The experiments with laboratory animals from a bioethical point of view--history, modern time, perspectives

The origin of laboratory animal science was called forth by violent development of experimental biology and medicine in the XIX century on the one hand, and on the other hand by the necessity to have standard healthy animals for experiments with strictly definite biological characteristics. With this aim in view management technology and animal use in experiments have been constantly improved. "Laboratory animal" notion has been formed by the end of the XIX century. At the beginning of laboratory animal science development ethical problems were not as urgent as they are now. It is established that the three Rs bioethical conception of W.M.S. Russel and R.L. Burch (1959) has influence on modern state and perspectives of the development of animal experimental methods. It is shown that the existence of laboratory animal protection laws and the reflection in them of compulsory ethical review of scientific project and statistics of used laboratory animals is absolutely necessary.     

Dental caries in nineteenth century Upper Canada

This study examines the presence of dental caries in a large sample of adult skeletons from the 19th century cemetery of St. Thomas' Anglican Church in Belleville, Ontario. The cemetery was used from 1821 to 1874. Caries prevalence and frequencies of diseased and missing teeth were calculated both by observing summary statistics of individual rates and by the total sample of teeth. Postmortem tooth loss is low in this sample and antemortem tooth loss is highest in first mandibular molars, all other molars and then premolars. Age at death, but not sex, was found to be significantly related to the overall Caries Rate while both age and sex were significantly associated with the Diseased-Missing Index. The increase in diseased and missing teeth in older individuals is expected while the sex difference is not explained by simple dietary factors. When compared to reports on British and American samples, caries and antemortem tooth loss in the St. Thomas' sample is most similar to a pre-1850 British group and higher than American samples. Although there is undoubtedly a complex of factors contributing to caries prevalence in this sample, more data are required from large historic samples, particularly from the American northeast and late 19th century Britain, to have a clearer understanding of the influence of diet, cultural, and environmental factors affecting caries rates in historic populations. Am J Phys Anthropol 104:71–87, 1997. © 1997 Wiley-Liss, Inc.     

Loss of identity in nineteenth-century Norway: Oslo's House of Correction

In Norway, there was considerable overlap between the poor house, the workhouse, and correctional facilities. An assemblage of human skeletal remains from Oslo's House of Correction cemetery, largely comprised of individuals disturbed during municipal development in 1989, consists of c. 314 individuals who died as inmates or residents; many were disabled or elderly. Although names were recorded at death, the cemetery lacked grave markers or a map, with no expectation families might one day reclaim relatives. All remains have been aged and sexed anthropologically, with samples collected for biomolecular investigations. Despite extensive archival documentation, including names recorded in the House of Correction church book, their identities are shrouded by time. Observations of age, injury, and disease can suggest why inmates ended up in the House of Correction cemetery. But there, individual stories end and identities are lost.     

Dietary behavior of the Middle Ages in Grenoble: application of isotopic biogeochemistry of the Saint-Laurent cemetery (XIIIth-XVth centuries, Isère, France)]

Isotopic analysis of 13C and 15N of 47 bones from Saint-Laurent de Grenoble cemetery (Grenoble, Isère), from the end of medieval period (XIIIth-XVth centuries AD) allowed to define the food status of animals with regard to the humans and to discuss the variability amongst adults. Adults who died young and those with small stature may have had diets poor in animal protein. The consumption of animal proteins was more important in the XVth than in the XIVth century and could illustrate a typical urban food economy providing its population with a more diversified diet than in rural areas.     

Demographic structure and microevolution of an Italian alpine isolated population

The isonymic method has been generally accepted in population genetic studies and surnames have been successfully used to investigate human populations as if they were genetic markers associated to the Y chromosome. In this study we analysed the microevolution dynamics of Postua, a mountain village of North Western Italy, by means of demographic methods. The uniqueness of this community is due to its past geographical and cultural isolation and to the high frequency of marriage between relatives. During the study period considered (1640–1989) the population underwent several fluctuations in size and other demographic parameters such as the endogamy, isonimy and consanguinity rates. Until the first half of the XIX century Postua appears to be a village characterised by a closed and isolated community, with high endogamy (80–90%) and isonimy (up to 34.4%). Only at the beginning of the XX century, when the population started to be subjected to significant immigration fluxes, data show a progressive reduction of the isolation. The population showed two demographic peaks, the first around the second half of the XVIII century (1639 inhabitants) and the second at the end of the XIX century (1464 inhabitants). The S/N ratio was low (0.2–0.3) and relatively constant until the beginning of the last century, and only in the last three decades of the XX century, when the population was subjected to immigration, Postua showed a significant increase in the S/N ratio values (to about 0.9). The surname frequency was constant until the 1850 when almost all surnames were the same as those already present in the XVII century. From the first half of the XIX century, the founder surnames decreased, whereas new surnames started to increase and became persistent in the population.     

Brief communication: A test and correction of the clavicle method of Stout and Paine for histological age estimation of skeletal remains

The histological method developed by Stout and Paine ([1992] Am. J. Phys. Antropol. 87:111–115) for estimating age at death using the clavicle is tested on a known age independent sample from a nineteenth century cemetery near Spitalfriedhof St. Johann in Basel, Switzerland. The mean absolute difference between reported ages and histologically predicted ages is 5.5 years. Mean predicted age for the sample is different from mean reported age. This difference is accounted for by differences in the age distributions between the original autopsy sample used to derive the histological age-predicting formula and the cemetery sample, and an inherent loss of reliability of histological age predictions for the skeletal remains of older individuals. A new formula based upon the combined original autopsy sample of Stout and Paine (1992) and the Swiss cemetery sample is presented. It is recommended that this formula be used when estimating ages for older individuals or archaeological skeletal samples. © 1996 Wiley-Liss, Inc.     

Polish pioneers in research on vertebrate circulatory systems at the Jagiellonian University of Krakow (Poland)

At the end of the XIX century and during the first half of the XX century, Henryk Hoyer and Zygmunt Grodzinski, two eminent Polish researchers, were carring out research on the lymphatic circulatory and blood circulatory systems. Later, these investigations were continued by their students and followers until 1966. Embryos from all groups of vertebrates were used for investigation and on that basis, models of the lymphatic and blood circulatory systems were proposed.     

The MAD legacy: How meaningful is mean age-at-death in skeletal samples

This study examines the representativeness of palaeodemographic reconstructions from human skeletal remains. Mean age-at-death (MAD) is the primary statistic used in interpretations of changing patterns of health and well being from palaeodemographic analyses. A series of sampling experiments were conducted on three documented 19th century samples representing the total cemetery population from which skeletal samples could be drawn. Comparisons of the age-at-death distributions of simulated skeletal samples to the parent population were made to assess the relative magnitude of deviation associated with different types of bias (age, sex, temporal). From the examples presented, variability in age-at-death distribution is high in samples of less than 100, suggesting that for samples of less than 100analyzable individuals, it is probable that the mortality profiles constructed are not an accurate reflection of the cemetery. It is proposed that whateverprocess mean age-at-death reflects for past populations (fertility or mortality), is irrelevant if the sample on which the statistic is calculated is not representative of the population. Given that most cemetery samples will be subject, differentially, to biases at a variety of levels, comparative studies based on palaeodemographic data cannot be considered reliablewithout careful control for those biases. It is suggested that representativeness is the primary theoretical obstacle for researches to overcome, and that it is necessary to shift our focus to rigorously exploring those factors that bias our samples. Without some direct quantification of the representativeness of a sample, palaeodemographic estimators such as mean age-at-death are meaningless and any subsequent interpretations regarding the past, dubious at best.     

Infectious disease among enslaved African Americans at Eaton's Estate, Warren County, North Carolina, ca. 1830-1850

The skeletal remains of 17 people buried in the Eaton Ferry Cemetery in northern North Carolina provide a means of examining health and infectious disease experience in the XIX century South. The cemetery appears to contain the remains of African Americans enslaved on the Eaton family estate from approximately 1830-1850, and thus offers a window into the biological impacts of North American slavery in the years preceding the Civil War. The sample includes the remains of six infants, one child, and one young and nine mature adults (five men, four women, and one unknown). Skeletal indices used to characterize health and disease in the Eaton Ferry sample include dental caries, antemortem tooth loss, enamel hypoplasia, porotic hyperostosis, periosteal lesions, lytic lesions, and stature. These indicators reveal a cumulative picture of compromised health, including high rates of dental disease, childhood growth disruption, and infectious disease. Specific diseases identified in the sample include tuberculosis and congenital syphilis. Findings support previous research on the health impacts of slavery, which has shown that infants and children were the most negatively impacted segment of the enslaved African American population.     

The environmental and cultural contexts of the late Iron Age and medieval settlement in the Mazurian Lake District,NE Poland: combined palaeobotanical and archaeological data

Pollen analysis of sediments from three lakes and analysis of plant macroremains including charcoal from archaeological sites in the Mazurian Lake District provide new data for the reconstruction of vegetation changes related to human activity between the 1st and 13th century ad. At that time settlements of the Bogaczewo culture (from the turn of the 1st century ad to the first part of the 5th century ad), the Olsztyn Group (second part of the 5th century ad to the 7th or beginning of the 8th century ad), and the Prussian Galinditae tribes (8th/9th–13th century ad) developed. The most intensive woodland clearing occurred between the 1st and 6th/7th century ad. Presence of Cerealia-type, Secale cereale and Cannabis-type pollen, as well as macroremains of Hordeum vulgare, S. cereale, Triticum spelta, T. cf. monococcum, T. cf. dicoccum, Avena sp. and Panicum miliaceum documented local agriculture. High Betula representation synchronous with microcharcoal occurrence suggests shifting agriculture. After forest regeneration between c. ad 650 and 1100, the area was strongly deforested due to the early medieval occupation by Prussian tribes. The archaeobotanical examination of samples taken in a cemetery and a large settlement of the Roman Iron Age revealed strong differences in the taxonomic composition of the fossil plant remains. An absolute dominance of birch charcoal in the samples from the cemetery indicates its selective use for funeral pyre construction. There is a difference between cereals found in both contexts: numerous grains of Triticum have been found in the cemetery, while in the settlement crops were represented mostly by Secale and Hordeum. Grass tubers, belonging probably to Phleum pratense, are among the particularly interesting plant remains found in the cemetery.     

Up-regulation of type XIX collagen in rhabdomyosarcoma cells accompanies myogenic differentiation

Rhabdomyosarcomas are known to recapitulate some of the early events in skeletal muscle embryogenesis, and cultures derived from these tumors have been extensively used to elucidate processes associated with the differentiation of primitive mesenchymal cells. These neoplasms have also provided important systems for studying different collagen types. This aspect is particularly relevant to type XIX collagen, which was originally identified from rhabdomyosarcoma cDNA clones. Although this collagen has been localized in vivo to basement membrane zones in a wide variety of tissues, including skeletal muscle, the tumor cells appear to be a unique source of its expression in vitro. We have found that one particular cell line-derived from a peritesticular embryonal rhabdomyosarcoma-produced relatively large amounts of type XIX collagen, especially in those rare instances in which these cells appear to spontaneously differentiate. To characterize this phenomenon, tumor cells were grown under conditions known to induce differentiation in normal myoblast cultures. In response to this treatment, the typical tumor cell morphology consistently and reproducibly switched from polygonal to round/spindle-shaped with the subsequent appearance of some structures resembling myotubes. Concurrently, the cultures commenced a dramatic up-regulation of type XIX collagen and skeletal muscle myosin heavy chain and alpha-actinin in a time-dependent fashion, whereas protein and mRNA levels of other matrix proteins were either decreased or unchanged. Moreover, immunocytochemical analysis revealed that only a subpopulation of the cells was responsible for the increased synthesis of type XIX collagen, alpha-actinin, and myosin, and that the same cells which stained positive for the collagen also stained positive for the muscle proteins. Taken together, the results suggested that type XIX collagen may be involved in the initial stages of skeletal muscle cell differentiation.     

Sanitary housing conditions modify the performance and behavioural response of weaned pigs to feed- and housing-related stressors

Pigs are confronted with changes in farming practices that may affect performance and animal well-being. The sanitary conditions of the farm can have an impact on the ability of pigs to adapt to these changes. This study aimed to analyse how weaned pigs respond to common farming practices of changes in diet and housing in terms of performance, health and behaviour, and how these responses are affected by the sanitary housing conditions, qualified here as good or poor. At weaning at 4 weeks of age, 20 piglets were assigned to 10 blocks of two littermates and each pig within a litter was randomly assigned to one of two sanitary conditions. Pigs were housed individually and received a starter diet. A diet change occurred on day 12 post weaning (starter to weaner diets) and pigs were transferred to the grower unit on day 33 post weaning and continued to receive the weaner diet. From 43 days post weaning, pigs were offered a grower diet and were vaccinated against swine influenza on day 47 and 61 post weaning. On the basis of this design, three post-weaning phases were identified: phase I from day 1 to 11 (post weaning), phase II from day 12 to 32 (after the diet change) and phase III from day 33 to 42 (after the housing change). Individual BW was measured every 3 days, and feed refusals and faecal scores were recorded on a daily basis. Behavioural observations were performed during 28 days by using the instantaneous scan sampling method. Individual blood samples were collected at the end of each phase to analyse the plasma concentration of haptoglobin and on day 68 post weaning to analyse the anti-influenza immunoglobulins G (IgG). Poor sanitary conditions resulted in a decrease in daily gain, feed intake and gain to feed ratio of, respectively, 11%, 5% and 7% (P < 0.05). Pigs in poor sanitary conditions had higher faecal scores (P < 0.05), tended to have higher plasma haptoglobin concentration in phase II (P = 0.06) and had a higher anti-influenza IgG titre (P = 0.11). The diet change affected performance and behavioural responses of pigs in poor but not in good sanitary conditions. Housing change resulted in a 30% decrease in growth and an increase in behaviour oriented towards exploration and excitement. The results of this study show an effect of sanitary conditions on the responses of pigs to a diet change, whereas those to a housing change were little affected by the sanitary conditions.     

Fluctuating asymmetry and stress in a medieval Nubian population

Fluctuating asymmetry is commonly used as a bioindicator of developmental stress. This study addresses asymmetry under nutritional/systemic stress in the human craniofacial skeleton and its utility as an indicator of developmental instability. Crania from the diachronic Christian cemeteries at Kulubnarti (Sudanese Nubia) were chosen as a model for nutrition/systemic stress. Previous studies indicate that individuals from the Early Christian cemetery were subjected to greater developmental stress when compared with individuals from the Late Christian cemetery. Therefore, crania from the Early Christian cemetery should display a greater magnitude of fluctuating asymmetry than crania from the Late Christian cemetery. Thirty adult crania of comparable age and sex were selected from each population. Landmark coordinates were digitized in two separate trials and averaged to minimize error. Euclidean distance matrix analysis (EDMA) was used to measure and compare the magnitude of fluctuating asymmetry in each sample. Results indicate that crania from the Early Christian cemetery display greater amounts of fluctuating asymmetry than those from the Late Christian cemetery, as predicted. The degree of fluctuating asymmetry for each linear distance is highly correlated between the cemeteries, suggesting that all humans may share common patterns of fluctuating asymmetry in the skull. In contrast, there is little correlation between magnitude of fluctuating asymmetry and length of linear distance, between-subject variability, or measurement error. These results support the hypothesis that poor nutrition/systemic stress increases developmental instability in the human skull and that increased fluctuating asymmetry constitutes morphological evidence of this stress.     

Prevalence and characterization of Clostridium perfringens from spices in Argentina

Spices can present high microbial counts and Clostridium perfringens, Bacillus cereus, Salmonella and Shigella, among others have been isolated from spices. C. perfringens is an important pathogen agent causing, among other diseases, enteritis in humans caused by C. perfringens enterotoxin (CPE) which causes human food poisoning and enterotoxemia in domestic animals. The aims of the present work were (i) to establish the hygienic sanitary quality of some spices in San Luis, Argentina; (ii) to determine the presence of C. perfringens in these spices by means of the most probable number (MPN) and count on plate methods; (iii) to characterize the enterotoxigenic strains of C. perfringens by PCR and immunological methods such as reverse passive latex agglutination (RPLA) and (iv) to type by PCR C. perfringens strains isolated. A total of 115 samples of spices, 67 of which were purchased in local retail stores and 48 domestically collected were analysed. Total aerobe counts on tryptone glucose yeast extract agar medium of the 115 samples were between <10 and 10(6) CFU/g. The colifecal counts using Mac Conkey broth of the 115 samples were <4-10(3)CFU/g, with 28 samples (24.34%) exceeding the limit established by the Spanish Alimentary Code (10 CFU/g) while 2 samples (1.73%) had a sulfite reducing anaerobe load above standard limits. A total of 14 C. perfringens strains (12.17%) were isolated and characterized from 115 samples by the standard biochemical tests. Four of which (28.60%) turned out to be enterotoxigenic by PCR and RPLA. In order to type C. perfringens strains based on their main toxins, the 14 strains were analysed by PCR. All strains belonged to type A. All RPLA positive strains were cpe(+) by PCR. The percentage of enterotoxigenic strains was more elevated that those reported in other studies for this type of sample. These results indicate that sanitary conditions in different production stages of species must be improved to reduce health hazards. The high percentage (24.34%) of samples with colifecal values above standard limits is an indication of deficient sanitary conditions. These results suggest the need to provide legislation on the sanitary and hygienic quality of spices in our country.     

The “proletarian disease” on stage. Theatrical anti-tuberculosis propaganda in the early Soviet Union

The present contribution analyses sanitary theatrical performances as a means of anti-tuberculosis propaganda in the early Soviet Union. Starting in the 1920s, sanitary theatrical performances were demonstrated in open-air theatres and clubhouses for workers and farmers. Since 1925, the newly founded Moscow Theatre for Sanitary Culture centrally managed the theatrical hygiene propaganda. It became a role model for other theatres of hygienic enlightenment and numerous sanitary amateur stages. Their anti-tuberculosis repertoire ranged from the so-called “mock trials” where a person or even Koch's bacillus must stand trial for the spreading of tuberculosis, to “living newspapers” which used entertainment elements such as music or acrobatics to provide a mass audience with the hygiene knowledge. The contribution describes in which images, figures and actions knowledge about tuberculosis was presented on stage, which genre traditions and communicative instruments were used and which changing political implications those performances were based on. To achieve this goal, the archive sources, selected texts of theatrical performances, reports and reviews in daily press have been evaluated.     

Problems in determination of skeletal lead burden in archaeological samples: an example from the First African Baptist Church population

Human bone lead content has been demonstrated to be related to socioeconomic status, occupation and other social and environmental correlates. Skeletal tissue samples from 135 individuals from an early nineteenth century Philadelphia cemetery (First African Baptist Church) were studied by electrothermal atomic absorption spectrometry and X-ray fluorescence for lead content. High bone lead levels led to investigation of possible diagenetic effects. These were investigated by several different approaches including distribution of lead within bone by X-ray fluorescence, histological preservation, soil lead concentration and acidity as well as location and depth of burial. Bone lead levels were very high in children, exceeding those of the adult population that were buried in the cemetery, and also those of present day adults. The antemortem age-related increase in bone lead, reported in other studies, was not evidenced in this population. Lead was evenly deposited in areas of taphonomic bone destruction. Synchrotron X-ray fluorescence studies revealed no consistent pattern of lead microdistribution within the bone. Our conclusions are that postmortem diagenesis of lead ion has penetrated these archaeological bones to a degree that makes their original bone lead content irretrievable by any known method. Increased bone porosity is most likely responsible for the very high levels of lead found in bones of newborns and children.     

Detection of SV40 DNA sequences in malignant mesothelioma specimens from the United States, but not from Turkey

The incidence of malignant mesothelioma (MM) shows a strong epidemiological association with exposure to asbestos fibers. Recently, simian virus 40 (SV40) DNA sequences have been reported in MM tumor specimens from the United States and several European countries, and the SV40 tumor virus has been implicated as a potential co-factor in the etiology of this disease. However, several large studies from the US, Finland, and Turkey did not detect SV40 sequences in MM samples. To address this discrepancy, MM specimens from Turkey and the US were analyzed in the same laboratory under identical conditions to detect the presence of SV40 DNA. We detected SV40 sequences in 4 of 11 specimens from the United States, but in none of the 9 Turkish samples examined. These findings suggest that geographical differences exist with regard to the involvement of SV40 in human tumors.     

PCR diagnosis of Entamoeba histolytica cysts in stool samples

Amebiasis is a protozoan disease caused by Entamoeba histolytica and a potential health threat in areas where sanitation and hygiene are inappropriate. Highly sensitive PCR methods for detection of E. histolytica in clinical and environmental samples are extremely useful to control amebiasis and to promote public health. The present study compared several primer sets for small subunit (SSU) rDNA and histone genes of E. histolytica cysts. A 246 bp of the SSU rDNA gene of pure cysts contained in phosphate-buffered saline (PBS) and in stool samples was successfully amplified by nested PCR, using the 1,147-246 bp primer set, of the primary PCR products which were pre-amplified using the 1,147 bp primer as the template. The detection limit of the nested PCR using the 1,147-246 primer set was 10 cysts in both groups (PBS and stool samples). The PCR to detect histone gene showed negative results. We propose that the nested PCR technique to detect SSU rDNA can be used as a highly sensitive genetic method to detect E. histolytica cysts in stool samples.     

Bacteriological Survey of the Frozen Prepared Foods Industry: IV. Frozen Breaded Fish

During sanitation-inspections of 23 breaded-fish processors, 573 finished product units and 604 line samples were collected and analyzed bacteriologically. Data for finished product units processed on the same data are presented in groups; most of the groups consisted of 10 units. Groups of frozen, raw-breaded fish produced under good and poor conditions of sanitation were studied. No more than 20% of the units in each group produced under good conditions of sanitation were positive for Escherichia coli or coagulase-positive staphylococci. Groups of frozen, fried-breaded fish produced under good and poor sanitary conditions were also studied. No more than 10% of the units in each good-sanitation group were positive for E. coli or coagulase-positive staphylococci. However, approximately 30% of the groups processed under poor sanitary conditions also did not exceed these viable bacterial counts because of the lethal effect of the terminal fry to which the product is subjected. It was found that line samples collected from the processing lines reflected the sanitary conditions of the plant and provided bacteriological support for inspectional evidence of plant insanitation.     

The Emergence of Mesolithic Cemeteries in SW Europe: Insights from the El Collado (Oliva,Valencia, Spain) Radiocarbon Record

Located on the Iberian Mediterranean coast, El Collado is an open-air site where a rescue excavation was conducted over two seasons in 1987 and 1988. The archaeological work excavated a surface area of 143m2 where 14 burials were discovered, providing skeletal remains from 15 individuals. We have obtained AMS dates for 10 of the 15 individuals by means of the direct dating of human bones. The ranges of the probability distribution of the calibrated dates suggest that the cemetery was used during a long period of time (781–1020 years at a probability of 95.4%). The new dates consequently set back the chrono-cultural attribution of the cemetery from the initial proposal of Late Mesolithic to an older date in the Early Mesolithic. Therefore, El Collado becomes the oldest known cemetery in the Iberian Peninsula, earlier than the numerous Mesolithic funerary contexts documented on the Atlantic façade such as the Portuguese shell-middens in the Muge and Sado Estuaries or the funerary sites on the northern Iberian coast.