Tentative indicators of malaria in archaeological skeletal samples, a pilot study testing different methods.

OBJECTIVE: This study attempts to integrate multiple methods to investigate the presence of malaria in human skeletal samples from an archaeological context. MATERIALS: 33 well preserved human remains originating from a 17th-century archaeological site in southeastern Romania. METHODS: The human bone samples were analyzed using rapid diagnostic tests for malaria antigens and PCR amplification of Plasmodium falciparum apical membrane antigen 1. A preliminary test was performed to identify and briefly characterize the presence of hemozoin using a combination of TEM imaging and diffraction. RESULTS: The rapid diagnostic tests indicated that more than half of the examined samples were positive for Plasmodium antigens, but no traces of the parasites' genetic material were detected despite repeated attempts. The TEM images indicated that hemozoin might be a promising diagnostic marker of malaria in ancient bones. CONCLUSIONS: The indisputable identification of malaria in the analyzed archaeological population was not possible as none of the applied methodological strategies turned out to be straightforward. SIGNIFICANCE: This study reinforces the intricacy and limitations of unequivocally identifying malaria in past populations and sets the stage for future studies on such life-threatening infectious disease in a geographical space, which is currently underrepresented in the bioarchaeological literature. LIMITATIONS: The low sample size and the lack of consistency across all assays hindered understanding the role of malaria in the studied population. SUGGESTIONS FOR FURTHER RESEARCH: Further thorough multidisciplinary approaches on malaria detection in ancient settlements would be appropriate to inform our knowledge of its origins, frequency, and pathogen changes over centuries.

Mitochondrial DNA Profiles of Individuals from a 12th Century Necropolis in Feldioara (Transylvania).

The genetic signature of modern Europeans is the cumulated result of millennia of discrete small-scale exchanges between multiple distinct population groups that performed a repeated cycle of movement, settlement, and interactions with each other. In this study we aimed to highlight one such minute genetic cycle in a sea of genetic interactions by reconstructing part of the genetic story of the migration, settlement, interaction, and legacy of what is today the Transylvanian Saxon. The analysis of the mitochondrial DNA control region of 13 medieval individuals from Feldioara necropolis (Transylvania region, Romania) reveals a genetically heterogeneous group where all identified haplotypes are different. Most of the perceived maternal lineages are of Western Eurasian origin, except for the Central Asiatic haplogroup C seen in only one sample. Comparisons with historical and modern populations describe the contribution of the investigated Saxon settlers to the genetic history of this part of Europe.

A probable case of infantile cortical hyperostosis in 2nd-4th centuries AD Romania.

OBJECTIVE: This study aims to discuss the differential diagnosis for the pathological alterations displayed on an infant skeleton from Romania. MATERIALS: One infant skeleton retrieved form the bathhouse of an abandoned Roman fort and dated between the 2nd and the 4th centuries AD. METHODS: All available skeletal elements were analyzed macroscopically. In addition, the isotopic signatures (δ13C and δ15N) and the control region of the human mitochondrial genome for this archaeological sample were analyzed. RESULTS: Based on dental development and long bone length, the skeleton was aged between birth and 2 months of age. Pathological lesions were noted on the mandible and diaphyses of long bones, but spared the metaphyses. CONCLUSIONS: The perinatal age of the individual, along with lesion morphology and location, suggests a diagnosis of infantile cortical hyperostosis. LIMITATIONS: The analysis would benefit from further stable isotope and mitochondrial genome analyses, which was limited due to the absence of comparative human and faunal remains from the site. SUGGESTIONS FOR FURTHER RESEARCH: Further multidisciplinary research on human archaeological remains from Romania would provide a clearer image of past disease and life histories in this geographic area.

Mitochondrial ancestry of medieval individuals carelessly interred in a multiple burial from southeastern Romania.

The historical province of Dobruja, located in southeastern Romania, has experienced intense human population movement, invasions, and conflictual episodes during the Middle Ages, being an important intersection point between Asia and Europe. The most informative source of maternal population histories is the complete mitochondrial genome of archaeological specimens, but currently, there is insufficient ancient DNA data available for the medieval period in this geographical region to complement the archaeological findings. In this study, we reconstructed, by using Next Generation Sequencing, the entire mitochondrial genomes (mitogenomes) of six medieval individuals neglectfully buried in a multiple burial from Capidava necropolis (Dobruja), some presenting signs of a violent death. Six distinct maternal lineages (H11a1, U4d2, J1c15, U6a1a1, T2b, and N1a3a) with different phylogenetic background were identified, pointing out the heterogeneous genetic aspect of the analyzed medieval group. Using population genetic analysis based on high-resolution mitochondrial data, we inferred the genetic affinities of the available medieval dataset from Capidava to other ancient Eurasian populations. The genetic data were integrated with the archaeological and anthropological information in order to sketch a small, local piece of the mosaic that is the image of medieval European population history.

Maternal DNA lineages at the gate of Europe in the 10th century AD.

Given the paucity of archaeogenetic data available for medieval European populations in comparison to other historical periods, the genetic landscape of this age appears as a puzzle of dispersed, small, known pieces. In particular, Southeastern Europe has been scarcely investigated to date. In this paper, we report the study of mitochondrial DNA in 10th century AD human samples from Capidava necropolis, located in Dobruja (Southeastern Romania, Southeastern Europe). This geographical region is particularly interesting because of the extensive population flux following diverse migration routes, and the complex interactions between distinct population groups during the medieval period. We successfully amplified and typed the mitochondrial control region of 10 individuals. For five of them, we also reconstructed the complete mitochondrial genomes using hybridization-based DNA capture combined with Next Generation Sequencing. We have portrayed the genetic structure of the Capidava medieval population, represented by 10 individuals displaying 8 haplotypes (U5a1c2a, V1a, R0a2'3, H1, U3a, N9a9, H5e1a1, and H13a1a3). Remarkable for this site is the presence of both Central Asiatic (N9a) and common European mtDNA haplotypes, establishing Capidava as a point of convergence between East and West. The distribution of mtDNA lineages in the necropolis highlighted the existence of two groups of two individuals with close maternal relationships as they share the same haplotypes. We also sketch, using comparative statistical and population genetic analyses, the genetic relationships between the investigated dataset and other medieval and modern Eurasian populations.

Release of Antibiotic Resistant Bacteria by a Waste Treatment Plant from Romania.

The occurrence and spread of bacterial antibiotic resistance are subjects of great interest, and the role of wastewater treatment plants has been attracting particular interest. These stations are a reservoir of bacteria, have a large range of organic and inorganic substances, and the amount of bacteria released into the environment is very high. The main purpose of the present study was to assess the removal degree of bacteria with resistance to antibiotics and identify the contribution of a wastewater treatment plant to the microbiota of Somesul Mic river water in Cluj county. The resistance to sulfamethoxazole and tetracycline and some of their representative resistance genes: sul1, tet(O), and tet(W) were assessed in this study. The results obtained showed that bacteria resistant to sulphonamides were more abundant than those resistant to tetracycline. The concentration of bacteria with antibiotic resistance changed after the treatment, namely, bacteria resistant to sulfamethoxazole. The removal of all bacteria and antibiotic-resistant bacteria was 98-99% and the degree of removal of bacteria resistant to tetracycline was higher than the bacteria resistant to sulfamethoxazole compared to total bacteria. The wastewater treatment plant not only contributed to elevating ARG concentrations, it also enhanced the possibility of horizontal gene transfer (HGT) by increasing the abundance of the intI1 gene. Even though the treatment process reduced the concentration of bacteria by two orders of magnitude, the wastewater treatment plant in Cluj-Napoca contributed to an increase in antibiotic-resistant bacteria concentrations up to 10 km downstream of its discharge in Somesul Mic river.

Allelic distribution of BDNF Val66Met polymorphism in healthy Romanian volunteers.

Population stratification of functional gene polymorphisms is a potential confounding factor in genetic association studies. The Val66Met (rs6265) single-nucleotide polymorphism in the brain-derived neurotrophic factor gene (BDNF) exhibits one of the highest variabilities in terms of allelic distribution between populations. The present study reports the distribution of BDNF Val66Met alleles in a sample of healthy volunteers (N = 1124) selected from the Romanian population. Frequencies were 80.74% for the Val allele and 19.26% for the Met allele. The data from this study extends efforts to map the allelic distribution of BDNF Val66Met in populations around the world and emphasizes that population stratification should be controlled for in future studies that report phenotypic associations in samples from different populations.

Can environment predict cryptic diversity? The case of Niphargus inhabiting Western Carpathian groundwater.

In the last decade, several studies have shown that subterranean aquatic habitats harbor cryptic species with restricted geographic ranges, frequently occurring as isolated populations. Previous studies on aquatic subterranean species have implied that habitat heterogeneity can promote speciation and that speciation events can be predicted from species' distributions. We tested the prediction that species distributed across different drainage systems and karst sectors comprise sets of distinct species. Amphipods from the genus Niphargus from 11 caves distributed along the Western Carpathians (Romania) were investigated using three independent molecular markers (COI, H3 and 28S). The results showed that: 1) the studied populations belong to eight different species that derive from two phylogenetically unrelated Niphargus clades; 2) narrow endemic species in fact comprise complexes of morphologically similar species that are indistinguishable without using a molecular approach. The concept of monophyly, concordance between mitochondrial and nuclear DNA, and the value of patristic distances were used as species delimitation criteria. The concept of cryptic species is discussed within the framework of the present work and the contribution of these species to regional biodiversity is also addressed.

Genes and dental disorders.

In recent decades with the advancement of molecular research, information regarding specific molecular mechanisms has exploded. In the present review we present the molecular basis of dental pathologies that are of particular interest to clinicians.

Isolation and characterization of the first microsatellite markers for the endangered relict mussel Hypanis colorata (Mollusca: Bivalvia: Cardiidae).

Hypanis colorata (Eichwald, 1829) (Cardiidae: Lymnocardiinae) is a bivalve relict species with a Ponto-Caspian distribution and is under strict protection in Romania, according to national regulations. While the species is depressed in the western Black Sea lagoons from Romania and Ukraine, it is also a successful invader in the middle Dniepr and Volga regions. Establishing a conservation strategy for this species or studying its invasion process requires knowledge about the genetic structure of the species populations. We have isolated and characterized nine polymorphic microsatellite markers in H. colorata. The number of alleles per locus ranged from 4 to 28 and the observed heterozygosity ranged from 0.613 to 1.000. The microsatellites developed in the present study are highly polymorphic and they should be useful for the assessment of genetic variation within this species.

Genetic variation at 15 polymorphic, autosomal, short tandem repeat loci of two Hungarian populations in Transylvania, Romania.

AIM: To determine allele distribution and genetic parameters for two populations living in the Romanian region of Transylvania: Hungarians from Cluj and Szeklers from Covasna county, and to compare the results between the two populations and with other Hungarian and Romanian populations. METHODS: Allele frequencies for 15 autosomal STR loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, and FGA), several forensic parameters, and paternity parameters were determined for Szekler Hungarians of Covasna county (CV-Sze, n=278) and non-Szekler Transylvanian Hungarians, who were represented by Hungarians from Cluj county (CJ-Hu, n=146). RESULTS: Average expected heterozygosity was above 70%. The combined power of discrimination and combined power of exclusion values were high. All tested loci were in agreement with Hardy-Weinberg equilibrium, with the exception of the CSF1PO locus for Covasna county. Pairwise population comparison tests and exact population differentiation tests showed no significant differences between the CJ-Hu and CV-Sze populations, and the CV-Sze group showed greater differences from other Romanian populations than did the CJ-Hu group. CONCLUSION: Hungarians from Cluj show greater genetic heterogeneity than Szeklers from Covasna. The loci tested are suitable for studying micro-differentiation between these two populations, and between these populations and other populations in Hungary and Romania.