Parentage Determination Research Articles

Multiple mating between black bears revealed by DNA fingerprinting

The reproductive strategies of large, solitary carnivores have seldom been studied because of the difficulties in obtaining detailed mating observations and reliable parentage determinations. Using DNA fingerprinting, the mating system of a natural black bear, Ursus americanus, population, occupying the Chapleau Crown Game Preserve, Ontario, Canada, was investigated. DNA fingerprints for paternity analysis were obtained using alkaline phosphatase-labelled multilocus probes and chemiluminescent detection. DNA samples were collected from four complete mother-offspring groups, and 45 males were tested as possible fathers. Fingerprinting demonstrated both single and multiple paternity within litters, and one male was identified as the father for cubs from two litters during 1 year. This evidence, coupled with field observations of pairs of adults, indicates that multiple mating is practised by both sexes.

RAPD Analysis for Parentage Determination in Prunus mume Sieb. et Zucc.

Application of RAPD analysis to determine the parentage of mume cultivars was utilized using 30 decamer primers.No difference was found in DNA fingerprints of the same sample under repeated polymerase chain reactions. The number of amplified products increased with increasing GC content of the primer from 40 to 60 %. The GC content of the primer may be a good measure of stable amplification.The polymorphic DNA fragments between parental plants were easily detected; nearly all segregated among their F1 plants. Southern blotting confirmed that the bands specific to parents plants were transmitted to F1 plants. Thus, it was possible to distinguish morphologically identical F1 plants from each other.As a consequence, RAPD analysis provided useful genetic markers to trace the parentage determination in mume cultivars.

Hybrid rice (Oryza sativa L.): identification and parentage determination by RAPD fingerprinting

DNA from three families of rice plants selected in Northern China (each comprising the male sterile, the restorer, the hybrid F1 and the maintainer lines) has been extracted and amplified by PCR with different random DNA primers (RAPD analysis). Then, DNA has been analysed by agarose gel electrophoresis and DNA bands scored as present or absent. The generated matrices are reproducible and amenable for identification of each single plant line. Thus, RAPD fingerprinting of the inbred parental lines and of the resulting hybrid is proposed as a convenient tool for the identification, protection and parentage determination of plant hybrids. Furthermore, by offering a molecular tool to verify the degree of dissimilarity between the parental lines, the RAPD analysis may also be used to search for new parental combinations.

Allele frequency data for VNTR locus D17S79: identification of an internal HaeIII polymorphism in the black population.

D17S79 is one of several highly polymorphic VNTR loci commonly used to generate DNA profiles for forensic identity testing and parentage determination. In this study, we used Southern hybridization analysis of HaeIII-digested genomic DNA to compile D17S79 allele frequency databases for the Caucasian, Southeast Asian, East Indian, and Black populations of Southern Ontario. In our Black population sample, low-molecular-weight D17S79 alleles are relatively common and approximately 7% of the individuals have three-band hybridization patterns due to a HaeIII restriction site polymorphism within a common D17S79 allele. The internal HaeIII site generates an allele consisting of two HaeIII fragments of approximately 980 and 420 bp. The estimated frequency of this "split-allele" in our Black population is 0.035. Since this allele was not observed in the other population groups, the internal HaeIII polymorphism probably originated within the Black population. Although the existence of low-molecular-weight alleles and the internal HaeIII polymorphism could affect the interpretation of some VNTR patterns, they are of little or no forensic consequence to the frequencies assigned to VNTR profiles.

Extrapair Fertilization in Monogamous Bull-Headed Shrikes Revealed by DNA Fingerprinting

-We analyzed parentage in a wild population of the Bull-headed Shrike (Lanius bucephalus) by DNA fingerprinting. Male Bull-headed Shrikes pair with a single female and defend a large all-purpose territory. Generally, both parents provide substantial parental care. Among 99 nestlings from 24 clutches, 10 (10%) nestlings from 4 (17%) clutches had extrapair paternity, but were offspring of the females. These results show that copulation frequency is not necessarily a good measure of reproductive success even in monogamous birds where no extrapair copulations are observed. The average band-sharing proportion between full siblings was the same as that between parent and offspring, but the variance was greater. This is likely the case because the band-sharing proportion reflects the gene-sharing proportion. Received 6 May 1991, accepted 9 February 1992. OvER 90% of birds are thought to be monogamous (Lack 1968), and males of these species employ mate guarding to avoid being cuckolded (M0ller 1985). Nevertheless, recent evidence from field observations and genetic analyses has clearly demonstrated that extrapair copulations (EPCs) and extrapair fertilizations (EPFs) occur in monogamous species. Among colonially breeding species of monogamous birds like Bank Swallows (Riparia riparia; Beecher and Beecher 1979) and Cattle Egrets (Bubulcus ibis; Fujioka and Yamagishi 1981), EPCs occur frequently. In Purple Martins (Progne subis; Morton et al. 1990) and Zebra Finches (Taeniopygia guttata; Birkhead et al. 1990), EPCs often result in fertilization, and intraspecific brood parasitism (ISBP) has been documented. In House Sparrows (Passer domesticus), which have a loosely colonial breeding system, up to 8% of the offspring were not fathered by their putative male parent (Wetton et al. 1987). EPCs have been recorded even in species with large all-purpose territories, where individuals are relatively difficult to observe. In Indigo Buntings (Passerina cyanea), EPCs often resulted in EPFs (Westneat 1987a, b), while in monogamous Willow Warblers (Phylloscopus trochilus), EPCs resulted in few EPFs (Gyllensten et al. 1990). These results show that copulation frequency is not necessarily a good measure of reproductive success. However, there are only a few genetic studies of strongly monogamous and territorial species; all have found low frequencies of EPFs. The Bull-headed Shrike (Lanius bucephalus) has a strongly monogamous breeding system and has a large all-purpose territory (radius ca. 100 m; Yamagishi 1982b). No EPCs have been observed (Yamagishi and Saito 1985). In monogamous birds for which no EPCs have been observed, there are few studies involving the determination of parentage; an exception is an analysis of Dunnocks (Prunella modularis) under a monogamous system (Burke et al. 1989). Their study showed that in monogamous pairs no EPFs occurred, so that the reproductive success of a pair was related to the number of fledged young. To test if this was also the case for Bull-headed Shrikes, we looked for evidence of EPFs and ISBP using DNA fingerprinting, which can be a powerful tool for determining parentage (Jeffreys et al. 1985b). STUDY AREA AND METHODS Study area and capture techniques.-The study was conducted from March to June 1989 in Oizumi city park, Osaka Prefecture, central Japan (34?34'N, 135?32'E). The park covers an area of about 70 ha, is about 20 m in elevation, and is planted mainly with oaks (Quercus). The incubating female and/or the female and the male that fed their nestlings were captured with a mist net, and were regarded as the putative father and mother of the brood. All males that produced extrapair offspring were captured during incubation or nestling period (i.e. when their female mate was not fertilizable, therefore, a capture of the male did not affect our results). Age determination was based on the coloration of the wing coverts (Yamagishi 1982a). DNA fingerprinting.-DNA was prepared from 100 ,ul blood samples collected by jugular venipuncture

DNA profilinf of Queensland Koalas reveals sufficient variability for individual identification and parentage determination.

M13 probe was used in combination with the restriction enzymes Msp I and Bam HI to produce DNA profiles of captive and free-range Queensland koalas (Phascolarctos cinereus). The Msp I-M13 combination resulted in profiles with an average of 22 clearly resolvable bands in the 1.5-7.6-kb range. When seven koalas (part of a 40-animal free-range population) were analysed, they exhibited 8-29% band polymorphism (average variation of 17%). The Bam HI-MI3 combination produced 28 resolvable bands with an average of 9% band polymorphism. The Msp I-M13 profiling system was also used to successfully determine paternity in two family groups. Of the 66 total bands produced when mother, father and offspring were profiled, 11 were common to all three family members, nine were unique to the mother and four were unique to the father. However, two maternal-specific and eight paternalspecific bands were inherited by the offspring. DNA profiling of koalas (at least of those from Queensland) should prove useful for assessing the degree of inbreeding in captive populations, solving disputed paternity cases in wildlife poaching, determining the social organisation of free-range koalas, identifying individual koalas and studying the genetics of the koala throughout its range.

Pollinator Effects on Crossing and Genetic Shift in a Three‐Flower‐Color Alfalfa Population

Understanding insect pollinator preference for different alfalfa (Medicago sativa L.) flower colors is important to maintain the genetic composition of plant introductions, germplasms, genetic stocks, and cultivars. This study was conducted to determine whether crossing and the relative proportion of hybrids in a model three‐flower‐color marker system were affected by kind or combination of insect pollinators [honey bees (Apis mellifera L.), leafcutter bees (Megachile rotundata F.), or both bee species combined). A secondary objective was to determine whether actual expression of compatibility between flower color lines, as determined by hand crosses in the greenhouse, was altered by insect pollinator behavior in the field. Noninbred (S0) and slightly inbred (S1) populations of cream‐ (CRM), white‐ (WHT), and yellow‐ (YEL) colored flower markers were grown at Fresno, CA, and Prosser, WA. When self‐ or sib‐pollinated, the flower colors of all parental strains were reproduced, while hybrids among the parental strains produced unique‐colored progeny that allowed determination of the flower color parentage of each line. All pollinator treatments showed similar preferences for flower color (CRM > WHT > YEL) at both locations. Slight inbreeding resulted in more hybrids with WHT seed parents (P ≤ 0.001), and increased hybrid production by CRM and WHT with pollen from YEL (P ≤ 0.06 and 0.08, respectively). Bee behavior altered the proportion of hybrids produced in the mixed flower‐color population from the expected line‐cross compatibility. The location of seed production also interacted with bee preference for CRM and WHT flowers. Level of inbreeding, insect pollinator behavior, and location of production may interact and change the described levels of different flower‐colored lines and result in genetic shifts in plant introductions, germplasms, genetic stocks, and cultivars.

Parentage determination in maize hybrids using the arbitrarily primed polymerase chain reaction (AP-PCR)

Using a novel procedure based on the polymerase chain reaction, we have developed a rapid, efficient, and economical method for identifying plant genotypes. The arbitrarily primed polymerase chain reaction (AP-PCR) generates reproducible fingerprints from any organism, without the need for DNA sequence information. These fingerprints include DNA fragment polymorphisms that can be (1) used for varietal identification and parentage determination, (2) followed in segregating populations produced by crosses, (3) used as markers for the construction of genetic maps, and (4) used to generate dendograms of phylogenetic relationships, especially at the intraspecific level. AP-PCR requires only minute quantities of DNA (10-25 ng per reaction) and therefore can be used in situations in which DNA is limiting. We demonstrate the use of AP-PCR to identify inbred parents of hybrid maize plants in double-blind experiments.

Parentage Identification in the Bovine Using “Deoxyribonucleic Acid Fingerprints”

Parentage determination in cattle by means of DNA fingerprints is examined at a theoretical level and its implementation illustrated by way of a practical example.

The Identification of Maternity in an Unusual Pregnancy-Related Homicide

A recent bizarre homicide which culminated in the delivery of a live-born infant necessitated careful determination of true maternal origin. A 23-year-old pregnant woman was abducted, subdued, strangled, and delivered of a term infant by a crude Cesarean section. The infant was stolen and subsequently presented to physicians by a woman posing as the mother. Methods used to help confirm the surviving infant's parentage involved red cell antigen and enzyme system evaluations as well as immunoglobulin allotyping, which ultimately proved to be the most effective serologic test performed. The forensic science investigation of this unusual case also used bite mark analysis and patterned injury interpretation. Immunoglobulin allotyping is specifically discussed as a forensic serology test which is currently available and particularly applicable in cases involving parentage determination.

Parental care and mating behaviour of polyandrous dunnocks Prunella modularis related to paternity by DNA fingerprinting

INDIVIDUALS are often assumed to behave so as to maximize their reproductive success1 but unambiguous determination of parentage is difficult, especially in species with complex social systems where a female may mate with several males and where there may also be intraspecific brood parasitism2,4. Even in apparently monogamous species, extra-pair paternity can be common5,7. DNA fingerprinting8,11 promises to revolutionize field studies by providing a powerful method for determining paternity and maternity12. Here we use this technique to link observations of mating behaviour and parental care with precise measurements of reproductive success. We show that in the dunnock Prunella modularis, a small passerine bird with a variable mating system13,14, males do not discriminate between their own young and those of another male in multiply-sired broods. Nevertheless, they increase their own reproductive success by feeding offspring in relation to their access to the female during the mating period, which is a good predictor of paternity.

Paternity Testing

There is evidence that parentage determination was a problem even in biblical times. The most significant developments in resolving these problems occurred in the 20th century starting with Landsteiner's description of the ABO blood group system in 1900. The utility of paternity testing increased as new blood group systems were identified. Major advances occurred between 1940 and 1970, starting with the discovery of the Rh system and reaching a highpoint with the application of HLA typing to paternity studies. Although the exclusion of falsely accused alleged fathers continues as the primary focus of paternity testing, statistical analysis has contributed significantly to the information that can be gleaned from the test results when exclusion has not been demonstrated. Paternal inclusion still cannot be conclusively proven but a mathematical estimation of the likelihood of paternity has found legal acceptance. New technology is now being directed toward identifying inherited DNA markers through the use of genetic probes. Segregation of restriction fragment length polymorphisms (RFLP) may provide the near ultimate test for paternity as it may supply indisputable evidence of family relationships.

The major histocompatibility complex of primates.

The major histocompatibility complex (MHC) encodes cell surface glycoproteins that function in self-nonself recognition and in allograft rejection. Among primates, the MHC has been well defined only in the human; in the chimpanzee and in two species of macaque monkeys the MHC is less well characterized. Serologic, biochemical and genetic evidence indicates that the basic organization of the MHC linkage group has been phylogenetically conserved. However, the number of genes and their linear relationship on the chromosomes differ between species. Class I MHC loci encode molecules that are the most polymorphic genes known. These molecules are ubiquitous in their tissue distribution and typically are recognized together with nominal antigens by cytotoxic lymphocytes. Class II MHC loci constitute a smaller family of serotypes serving as restricting elements for regulatory T lymphocytes. The distribution of class II antigens is limited mainly to cell types serving immune functions, and their expression is subject to up and down modulation. Class III loci code for components C2, C4 and Factor B (Bf) of the complement system. Interspecies differences in the extent of polymorphism occur, but the significance of this finding in relation to fitness and natural selection is unclear. Detailed information on the structure and regulation of MHC gene expression will be required to understand fully the biologic role of the MHC and the evolutionary relationships between species. Meanwhile, MHC testing has numerous applications to biomedical research, especially in preclinical tissue and organ transplantation studies, the study of disease mechanisms, parentage determination and breeding colony management. In this review, the current status of MHC definition in nonhuman primates will be summarized. Special emphasis is placed on the CyLA system of M. fascicularis which is a major focus in our laboratory. A highly polymorphic cynomolgus MHC has been partially characterized and consists of at least 14 A locus, 11 B locus, 7 C locus class I allelic specificities, 9 Ia-like class II antigens and 6 Bf (class III) variants.

Genetic polymorphism in varietal identification and genetic improvement

New sources of genetic polymorphisms promise significant additions to the number of useful genetic markers in agricultural plants and animals, and prompt this review of potential applications of polymorphic genetic markers in plant and animal breeding. Two major areas of application can be distinguished. The first is based on the utilization of genetic markers to determine genetic relationships. These applications include varietal identification, protection of breeder's rights, and parentage determination. The second area of application is based on the use of genetic markers to identify and map loci affecting quantitative traits, and to monitor these loci during introgression or selection programs. A variety of breeding applications based on these possibilities can be envisaged for Selfers, particularly for those species having a relatively small genome size. These applications include: (i) screening genetic resources for useful quantitative trait alleles, and introgression of chromosome segments containing these alleles from resource strain to commercial variety; (ii) development of improved pure lines out of a cross between two existing commercial varieties; and (iii) development of crosses showing increased hybrid vigor. Breeding applications in segregating populations are more limited, particularly in species with a relatively large genome size. Potential applications, however, include: (i) preliminary selection of young males in dairy cattle on the basis of evaluated chromosomes of their proven sire; (ii) genetic analysis of resource strains characterized by high values for a particular quantitative trait, and introgression of chromosome segments carrying alleles contributing to the high values from resource strain to recipient strain.

A Review of Probability of Inclusion in Paternity Testing

Abstract In 1978, the American Association of Blood Banks released its first comprehensive publication on the laboratory aspects of disputed paternity (reviewed in this Journal, Vol. 24, No. 3, July 1979, pp. 705). The primary emphasis of this book was on testing for the purpose of exclusion in paternity cases. However, the issue of probability analysis of inclusion of paternity was also addressed by several of the contributors. This generated a great deal of interest and debate in the legal and medical community because of current pressing social and economic considerations. The controversy over this new approach to parentage determination finally led to a meeting in May 1982 in Airlie, VA of an International Conference on Inclusion Probabilities in Parentage Testing. The result was the establishment of accepted principles for determining and expressing the probability of a paternal relationship. In part, these include the logic involved in probability determinations, a more precise terminology, and such legal issues as reliability and interpretation of information.

Use of odds of paternity computations in determining the reliability of single exclusions in paternity testing.

In parentage determination with genetic markers the concept of paternity index is used to evaluate the relative likelihood of fathership of an accused male individual. When a battery of genetic tests are performed, the event of inadvertant technical errors or the occurrence of suppressor genes that cause an apparent paternity exclusion of the male concerned reduces this likelihood to zero. However, through a study of statistical properties of the paternity index statistic based on the other genetic systems, it is possible to examine the reliability of one or a few such apparent exclusions. The details of this approach is examined here by analytical methods following the statistical principles of likelihood ratio test procedures. Some illustrative data are also analyzed to show the practical utility of this approach.

Incorporation of cholesterol by membranes of bacterial L-phase variants with an appendix on the determination of the L-phase parentage by the electrophoretic patterns of cell proteins.

SUMMARY L-phase variants of Streptobacillus moniliformis and Proteus mirabilis resembled mycoplasmas in their ability to incorporate considerable amounts of exogenous cholesterol into their cell membranes. Much smaller quantities were incorporated by L-phase variants of Streptococcus pyogenes and Staphylococcus aureus, by the parent bacteria of all L-phase variants except S. moniliformis, and by a series of wall-covered bacteria, including their protoplasts and spheroplasts. Cholesterol-binding ability corresponded roughly to sensitivity to growth inhibition and lysis by digitonin. Like myco-plasma, the S. moniliformis L-phase had a non-enzymic cholesterol uptake mechanism, but its growth in a cholesterol-free medium was not improved by adding cholesterol. It is concluded that a bacterium, on transforming to its stable L-phase, does not necessarily develop the ability to bind cholesterol; if it does, the cholesterol bound is not essential for growth of the L-phase, unlike cholesterol-requiring mycoplasmas.

Determination of Parentage by the Use of Polyvalent Immune Sera in Chickens

Abstract IT IS well known that blood groups, characterized as polymorphism, can be applied to various problems of identity and parentage, because of the antigenic individuality of red cells and sera of man and other animals, and because of the rigid manner of their inheritance (Race and Sanger, 1958). In chickens, also the chance that even two closely related individuals will inherit the same blood groups is small because the number of genes determining blood groups is very large (Briles, McGibbon and Irwin, 1950; Briles, Allen and Millen, 1957; Gilmour, 1959). Although blood groups are effective as a means of solving disputed parentage cases, a large number of reagents must be available for carrying out blood group tests. Furthermore, a considerable volume of each reagent is necessary. Simplified techniques of blood grouping would be desirable in order to test a large number of offspring of questionable parentage. An example is the…

Distribution and Inheritance of Hemoglobin Variants in American Cattle

Summary The relative frequencies and genetic determination of the two electrophoretically distinguishable adult bovine hemoglobin types were investigated in a large sample drawn from a number of different breeds and sources. Electrophoretic studies of hemoglobins from 121 males and 646 females, representing nine cattle breeds, indicate that the Ayrshire, Holstein, Angus, Hereford, and Milking Shorthorn breeds probably have only Type A hemoglobin; whereas, the Brown Swiss, Guernsey, Jersey, and Sindhi breeds have both Type A and Type B. An analysis of hemoglobin types among progeny from parents of known types substantiates the theory that the occurrence of these two types of hemoglobin is genetically determined by a pair of codominant alleles—HbA, determining Type A, and HbB, determining Type B. Estimates of the frequeney of HbB among the major dairy breeds are: Ayrshire—0.000, Holstein—0.000, Guernsey—0.116, Brown Swiss—0.117, Jersey—0.333. No gross physiological differences have been observed to be correlated with differences in hemoglobin type. The trait may be of some practical value in parentage determination and in the tracing of breed origins. The most significant feature of these results is that they represent a further demonstration of single-gene determination of a protein specificity.

Serological test for determination of parentage in cattle.

THE antigenic individuality of red cells of cattle was noted in 19101, and studies conducted since 1940 by many workers have made it possible to identify more than fifty blood factors. The number of blood-group combinations is very large, and in practice it is difficult to find two individuals with the same combination. Identification of red cell antigens in cattle and knowledge of the mode of inheritance of these characters have been found to have a practical application among others in the exclusion of paternity and the control of pedigree. A condition for the efficacy of these tests is the possession by the laboratory of a large number of immune sera (reagents).