Identification of blood-feeding sources in Panstrongylus, Psammolestes, Rhodnius and Triatoma using amplicon-based next-generation sequencing.

BACKGROUND: Triatomines are hematophagous insects that play an important role as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. These insects have adapted to multiple blood-feeding sources that can affect relevant aspects of their life-cycle and interactions, thereby influencing parasitic transmission dynamics. We conducted a characterization of the feeding sources of individuals from the primary circulating triatomine genera in Colombia using amplicon-based next-generation sequencing (NGS). METHODS: We used 42 triatomines collected in different departments of Colombia. DNA was extracted from the gut. The presence of T. cruzi was identified using real-time PCR, and discrete typing units (DTUs) were determined by conventional PCR. For blood-feeding source identification, PCR products of the vertebrate 12S rRNA gene were obtained and sequenced by next-generation sequencing (NGS). Blood-meal sources were inferred using blastn against a curated reference dataset containing the 12S rRNA sequences belonging to vertebrates with a distribution in South America that represent a potential feeding source for triatomine bugs. Mean and median comparison tests were performed to evaluate differences in triatomine blood-feeding sources, infection state, and geographical regions. Lastly, the inverse Simpson's diversity index was calculated. RESULTS: The overall frequency of T. cruzi infection was 83.3%. TcI was found as the most predominant DTU (65.7%). A total of 67 feeding sources were detected from the analyses of approximately 7 million reads. The predominant feeding source found was Homo sapiens (76.8%), followed by birds (10.5%), artiodactyls (4.4%), and non-human primates (3.9%). There were differences among numerous feeding sources of triatomines of different species. The diversity of feeding sources also differed depending on the presence of T. cruzi. CONCLUSIONS: To the best of our knowledge, this is the first study to employ amplicon-based NGS of the 12S rRNA gene to depict blood-feeding sources of multiple triatomine species collected in different regions of Colombia. Our findings report a striking read diversity that has not been reported previously. This is a powerful approach to unravel transmission dynamics at microgeographical levels.

[Results of a study of parvovirus B19 (Parvoviridae, Parvovirinae, Erythroparvovirus, Primate erythroparvovirus 1) prevalence and circulation activity in socially significant categories of the population].

Currently, along with the increasing need of medical organizations for blood preparations, algorithms for laboratory testing of blood donors are not available for all infections with hemo-contact mechanism of transmission. A representative example is infection caused by parvovirus В19. PURPOSE OF THE STUDY: The article presents the results of the original study, the purpose of which was to study the prevalence of antibodies to parvovirus B19 and the activity of the circulation of this virus in socially important categories of the population. MATERIAL AND METHODS: The materials of the study were blood samples from blood donors of Saint Petersburg, as well as parvovirus В19 sequences isolated from DNA-positive plasma samples. RESULTS AND DISCUSSION: According to the results of the laboratory examination, a high proportion of carriers of virus-specific IgG antibodies was found in studied group of donors, which confirms the previous infection of parvovirus B19 in them and illustrates the high prevalence of infection in this socially significant group. Based on the results of the blood preparations testing, the presence of parvovirus DNA В19 in a significant number of samples was determined by polymerase chain reaction method. This indicates an current parvovirus infection in the examined donors and points to a high epidemiological risk of the blood products obtained from them. Sequencing and phylogenetic analysis of a fragment of the VP1 gene demonstrated that the studied isolates belonged to А1 genotype and its subtype 1А2, which correlates with the genotypes of parvovirus В19 circulating in the European Union and Asia. In addition, two previously unknown В19 parvovirus isolates were isolated, the nucleotide sequences of which were deposited into the international GenBank database. CONCLUSION: Based on the results of the study, it is justified to include testing of blood samples for markers of В19 parvovirus infection in existing algorithms of laboratory examination of donors, which will ensure prevention of hemo-contact infection of blood recipients with parvovirus В19.

SEROLOGICAL DIAGNOSIS OF BAYLISASCARIS PROCYONIS IN PRIMATES USING A HUMAN ELISA TEST.

The usefulness of a human enzyme-linked immunosorbent assay (ELISA) for serological diagnosis of Baylisascaris procyonis larva migrans was assessed in nonhuman primates (NHP). The test was originally developed as an assay performed on human samples at Purdue University. Six participating zoos submitted 258 NHP serum samples, spanning these major phylogenetic groups: 1) great apes (n = 84), 2) lesser apes (n = 17), 3) Old World monkeys (n = 84), 4) New World monkeys (n = 20), and 5) prosimians (n = 53). Sera were tested in duplicate using a microtiter-well ELISA with B. procyonis larval excretory-secretory proteins as antigen, and serum from an experimentally infected baboon (Papio anubis) served as positive control. The ELISA clearly identified seropositive animals in all zoos. With putative cutoffs of optical density (OD) measured at 405 nm (OD405) of <0.150 = negative, 0.150-0.250 = indeterminate, and >0.250 = positive, 149 of 258 (57.8%) were clearly negative (mean OD 0.046), and 78 of 258 (30.2%) were clearly positive (mean OD 0.657, range 0.253-1.773), the rest being indeterminate. Of these, 15 were high positive with OD 1.095-1.773 (mean 1.314). Positive animals were seen from all zoos; 76 (97.4%) were great apes, lesser apes, or Old World monkeys. The four highest ODs were in a siamang (Symphalangus syndactylus), lion-tailed macaque (Macaca silenus), Sumatran orangutan (Pongo abelii), and western lowland gorilla (Gorilla gorilla gorilla), all from different zoos. Prosimians had a mean OD of 0.039 and New World monkeys 0.021, indicating that human reagents either did not work for these groups or few infected animals were represented. These results indicate that the human ELISA for B. procyonis works well for at least higher phylogeny NHP and that serologic evidence of infection is surprisingly common, correlating with what is known for exposure to this parasite in zoos.

Nonhuman Primates with Acute Radiation Syndrome: Results from a Global Serum Metabolomics Study after 7.2 Gy Total-Body Irradiation.

Threats of nuclear terrorism coupled with potential unintentional ionizing radiation exposures have necessitated the need for large-scale response efforts of such events, including high-throughput biodosimetry for medical triage. Global metabolomics utilizing mass spectrometry (MS) platforms has proven an ideal tool for generating large compound databases with relative quantification and structural information in a short amount of time. Determining metabolite panels for biodosimetry requires experimentation to evaluate the many factors associated with compound concentrations in biofluids after radiation exposures, including temporal changes, pre-existing conditions, dietary intake, partial- vs. total-body irradiation (TBI), among others. Here, we utilize a nonhuman primate (NHP) model and identify metabolites perturbed in serum after 7.2 Gy TBI without supportive care [LD70/60, hematologic (hematopoietic) acute radiation syndrome (HARS) level H3] at 24, 36, 48 and 96 h compared to preirradiation samples with an ultra-performance liquid chromatography quadrupole time-of-flight (UPLC-QTOF) MS platform. Additionally, we document changes in cytokine levels. Temporal changes observed in serum carnitine, acylcarnitines, amino acids, lipids, deaminated purines and increases in pro-inflammatory cytokines indicate clear metabolic dysfunction after radiation exposure. Multivariate data analysis shows distinct separation from preirradiation groups and receiver operator characteristic curve analysis indicates high specificity and sensitivity based on area under the curve at all time points after 7.2 Gy irradiation. Finally, a comparison to a 6.5 Gy (LD50/60, HARS level H2) cohort after 24 h postirradiation revealed distinctly increased separations from the 7.2 Gy cohort based on multivariate data models and higher compound fold changes. These results highlight the utility of MS platforms to differentiate time and absorbed dose after a potential radiation exposure that may aid in assigning specific medical interventions and contribute as additional biodosimetry tools.

Utility of arterial blood gas, CBC, biochemistry and cardiac hormones as evaluation parameters of cardiovascular disease in nonhuman primates.

Cardiovascular disease (CVD) has a tremendous impact on the quality of life of humans. While experimental animals are valuable to medical research as models of human diseases, cardiac systems differ widely across various animal species. Thus, we examined a CVD model in cynomolgus monkeys. Laboratory primates are precious resources, making it imperative that symptoms of diseases and disorders are detected as early as possible. Thus, in this study we comprehensively examined important indicators of CVD in cynomolgus monkeys, including arterial blood gas, complete blood count (CBC), biochemistry and cardiac hormones. The control group included 20 healthy macaques showing non-abnormal findings in screening tests, whereas the CVD group included 20 macaques with valvular disease and cardiomyopathy. An increase of red blood cell distribution width was observed in the CBC, indicating chronic inflammation related to CVD. An increase of HCO3 was attributed to the correction of acidosis. Furthermore, development of the CVD model was supported by significant increases in natriuretic peptides. It is suggested that these results indicated a correlation between human CVD and the model in monkeys. Moreover, blood tests including arterial blood gas are non-invasive and can be performed more easily than other technical tests. CVD affected animals easily change their condition by anesthesia and surgical invasion. Pay attention to arterial blood gas and proper respond to their condition are important for research. This data may facilitate human research and aid in the management and veterinary care of nonhuman primates.

Scaling of bony canals for encephalic vessels in euarchontans: Implications for the role of the vertebral artery and brain metabolism.

Supplying the central nervous system with oxygen and glucose for metabolic activities is a critical function for all animals at physiologic, anatomical, and behavioral levels. A relatively proximate challenge to nourishing the brain is maintaining adequate blood flow. Euarchontans (primates, dermopterans and treeshrews) display a diversity of solutions to this challenge. Although the vertebral artery is a major encephalic vessel, previous research has questioned its importance for irrigating the cerebrum. This presents a puzzling scenario for certain strepsirrhine primates (non-cheirogaleid lemuriforms) that have reduced promontorial branches of the internal carotid artery and no apparent alternative encephalic vascular route except for the vertebral artery. Here, we present results of phylogenetic comparative analyses of data on the cross-sectional area of bony canals that transmit the vertebral artery (transverse foramina). These results show that, across primates (and within major primate subgroups), variation in the transverse foramina helps significantly to explain variation in forebrain mass even when variation in promontorial canal cross-sectional areas are also considered. Furthermore, non-cheirogaleid lemuriforms have larger transverse foramina for their endocranial volume than other euarchontans, suggesting that the vertebral arteries compensate for reduced promontorial artery size. We also find that, among internal carotid-reliant euarchontans, species that are more encephalized tend to have a promontorial canal that is larger relative to the transverse foramina. Tentatively, we consider the correlation between arterial canal diameters (as a proxy for blood flow) and brain metabolic demands. The results of this analysis imply that human investment in brain metabolism (∼27% of basal metabolic rate) may not be exceptional among euarchontans.

Enzootic Circulation of Chikungunya Virus in East Africa: Serological Evidence in Non-human Kenyan Primates.

Chikungunya virus (CHIKV) is a globally emerging pathogen causing debilitating arthralgia and fever in humans. First identified in Tanzania (1953), this mosquito-borne alphavirus received little further attention until a 2004 re-emergence in Kenya from an unknown source. This outbreak subsequently spread to the Indian Ocean, with adaptation for transmission by a new urban vector. Under the hypothesis that sylvatic progenitor cycles of CHIKV exist in Kenya (as reported in West Africa, between non-human primates (NHPs) and arboreal Aedes spp. mosquitoes), we pursued evidence of enzootic transmission and human spillover events. We initially screened 252 archived NHP sera from Kenya using plaque reduction neutralization tests. Given an overall CHIKV seroprevalence of 13.1% (marginally higher in western Kenya), we sought more recent NHP samples during 2014 from sites in Kakamega County, sampling wild blue monkeys, olive baboons, and red-tailed monkeys (N = 33). We also sampled 34 yellow baboons near Kwale, coastal Kenya. Overall, CHIKV seropositivity in 2014 was 13.4% (9/67). Antibodies reactive against closely related o'nyong-nyong virus (ONNV) occurred; however, neutralization titers were too low to conclude ONNV exposure. Seroprevalence for the flavivirus dengue was also detected (28%), mostly near Kwale, suggesting possible spillback from humans to baboons. CHIKV antibodies in some juvenile and subadult NHPs suggested recent circulation. We conclude that CHIKV is circulating in western Kenya, despite the 2004 human outbreaks only being reported coastally. Further work to understand the enzootic ecology of CHIKV in east Africa is needed to identify sites of human spillover contact where urban transmission may be initiated.

Hematocrit of mammals (Artiodactyla, Carnivora, Primates) at 1500m and 2100m altitudes.

The rise in hematocrit (Hct) is one of the hallmarks of human acclimatization to high altitude and, in chronic conditions, reflects the hypoxia-induced polycythemia. However, it is not a uniform response among domestic species and it is not found in Andean camelids, species long adapted to high altitudes. Hence, we asked to what extent the polycythemia of humans is common among mammals. Hct data were collected from captive mammals of three orders (Primates, Artiodactyla, Carnivora), 70 specimens of 33 species at ∼1500m altitude (barometric pressure Pb=635mmHg) and 296 specimens of 64 species at ∼2100m (Pb=596mmHg), long-term residents at those altitudes. Sea level values and data in men and women at the corresponding altitudes were from a compilation of literature sources. At either altitude Hct was significantly higher than at sea level both in men and women; the increase (ΔHct) for genders combined averaged 3.4% (±0.7 SEM) at 1500m and 5.4% (±0.3) at 2100m. Differently, among the three mammalian orders studied a significant increase in Hct occurred only in females of Carnivora (at 1500m) and in males of Primates (at 2100m). The average ΔHct of all species combined was 0.8% (±0.7) at 1500m and 1.5% (±0.4) at 2100m, both significantly less than in humans (P<0.001). At 2100m the average ΔHct of nine species long adapted to high altitude was 0.4% (±1), significantly less than in non-adapted species (P<0.001). A polycythemic response like that of men and women at 2100m occurred in less than 10% of the mammals examined. We conclude that, at least for the altitudes studied, a minimal polycythemia is a general feature of both high-altitude adapted and non-adapted species, and the magnitude of the human response is exceptional among mammals.

Avaliação da infecção por leptospiras patogênicas em primatas silvestres mantidos em cativeiro em Salvador, Bahia, Brasil / Evaluation of infection by pathogenic Leptospira in wild primates kept in captivity in Salvador, Bahia, Brazil

Estima-se que 70% das doenças infecciosas emergentes em humanos estão relacionados a exposição a animais silvestres. Estudos envolvendo infecções em primatas não humanos e o seu papel na epidemiologia da leptospirose são escassos.Uma instrução normativa do Ministério do Meio Ambiente(179 de 25/06/2008)recomenda o uso de teste de microaglutinação (MAT) para detecção de anticorpos aglutinantes anti-Leptospira em espécies silvestres em programas de reintrodução à vida livre. Neste contexto, o presente projeto propôs investigar a evidência de prévia exposição e a ocorrência do estado de portador de leptospira patogênicas em primatas silvestres residentes no Parque Zoobotânico Getúlio Vargas de Salvador (PZBGV) e recebidos pelo Centro de Triagem de Animais Silvestres Chico Mendes (CETAS). Foram analisados 42 amostras de soro e sangue total de 42 primatas do PZBGV e 16 amostras de soros de 16 primatas do CETAS pelo teste de aglutinação microscópica (MAT) e PCR respectivamente. Amostras de urina de 3 de primatas do PZBGV e 13 do CETAS também foram submetidas à PCR. A soroprevalência encontrada no PZGV foi de 2%(1/42)enquanto que no CETAS foi de 31%(5/16). O único primata do PZBGV positivo na MAT foi um Allouata caraya (bugio preto) fêmea, adulta, que apresentou reação mista para os sorogrupos Australis e Icterohaemorrhagiae. As cinco amostras positivas do CETAS ocorreram em macacos-prego(Cebus sp.)e foram distribuídos nos sorogrupos: Ballum(1:100), Semaranga (1:200), Grippotyphos (1:100),Cynopeteri(1:100) e uma reação mista Tarassovi / Autumnalis (1:100). Todas as amostras de sangue total e urina analisadas por PCR foram negativas. Conclui-se que a soroprevalência de anticorpos anti-Leptospira foi baixa no PZBGV de Salvador, apesar da alta frequência de roedores na área e endemicidade da leptospirose humana em Salvador. A prevalência elevada foi observada entre os animais resgatados do comércio ilegal no estado da Bahia e esta evidência sorológica de exposição sugere um risco potencial de transmissão da leptospirose ao se adotar estes primatas como animais de estimação

It is estimated that 70% of emerging infectious diseases in humans are related to exposure to wild animals. Studies involving infections in nonhuman primates and their role in the epidemiology of leptospirosis are scarce. A normative statement of the Ministry of the Environment (179 of 25/06/2008) recommends the use of microscopic agglutination test (MAT) for antibodies binding anti-Leptospira in wild species reintroduction programs in the wild. Positive tests indicate the need for quarantine and antimicrobial treatment. In this context, this project proposes to investigate the evidence of prior exposure and the occurrence of carrier status of pathogenic Leptospira in wild primates living in the Parque Vargas Zoobotânico Salvador (PZBGV) and received by the Center for Wildlife Screening Chico Mendes (CETAS). We analyzed 42 serum samples and 42 whole blood PZBGV primates and 16 sera from 16 primates CETAS by microscopic agglutination test (MAT) and PCR respectively. Urine samples of 3 primates PZBGV CETAS and 13 were also subjected to PCR. The seroprevalence in PZBGV was 2% (1/42) while the CETAS was 31% (5/16). The only positive was a primate Allouata caraya (black howler monkey) female, adult, which showed mixed reaction to serogroups Australis and Icterohaemorrhagiae. The five samples positive from CETAS occurred in monkeys (Cebus sp.) And were divided into serogroups: Ballum (1:100), Semaranga (1:200), Grippotyphosa (1:100), Cynopeteri (1:100) and mixed reaction Tarassovi / Autumnalis (1:100). All blood samples and urine samples were analyzed by PCR negative. It is concluded that the seroprevalence of anti- Leptospira antibodies was low in the Zoo of Salvador, despite the high frequency of rodents in the area and endemicity of leptospirosis in Salvador...

Leaky gut, clotting, and vasculopathy in SIV.

In this issue of Blood, Pandrea et al provide a mechanistic link between SIV-associated microbial translocation and both thrombotic and cardiovascular disease in the pigtail macaque model.

Coagulation biomarkers predict disease progression in SIV-infected nonhuman primates.

HIV infection is associated with increased risk of cardiovascular complications, the underlying mechanism of which remains unclear. Plasma levels of the coagulation biomarker D-dimer (DD) correlate with increased mortality and cardiovascular events in HIV-infected patients. We compared the incidence of cardiovascular lesions and the levels of the coagulation markers DD and thrombin antithrombin in pathogenic SIV infections of rhesus and pigtailed macaques (PTMs) and in nonpathogenic SIV infection of African green monkeys (AGMs) and sooty mangabeys. Hypercoagulability and cardiovascular pathology were only observed in pathogenic SIV infections. In PTMs infected with SIV from AGMs (SIVagm), DD levels were highly indicative of AIDS progression and increased mortality and were associated with cardiovascular lesions, pointing to SIVagm-infected PTMs as an ideal animal model for the study of HIV-associated cardiovascular disease. In pathogenic SIV infection, DD increased early after infection, was strongly correlated with markers of immune activation/inflammation and microbial translocation (MT), and was only peripherally associated with viral loads. Endotoxin administration to SIVagm-infected AGMs (which lack chronic SIV-induced MT and immune activation) resulted in significant increases of DD. Our results demonstrate that hypercoagulation and cardiovascular pathology are at least in part a consequence of excessive immune activation and MT in SIV infection.

Plasma proteomic alterations in non-human primates and humans after chronic alcohol self-administration.

Objective diagnostics of excessive alcohol use are valuable tools in the identification and monitoring of subjects with alcohol use disorders. A number of potential biomarkers of alcohol intake have been proposed, but none have reached widespread clinical usage, often due to limited diagnostic sensitivity and specificity. In order to identify novel potential biomarkers, we performed proteomic biomarker target discovery in plasma samples from non-human primates that chronically self-administer high levels of ethanol. Two-dimensional difference in-gel electrophoresis (2D-DIGE) was used to quantify plasma proteins from within-subject samples collected before exposure to ethanol and after 3 months of excessive ethanol self-administration. Highly abundant plasma proteins were depleted from plasma samples to increase proteomic coverage. Altered plasma levels of serum amyloid A4 (SAA4), retinol-binding protein, inter-alpha inhibitor H4, clusterin, and fibronectin, identified by 2D-DIGE analysis, were confirmed in unmanipulated, whole plasma from these animals by immunoblotting. Examination of these target plasma proteins in human subjects with excessive alcohol consumption (and control subjects) revealed increased levels of SAA4 and clusterin and decreased levels of fibronectin compared to controls. These proteins not only serve as targets for further development as biomarker candidates or components of biomarker panels, but also add to the growing understanding of dysregulated immune function and lipoprotein metabolism with chronic, excessive alcohol consumption.

Parameters of the cytokine system functioning in laboratory primates.

The cytokine status (IFN, IL, etc.) of different monkey species (M. mulatta, P. hamadryas, C. aethiops) was studied. The interferon status is determined by the following parameters: IFN content in circulating blood and production of IFN-alpha and IFN-gamma by lymphocytes after appropriate in vitro induction. The interferon status of monkeys is similar to that of humans. The capacity to produce IFN reduces with age. It was found that genes of virtually all studied cytokines are expressed in blood cells and hence, in immune system cells.

Development, application, and quality control of serology assays used for diagnostic monitoring of laboratory nonhuman primates.

The careful development, validation, and implementation of serodiagnostic assays can provide reliable results that make them a valuable tool in microbial quality control for nonhuman primates. This article includes identification and description of the components of assay development, including formulas for calculating the number of positive serum samples needed for assay validation and methods for calculating their diagnostic sensitivity and specificity. To ensure that assays are performing within predetermined specifications, there must be a quality control system that includes appropriate system and sample suitability controls as well as mechanisms to track assay performance over time. The section on quality assurance includes definitions of precision and accuracy in assay performance, and how to interpret these two factors using the Levey-Jennings chart, Westgard's rules, and other monitoring methods. Because all serologic assays are prone to false positive and false negative results, it is essential to interpret all diagnostic test results using both the expected prevalence of disease in the population and the population-specific assay performance characteristics that are determined during assay validation. The discussion on interpreting diagnostic test results also includes guidelines for calculating the positive and negative predictive values of an assay and for interpreting results based on the disease prevalence of the test population. A glossary provides definitions of commonly used terms.

Divergent expression of cellular prion protein on blood cells of human and nonhuman primates.

BACKGROUND: Four recent transmissions of variant Creutzfeldt-Jakob disease infection by transfusion highlight the need for detailed understanding of blood-related prion pathogenesis. Nonhuman primates are the most relevant models of human prion diseases. STUDY DESIGN AND METHODS: Quantitative flow cytometry with monoclonal antibodies FH11, 3F4, and 6H4 against different parts of the normal cellular form of the prion protein (PrP(C)) was used to evaluate its expression on blood cells of humans, chimpanzees, cynomolgus macaques, rhesus macaques, squirrel monkeys, and microcebe lemurs. RESULTS: Chimpanzees, rhesus macaques, and squirrel monkeys displayed a much higher quantity of total blood cell membrane PrP(C) than humans, due to a markedly higher expression of PrP(C) on their red blood cells (RBCs). In contrast, cynomolgus macaques and lemurs demonstrated substantially lower levels of membrane PrP(C) due to the lack of significant PrP(C) expression on RBCs and platelets (PLTs). All species displayed PrP(C) on white blood cells (WBCs), with the highest levels found on human cells. Only humans, chimpanzees, and to a lesser degree rhesus macaques expressed PrP(C) on PLTs. CONCLUSION: If PrP(C) contributes to the propagation or transport of prion infectivity in blood, the differences reported here need to be considered when extrapolating results of transmission studies in primate models to blood and blood components in humans.

Bare skin, blood and the evolution of primate colour vision.

We investigate the hypothesis that colour vision in primates was selected for discriminating the spectral modulations on the skin of conspecifics, presumably for the purpose of discriminating emotional states, socio-sexual signals and threat displays. Here we show that, consistent with this hypothesis, there are two dimensions of skin spectral modulations, and trichromats but not dichromats are sensitive to each. Furthermore, the M and L cone maximum sensitivities for routine trichromats are optimized for discriminating variations in blood oxygen saturation, one of the two blood-related dimensions determining skin reflectance. We also show that, consistent with the hypothesis, trichromat primates tend to be bare faced.

Analysis in non-human primates reveals that the ancestral Band 3 gene encodes Dib and the Band 3-Memphis phenotype.

BACKGROUND: The anion exchanger, Band 3, carries antigens in the Diego blood group system, and can carry the Band 3-Memphis phenotype. Although Di(b) is of high prevalence and Band 3-Memphis is of low prevalence in humans, it has been suggested that both are on the ancestral gene. We determined the orthologue nucleotide sequences corresponding to these two polymorphic sites, Di(a)/Di(b) (2561T > C; Leu854Pro) and Band 3-Memphis(166A > G; Lys56Glu) in several nonhuman primates. METHODS: Genomic DNA was extracted from blood samples of great apes, lesser apes, old world monkeys, new world monkeys and prosimians. PCR amplifications were done with primer pairs that were located in the flanking intronic regions of Exon 4 and Exon 19; and the amplified products were sequenced. RESULTS: Amino acid sequence alignment of nonhuman primates band 3 with that of human showed extensive homologies. In exon 4, Glu56Lys polymorphic site showed Glu similar to Band 3-Memphis type and in exon 19, Leu854Pro polymorphic site showed Pro indicating Di(b) phenotype. CONCLUSIONS: The nonhuman primates have nucleotide sequences of Di(b)(2561C) in cis to Band 3-Memphis (166G), which is consistent with the assertion that the Di(b) and Band 3-Memphis phenotype represents the ancestral Band 3 gene.

Micronucleated erythrocyte frequencies in old and new world primates: measurement of micronucleated erythrocyte frequencies in peripheral blood of Callithrix jacchus as a model for evaluating genotoxicity in primates.

Nonhuman primates are of particular relevance in evaluating the potential toxicity of drugs and environmental agents. We have used previously published information and data from the present study to establish a relationship for New World (NW) and Old World (OW) primates on the basis of the frequency of spontaneous micronucleated erythrocytes (MNEs) observed in peripheral blood. Data on spontaneous MNEs in peripheral blood from 15 species of primates, including humans, indicate that NW primates have significantly (P < 0.01) higher MNE frequencies (group mean, 9.5 +/- 7.3 MNEs/10,000 erythrocytes; range, 0.7-20.5/10,000 erythrocytes) than OW primates (group mean, 1.0 +/- 0.9 MNEs/10,000 erythrocytes; range, 0.0-2.6 MNEs/10,000 erythrocytes). Humans are believed to have developed in the OW, and human MNE frequencies were similar to those described for OW primate species. We selected the common marmoset (Callithrix jacchus), a NW primate, to determine whether therapeutic pediatric doses of Metotrexate (MTX; 2.5 mg/kg), Cyclophosphamide (CP; 5 mg/kg), Cytosine-arabinoside (Ara-C; 3 mg/kg), or 5-Fluorouracil (5-FU; 10 mg/kg), administered daily for two consecutive days, increase the frequency of micronuclei. Micronucleated polychromatic erythrocyte frequencies were increased significantly in groups receiving MTX, CP and Ara-C, while MNE frequencies were increased by the Ara-C treatment. The results of this study indicate that NW primates have higher spontaneous MNE frequencies than OW primates, and because of this, NW primates like the common marmoset, may be suitable for evaluating the genotoxicity of chemical agents.