of ten microsatellite loci and two fragments of mitochondrial DNA (control region and cyt‐b gene), the evolutionary history of three charr species from Lake El'gygytgyn was reconstructed, and phylogenetic relationships between the main representatives of the genus were revealed. Three species from Lake El'gygytgyn were strongly reproductively isolated. Long‐finned charr described previously as Salvethymus svetovidovi, an ancient endemic form in the lake, originated 3.5 Mya (95% Bayesian credible intervals: 1.7, 6.1). Placement of this species in the phylogenetic tree of Salvelinus was not determined strictly, but it should be located in the basal part of the clade Salvelinus alpinus – S. malma species complex. The origin of small‐mouth charr S. elgyticus and Boganida charr S. boganidae in Lake El'gygytgyn was related to allopatric speciation. Their ancestors were represented by two glacial lineages of Taranets charr S. alpinus taranetzi from Asia. In Lake El'gygytgyn, these lineages entered into secondary contact postglacially. A revision of the main phylogenetic groups within the Salvelinus alpinus – S. malma complex is conducted. The Boganida charrs from Lakes El'gygytgyn and Lama (Taimyr) belong to different phylogenetic groups of Arctic charr and should not be regarded as a single species S. boganidae. Using the charrs from Lakes El'gygytgyn and Lama as a case study, we show that a model of sympatric speciation, which seemed more probable based on previous empirical evidence, was rejected by other data.     

Arthropod Diversity in Lama Forest Reserve (South Benin), a Mosaic of Natural, Degraded and Plantation Forests

Arthropod assemblages were examined in Lama forest reserve, a protected area situated in the Dahomey gap, southern Benin, composed of plantations, degraded forest and remnants of natural forest. The objectives were to compare assemblages in relation to forest type and use, to elucidate the value of forest plantations for biodiversity conservation and to identify indicator species for specific forest habitats. Arthropods were collected over an 11-month period, using standardized sets of traps (pitfall, emergence, Malaise and flight intercept traps). Nine different habitats were studied, including natural and degraded forest, forest plantations (Tectona grandis and Senna siamea) of different age, and isolated forest fragments. Our analysis focused on detritivorous and xylophagous arthropods but also included ground beetles and heteropterans, totalling 393 species. We found no differences in species richness among natural and degraded forest habitats in the centre of the reserve (Noyau central). Outside of the Noyau central, species richness was highest in old teak plantations and isolated forest fragments and lowest in young teak and fuelwood plantations. Detrended correspondence analysis (DCA) separated three main groups: (1) natural forest, (2) degraded forest and young plantations, and (3) old plantations and isolated forest fragments. Multiple regression of DCA scores of the first two axes on environmental variables identified one natural and three disturbance-related predictors of arthropod assemblages in Lama forest: soil type (texture), canopy height, naturalness (proportion of Guineo-Congolian plant species) and understorey vegetation cover. We identified 15 indicator species for six different forest habitats. The highest numbers were found in abandoned settlements and old teak plantations. β-diversity was similar among the three DCA ordination groups (degraded forest excluded). Values for β-diversity were relatively high, suggesting that all major forest habitats contribute significantly to regional species pools and should therefore be protected. To enhance arthropod diversity, we propose that management practices in Lama forest should aim to encourage the development of species-rich understorey vegetation of the Guineo-Congolian phytogeographical region.     

Die Halsäste des Nervus vagus beim Alpaka,Lama guanicoe pacos (Linné 1758)

Demmel, U. 1980. Die Halsäste des Nervus vagus beim Alpaka, Lama guanicoe pacos (Linné 1758). [The branches of the vagus nerve in the neck of Alpaka, Lama guanicoe pacos (Linné 1758).] (Anatomisches Institut der Universität Köln, Bundesrepublik Deutschland.) — Acta zool. (Stockh.) 61(3): 141–146. Dissections of two newborn and two adult alpacas have shown: that the trapezius musculature of either side is supplied by branches of seven cervical spinal nerves and not by the external ramus of truncus nervi accessorii as in mammals in general; that the nerves which innervate the inner laryngic muscles as well as the mucous membrane of the lower part of the larynx are given off from the main trunk of the vagus nerve just outside the jugular foramen; that a nerve corresponding to the n. laryngeus recurrens of other mammals is absent. It is concluded that the lack of recurvation of the inferior laryngic nerve and the absence of the spinal part of the accessory nerve are character states of tylopod artiodactyls.     

Clinico-pathological findings and cerebrospinal fluid analysis in llamas (<Emphasis Type="Italic">Lama glama</Emphasis>) experimentally infected with the meningeal worm <Emphasis Type="Italic">Parelaphostrongylus tenuis</Emphasis>

In this study, the clinical progression of experimental infection of llamas ( Lama glama) with Parelaphostrongylus tenuis is described. The onset and characterization of neurologic deficits and changes in the hematology, serum biochemical, and cerebrospinal fluid (CSF) in affected llamas are compared before and after appearance of clinical signs. Sixteen apparently healthy, male llamas (median age; 12 months) were used. Five llamas were maintained as uninfected controls, and five, three, and three llamas were orally inoculated with five, ten, and 25 third-stage larvae (L3) of P. tenuis, respectively. The onset, degree, and characterization of neurologic deficits were recorded. Hematology, serum biochemistry, and CSF were analyzed before exposure (Pre) and after neurologic signs appeared (Post). Necropsy and worm recovery from the central nervous system (CNS) were also carried out to confirm infection. Exposure of llamas to 25 L3 of P. tenuis orally resulted in consistent infection and development of disease. Overall, ten of the 11 animals exposed to P. tenuis L3 developed varying degrees of neurologic signs between 45 and 82 days post-exposure (dpe). P. tenuis was recovered from the CNS of seven of the 11 exposed llamas. Neurologic signs progressed from ataxia to paresis, and eventually paralysis. In the hematologic and serum biochemical analyses, only eosinophil count was significantly elevated in the post-exposure samples in the infected groups. Protein was significantly elevated and Pandy test was positive in CSF of infected llamas. This study comprehensively describes the clinical progression of P. tenuis infection in llamas that will prove helpful in future studies to investigate the possibilities of developing antemortem diagnostic tests and new therapeutic programs.     

Diversity of xylophagous beetles and vulnerability of debarked-medicinal plants in southern of Benin

We studied the impact of debarking of medicinal tree species on the diversity of xylophagous beetles in Lama Forest Reserve, in Lokoli swampy forest and in crop fields surrounded Lokoli forest. A total of 108 interception traps were set up on debarked and nondebarked trees covering nine medicinal plants that are Anogeissus leiocarpa, Dialium guineense, Khaya senegalensis in Lama Forest; Nauclea diderrichii, Ficus trichopoda, Syzygium owariense in swampy forest; and Parkia biglobosa, Bridelia ferruginea, Pterocarpus erinaceus in crop fields. A total of 116 beetle species were collected belonging to 19 coleoptera families with higher xylophagous beetles than predators. Specific richness and individual abundance of xylophagous beetles were significantly higher in crop fields than in forests. Furthermore, in all habitats, debarked-tree species were more attractive to xylophagous beetles than control trees and were significantly more attacked in crop fields than forests. The most vulnerable medicinal trees to debarking were P. biglobosa, D. guineense, F. trichopoda and P. erinaceus. Three groups of indicator insects according to habitat type, debarking and tree species have been distinguished. Our results clearly imply that by exploiting natural resources humans can impact on the abundance and specific richness of xylophagous beetles by modulating their resources.     

Eimeria macusaniensis n. sp. (Protozoa: Eimeriidae) of the Alpaca Lama pacos

SYNOPSIS. A new species of coccidium, Eimeria macusaniensis n. sp., is described from the alpaca Lama pacos in Macusani, Peru. The oocysts are ovoid to piriform, 81–107 by 61–80 μ (mean 93.6 by 67.4 μ) with a micropylar cap and elongate ovoid sporocysts 33–40 by 16–20 μ (mean 36.3 by 18.3 μ).     

Patterns of development of the coldwater lacustrine-riverine form of the Drjagin charr (genus <Emphasis Type="Italic">Salvelinus</Emphasis>) from Lake Lama (the Taimyr Peninsula) during the larval period

Reproduction of the lacustrine-riverine form of the Drjagin charr (genus Salvelinus) from Lake Lama (the Noril’sk-Pyasina aquatic system on the Taimyr Peninsula) in a mountain tributary, the Bunisyak River, as well as the formation and development of skeletal elements in embryos and larvae reared under different experimental conditions are described. Patterns of the development and the heterochrony between the rates of somatic growth and osteogenesis are an adaptation to the extremely low water temperature and oligotrophic spawning habitat of the mountain stream. Still, the formation of vertebral centrae is retarded even at rather high rearing temperatures. As compared to larvae of the small normal (nondwarf) planktivorous lacustrine form of Arctic charr from Lake Davatchan (Transbaikalia) reared at 3°C, larvae of Drjagin charr from Lake Lama reared at 1°C are characterized by accelerated growth and heterochronies of osteogenesis. The divergence of the lacustrine-riverine form of Drjagin charr from the generalized form of Salvelinus alpinus in morphology begins during the larval period of the ontogeny.     

Issue Information

ON THE COVER: Overlooked by Andean volcanos, the Bofedeles de Parinacota is an ecologically important wetland in the Chilean altiplano, where Nostoc sheets bearing diverse epimicrobiota occur in pools surrounded by bird flocks and grazing herds of camelids such as guanaco (Lama guanaco), llama (Lama glama), vicuna (Vicugna vicugna), and alpaca (Vicugna pacos).

     

Lama1 mutations lead to vitreoretinal blood vessel formation, persistence of fetal vasculature, and epiretinal membrane formation in mice

Background

Valuable insights into the complex process of retinal vascular development can be gained using models with abnormal retinal vasculature. Two such models are the recently described mouse lines with mutations in Lama1, an important component of the retinal internal limiting membrane (ILM). These mutants have a persistence of the fetal vasculature of vitreous (FVV) but lack a primary retinal vascular plexus. The present study provides a detailed analysis of astrocyte and vascular development in these Lama1 mutants.

Results

Although astrocytes and blood vessels initially migrate into Lama1 mutant retinas, both traverse the peripapillary ILM into the vitreous by P3. Once in the vitreous, blood vessels anastomose with vessels of the vasa hyaloidea propria, part of the FVV, and eventually re-enter the retina where they dive to form the inner and outer retinal capillary networks. Astrocytes continue proliferating within the vitreous to form a dense mesh that resembles epiretinal membranes associated with persistent fetal vasculature and proliferative vitreoretinopathy.

Conclusions

Lama1 and a fully intact ILM are required for normal retinal vascular development. Mutations in Lama1 allow developing retinal vessels to enter the vitreous where they anastomose with vessels of the hyaloid system which persist and expand. Together, these vessels branch into the retina to form fairly normal inner retinal vascular capillary plexi. The Lama1 mutants described in this report are potential models for studying the human conditions persistent fetal vasculature and proliferative vitreoretinopathy.     

Functional role of laminin α1 chain during cerebellum development

We had developed a conditional Laminin α1 knockout-mouse model (Lama1^cko) bypassing embryonic lethality of Lama1 deficient mice to study the role of this crucial laminin chain during late developmental phases and organogenesis. Here, we report a strong defect in the organization of the adult cerebellum of Lama1^cko mice. Our study of the postnatal cerebellum of Lama1^cko animals revealed a disrupted basement membrane correlated with an unexpected excessive proliferation of granule cell precursors in the external granular layer (EGL). This was counteracted by a massive cell death occurring between the postnatal day 7 (P7) and day 20 (P20) resulting in a net balance of less cells and a smaller cerebellum. Our data show that the absence of Lama1 has an impact on the Bergmann glia scaffold that aberrantly develops. This phenotype is presumably responsible for the observed misplacing of granule cells that may explain the overall perturbation of the layering of the cerebellum and an aberrant folia formation.Key words: cerebellum, development, laminin, cell migration, laminin-111     

Mutations in Lama1 Disrupt Retinal Vascular Development and Inner Limiting Membrane Formation

The Neuromutagenesis Facility at the Jackson Laboratory generated a mouse model of retinal vasculopathy, nmf223, which is characterized clinically by vitreal fibroplasia and vessel tortuosity. nmf223 homozygotes also have reduced electroretinogram responses, which are coupled histologically with a thinning of the inner nuclear layer. The nmf223 locus was mapped to chromosome 17, and a missense mutation was identified in Lama1 that leads to the substitution of cysteine for a tyrosine at amino acid 265 of laminin α1, a basement membrane protein. Despite normal localization of laminin α1 and other components of the inner limiting membrane, a reduced integrity of this structure was suggested by ectopic cells and blood vessels within the vitreous. Immunohistochemical characterization of nmf223 homozygous retinas demonstrated the abnormal migration of retinal astrocytes into the vitreous along with the persistence of hyaloid vasculature. The Y265C mutation significantly reduced laminin N-terminal domain (LN) interactions in a bacterial two-hybrid system. Therefore, this mutation could affect interactions between laminin α1 and other laminin chains. To expand upon these findings, a Lama1 null mutant, Lama1^tm1.1Olf, was generated that exhibits a similar but more severe retinal phenotype than that seen in nmf223 homozygotes. The increased severity of the Lama1 null mutant phenotype is probably due to the complete loss of the inner limiting membrane in these mice. This first report of viable Lama1 mouse mutants emphasizes the importance of this gene in retinal development. The data presented herein suggest that hypomorphic mutations in human LAMA1 could lead to retinal disease.     

Forest degradation and invasive species synergistically impact Mimusops andongensis (Sapotaceae) in Lama Forest Reserve,Benin

Harvesting of Non‐Timber Forest Products (NTFPs) can threaten target species, especially those with limited distribution and density. Exploited species also face threats from habitat fragmentation, fire, and invasive species. We assessed the impact of human disturbances and invasive species on the population of a key multipurpose NTFP species, Mimusops andongensis, in Lama Forest reserve (Benin). The densities of adult trees and regenerative stems decreased with increasing degradation. Mimusops andongensis contributed less to total tree density with increasing human disturbance. There were significantly fewer M. andongensis recruits with increasing cover of invasive Chromolaena odorata. Smaller diameter individuals predominated in non‐degraded and moderately degraded sites while in degraded sites, the structure showed a negative exponential trend with the density of small diameter individuals being less than two trees/ha. Larger individuals were also rare in degraded sites. The low density of both mature trees and seedlings in degraded sites may undermine the long‐term viability of M. andongensis, despite existing protection against NTFP harvesting and other anthropogenic pressures. Management should emphasize facilitating recruitment subsidies and limiting the presence of C. odorata.     

Prevalence of Eimeria macusaniensis (Apicomplexa: Eimeriidae) in midwestern Lama spp

To compare the prevalence of Eimeria macusaniensis among midwestern llamas (Lama glama), alpacas (Lama pacos), and guanacos (Lama guanicoe), feces were obtained from Lama spp. in 10 states between October 1989 and February 1996. Feces were examined by centrifugal flotation in sugar solution (specific gravity--1.28-1.30), and oocysts were quantified by a modified McMaster method. Data were compared by host species and age classifications. Typical oocysts occurred in samples from 28% of 76 herds and 10.4% of 443 animals including 12% of 301 llamas, 7% of 115 alpacas, and 7.4% of 27 guanacos. Prevalence was significantly greater (P = 0.009) in animals < 1 yr of age in comparison to older animals for llams (22.1 v.s. 8.5%) and for all Lama spp. combined (17.1 vs. 8.4%). Fecal oocyst abundance was significantly greater (P = 0.001) in llamas < 1 yr of age in comparison to older llamas (30 vs. 16 oocysts per g of feces). Fecal oocyst intensities did not differ significantly. Prevalence in both age groups of midwestern llamas was greater than previously reported for llamas in the western United States. Prevalence in midwestern alpacas < 1 yr of age was lower than reported for alpacas of similar age in South America, but oocyst intensities were similar. These results indicate that infection with E. macusaniensis is more common in Lama spp. in North America than previously recognized.     

Location of the 9257 and ataxia mutations on mouse Chromosome 18

The location of three mutations on proximal Chromosome (Chr) 18 was determined by analysis of the offspring of several backcrosses. The results demonstrate that ataxia and the insertional mutation TgN9257Mm are separated by less than 1 cM and are located approximately 3 cM from the centromere, while the balding locus is 7 cM more distal. Previous data demonstrated that the twirler locus also maps within 1 cM of ataxia. The corrected locations will contribute to identification of appropriate candidate genes for these mutations. Two polymorphic microsatellite markers for proximal Chr 18 are described, D18Umi1 and D18Umi2. The Lama3 locus encoding the α3 subunit of nicein was mapped distal to ataxia and did not recombine with Tg9257. Received: 19 December 1995 / Accepted: 29 January 1996     

The influence of the arid Andean high plateau on the phylogeography and population genetics of guanaco (Lama guanicoe) in South America

A comprehensive study of the phylogeography and population genetics of the largest wild artiodactyl in the arid and cold‐temperate South American environments, the guanaco (Lama guanicoe) was conducted. Patterns of molecular genetic structure were described using 514 bp of mtDNA sequence and 14 biparentally inherited microsatellite markers from 314 samples. These individuals originated from 17 localities throughout the current distribution across Peru, Bolivia, Argentina and Chile. This confirmed well‐defined genetic differentiation and subspecies designation of populations geographically separated to the northwest (L. g. cacsilensis) and southeast (L. g. guanicoe) of the central Andes plateau. However, these populations are not completely isolated, as shown by admixture prevalent throughout a limited contact zone, and a strong signal of expansion from north to south in the beginning of the Holocene. Microsatellite analyses differentiated three northwestern and 4–5 southeastern populations, suggesting patterns of genetic contact among these populations. Possible genetic refuges were identified, as were source‐sink patterns of gene flow at historical and recent time scales. Conservation and management of guanaco should be implemented with an understanding of these local population dynamics while also considering the preservation of broader adaptive variation and evolutionary processes.