On the PhyloQuant protein expression profiles approach to the taxonomic problem.

Inferring evolutionary relationships among organisms has been a fundamental problem in evolutionary biology. The current phylogenetic molecular methods serve as the baseline model to test new evolutionary hypotheses with taxonomic purposes. Leishmaniinae trypanosomatids subfamily includes protozoan parasites of clinical importance to humans. They have an intricate taxonomic history defined by morphological elements, host range, and molecular phylogenies. Unraveling the increasing diversity of this group has shown limitations in reconstructing the true evolutionary relationships among Trypanosomatidae species. Toward the goal of inferring evolutionary relationships that help to resolve phylogenetic and taxonomic controversies among parasites of the subfamily Leishmaniinae, Mule et al. propose the method PhyloQuant as a valuable approach, based on differential protein expression obtained from high throughput mass spectrometry data. Employing a pioneering methodological approach, Mule et al. assess the taxonomic problem for species hypothesis within Leishmaniinae, from quantitative phenetic protein expression profiles, in contrast to the standard multilocus phylogenetic approaches.

Neutral evolution of human enamel-dentine junction morphology.

Teeth have been studied for decades and continue to reveal information relevant to human evolution. Studies have shown that many traits of the outer enamel surface evolve neutrally and can be used to infer human population structure. However, many of these traits are unavailable in archaeological and fossil individuals due to processes of wear and taphonomy. Enamel-dentine junction (EDJ) morphology, the shape of the junction between the enamel and the dentine within a tooth, captures important information about tooth development and vertebrate evolution and is informative because it is subject to less wear and thus preserves more anatomy in worn or damaged specimens, particularly in mammals with relatively thick enamel like hominids. This study looks at the molar EDJ across a large sample of human populations. We assessed EDJ morphological variation in a sample of late Holocene modern humans (n = 161) from archaeological populations using µ-CT biomedical imaging and geometric morphometric analyses. Global variation in human EDJ morphology was compared to the statistical expectations of neutral evolution and "Out of Africa" dispersal modeling of trait evolution. Significant correlations between phenetic variation and neutral genetic variation indicate that EDJ morphology has evolved neutrally in humans. While EDJ morphology reflects population history, its global distribution does not follow expectations of the Out of Africa dispersal model. This study increases our knowledge of human dental variation and contributes to our understanding of dental development more broadly, with important applications to the investigation of population history and human genetic structure.

Who were the Nataruk people? Mandibular morphology among late Pleistocene and early Holocene fisher-forager populations of West Turkana (Kenya).

Africa is the birthplace of the species Homo sapiens, and Africans today are genetically more diverse than other populations of the world. However, the processes that underpinned the evolution of African populations remain largely obscure. Only a handful of late Pleistocene African fossils (∼50-12 Ka) are known, while the more numerous sites with human fossils of early Holocene age are patchily distributed. In particular, late Pleistocene and early Holocene human diversity in Eastern Africa remains little studied, precluding any analysis of the potential factors that shaped human diversity in the region, and more broadly throughout the continent. These periods include the Last Glacial Maximum (LGM), a moment of extreme aridity in Africa that caused the fragmentation of population ranges and localised extinctions, as well as the 'African Humid Period', a moment of abrupt climate change and enhanced connectivity throughout Africa. East Africa, with its range of environments, may have acted as a refugium during the LGM, and may have played a critical biogeographic role during the heterogene`ous environmental recovery that followed. This environmental context raises a number of questions about the relationships among early Holocene African populations, and about the role played by East Africa in shaping late hunter-gatherer biological diversity. Here, we describe eight mandibles from Nataruk, an early Holocene site (∼10 Ka) in West Turkana, offering the opportunity of exploring population diversity in Africa at the height of the 'African Humid Period'. We use 3D geometric morphometric techniques to analyze the phenotypic variation of a large mandibular sample. Our results show that (i) the Nataruk mandibles are most similar to other African hunter-fisher-gatherer populations, especially to the fossils from Lothagam, another West Turkana locality, and to other early Holocene fossils from the Central Rift Valley (Kenya); and (ii) a phylogenetic connection may have existed between these Eastern African populations and some Nile Valley and Maghrebian groups, who lived at a time when a Green Sahara may have allowed substantial contact, and potential gene flow, across a vast expanse of Northern and Eastern Africa.

Dental size variability in Central African Pygmy hunter-gatherers and Bantu-speaking farmers.

OBJECTIVES: Odontometric studies of African populations show high within-group variation in tooth size. Overall, North Africans exhibit smaller dimensions than groups from eastern and southern sub-Saharan regions, but no previous studies have analyzed the full dental metrics among extant African Pygmy hunter-gatherers and Bantu-speaking farmers. Furthermore, the population variability in tooth crown sizes from equatorial rainforest regions remains to be elucidated. MATERIALS AND METHODS: The mesiodistal and buccolingual diameters of the permanent teeth (I1-M2) were measured in vivo using high-resolution replicas from Baka Pygmies and Mvae and Yassa Bantu-speakers from Cameroon (western Africa). Analyses of variance were used to record sex-related and population-level differences in tooth sizes, and a principal component analysis of geometrically scaled measures was used to plot the odontometric variability among groups. RESULTS: Cameroonian Baka Pygmies differ in dental size from their Bantu-speaking neighbors. Molar teeth are larger in Pygmies than in Bantu individuals, while the anterior dentition is larger in the Bantu. Baka males exhibit significantly larger teeth than females, whereas sexual dimorphism in non-Pygmies is only present in the anterior dentition. DISCUSSION: Odontometric patterns and the degree of sexual dimorphism in dental size differ among Central African groups, indicating adaptation to their different forager and farmer lifestyles. In particular, the admixture of Bantu-speakers in Baka populations is smaller than that in other western Pygmy groups. The greater dental phenetic diversity in Baka compared to that of the smaller-toothed farmers suggests that ecogenetic and microevolutionary factors are influencing a particular divergence scenario.

The apportionment of tooth size and its implications in Australopithecus sediba versus other Plio-pleistocene and recent African hominins.

OBJECTIVES: Australopithecus sediba is characterized further by providing formerly unpublished and refined mesiodistal and buccolingual crown measurements in the MH1 and MH2 specimens. After size correction, these data were compared with those in other fossil and recent samples to facilitate additional insight into diachronic hominin affinities. MATERIALS AND METHODS: Six comparative samples consist of fossil species: A. africanus, A. afarensis, Homo habilis, Paranthropus robustus, P. boisei, and H. erectus. Others comprise H. sapiens and Pan troglodytes. Re-estimates of "actual" dimensions in damaged A. sediba teeth were effected through repeated measurements by independent observers. X-ray synchrotron microtomography allowed measurement of crowns obscured by matrix and noneruption. Tooth size apportionment analysis, an established technique for intraspecific comparisons, was then applied at this interspecific level to assess phenetic affinities using both within- and among-group data. RESULTS: Comparison of these highly heritable dimensions identified a general trend for smaller posterior relative to larger anterior teeth (not including canines), contra Paranthropus, that allies A. sediba with other australopiths and Homo; however, specific reductions and/or shape variation in the species' canines, third premolars, and anterior molars relative to the other teeth mirror the patterning characteristic of Homo. DISCUSSSION: Of all samples, including east African australopiths, A. sediba appears most like H. habilis, H. erectus and H. sapiens regarding how crown size is apportioned along the tooth rows. These findings parallel those in prior studies of dental and other skeletal data, including several that suggest A. sediba is a close relative of, if not ancestral to, Homo.

Who were they really? model-free and model-bound dental nonmetric analyses to affirm documented population affiliations of seven South African "Bantu" samples.

OBJECTIVES: For bioarchaeological biodistance analyses it is common to "assume" that skeletal samples are representative of the populations to which they are attributed. Here, alternatively, samples with "known" attribution in the Raymond A. Dart Collection are assessed regarding their suitability for use in such analyses. Prior curation issues may call their ascribed identities into question. MATERIALS AND METHODS: These 20th century samples ostensibly derive from South African Ndebele, Sotho, Swazi, Tswana, Venda, Xosa, and Zulu populations. First, the mean measure of divergence (MMD) is used to obtain among-sample dental phenetic distances for comparison with documented population relationships. Second, the Mantel test evaluates fit of the isolation-by-distance model between MMD and geographic distances, i.e., among the historic homelands. Third, R-matrices and minimum and estimated Fst from MMD distances give an indication of genetic micro-differentiation. RESULTS: Output from these model-free and model-bound analyses suggest that five and perhaps six samples are representative of their attributed populations-presenting differences along population lines and evidence of more ancient ancestry. DISCUSSION: Other than the Swazi and perhaps Nedebele, the among-sample variation: 1) mirrors documented population history, 2) reveals a moderately positive correlation between phenetic and geographic distances, and 3) although evidencing much homogeneity, provides measures of genetic distance in support of the phenetic distances. Therefore, with the two noted exceptions-perhaps from collection issues, swamping of past genetic structure, or both, most samples appear suitable for bioarchaeological analyses. On this basis, results are offered to supplement published findings concerning the biological relationships of these peoples.

Questions of Khoesan continuity: dental affinities among the indigenous Holocene peoples of South Africa.

The present report follows up on the findings of previous research, including recent bioarchaeological study of well-dated Khoesan skeletal remains, that posits long term biological continuity among the indigenous peoples of South Africa after the Pleistocene. The Arizona State University Dental Anthropology System was used to record key crown, root, and intraoral osseous nonmetric traits in six early-through-late Holocene samples from the Cape coasts. Based on these data, phenetic affinities and an identification of traits most important in driving intersample variation were determined using principal components analysis and the mean measure of divergence distance statistic. To expand biological affinity comparisons into more recent times, and thus preliminarily assess the dental impact of disproportionate non-Khoesan gene flow into local peoples, dental data from historic Khoekhoe and San were also included. Results from the prehistoric comparisons are supportive of population continuity, though a sample from Matjes River Rockshelter exhibits slight phenetic distance from other early samples. This and some insignificant regional divergence among these coastal samples may be related to environmental and cultural factors that drove low-level reproductive isolation. Finally, a close affinity of historic San to all samples, and a significant difference of Khoekhoe from most early samples is reflective of documented population history following immigration of Bantu-speakers and, later, Europeans into South Africa.

Population affinities in pre-colonial West Africa: The case of the burial cave Iroungou (Gabon, 14th-15th century CE).

INTRODUCTION: Our knowledge of the populations of sub-Saharan Africa in the periods before European colonization is limited. Few archeological sites containing human remains have been identified, and written sources for these periods are rare. The discovery in 2018 of the Iroungou sepulchral cave (Gabon), whose use predates the arrival of the Portuguese (14th-15th centuries CE), is an exceptional source of information: at least 28 individuals associated with numerous metal artifacts were found there. The anthropobiological remains were left in situ, but the eight best preserved crania were digitized. OBJECTIVES: This study focuses on the population affinities of these crania, whose morphology was described using 237 landmarks. MATERIALS AND METHODS: Geometric morphometric analyses were used to compare the eight Iroungou specimens with 154 individuals representing 12 well-defined African populations. After alignment (Generalized Procrustes Analysis), morphological affinity was assessed using Euclidean and Mahalanobis distances, and posterior probabilities of population membership (discriminant analysis). RESULTS: Results indicate that the eight Iroungou crania have, on average, more affinity with Bayaka Pygmy, followed by Central African Bantu. Nevertheless, individually, the Iroungou specimens show an important morphological variation and the eight crania can be separated into different affinity groups: Bayaka and Central African Bantu, KhoeSan, and East-African Bantu. Finally, one individual presents strong affinity with Somalis. CONCLUSION: This phenetic mapping of the Iroungou sample raises questions about the profile of the individuals deposited in the cave in a geographical area known for the Loango pre-colonial kingdom, which ruling class seemed to have had privileged relationships with the Pygmy populations.

INTRODUCTION: Nos connaissances sur les populations d'Afrique sub­saharienne des périodes précédant la colonisation européenne sont limitées. Peu de sites archéologiques contenant des restes humains ont été identifiés, et les sources écrites pour ces périodes sont rares. La découverte en 2018 de la grotte sépulcrale d'Iroungou (Gabon), dont l'utilisation remonte au XIVe­XVe siècles de notre ère, avant l'arrivée des Portugais, constitue une source d'information exceptionnelle: au moins 28 individus associés à de nombreux artefacts métalliques y ont été retrouvés. Les restes anthropobiologiques ont été laissés in situ, néanmoins, les huit crânes les mieux préservés ont été numérisés. OBJECTIFS: Cette étude s'intéresse aux affinités populationnelles de ces crânes, dont la morphologie a été décrite à l'aide de 237 points­repères répartis sur le massif facial supérieur et le calvarium. MATÉRIEL ET MÉTHODES: Des analyses par morphométrie géométrique ont permis de comparer les spécimens d'Iroungou avec 154 individus représentants 12 populations africaines bien définies. Après alignement des conformations (analyse Procrustes généralisée), l'affinité morphologique a été évaluée à l'aide des distances procrustes, euclidienne et mahalanobis, ainsi que des probabilités postérieures d'appartenance à une population (analyse discriminante). RÉSULTATS: Les résultats indiquent que les huit crânes d'Iroungou présentent en moyenne plus d'affinités avec les populations pygmées Bayaka, suivies des Bantus d'Afrique centrale. DISCUSSION: Cette affinité moyenne recouvre une réalité complexe: la population d'Iroungou est la plus hétérogène de notre échantillon et les huit crânes peuvent être séparés en différents groupes d'affinités: avec les Bayakas et les Bantus d'Afrique Centrale, les KhoeSan, et avec les Bantus d'Afrique de l'est. Enfin, un individu présente une affinité très forte avec les Somalis de notre échantillon. Cette cartographie phénétique de la population d'Iroungou interroge sur le profil des individus déposés dans la grotte dans une zone géographique connue le royaume précolonial Loango dont la classe dirigeante semble avoir eu des relations privilégiées avec les populations pygmées.

Dental morphological variation in Chalcolithic and Bronze Age human populations from North-Eastern Romania.

BACKGROUND: This study aims to evaluate the dental variability and the phenetic relationships among archaeological human populations of the Chalcolithic and Bronze Age in North-Eastern Romania. METHODS: The evaluation of phenotypic variability was performed using the 2D Geometric Morphometrics on the second molars (upper - M2, and lower - M2) belonging to adult skeletons discovered in archaeological sites of the Chalcolithic and Bronze Age in North-Eastern Romania. RESULTS: The size (Centroid Size, CS) and shape analysis of the second molar highlighted specific characteristics for each prehistoric period. The CS doesn't achieve any statistical significance for two of the comparison criterium used (i.e., period, archaeological site). Instead, statistically significant differences were recorded between males and females. The greater variability in the shape of M2 compared to M2 was noticed. The M2 model of the Bronze Age is flattened mesiodistal compared to the Chalcolithic one, while differences in M2 morphotypes are associated with the hypocone shape, particularly. CONCLUSION: This study of dental morphology provides new data that could contribute as evidence of interactions between biological human groups in different regional prehistoric contexts.

Evaluating relationships between seed morphological traits and seed dormancy in Chenopodium quinoa Willd.

Introduction: Quinoa is a high-value, nutritious crop that performs well in variable environments, marginal soils, and in diverse crop rotations. Quinoa's many attributes make it an ideal crop for supporting human health in global communities and economies. To date, quinoa research has largely focused on traits in adult plants important for enhancing plant phenotypic plasticity, abiotic stress, disease resistance, and yield. Fewer studies have evaluated quinoa seed dormancy and suggest that most modern quinoa varieties have weak or no seed dormancy, and a narrow window of seed viability post-harvest. In other crops, diminished seed dormancy is a major risk factor for preharvest sprouting (PHS; germination on the panicle due to rain prior to harvest) and may also pose a similar risk for quinoa. Methods: This study (1) developed a dormancy screening assay to characterize seed dormancy strength in a large collection of quinoa varieties, (2) investigated if morphological variables including seed coat color, seed coat thickness, seed shape including eccentricity which evaluates the roundness or flatness of a seed, and other agronomic traits like crude protein content and seed moisture, contribute to quinoa seed dormancy, and (3) evaluated the use of a phenetic modeling approach to explore relationships between seed morphology and seed dormancy. Results: Dormancy screening indicated seed dormancy ranges in quinoa varieties from none to strong dormancy. Further, phenetic modeling approaches indicate that seed coat thickness and eccentricity are important morphological variables that impact quinoa seed dormancy strength. Conclusions: While dormancy screening and phenetic modeling approaches do not provide a direct solution to preventing PHS in quinoa, they do provide new tools for identifying dormant varieties as well as morphological variables contributing to seed dormancy.

The comparative palynomorphological studies on Turkish Muscari genera and close relatives (Asparagaceae-scilloideae) with their taxonomic implications.

By this study, the palynomorphological traits of 49 taxa (53 plant samples) of the genus Muscari which has been the main research topic and also its relatives, Pseudomuscari, Hyacinthella and Bellevalia have been studied in details of both using light and scanning electron microscope. In the end of palynologic works, it was suggested the descriptive or symbolized main pollen types to make easier the separation or identification of the species of Muscari and its relative groups. According to this symbolization, the pollen types of species may be changeable as D-shaped, elliptical, triangular, and spindle shaped and all types could be seen in different members of the studied taxonomic groups in this paper. One of the most important findings of the study is that the taxa of the genus Muscari, Hyacinthella and Bellevalia do not show a main difference in terms of pollen structures and especially they exhibit harmony. Therefore, it does not possible to mention about the special pollen shape own to one species but it is possible to see common pollen types for closely related taxa or taxon groups. As a result, it could be commented that there is not evolved the close taxonomical groups within Asparagaceae in terms of pollen shape and morphologies and there is considerably a high suitability in point of theirs germ cells or microspores among them. RESEARCH HIGHLIGHTS: A detailed palynological description is provided for Muscari and close relative genus using scanning electron microscopy (SEM) and light microscopy (LM). Pollen size and sulcus length are been determined as the best informative pollen characters to understand Muscari and other studied species.

Different Traits, Different Evolutionary Pathways: Insights from Salamandrina (Amphibia, Caudata).

Species delimitation is often based on a single or very few genetic or phenetic traits, something which leads to misinterpretations and often does not provide information about evolutionary processes. Here, we investigated the diversity pattern of multiple phenetic traits of the two extant species of Salamandrina, a genus split only after molecular traits had been studied but the two species of which are phenetically very similar. The phenetic traits we studied are size, external body shape and head colour pattern, in a model comparison framework using non-linear mixed models and unsupervised and supervised clustering. Overall, we found high levels of intra-specific variability for body size and shape, depending on population belonging and habitat, while differences between species were generally lower. The habitat the salamanders dwell in also seems important for colour pattern. Basing on our findings, from the methodological point of view, we suggest (i) to take into account the variability at population level when testing for higher level variability, and (ii) a semi-supervised learning approach to high dimensional data. We also showed that different phenotypic traits of the same organism could result from different evolutionary routes. Local adaptation is likely responsible for body size and shape variability, with selective pressures more similar across species than within them. Head colour pattern also depends on habitat, differently from ventral colour pattern (not studied in this paper) which likely evolved under genetic drift.

Phenotypic Characterization and Identification of Potential L-Asparaginase-Producing Thermohalophilic Bacteria from Wawolesea Hot Spring, North Konawe, Southeast Sulawesi, Indonesia.

<b>Background and Objective:</b> L-asparaginase-producing thermohalophilic bacteria have the potential of producing an enzyme tolerant to high heat and salt levels. This enzyme, L-asparaginase, can be used as a biological agent for the cancer therapy of acute lymphoblastic leukemia and melanosarcoma as it has a specific ability to inhibit the formation of nutrients for cancer cells. This enzyme is also used effectively in food industries operating at high temperatures due to its ability to reduce acrylamide, a trigger of cancer cells. This study sought to figure out the phenotypic characters of and identify potential L-asparaginase-producing thermohalophilic bacteria from Wawolesea Hot Spring, North Konawe, Southeast Sulawesi. <b>Materials and Methods:</b> The characterization conducted on potential L-asparaginase-producing thermohalophilic bacterial isolates consisted of the following: Colony morphological characterization, covering the shapes, edges, internal structures, elevations and colours of the colonies, cell morphological characterization, covering gram staining, endospore formation and motility, biochemical characterization, covering catalase, Methyl Red and Voges Proskauer (MR-VP), gelatin hydrolysis, citrate, indole and carbohydrate fermentation tests and physiological characterization, covering pH effect, salinity, oxygen demand and temperature effect tests. Bacterial isolate identification was carried out in two stages, namely phenetic identification based on the phenotypic characterization data determine through a preliminary identification and numeric-phenetic identification. <b>Results:</b> The characterization results showed that the bacterial isolates AAT 1.4, AAT 3.2 and CAT 3.4 were <i>bacillus</i>-shaped, Gram-positive, motile, catalase-positive and aerobic. Based on the numeric-phenetic analysis results, the isolates AAT 1.4 and CAT 3.4 had a 92.9% similarity to <i>Bacillus subtilis</i>, while isolate AAT 3.2 had a 92.9% similarity to <i>Brevibacillus limnophilus</i>. <b>Conclusion:</b> According to the numeric-phenetic analysis results, the isolates AAT 1.4 and CAT 3.4 belong to the species <i>Bacillus subtilis</i>, while isolate AAT 3.2 belongs to the species <i>Brevibacillus limnophilus</i>.

Common pattern of distribution for Mesoamerican Triatoma dimidiata suggest geological and ecological association.

The phylogeny of the Triatoma dimidiata complex has been widely assessed with different genetic and morphological data, which has allowed to reach the consensus that the complex consists of at least three taxonomic units. However, these taxonomic units seem to have a distribution related to geography throughout Mesoamerica, with different groupings depending on the source of information used. In the present study, we aimed to determine if there is a common biogeographical, genetic and phenetic distribution pattern among the T. dimidiata species in Mesoamerica and if this pattern is related to ecological and geological variability of the region. We found that panbiogeographical analysis showed three generalized tracks that coincide with genetic/phenetic data which showed a general pattern of distribution in two big clusters to the north and south of Mesoamerica. We also found that these clusters were significantly related to geological tectonic plates and ecotypes. We conclude that the geological history may be a plausible explanation for the greater differentiation observed in the T. dimidiata complex, but that the current ecological characteristics of the morphotectonic units or ecotypes may be responsible for the additional variation observed and therefore differential control strategies for each cluster considering geological history and ecotype should be used. Further, more detailed biogeographical and landscape genetic analyses are necessary with the goal to elucidate T. dimidiata differentiation related with ecological and geological variables in the region and the possible epidemiological and evolutionary consequences.

Determination of yeast diversity in fermented Sumbawa mare’s milk using internal transcribed spacers (ITS) fragment analysis

Aims@#Molecular identification of yeast has been conducted on various fermentation products. However, the identification of yeast in fermented Sumbawa mare’s milk based on the genotyping method has not been carried out. This study was aimed to determine the diversity profile of yeasts in fermented Sumbawa mare’s milk using phenetic characters and PCR-RFLP analysis technique based on the ITS region.@*Methodology and results@#Yeast isolates were phenotypically characterized and visualized in a dendrogram using CLAD97 software. Then, the yeast DNA was extracted using heat treatment and amplified using ITS1 and ITS4 primers. The amplicons were analyzed by RFLP using HindIII and HaeIII enzymes. The phylogenetic tree was constructed using MEGA 7.0. Based on the result of grouping by phenetic analysis and PCR-RFLP, the 12 isolates were divided into four groups with different members. The results of the phenetic analysis were divided into group I (all isolates of Dompu), group II (isolate B3, B4, S3), group III (isolate B5) and group IV (isolate S1). The types of yeast that were identified molecularly and represented each group of PCR-RFLP results included in group I were Kluyveromyces marxianus D1A and K. marxianus D1B, group II: K. marxianus D7, group III: Kazachstania humilis D4, while milk from Bima and Sumbawa has one yeast species as a member of group IV, namely Pichia kudriavzevii B3. Kluyveromyces marxianus was the yeast frequently found in Sumbawa fermented mare’s milk.@*Conclusion, significance and impact of study@#Various yeast species as a consortium of the milk samples can contribute to the increasing quality of fermented Sumbawa mare’s milk.

Guinea Pigs Naturally Infected by Leishmania enriettii: Clinical Analyses, Parasite Isolation and Identification

Abstract Leishmania enriettii has only been found in Curitiba metropolitan region, southern Brazil were it was first observed in Cavia porcellus from the vivarium of Instituto de Biologia e Pesquisas Tecnológicas (IBPT - today named TECPAR) by Medina, 1944. Despite more than a half century from its discovery and several research articles on this species, the natural clinical signs in guinea pigs and the parasite genetic variability is still unclear. The aims of this study were to describe the clinical features, investigate the potential wild reservoirs and, in addition, we intended to understand the polymorphism trait of the species. We analyzed 26 naturally infected guinea pigs from eight Paraná state cities. All animals showed lesions compatible with leishmaniosis, such as skin nodules or ulcers on body extremities. Direct examination of the lesion samples obtained by fine-needle aspiration or punch biopsy was conducted followed by isolation and identification of parasite DNA by random amplification of polymorphic DNA (RAPD)-PCR. Through the direct exam, a large number of intracellular amastigote forms were observed in the lesions. Different strains of the parasite, isolated from the 26 animals, were grouped in 5 clusters of approximately 65% similarity. We looked for L. enriettii in other potential reservoir hosts but the parasite was not observed. These results confirm that distinct strains of L. enriettii circulate in guinea pigs from Paraná state, more specifically in the Atlantic forest region, where we believe it serves as the center for dispersion of the species.

GENETIC DIVERSITY OF Lutzomyia (Nyssomyia) intermedia IN AN ENDEMIC AREA OF AMERICAN CUTANEOUS LEISHMANIASIS, STATE OF PARANÁ, BRAZIL / Diversidad genética de Lutzomyia (Nyssomyia) intermedia en un área endémica de leishmaniasis cutánea americana del estado de Paraná, Brasil

ABSTRACT Lutzomyia intermedia (Diptera: Psychodidae) features as one of the main vectors that are involved in the transmission of American cutaneous leishmaniasis (ACL) in the Neotropical region. However, genetic studies involving this taxon are still incipient and important for understanding the level of variability of different populations, their role, and implications as vectors. The aim of this study was to determine the level of genetic diversity of L. intermedia present in the Ribeira River Valley, an area of ACL transmission in the state of Paraná, Brazil, through the Random amplified polymorphic DNA (RAPD). Two municipalities were chosen to collect sand flies: Cerro Azul (new transmission area of the ACL) and Adrianópolis (endemic area of the ACL). The insects were captured in the house, in the peridomicile and in the wild (forest). Two of the used markers made it possible to estimate the polymorphism of the studied populations, resulting in 40 genotypes, most of them from peridomicile. The dendrogram generated by the analysis with the primer A10 showed different degrees of similarity, suggesting that there may be gene flow in the studied populations. The Principal Coordinate Analysis (PCO) with the A2 primer, was useful in grouping L. intermedia according to its ecological and geographical origin. There was no distinction between the lineages composing the L. intermedia complex. The results of this study, with the record of great genotypic diversity in L. intermedia, may contribute to explain the maintenance of the life cycle of Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) in the region.

RESUMEN Lutzomyia intermedia (Diptera: Psychodidae) es uno de los principales vectores que participan en la transmisión de leishmaniasis cutánea americana (LCA) en la región Neotropical. A pesar de que aún los estudios genéticos que involucran a este taxón son incipientes, tienen una gran importancia para comprender el nivel de variabilidad de las diferentes poblaciones y sus implicaciones en su papel vectorial. El objetivo de este estudio fue determinar el nivel de diversidad genética de L. intermedia presente en el Valle del Río Ribeira, área de transmisión de LCA en el estado de Paraná, Brasil, mediante RAPD (ADN polimórfico amplificado aleatoriamente). Los flebótomos fueron recolectados en los municipios Cerro Azul (nueva área de transmisión de LCA) y Adrianópolis (área endémica de LCA), donde fueron capturados en ambientes residenciales, en el peridomicilio y en el bosque. Dos de los marcadores utilizados permitieron estimar el polimorfismo en las poblaciones estudiadas con la obtención de 40 genotipos, la mayoría de ellos en el peridomicilio. El dendrograma generado por el análisis con el cebador A10 mostró diferentes grados de similitud, lo que sugiere que puede haber flujo gènico en las poblaciones. El Análisis de Coordenadas Principales (PCO) con el cebador A2 fue útil para agrupar L. intermedia según su origen ecológico y geográfico. No hubo distinción entre los linajes que componen el complejo L. intermedia. Los resultados de este estudio, con el registro de gran diversidad genotipica en L. intermedia, pueden contribuir a explicar el mantenimiento del ciclo biológico de Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) en la región.

In situ morphometric characterization of Aframomum melegueta accessions in Ghana.

In spite of the huge economic importance of Aframomum melegueta in the herbal and pharmaceutical industries, its production is limited by lack of planting materials (propagules). The plant also lacks scientific descriptors, which has often led to misidentification with adverse health implications. We therefore aimed at developing a descriptor list to facilitate the identification of A. melegueta using 34 morphometric traits comprising 18 quantitative and 16 qualitative characters. The morphological traits showed that A. melegueta has a characteristic stolon that produces tillers instead of rhizomes. The unweighted pair group method with arithmetic mean using both the nearest-neighbour and complete-linkage methods based on the 34 morphometric traits clustered the eight accessions into two main groups based on ecological location. The accessions from the Eastern and Ashanti regions were separated at similarity coefficients of 0.822 and 0.644, respectively, with a highly significant discriminant function. The Eastern accessions were further clustered into red or yellow fruits at similarity indexes of 0.936 and 0.865 using the nearest-neighbour and complete-linkage methods, respectively. The present study has shown that morphometric traits of A. melegueta are greatly influenced by its ecological habitat. It is envisaged that the descriptor list developed coupled with a morphometric description would enhance its identification and utilization.

Relationship between species in the genus Rosa, section Pimpinellifoliae.

The morphology of twelve species of Rosa is described and similarities between these species are assessed. Possible origins of the tetraploid species from diploid species are indicated on grounds of comparative morphology. The wild origins of living and herbarium specimens are given in order to supplement published data on geographical distribution. Meiosis in pollen mother cells, viability of pollen grains at anthesis and ability to set seed was studied in several F1 hybrids: no indication of complete or even partial sterility was found. Reproductive isolation is therefore unlikely to be maintained by reduced fertility of interspecific hybrids. Three species are reduced to synonymy with three other species, being retained as subspecific taxa. Two species are transferred from section Pimpinellifoliae to section Cinnamomeae.