Darwin was right: inbreeding depression on male fertility in the Darwin family

Charles Darwin, who was married to his first cousin Emma Wedgwood, was the first experimentalist to demonstrate the adverse effects of inbreeding. He documented the deleterious consequences of self‐fertilization on progeny in numerous plant species, and this research led him to suspect that the health problems of his 10 children, who were very often ill, might have been a consequence of his marriage to his first cousin. Because Darwin's concerns regarding the consequences of cousin marriage on his children even nowadays are considered controversial, we analyzed the potential effects of inbreeding on fertility in 30 marriages of the Darwin–Wedgwood dynasty, including the marriages of Darwin's children, which correspond to the offspring of four cousin marriages and three marriages between unrelated individuals. Analysis of the number of children per woman through zero‐inflated regression models showed a significantly adverse effect of the husband inbreeding coefficient on family size. Furthermore, a statistically significant adverse effect of the husband inbreeding coefficient on reproductive period duration was also detected. To our knowledge, this is the first time that inbreeding depression on male fertility has been detected in humans. Because Darwin's sons had fewer children in comparison to non‐inbred men of the dynasty, our findings give empirical support to Darwin's concerns on the consequences of consanguineous marriage in his own progeny. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114 , 474–483.     

Evidence-based palaeopathology: Meta-analysis of PubMed-listed scientific studies on ancient Egyptian mummies

There is a plethora of published scientific studies on ancient Egyptian mummies. Surprisingly, hitherto there is no systematic review of this research, which would help to assess the quality of this vast body of published literature and thus to increase “evidence” in palaeopathological research. The aim of this study was to review all PubMed-listed scientific studies performed on Ancient Egyptian mummies. A total of 131 studies were found in the database for the selected time period, 1977-2005. Our “meta-analysis” showed that the number of publications per year varies enormously. The majority of mummies examined date to the third intermediate and Ptolemaic periods; data from other time periods were lacking. Identification of the cause of death and ¹⁴C-dating of the mummy or funeral goods were rarely addressed. There was a tendency towards an increased use of non-invasive examination methods in more modern times. Our meta-analysis addressed both scientific content (e.g. palaeopathological findings/examination methods) and publication issues (e.g. location of the first author or year of publication) in these studies. Based on our experience, we recommend some minimum publication standards for palaeopathologic studies on ancient mummies, which shall improve evidence-based research in palaeopathology in general.     

Technical note: Terahertz imaging of ancient mummies and bone

Ancient mummified soft‐tissues are a unique source to study the evolution of disease. Diagnostic imaging of such historic tissues is of foremost interest in paleoanthropology or paleopathology, with conventional x‐ray and computed tomography (CT) being the gold‐standard. Longer wavelength radiation in the far‐infrared or Terahertz region allows diagnostic close‐to‐surface tissue differentiation of bone morphology while being harmless to human cells. The aim of this study is to show the feasibility and the morpho‐diagnostic impact of THz imaging of historic remains. Images of an artificially embalmed ancient Egyptian human mummy hand, an artificially embalmed ancient Egyptian mummified fish and a macerated human lumbar vertebra were obtained by THz‐pulse imaging and compared with conventional X‐ray and CT images. Although conventional x‐ray imaging provides higher spatial resolution, we found that THz‐imaging is well‐suited for the investigation of ancient mummified soft tissue and embalming‐related substances / wrappings. In particular, bone and cartilaginous structures can be well differentiated from surrounding soft‐tissues and bandage‐wrappings by THz imaging. Furthermore, THz‐pulse imaging also measures the time‐delay of the pulsed signal when passing through the sample, which provides supplementary information on the optical density of the sample that is not obtained by X‐ray and CT. Terahertz radiation provides a completely non‐invasive diagnostic imaging modality for historic dry specimens. We anticipate this modality also to be used for detection of hidden objects in historic samples such as funerary amulets still in situ in wrapped mummies, as well as potentially for the identification of spectral signatures from chemical substances, e.g., in embalming essences.. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

First insights into the metagenome of Egyptian mummies using next-generation sequencing

We applied, for the first time, next-generation sequencing (NGS) technology on Egyptian mummies. Seven NGS datasets obtained from five randomly selected Third Intermediate to Graeco-Roman Egyptian mummies (806 BC–124AD) and two unearthed pre-contact Bolivian lowland skeletons were generated and characterised. The datasets were contrasted to three recently published NGS datasets obtained from cold-climate regions, i.e. the Saqqaq, the Denisova hominid and the Alpine Iceman. Analysis was done using one million reads of each newly generated or published dataset. Blastn and megablast results were analysed using MEGAN software. Distinct NGS results were replicated by specific and sensitive polymerase chain reaction (PCR) protocols in ancient DNA dedicated laboratories. Here, we provide unambiguous identification of authentic DNA in Egyptian mummies. The NGS datasets showed variable contents of endogenous DNA harboured in tissues. Three of five mummies displayed a human DNA proportion comparable to the human read count of the Saqqaq permafrost-preserved specimen. Furthermore, a metagenomic signature unique to mummies was displayed. By applying a “bacterial fingerprint”, discrimination among mummies and other remains from warm areas outside Egypt was possible. Due to the absence of an adequate environment monitoring, a bacterial bloom was identified when analysing different biopsies from the same mummies taken after a lapse of time of 1.5 years. Plant kingdom representation in all mummy datasets was unique and could be partially associated with their use in embalming materials. Finally, NGS data showed the presence of Plasmodium falciparum and Toxoplasma gondii DNA sequences, indicating malaria and toxoplasmosis in these mummies. We demonstrate that endogenous ancient DNA can be extracted from mummies and serve as a proper template for the NGS technique, thus, opening new pathways of investigation for future genome sequencing of ancient Egyptian individuals.     

Royal dynasties as human inbreeding laboratories: the Habsburgs

The European royal dynasties of the Early Modern Age provide a useful framework for human inbreeding research. In this article, consanguineous marriage, inbreeding depression and the purging of deleterious alleles within a consanguineous population are investigated in the Habsburgs, a royal dynasty with a long history of consanguinity over generations. Genealogical information from a number of historical sources was used to compute kinship and inbreeding coefficients for the Habsburgs. The marriages contracted by the Habsburgs from 1450 to 1750 presented an extremely high mean kinship (0.0628±0.009), which was the result of the matrimonial policy conducted by the dynasty to establish political alliances through marriage. A strong inbreeding depression for both infant and child survival was detected in the progeny of 71 Habsburg marriages in the period 1450–1800. The inbreeding load for child survival experienced a pronounced decrease from 3.98±0.87 in the period 1450–1600 to 0.93±0.62 in the period 1600–1800, but temporal changes in the inbreeding depression for infant survival were not detected. Such a reduction of inbreeding depression for child survival in a relatively small number of generations could be caused by elimination of deleterious alleles of a large effect according with predictions from purging models. The differential purging of the infant and child inbreeding loads suggest that the genetic basis of inbreeding depression was probably very different for infant and child survival in the Habsburg lineage. Our findings provide empirical support that human inbreeding depression for some fitness components might be purged by selection within consanguineous populations.     

Inbreeding in Finland

We have compiled data on the frequency of first-cousin marriages in Finland using royal dispensation records for the time period 1810-1872 and national population statistics for the time period 1878-1920. For the earlier period, 0.315% of Finland's marriages were contracted between first cousins (2,331 of 739,387). During the second time period, 0.174% of Finland's marriages took place between first cousins (1,325 of 761,976). These figures, which yield average kinship coefficients of 0.00020 and 0.00011, respectively, show that the level of inbreeding in Finland due to first-cousin marriage has been quite low. An analysis of individual parishes shows that first-cousin marriages are, on average, substantially less frequent than predicted by a random-mating model. In order to evaluate determinants of first-cousin marriage, several predictive variables have been examined: parish ethnic composition (proportion of Swedish and Finnish speakers), husband's occupation (graded into 6 socioeconomic levels), geographic distance between spouses' premarital residences, population density, parish endogamy, and urban vs. rural residence. Various logistic and linear regression models were analyzed in which consanguinity was the dependent variable. The best predictors of consanguinity were ethnic composition and occupation. The other variables were not in general significant predictors. These results show that many of the "mate availability" factors that would be predicted theoretically to account for consanguinity variation (population density, geographic isolation, urban vs. rural residence) do not. Instead, the best predictors of consanguinity at the first-cousin level are cultural factors such as ethnicity and occupation. Evaluation of cultural variables can provide a greatly enriched interpretation of complex biosocial phenomena such as inbreeding.     

Evidence based palaeopathology: Meta-analysis of Pubmed-listed scientific studies on pre-Columbian,South American mummies

The aim of this study was to review all Pubmed^®-listed palaeopathological studies performed on pre-Columbian South American mummies. A total of 61 studies were found (1977–2005). Review criteria included e.g. method of examination, method of mummification, palaeopathological diagnoses and individual age of mummies as well as dating, which ranged from 7500 to 500 years BP, mainly (if reported) originating from the Chiribaya and Chinchorro cultures. The average age of the 99 individually reported mummies was about 25 years. Only six studies included computed tomography, thirteen studies used classical radiography as an examination method. Three studies analysed parasite related diseases, especially caused by Trypanosoma cruzi. Among all of the reported infectious diseases (n = 9), there were seven studies presenting cases of tuberculosis. The results were also compared inter-culturally. In 61 studies (37 epidemiological and 24 case reports) more than 6400 mummified individuals were analysed. By contrast, meta-analytic data for ancient Egyptian mummies ( Zweifel et al., 2009) included about 3000 analysed individuals in 131 studies (85 case reports and 46 epidemiological studies). In general, ancient Egyptian mummies were shown to be intentionally mummified, whereas the Pre-Columbian American mummies showed a great diversity of spontaneous mummification. However, ritualistic mummification methods were also practised (n = 2). This study's results shall assist to improve evidence-based research in palaeopathology.     

Brief communication: Twentieth-century replication of an Egyptian mummy: Implications for paleopathology

Replication in a modern human cadaver of ancient Egyptian mummification focused on tools used by ancient Egyptian embalmers, the use of natron (a mixture of sodium carbonate, bicarbonate, and chloride) in the preparation of the mummy, surgical procedures in the removal of the viscera and brain, and histologic examination of the viscera. The first three areas have been reported separately (Brier and Wade [1997] ZAS 124:89–100). In this paper, we demonstrate a degree of histologic preservation comparable to that seen in Egyptian mummies, indicating the effectiveness of ancient mummification and that the histologic appearance of such mummies is little altered by the passage of millennia. Am J Phys Anthropol 107:417–420, 1998. © 1998 Wiley-Liss, Inc.     

What Is the Accurate Prevalence of Obesity in Sweden in the 21st Century? Methodological Experiences From the Skaraborg Project

Objective: To examine the impact of non‐response to self‐reported body weight and height in health questionnaires for the estimation of obesity prevalence. Methods and Procedures: A cross‐sectional population‐based health survey in the community of Vara with 16,009 residents (in year 2002) in South‐western Sweden. Participants randomly selected in strata by sex and age among residents being 30–74 years old were consecutively invited to the local health care center for a health examination, including two visits. Self‐reported information on body weight and height were obtained by health questionnaires at the first visit, and measured information on both variables at the second visit. For this study 1,809 subjects (904 men and 905 women) completed both visits (participation rate 81%), and a nurse measured body weight and height of all at visit two. Participants not self‐reporting body weight and/or height at the first visit were defined as non‐responders. Results: Both male and female non‐responders were significantly older than responders. Female non‐responders had significantly higher BMI (29.8 ± 5.8 kg/m²) than female responders (26.6 ± 5.3 kg/m²), (P < 0.001). No similar findings were seen in men. Non‐responders were more likely to be obese than responders both in men (odds ratio (OR) 2.06, 95% confidence interval (CI) 1.03–4.11) and in women (OR 2.24, 95% CI 1.25–4.02). Discussion: Non‐responders to self‐reported body weight and height in health questionnaires contribute to the underestimation of obesity. Measured body weight and height are to prefer when describing the accurate prevalence of obesity in populations.     

Parental consanguinity as a cause for increased incidence of births defects in a study of 238,942 consecutive births.

The risk for birth defects in the offspring of first-cousin matings has been estimated to increase sharply compared to non consanguineous marriages. As a general decline in the frequency of consanguineous marriages was observed in this century, one wonders whether consanguinity is still a factor in the appearance of birth defects in developed countries. Based on our registry of congenital anomalies we tried to answer to this question. In the population studied in North-Eastern France a consanguineous mating was known in 1.21% of the cases with congenital anomalies, vs. 0.27% in controls, (p < 0.001). The frequency of the malformations recorded paralleled the degree of consanguinity: out of 89 malformed children, 51 were seen in first-cousins mating (10.3 times more frequent than in offspring of non consanguineous couples), 17 in second-cousins marriages and 18 in more distant relatives mating. Three were uncle-niece marriage. Excluding known mendelian conditions these numbers were 73, 36, 17 and 17 respectively and the corresponding relative risk were 3.68, 3.01, 3.41 and 4.89 respectively. Therefore there is a negative dose-response effect between level of inbreeding and risk of congenital malformations. Consanguineous mothers were more often pregnant than non consanguineous mothers (p < 0.01) and they had more stillbirths than non consanguineous mothers. These results show that consanguinity is still a factor of birth defects and they must be taken into account for genetic counseling of inbred marriages, in developed countries.     

Inbreeding in the Samarkand region]

Data on the frequency of inbreeding in some isolated populations of Samarkand region are given. Statistically significant differences were established in the frequency of inbreeding in separate groups and in the general population of the region. Inbreeding marriages were more observed between paralel cousins, occasionally they were made between secon cousins. Strict time dependence of inbreeding frequency was not found, but it was noticed that the level of inbreeding was reduced since 1930 to 1949.     

Consanguinity and prereproductive mortality in the Utah Mormon population

To test the effects of parental consanguinity on mortality among offspring, inbreeding coefficients were estimated for 303,675 members of the Utah Mormon population who were born between 1847 and 1945. Although consanguinity has been relatively rare in this population, the large sample size permitted the identification of more than 3,500 inbred offspring. Among the offspring of unrelated parents, 13.2% died before the age of 16. Significant elevations in prereproductive mortality were seen among the offspring of first-cousin marriages (22%) and among the offspring of closer unions (32%). The cor- responding relative risks are 1.70 (95% confidence limits = 1.52, 1.91) and 2.41 (95% confidence limits = 1.59, 3.41), respectively. Other categories of relationship did not produce significant elevations in offspring mortality. Similar results were obtained when a case-control approach was used to remove the effects of socioeconomic variation. Consistent with many other studies of populations with low consanguinity rates, this population experienced a relatively high absolute increase in mortality among the offspring of first-cousin marriages (9%). Preliminary evidence is offered for the hypothesis that mortality differentials are larger in populations with low inbreeding and low mortality because nongenetic causes of death do not obscure the effects of consanguinity.     

Technical note: PCR analysis of minimum target amount of ancient DNA

The study of ancient DNA plays an important role in archaeological and palaeontological research as well as in pathology and forensics. Here, we present a new tool for ancient DNA analysis, which overcomes contamination problems, DNA degradation, and the negative effects of PCR inhibitors while reducing the amount of starting target material in the picogram range. Ancient bone samples from four Egyptian mummies were examined by combining laser microdissection, conventional DNA extraction, and low‐volume PCR. Initially, several bone particles (osteons) in the micrometer range were extracted by laser microdissection. Subsequently, ancient DNA amplification was performed to verify our extraction method. Amelogenin and β‐actin gene specific fragments were amplified via low‐volume PCR in a total reaction volume of 1 μl. Results of microdissected mummy DNA samples were compared to mummy DNA, which was extracted using a standard DNA extraction method based on pulverization of bone material. Our results highlight the combination of laser microdissection and low‐volume PCR as a promising new technique in ancient DNA analysis. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

The prevalence of consanguineous marriages in an underserved area in Lebanon and its association with congenital anomalies

Background: Consanguinity is a recognized common practice among marriages in the Middle East. Many studies have suggested a strong association between first cousin marriages and the incidence of autosomal recessive diseases and congenital anomalies. The objectives of this study were to study the prevalence of consanguinity among the marriages of Bekaa (a region in Lebanon) with its sociodemographic correlates, and to assess the prevalence of congenital anomalies associated with these marriages. Methods: This study was a cross-sectional study done in three of the major areas of the Bekaa region. The sample size consisted of 552 households chosen based on proportionate random sampling according to population size in each area. The survey was conducted based on face-to-face interview with a member of the couples of each household. Results: The overall prevalence of consanguineous marriages was reported to be 42% with first cousin marriage constituting around 31% of the total marriages. No association was found between different socioeconomic status (SES) correlates and first cousin marriages. Results showed a significant association between first cousin marriage and mental retardation, physical retardation, bilateral cleft lip +/- cleft palate, cystic fibrosis, and congenital blindness. Conclusion: In a population with a high degree of inbreeding, the formulation of a public health program with multiapproach strategy, including education about the anticipated genetic consequences, prenatal diagnosis, neonatal screening, and genetic counseling, is a necessity.     

The mummy returns… and sheds new light on old questions

Whether as the ancient Egyptian crocodile‐god Sobek, a terrifying predator of African waterways, or simply as a premium handbag leather, the Nile crocodile (Crocodylus niloticus) has long held the fascination of mankind. Despite 200 years of study, however, uncertainty remains as to its taxonomy. While resolving such issues are key to understanding the origins and biogeography of the so‐called true crocodiles of genus Crocodylus, given widespread ongoing range contraction, such issues are paramount for design of future conservation strategies. In this issue of Molecular Ecology, Hekkala et al. (2011) apply analysis of modern, historic and ancient DNA (aDNA) to the questions, with far‐reaching implications. First they demonstrate that, as currently described, the Nile crocodile is paraphyletic, with individuals from the east and western clades separated by a number of New World crocodile species. The consequences of this finding are as important for conservation efforts as for their impact on crocodile taxonomy. Furthermore, they strike at the heart of the long‐standing debate over whether aDNA analysis of ancient Egyptian mummies is scientifically sound.     

An ancient relation between units of length and volume based on a sphere

The modern metric system defines units of volume based on the cube. We propose that the ancient Egyptian system of measuring capacity employed a similar concept, but used the sphere instead. When considered in ancient Egyptian units, the volume of a sphere, whose circumference is one royal cubit, equals half a hekat. Using the measurements of large sets of ancient containers as a database, the article demonstrates that this formula was characteristic of Egyptian and Egyptian-related pottery vessels but not of the ceramics of Mesopotamia, which had a different system of measuring length and volume units.     

Growth and fluctuating asymmetry of human newborns: Influence of inbreeding and parental education

Historically, medical concerns about the deleterious effects of closely inbred marriages have focused on the risk posed by recessive Mendelian disease, with much less attention to developmental instability. We studied the effects of inbreeding (first‐cousin marriage) on growth and fluctuating asymmetry of 200 full‐term infants (101 inbred and 99 outbred) whose parents were of similar socioeconomic status in Sivas Province, Turkey. In addition to differences in their mean inbreeding coefficients (f = 1/16 for first cousins and f < 1/1,024 for unrelated parents), the consanguineous parents were less well educated (3 years, on average for both husbands and wives). We measured weight, height, head circumference, and chest circumference of the newborns, as well as four bilateral traits (ear width, ear length, and second and fourth digit lengths). After taking education into account, none of the measures of size (weight, height, head circumference, and chest circumference) and fluctuating asymmetry differed between the inbred and outbred groups. Male children of well‐educated parents, however, were larger and had less fluctuating asymmetry. Female children of well‐educated parents weighed more than those of less well‐educated parents, but were otherwise indistinguishable for height, head circumference, chest circumference, and fluctuating asymmetry. We conclude that inbreeding depression causes neither an increase in fluctuating asymmetry of full‐term newborns, nor a decrease in body size. Unmeasured variables correlated with education appear to have an effect on fluctuating asymmetry and size of male children and only a weak effect on size (weight) of female children. Am J Phys Anthropol 153:45–51, 2014. © 2013 Wiley Periodicals, Inc.     

Inbreeding in Southeastern Spain

In this paper, the structure of a southeastern Spanish population was studied for the first time with respect to its inbreeding patterns and its relationship with demographic and geographic factors. Data on consanguineous marriages (up to second cousins) from 1900 to 1969 were taken from ecclesiastic dispensations. Our results confirm that the patterns and trends of inbreeding in the study area are consistent with those previously observed in most non-Cantabrian Spanish populations. The rate of consanguineous marriages was apparently stable between 1900 and 1935 and then sharply decreased since 1940, which coincides with industrialization in Spain. A marked departure from Hardy-Weinberg expectations (0.25) in the ratio of first cousin (M22) to second cousin (M33) marriages in the study population (0.88) was observed. The high levels of endogamy (>80%) and its significant steadiness throughout the twentieth century is noteworthy. Accordingly, our results show that exogamous marriages were not only poorly represented but also that this reduced mobility (<6 km) suggests that the choice of a mate was preferentially local. We found higher mobility in M22 with respect to M33 cousin mating. The relationships between population size and consanguinity rates and inbreeding fit power-law distributions. A significant positive correlation was observed between inbreeding and elevation. Many Spanish populations have experienced a prolonged and considerable isolation across generations, which has led to high proportions of historical and local endogamy that is associated, in general, with high \( \overline{F} \) values. Thus, assessing genomic inbreeding using runs of homozygosity (ROH) in current Spanish populations could be an additional pertinent strategy for obtaining a more refined perspective regarding the population history inferred from the extent and frequency of ROH regions.     

Cold‐induced expression of diapause in Praon volucre: fitness cost and morpho‐physiological characterization

Cold exposure (2°C for 7 days) in constant darkness at mummy stage induces diapause expression in 9% of the Praon volucre Haliday population. Diapausing parasitoids show a significant delay in emergence time compared with nondiapausing counterparts. A diapause‐mediated polyphenism is observed in mummy colour, with diapausing mummies being clearly darker than nondiapausing ones. The diapause status of dark mummies is confirmed by a significant reduction in metabolic rate. Diapausing parasitoids also display specific morphological characteristics: they are heavier (fresh and dry mass) and accumulate larger fat reserves than nondiapausing counterparts. The diapause status is associated with a fitness cost in terms of adult longevity. There is no evidence of diapause‐related change in supercooling ability.     

Temporal trends in marital structure and isonymy in S. Paolo Albanese, Italy

In the fifteenth century, after the Turkish conquest of the Balkan area, Albanian communities migrated to Southern Italy. I investigated temporal trends in isolation from 1820 to 1982 in one of these communities, the population of S. Paolo Albanese, Basilicata, which still uses the original language and religious rites. Marital structure is characterized by a high average frequency of village endogamy (75.2%). Among the exogamous marriages there is a preference for mates from Italo-Albanian settlements, with higher values in the 1800s. The distribution of marital distances reflects the positive assortative mating by ethnic community. The mean frequency of isonymous marriages was 9.01% from 1820 to 1982. These results indicate that total inbreeding from isonymy is a reliable indicator of isolation, showing temporal trends related to changes in endogamy. Fr accounts for the greater percentage of Ft in relation to the small population size and regularly decreases with time. The breakdown of isolation, as documented by the decrease in population size, endogamy, and inbreeding, is a recent feature (since 1960).