Actinomycetes in Karstic caves of northern Spain (Altamira and Tito Bustillo).

A variety of isolation procedures were carried out to study the involvement of bacteria in the colonisation and biodeterioration of Spanish caves with paleolithic rock art (Altamira and Tito Bustillo). The applied techniques mainly aimed to isolate heterotrophic bacteria such as streptomycetes, nocardioform and coryneform actinomycetes, and other gram-positive and gram-negative bacteria. The results demonstrated that actinomycetes were the most abundant gram-positive bacteria in the caves. Actinomycetes revealed a great taxonomic diversity with the predominant isolates belonging to the genus Streptomyces. Members of the genera Nocardia, Rhodococcus, Nocardioides, Amycolatopsis, Saccharothrix, Brevibacterium, Microbacterium, and coccoid actinomycetes (family Micrococcaceae) were also found.     

The biogeochemical role of Actinobacteria in Altamira Cave, Spain

The walls and ceiling of Altamira Cave, northern Spain, are coated with different coloured spots (yellow, white and grey). Electron microscopy revealed that the grey spots are composed of bacteria and bioinduced CaCO(3) crystals. The morphology of the spots revealed a dense network of microorganisms organized in well-defined radial and dendritic divergent branches from the central area towards the exterior of the spot, which is coated with overlying spheroidal elements of CaCO(3) and CaCO(3) nest-like aggregates. Molecular analysis indicated that the grey spots were mainly formed by an unrecognized species of the genus Actinobacteria. CO(2) efflux measurements in rocks heavily covered by grey spots confirmed that bacteria-forming spots promoted uptake of the gas, which is abundant in the cave. The bacteria can use the captured CO(2) to dissolve the rock and subsequently generate crystals of CaCO(3) in periods of lower humidity and/or CO(2). A tentative model for the formation of these grey spots, supported by scanning electron microscopy and transmission electron microscopy data, is proposed.     

Early Pleistocene human mandible from Sima del Elefante (TE) cave site in Sierra de Atapuerca (Spain): a palaeopathological study

Here we present a detailed palaeopathological study of the hominin mandible ATE9-1 found at the Sima del Elefante site (TE), Sierra de Atapuerca, Spain. This fossil represents the earliest hominin remains from Western Europe with an age of ca. 1.3 Ma. The specimen displays several dento-gnathic lesions; the antiquity and geographic location of this fossil justifies a detailed palaeopathological study to determine if the pathologies have significantly altered taxonomically relevant features. Our study reveals severe dental attrition combined with generalized hypercementosis, alveolar root exposure, mild periodontal disease, tooth dislocation, and an anomalous occlusal plane. We have also observed calculus deposits, two cystic lesions and an anomalous wear facet compatible with tooth picking. The majority of these pathological signs can be explained by compensatory eruption. We propose that these lesions are associated as causes, consequences, and amplifiers of one another within the framework of heavy and even traumatic occlusion, masticatory habits, or both traumatic occlusion and masticatory habits. Despite the severity of these lesions, occlusion was at least partially functional so it was unlikely to influence the survival of this individual. In addition, the lesions do not prohibit the taxonomic assessment of the mandible.     

Comparing bacterial community fingerprints from white colonizations in Altamira Cave (Spain)

Monitoring bacterial communities is critical for assessing biodeterioration among other processes. This study presents a strategy and an example of comparative analysis of bacterial communities developing in a cave environment, Altamira Cave which contains unique paleolithic paintings. The analyzed question was whether white colonizations discovered throughout the cave corresponded to similar or different bacterial communities. Molecular fingerprints were obtained by PCR–DGGE from DNA and RNA and statistically compared. Results based on DNA analysis showed that a similar bacterial community was present in white colonizations throughout the cave. Fingerprints based on RNA confirmed the similarity of the major metabolically active components of these communities. The proposed procedure confirmed that white colonizations in Altamira Cave were a consequence of the development of a single complex bacterial community, and the method proves to be highly useful for comparative analysis of microbial communities, including biodeteriorating processes and any other comparative analysis of bacterial communities.     

Mesolithic burials in los canes cave (Asturias,Spain)

Los Canes is a small cave situated half way up the abrupt southern slope of Cuera range, near the village of Arangas (Eastern Asturias, Spain). Three funerary structures have been discovered in the entrance of that cave. They consisted of burials (one of them, number II, reused) where the corpses of three individuals had been placed in different positions, with the legs strongly bent. Various grave goods (animal bones, pierced teeth, a perforated antler, etc.) accompanied them. The individuals found in these burials have been directly dated by AMS. Their age ranges between 6930±95 and 6265±75 BP (5930–5680 and 5330–5010 cal. BC). The present paper describes in detail the burials and discusses the questions raised by their datations and characteristics. Fourthermore the authors compare them with other Cantabrian funerary structures, taken into account the different mesoecological factors and living conditions of these last hunter-gatherer populations. Some preliminary anthropological data about Los Canes individual n^oI are also considered.     

Early Pleistocene human mandible from Sima del Elefante (TE) cave site in Sierra de Atapuerca (Spain): a comparative morphological study

We present a detailed morphological comparative study of the hominin mandible ATE9-1 recovered in 2007 from the Sima del Elefante cave site in Sierra de Atapuerca, Burgos, northern Spain. Paleomagnetic analyses, biostratigraphical studies, and quantitative data obtained through nuclide cosmogenic methods, place this specimen in the Early Pleistocene (1.2-1.3 Ma). This finding, together with archaeological evidence from different European sites, suggests that Western Europe was colonised shortly after the first hominin expansion out of Africa around the Olduvai subchron. Our analysis of the ATE9-1 mandible includes a geometric morphometric analysis of the lower second premolar (LP₄), a combined and detailed external and internal assessment of ATE9-1 roots through CT and microCT techniques, as well as a comparative study of mandibular and other dental features. This analysis reveals some primitive Homo traits on the external aspect of the symphysis and the dentition shared with early African Homo and the Dmanisi hominins. In contrast, other mandibular traits on the internal aspect of the symphysis are derived with regard to African early Homo, indicating unexpectedly large departures from patterns observed in Africa. Reaching the most occidental part of the Eurasian continent implies that the first African emigrants had to cross narrow corridors and to overcome geographic barriers favouring genetic drift, long isolation periods, and adaptation to new climatic and seasonal conditions. Given these conditions and that we are dealing with a long time period, it is possible that one or more speciation events could have occurred in this extreme part of Eurasia during the Early Pleistocene, originating in the lineages represented by the Sima del Elefante-TE9 hominins and possibly by the Gran Dolina-TD6 hominins. In the absence of any additional evidence, we prefer not include the specimen ATE9-1 in any named taxon and refer to it as Homo sp.     

Comparison of intradiurnal variation of airborne pollen in Mar del Plata (Argentina). Part I. Non-arboreal pollen

Intradiurnal variation of non-arboreal pollen (NAP) aredescribed in relation to the associated weather using the data obtainedwith a Burkard trap in Mar del Plata city, for a three year data set(1988, 1992 and 1995). The daily pattern of concentration was quitehomogeneous during the years analysed except for 1995, when it showed avariation due to the change of position of the station. Qualitativecomposition of the NAP airborne pollen was mainly represented byPoaceae, Plantago and Cyperaceae during the three years. The daily trendshowed high concentrations during daylight hours, when instability andconvective movements prevail, and low concentrations in the night, whenmeteorological conditions favour the development of a stable stratifiedlayer.     

Pigment spectroscopy analyses in Maltravieso cave,Spain

Maltravieso cave (Estremadura, Spain) was recently the subject of ATR-FTIR spectroscopy of 8 figures (four hand stencils, a red horse figure, a black bull figure, a pair of black lines, and a pair of brown lines). These artistic representations are composed of different colors including red, brown, and black figures. The analyses showed that different naturally occurred material compounds in the cave (such as kaolinite) were used in the creation of the ochre to obtain different hues. Hand stencils are made of earth pigment that is naturally present in the cave which contained hematite, magnetite, and goethite as chromophores material. The red horse figure was made with kaolinite-based ochre. The black bull figure was made with charcoal (identified with Raman spectroscopy). The pigmental composition analyses show that they are composed of the same natural matter found inside the cave. Our results suggest that different paintings in the cave have been produced by different techniques and these techniques are linked to different chronological periods.     

Moonmilk Deposits Originate from Specific Bacterial Communities in Altamira Cave (Spain)

The influence of bacterial communities on the formation of carbonate deposits such as moonmilk was investigated in Altamira Cave (Spain). The study focuses on the relationship between the bacterial communities at moonmilk deposits and those forming white colonizations, which develop sporadically throughout the cave. Using molecular fingerprinting of the metabolically active bacterial communities detected through RNA analyses, the development of white colonizations and moonmilk deposits showed similar bacterial profiles. White colonizations were able to raise the pH as a result of their metabolism (reaching in situ pH values above 8.5), which was proportional to the nutrient supply. Bacterial activity was analyzed by nanorespirometry showing higher metabolic activity from bacterial colonizations than uncolonized areas. Once carbonate deposits were formed, bacterial activity decreased drastically (down to 5.7% of the white colonization activity). This study reports on a specific type of bacterial community leading to moonmilk deposit formation in a cave environment as a result of bacterial metabolism. The consequence of this process is a macroscopic phenomenon of visible carbonate depositions and accumulation in cave environments.     

Seasonal airborne pollen pattern in Mar del Plata City, Argentina

Continuous aerobiological survey of the atmosphere of Mar del Plata was carried out from December 1991 to November 1993 with a Burkard volumetric spore trap. Daily slides were prepared and studied every 2 h with standard techniques. Weekly records were kept for 27 relevant pollen types selected either by their prevalence or relative high atmospheric concentration. Quantitative multivariate analysis enabled to distinguish three major pollen seasons, related to atmospheric dominance either arboreal pollen (AP) or non-arboreal pollen (NAP). June to October is the richest period in number of pollen types, mainly dominated by AP; while from November to May, there is an overwhelming dominance of NAP types, represented by grass, herb and weed pollen. The study and prediction of this phenomenon is of great interest not only from the ecosystem point of view, but in relation to human disease as well.     

Metabolically active microbial communities of yellow and grey colonizations on the walls of Altamira Cave, Spain

Aims: To determine the major components of total and metabolically active microbial communities of yellow and grey colonizations threatening the conservation of palaeolithic paintings in Altamira Cave (Spain). Methods and Results: Micro‐organisms present in yellow and grey colonizations were determined from DNA analysis with those showing metabolic activity determined from RNA analysis. Microbial community fingerprints were obtained by denaturing gradient gel electrophoresis (DGGE) and 16S rDNA libraries were constructed from PCR amplified products. Proteobacteria was the most frequent bacterial phylum. Other phyla detected from RNA‐based microbial surveys were Acidobacteria, Actinobacteria, Firmicutes, Nitrospirae and Gemmatimonadetes. The detected metabolically active micro‐organisms represented only a fraction of the total bacterial community present in the studied colonizations as compared from DGGE analysis. Conclusions: The major bacterial participants in the development of yellow and grey colonizations in Altamira Cave were determined using RNA‐based molecular techniques. Micro‐organisms showing undetectable activity represent a potential risk for the conservation of these paintings if environmental conditions experience variations. Significance and Impact of the Study: Caves with palaeolithic paintings are affected by microbial deterioration. Assessing the composition of the microbial communities colonizing these caves represents a first stage to understand and control these colonizations.     

Comparison of diurnal variation of airborne pollen in Mar del Plata (Argentina)

Intradiurnal variation of arboreal pollen (AP) in Mar del Plata city is compared during three non - consecutive years of survey and described in relation to the associated weather. The daily pattern of pollen abundance has a maximum between 10:00 and 12:00?h, while a minimum occurs at 18:00?h. The first two years of survey showed homogeneous daily trends, but in 1995 the maximum and minimum concentrations were delayed because of the change in position of the collecting station. Arboreal pollen spectrum presented qualitative and quantitative changes in the three years analysed. Results indicate optimal conditions for diurnal dispersion of arboreal pollen are high temperatures and low relative humidity. Also interaction between source position and wind direction has important effects on the timing of the peaks of some pollen types.     

Luminescence chronology of cave sediments at the Atapuerca paleoanthropological site, Spain

Ascertaining the timing of the peopling of Europe, after the first out-of-Africa demographic expansion at the end of the Pliocene, is of great interest to paleoanthropologists. One of the earliest direct evidences for fossil hominins in western Europe comes from an infilled karstic cave site called Gran Dolina at Atapuerca, in a stratum approximately 1.5m below the Brunhes-Matuyama (B-M) geomagnetic boundary (780ka) within lithostratigraphic unit TD6. However, most of the meters of fossil- and tool-bearing strata at Gran Dolina have been difficult to date. Therefore, we applied both thermoluminescence (TL) and infrared-stimulated-luminescence (IRSL) multi-aliquot dating methods to fine-silt fractions from sediment samples within Gran Dolina and the nearby Galería cave site. We also applied these methods to samples from the present-day surface soils on the surrounding limestone hill slopes to test the luminescence-clock-zeroing-by-daylight assumption. Within the uppermost 4m of the cave deposits at Gran Dolina, TL and paired TL and IRSL ages range stratigraphically from 198+/-19ka to 244+/-26ka. Throughout Gran Dolina, all luminescence results are stratigraphically self-consistent and, excepting results from two stratigraphic units, are consistent with prior ESR-U-series ages from progressively deeper strata. Thermoluminescence ages culminate at 960+/-120ka approximately 1m below the 780ka B-M boundary. At Galería, with one exception, TL and IRSL ages range stratigraphically downward from 185+/-26ka to 503+/-95ka at the base of the lowermost surface-inwash facies. These results indicate that TL and (sometimes) IRSL are useful dating tools for karstic inwash sediments older than ca. 100ka, and that a more accurate chronostratigraphic correlation is now possible among the main Atapuerca sites (Gran Dolina, Galería, Sima de los Huesos). Furthermore, the oldest TL age of ca. 960ka from Gran Dolina, consistent with biostratigraphic and paleomagnetic evidence, implies a probable numeric age of 900-950ka for the oldest hominin remains ( approximately 0.8m below the TL sample). This age window suggests a correspondence to Marine Isotope Stage (MIS) 25, a relatively warm and humid interglaciation.     

A Neandertal mandible from the Cova del Gegant (Sitges, Barcelona, Spain)

A human mandible from the site of Cova del Gegant is described here for the first time and compared with other Middle and Upper Pleistocene representatives of the genus Homo from Europe and Southwest Asia. The specimen was recovered from sediments which also yielded Mousterian stone tools and Pleistocene fauna. The preserved morphology of the mandible, particularly in the region of the mental foramen, clearly aligns it with the Neandertals, making the Cova del Gegant the only known site in Catalonia documenting diagnostic human skeletal remains in association with Middle Paleolithic stone tools. This represents an important new addition to the human fossil record from the Iberian Peninsula and joins the Ba?olas mandible in documenting the course of human evolution in the northern Mediterranean region of Spain.     

Comparative biometrical study of the Mousterian mandible from Cueva del Boquete de Zafarraya (Málaga, Spain)

The Zafarraya Neanderthal mandible has a very late date for Neanderthals and biometrical analysis shows it shares some specific traits with modern humans.     

Microbiological characteristics of Pacific shrimp (Pandalus jordani).

Microorganisms associated with Pacific shrimp (Pandalus jordani) were isolated and identified. Those on the iced raw shrimp, which yielded an average count of 1.6 x 10(6), were predominantly Moraxella, Pseudomonas, Acinetobacter, Arthrobacter, and Flavobacterium-Cytophaga spp. The blanching and peeling reduced the microbial level to 3.3 x 10(4) and also selectively eliminated Moraxella spp. The microbial flora changed after each processing sequence, and the heat sensitivity and growth characteristics of the representative microbial groups suggested that the presence of Arthrobacter and Acinetobacter spp. in peeled shrimp may indicate inadequate cleaning of raw shrimp or a shorter blanching time. The presence of Moraxella and Flavobacterium-Cytophaga spp. would indicate the degree of secondary contamination, and the presence of Pseudomonas spp. would indicate the shelf-age of the processed shrimp.     

Blood groups of the Cantabrian population (Spain).

The genetic polymorphism of eight red cell antigenic systems was examined in two population samples from Cantabria (north Spain). No significant differences were found between the autochthonous and the resident populations. The results obtained are compared with those for other populations in the north of Spain. The low frequency of the B allele of the AB0 system and the high frequency of the 'cde' haplotype of the Rh system in the Cantabrain and Asturian population suggests that the anthropological characteristics of these two systems, which hitherto have been attributed to the Basques, also apply to the western neighbouring regions.