DNA based method for determining source country of the short beaked echidna (Tachyglossus aculeatus) in the illegal wildlife trade.

The illegal trade in wild animals being sold as 'captive bred' is an emerging issue in the pet and zoo industry and has both animal welfare and conservation implications. DNA based methods can be a quick, inexpensive, and definitive way to determine the source of these animals, thereby assisting efforts to combat this trade. The short beaked echidna (Tachyglossus aculeatus) is currently one of the species suspected to be targeted in this trade. As this species is distributed throughout Australia and in New Guinea (currently comprising of five recognised sub-species), this project aimed to develop a DNA based method to definitively determine the source country of an echidna and explore the use of non-invasive sampling techniques. Here we use non-invasively sampled echidna quills and demonstrate the extraction of mitochondrial DNA and amplification of a region of the mitochondrial genome. Phylogenetically informative markers for analysis of a 322bp segment of the D-loop region were developed, and subsequently validated, using animals with known source locations allowing us to reliably distinguish between echidnas from New Guinea, and Australia. This research presents the first validated forensic protocols for short beaked echidnas and will be an integral tool in understanding the movement of animals in this emerging trade.

Investigation into ethylene oxide treatment and residuals on DNA and downstream DNA analysis.

Recent years have seen a significant increase in the sensitivity of DNA testing, enabling the determination of DNA profiles from low levels of cellular material. However, the increased sensitivity is in many ways a double-edged sword as background contaminating DNA generated during the manufacture of consumables and sampling devices is now being detected and may compromise the interpretation of the DNA profile results. This study initially demonstrated the effectiveness of ethylene oxide (EO) as a post-production treatment to eliminate DNA on swabs, used as a sampling device for the recovery of cellular material. Subsequently, the potential adverse effects of any residual EO remaining on the swabs on the downstream DNA analysis on both rayon and cotton swabs were investigated and the levels of remaining EO measured. Two main variables were tested: the amount of time elapsed since EO treatment of the swabs prior to use, and the time elapsed between cellular material collection and DNA analysis. Residual levels of EO were found to be below quantitation levels and therefore also international standards. The results indicated that while there was a negligible effect of EO treatment on DNA recovered from rayon swabs, there was however an adverse effect on the DNA profiles recovered from cotton swabs. The adverse effect was negatively correlated with time since EO treatment and positively correlated with time to DNA analysis.

Evaluation of forage soybean, with and without pearl millet, as an alternative for beef replacement heifers.

Apparent ruminal digestibility of forage soybean-based silages, with and without pearl millet, was determined along with evaluation of silages on heifer performance and reproductive function. Fermenters were utilized in a Latin square design and randomly assigned to 1 of the following treatments: 1) control diet of alfalfa haylage (CON), 2) soybean silage (SB) or 3) soybean and pearl millet silage (SB×PM). All diets were formulated to meet or exceed nutrient requirements of replacement beef heifers targeted to gain 0.79 kg/d. These same diets were fed to 90 Angus-Simmental beef replacement heifers [body weight (BW) = 366 kg; body condition score (BCS) = 5.53; age = 377 ± 11 d] 65 d prior to timed artificial insemination (TAI). Heifers were randomly allotted by breed, BCS and BW to 1 of the 3 treatments, with 3 reps/treatment. Diets were terminated 21 d post-TAI and heifers were commingled and placed on a common diet. Pubertal status was determined by progesterone concentrations of 2 blood samples taken 10 d apart prior to both trial initiation as well as initiation of estrous synchronization. Ovulatory follicle diameter was determined at time of breeding by ultrasonography. Pregnancy diagnosis was accomplished 35 and 66 d post-TAI, respectively, to calculate TAI and end of season pregnancy rates. Neither SB nor SB×PM had an effect (P > 0.37) on apparent ruminal digestion of nutrients compared to the CON. Final BW (414 kg; P ≥ 0.10) and BCS (5.28; P ≥ 0.26) for the heifers were similar among treatments. Likewise, there were no differences in TAI (48%; P > 0.43) or overall breeding season (93%; P > 0.99) pregnancy rates. Ovulatory follicle diameters (11.7 mm) was not different (P > 0.19) among treatments. In summary, forage soybean-based silages, with and without pearl millet, was an acceptable alternative forage for developing replacement beef heifers.

Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 1. Excessive supply from corn gluten meal or soybean meal.

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, the source from which excess CP is derived and how it affects reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of feeding excess metabolizable protein (MP) from feedstuffs differing in rumen degradability on ovulatory follicular dynamics, subsequent corpus luteum (CL) development, steroid hormone production and circulating amino acids (AA) in beef cows. Non-pregnant, non-lactating mature beef cows (n=18) were assigned to 1 of 2 isonitrogenous diets (150% of MP requirements) designed to maintain similar BW and body condition score (BCS) between treatments. Diets consisted of ad libitum corn stalks supplemented with corn gluten meal (moderate rumen undegradable protein (RUP); CGM) or soybean meal (low RUP; SBM). After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, gonadotropin releasing hormone (GnRH) was administered to reset ovarian follicular growth. Starting at GnRH administration and daily thereafter until spontaneous ovulation, transrectal ultrasonography was used to diagram ovarian follicular growth, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of visual detection of estrus, CL size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. As designed, cow BW and BCS were not different (P⩾0.33). Ovulatory follicular wavelength, antral follicle count, ovulatory follicle size at dominance and duration of dominance were not different (P>0.13) between treatments. Cows supplemented with CGM had greater post-dominance ovulatory follicle growth, larger dominant follicles at spontaneous luteolysis, shorter proestrus, and larger ovulatory follicles (P⩽0.03) than SBM cows. No differences (P⩾0.44) in peak estradiol, ratio of estradiol to ovulatory follicle volume, or plasma urea nitrogen were observed. While CL volume and the ratio of progesterone to CL volume were not affected by treatment (P⩾0.24), CGM treated cows tended to have decreased (P=0.07) circulating progesterone 7 days post-estrus compared with SBM cows. Although total circulating plasma AA concentration did not differ (P=0.70) between treatments, CGM cows had greater phenylalanine (P=0.03) and tended to have greater leucine concentrations (P=0.07) than SBM cows. In summary, these data illustrate that excess MP when supplemented to cows consuming a low quality forage may differentially impact ovarian function depending on ruminal degradability of the protein source.

Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 2. Excessive supply in varying concentrations from corn gluten meal.

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, amount of excess CP effects on reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of excess metabolizable protein (MP) supplementation from a moderately abundant rumen undegradable protein (RUP) source (corn gluten meal: 62% RUP) on ovarian function and circulating amino acid (AA) concentrations in beef cows consuming low quality forage. Non-pregnant, non-lactating beef cows (n=16) were allocated by age, BW and body condition score (BCS) to 1 of 2 isocaloric supplements designed to maintain BW for 60 days. Cows had ad libitum access to corn stalks and were individually offered a corn gluten meal-based supplement daily at 125% (MP125) or 150% (MP150) of National Research Council (NRC) MP requirements. After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, follicular growth was reset with gonadotropin releasing hormone. Daily thereafter, transrectal ultrasonography was performed to diagram ovarian follicular waves, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of observation of estrus, corpus luteum (CL) size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. No differences (P⩾0.21) in BW and BCS existed throughout the study; however, plasma urea N at ovulation was greater (P=0.04) in MP150. Preovulatory ovarian follicle size at dominance, duration of dominance, size at spontaneous luteolysis, length of proestrus and wavelength were not different (P⩾0.11) between treatments. However, ovulatory follicles were larger (P=0.04) and average antral follicle count was greater (P=0.01) in MP150 than MP125. Estradiol concentration and ratio of estradiol to ovulatory follicle volume were not different due to treatment (P⩾0.25). While CL volume 7 days post-estrus was greater (P<0.01) in MP150 than MP125, circulating progesterone 7 days post-estrus and ratio of progesterone to CL volume were not different (P⩾0.21). Total AA were not different (P⩾0.76) at study initiation or completion; however, as a percent of total AA, branched-chain AA at ovulation were greater (P=0.02) in MP150. In conclusion, supplementation of CP at 150% of NRC MP requirements from a moderately undegradable protein source may enhance growth of the ovulatory follicle and subsequent CL compared with MP supplementation at 125% of NRC MP requirements.

Feeding distiller's grains as an energy source to gestating and lactating beef heifers: Impact on female progeny growth, puberty attainment, and reproductive processes.

This study compared postweaning growth, puberty attainment, and reproductive processes of female progeny (n = 33) born to Angus-Simmental beef heifers treated with either a control diet or a diet in which dried distiller's grains with solubles (DDGS) were fed as an energy source during late gestation and early lactation. From 192 d of gestation through 118 ± 4 d in lactation, dams were fed either a corn silage­based control diet (CON) orcorn residue with DDGS, where DDGS were supplemented as an energy source (DG). Diets were formulated to provide similar daily NEg between diets, but CP requirements were drastically exceeded in the DG treatment. Heifer progeny (n = 33) were weaned, commingled at 191 ± 4 d of age, and similarly managed for the remainder of the project. Heifer BW and blood samples for progesterone assessment to determine onset of puberty were collected weekly beginning at weaning. At 255 ± 4 d of age, a single follicular wave was mapped via ultrasonography in 10 prepubertal heifers per treatment. Prepubertal antral follicle count and ovarian size were determined at 253 ± 4 d of age. Hip height was recorded at 213,297, and 437 ± 4 d of age. Estrous synchronization and AI was initiated at 447 ± 4 d of age. Binary data were analyzed with the GLIMMIX procedures of SAS and all other data were analyzed with the MIXED procedures of SAS. Progeny from DG-treated dams tended to be heavier (P = 0.08) than progeny from CON-treated dams from weaning until breeding. In addition, DG progeny had a greater (P < 0.01) frame score than CON throughout the developmental period. Ovarian size, antral follicle count, and follicular growth parameters did not differ between treatments. Age at puberty did not differ between CON (303 ± 10 d) and DG (320 ± 10 d) progeny; however, BW at puberty was greater (P = 0.01) for DG (326 ± 7 kg) than CON (298 ± 8 kg) progeny. Pregnancy rates to AI were greater (P = 0.05) in DG progeny (70.6%) than CON (33.3%), but overall breeding season pregnancy rate did not differ (P = 0.97). Moreover, rate of dystocia in female progeny at first parturition and grand-offspring birth BW did not differ due to treatment (P ≥ 0.74). In summary, feeding DDGS as an energy source during late gestation and early lactation to first-parity heifers resulted in female progeny with greater skeletal growth that were heavier at onset of puberty and had increased AI pregnancy rates.

Determination of the appropriate delivery of prostaglandin F2α in the five-day CO-Synch + controlled intravaginal drug release protocol in suckled beef cows.

The objective of this experiment was to determine if 2 doses of prostaglandin F(2α) (PGF) administered concurrently at controlled intravaginal drug release (CIDR) removal was an efficacious method for delivery of PGF in the 5-d CO-Synch + CIDR protocol. Postpartum beef cows (n = 2,465) from 13 herds in 8 states were enrolled in the 5-d CO-Synch + CIDR protocol and assigned to receive 2 doses of PGF (25 mg/dose) 8 h apart with the initial injection given at CIDR insert removal (8h-PGF), 2 doses (25 mg/dose) of PGF delivered in 2 injection sites, both administered at CIDR insert removal (Co-PGF), or a single 25-mg dose of PGF at CIDR insert removal (1x-PGF). Cows were fixed timed-artificially inseminated (FTAI) 72 h after CIDR removal concurrent with GnRH administration. Estrus-cycling status (54% cyclic) was determined by evaluation of progesterone in 2 blood samples collected before CIDR insertion. Determination of pregnancy was performed by transrectal ultrasonography 39 ± 0.1 d after FTAI and at least 35 d after the conclusion of the breeding season. Fixed timed-AI pregnancy rates were greater (P < 0.05) for the 8h-PGF (55%) than the 1x-PGF (48%) treatment, with the Co-PGF (51%) treatment intermediate and not different (P > 0.10) from the other treatments. Contrast analysis demonstrated that cows receiving 50 mg of PGF (8h-PGF and Co-PGF) had greater (P < 0.05) FTAI pregnancy rates than those receiving 25 mg (1x-PGF). Pregnancy rates to FTAI were greater (P < 0.05) in cyclic (55%) than noncyclic (47%) and greater (P < 0.05) in multiparous (≥3 yr of age; 54%; n = 1,940) than primiparous cows (40%; n = 525). Luteolysis after PGF treatment was assessed in a subset of cows (n = 277) and treatment tended (P = 0.09) to affect the proportion of cows having luteolysis. The percentage of cows that had luteolysis was least in the 1x-PGF treatment (89%) and greatest in the 8h-PGF treatment (97%), with the Co-PGF treatment (94%) being intermediate. Breeding season pregnancy rate (88%) did not differ (P > 0.10) among treatments but was greater (P < 0.01) in multiparous (90%) than primiparous (78%) cows. In summary, 50 mg of PGF was required in the 5-d CO-Synch + CIDR protocol to maximize pregnancy rates; however, pregnancy rate did not differ when 50 mg of PGF was administered simultaneously with CIDR removal or split with 25 mg administered at 0 and 8 h after CIDR removal.

Waist-to-height ratio is a better screening tool than waist circumference and BMI for adult cardiometabolic risk factors: systematic review and meta-analysis.

Our aim was to differentiate the screening potential of waist-to-height ratio (WHtR) and waist circumference (WC) for adult cardiometabolic risk in people of different nationalities and to compare both with body mass index (BMI). We undertook a systematic review and meta-analysis of studies that used receiver operating characteristics (ROC) curves for assessing the discriminatory power of anthropometric indices in distinguishing adults with hypertension, type-2 diabetes, dyslipidaemia, metabolic syndrome and general cardiovascular outcomes (CVD). Thirty one papers met the inclusion criteria. Using data on all outcomes, averaged within study group, WHtR had significantly greater discriminatory power compared with BMI. Compared with BMI, WC improved discrimination of adverse outcomes by 3% (P < 0.05) and WHtR improved discrimination by 4-5% over BMI (P < 0.01). Most importantly, statistical analysis of the within-study difference in AUC showed WHtR to be significantly better than WC for diabetes, hypertension, CVD and all outcomes (P < 0.005) in men and women. For the first time, robust statistical evidence from studies involving more than 300 000 adults in several ethnic groups, shows the superiority of WHtR over WC and BMI for detecting cardiometabolic risk factors in both sexes. Waist-to-height ratio should therefore be considered as a screening tool.

Effects of natural (RRR alpha-tocopherol acetate) or synthetic (all-rac alpha-tocopherol acetate) vitamin E supplementation on reproductive efficiency in beef cows.

The objective was to determine the effects of natural- or synthetic-source vitamin E on reproductive efficiency in Angus-cross beef cows. In Exp. 1, one hundred fifty-two cows were fed hay and corn silage based diet and assigned to 1 of 3 dietary supplements (3 pens/treatment): 1) containing no additional vitamin E (CON), 2) formulated to provide 1,000 IU x d(-1) of synthetic-source vitamin E (SYN; all-rac or dl-alpha-tocopherol acetate), or 3) formulated to provide 1,000 IU x d(-1) of natural-source vitamin E (NAT; RRR or D-alpha-tocopherol acetate). In Exp. 2, seventy-five cows (2 reps/treatment) were assigned to similar treatments as Exp. 1; however, a vitamin-mineral supplement was offered for ad libitum intake and vitamin intake was calculated from predicted mineral intakes. Cows grazed pastures rather than being fed hay and corn silage as in Exp. 1. In Exp. 1 and 2, supplementation began 6 wk prepartum and continued until initiation of the breeding season. Blood samples were collected at calving (Exp. 1) or breeding (Exp. 2) to determine alpha-tocopherol concentration and weekly beginning 4 wk postpartum (Exp. 1) or 7 and 14 d before estrus synchronization (Exp. 2) to determine return to estrus via progesterone concentration. Cows were synchronized and bred by AI based on heat detection; nonresponding cows were time bred (AI) 66 h after PGF(2 alpha) injection, and cows returning to estrus after AI were bred by natural service. In Exp. 1, cows supplemented with NAT and SYN had greater (P < 0.001) serum concentrations of alpha-tocopherol at calving compared with CON cows. Dietary supplement did not affect (P >or= 0.55) the percentage of cows cycling before synchronization or the number of days to return to estrus by cows that resumed estrus before synchronization. Cows supplemented with SYN tended to have greater first service conception rates compared with CON and NAT (P = 0.09); however, first plus second services combined and overall conception rates were not affected (P >or= 0.23). In Exp. 2, NAT cows had greater (P = 0.002) concentrations of alpha-tocopherol at breeding, whereas there was no difference (P > 0.05) between SYN and CON. Supplementation of SYN or NAT did not affect (P >or= 0.17) days to resumption of estrus before breeding, first service, first plus second services combined, or overall conception rates. These data suggest that supplementation of SYN or NAT source vitamin E increased alpha-tocopherol concentration in cows; however, effects on reproductive efficiency are minimal.

Effects of maternal natural (RRR alpha-tocopherol acetate) or synthetic (all-rac alpha-tocopherol acetate) vitamin E supplementation on suckling calf performance, colostrum immunoglobulin G, and immune function.

The objective of this study was to determine the effects of maternally supplemented natural- or synthetic-source vitamin E on suckling calf performance and immune response. In a 2-yr study, one hundred fifty-two 2- and 3-yr-old, spring-calving, Angus-cross beef cows were blocked by age, BW, and BCS into 1 of 3 isocaloric, corn-based dietary supplements containing 1) no additional vitamin E (CON), 2) 1,000 IU/d of synthetic-source vitamin E (SYN), or 3) 1,000 IU/d of natural-source vitamin E (NAT). Maternal supplementation began approximately 6 wk prepartum and continued until the breeding season. Colostrum from cows and blood from calves was collected 24 h postpartum for analysis of IgG concentration as an indicator of passive transfer and circulating alpha-tocopherol concentration. At 19 d of age, blood was collected from calves to determine the expression of CD14 and CD18 molecules on leukocytes. At 21 and 35 d of age, humoral immune response was measured by a subcutaneous injection, in the neck, with ovalbumin (20 mg; OVA) and blood samples collected weekly until d 63 of age to determine antibodies produced against OVA. At d 63 of age, calves were administered an intradermal injection of OVA (1 mg) in the neck to assess cell-mediated immunity, which was determined on d 65 of age by measuring nodule size with calipers. Circulating alpha-tocopherol concentrations were increased at both 24 h (P = 0.001) and at the day of initial OVA challenge (P < 0.001) in SYN and NAT compared with CON calves. No differences were detected (P > 0.05) for calf birth BW, ADG, or weaning BW. There were no differences (P > 0.05) in calf serum total IgG or cow colostrum total IgG at 24 h or presence of CD14 and CD18 receptors at d 19 of age. The NAT calves had a greater antigen response to OVA at d 63 than SYN calves (P = 0.01; treatment x day interaction). As an indicator of cell-mediated immunity to OVA, nodule size at 65 d of age was not affected (P = 0.92) by maternal dietary supplementation. In conclusion, calves suckling cows supplemented with natural- and synthetic-source vitamin E had increased circulating concentrations of alpha-tocopherol at 24 h, which appeared to continue throughout maternal supplementation; however, calf immune function and performance were not affected.

Effects of crude glycerin on performance and carcass characteristics of finishing wether lambs.

The objective of this study was to evaluate the effects of crude glycerin on performance and carcass characteristics in finishing wether lambs. Thirty black-faced, Suffolk-cross wether lambs (44.1 +/- 5.6 kg initial BW) were stratified and blocked by BW to 1 of 5 individually fed, isocaloric, isonitrogenous dietary treatments containing 0, 5, 10, 15, or 20% crude glycerin (88% pure) on a DM basis. Diets were fed once daily for ad libitum consumption and contained 15% chopped hay, approximately 25% dried distillers grains with solubles, and the specified treatment combination of cracked corn replaced with increasing amounts of crude glycerin. Wethers were weighed on 14-d intervals and were selected for slaughter when they reached an approximate 12th-rib fat depth of 0.51 cm (28 to 84 d on trial). Carcass characteristics were collected after a 48-h chill. Dry matter intake (linear, P = 0.004) and ADG (quadratic, P = 0.05) increased with increasing concentrations of glycerin in the diet during the first 14 d of the feeding period. Similarly, G:F tended to increase quadratically (P = 0.06) with increasing concentrations of crude glycerin in the diet during the first 14 d. However, there were no differences among treatments for final BW, days on feed, or cumulative DMI, cumulative ADG, and cumulative G:F (P >or= 0.11). Body wall thickness, dressing percent, HCW, LM area, flank streaking, leg score, conformation score, quality grade, yield grade, and percent boneless, closely trimmed retail cuts did not differ (P >or= 0.21). Adding up to 15% crude glycerin to finishing wether diets improved feedlot performance, particularly during the first 14 d, without any concomitant effect on carcass characteristics.

Effects of dietary fat and crude protein on feedlot performance, carcass characteristics, and meat quality in finishing steers fed differing levels of dried distillers grains with solubles.

The objective of this study was to evaluate the influence of dietary protein and fat from distillers dried grains with solubles (DDGS) on feedlot performance, carcass characteristics, and meat quality in finishing steers. Angus-cross steers (n = 105; 443 +/- 20 kg of BW) were blocked by BW and randomly assigned to 1 of 5 dietary treatments: 1) corn-based diet with DDGS included at 25% of DM (CON), 2) CON with DDGS included at twice the amount of CON (50% of DM; 50DDGS), 3) CON with added corn protein to equal the CP in the 50DDGS diet (CON+CP), 4) CON with added vegetable oil to equal the fat in the 50DDGS diet (CON+VO), and 5) CON with protein and fat added to equal the CP and fat in the 50DDGS diet (CON+CPVO). Steers were fed to a common 12th-rib fat depth endpoint (1.3 +/- 0.2 cm; 68 to 125 d on trial). Loins and rounds were collected from 44 carcasses for Warner-Bratzler shear force (WBSF), ether extract, and case-life analyses. Data were analyzed using the MIXED procedure of SAS. Contrasts between 1) CON vs. elevated CP diets (50DDGS, CON+CP, and CON+CPVO; EP), 2) CON vs. elevated fat diets (50DDGS, CON+VO, and CON+CPVO; EF) and 3) CON vs. diets with elevated CP and fat (50DDGS and CON+CPVO; EPF) were analyzed. There were no differences in days on feed or DMI among treatments. Steers fed CON had greater ADG (P or= 0.06). Steers fed the CON diet had greater marbling scores (P or= 0.44). However, CON steers had greater (P = 0.02) L* values than EF-fed steers and greater b* values than EP, EF, and EPF steers (P

Adjustment of children who have a sibling with Down syndrome: perspectives of mothers, fathers and children.

BACKGROUND: A number of methodological weaknesses have contributed to our relatively poor understanding of the impact on children of having a brother or sister with a disability. These include a focus on poor adjustment, using multidiagnostic groups, inadequate matching, and a failure to consider the perspectives of children and parents together. METHOD: This study compared the adjustment of 53 siblings of a child with Down syndrome with a comparison group of siblings of children who were developing typically. Children were matched on a case-by-case basis for gender, age and position in family. Families were matched for family size and father's occupation. The age range of the target siblings was 7-14 years. Data were gathered from mothers, fathers and siblings. RESULTS: There were no significant differences between the groups on adjustment measures. These included parent perceptions of externalizing and internalizing behaviours, parent perceptions of sibling competence, and sibling perceptions of their own competence and self-worth. Associations between measures of adjustment and child reports of their contribution to household functioning depended on sex rather than group membership. There was an association between parental reports of externalizing behaviour and sibling relationships with the brother/sister closest in age. CONCLUSIONS: Having a brother or sister with Down syndrome does not inevitably lead to poor adjustment. Examination of within-family processes would appear to be more useful in identifying children at risk than merely group membership.

DNA analysis in disputed parentage: the occurrence of two apparently false exclusions of paternity, both at short tandem repeat (STR) loci, in the one child.

In cases of disputed paternity investigated by DNA analysis, it is generally accepted that at least two independent exclusions be observed before concluding that a putative father is not the biological father of the child. We report here a case in which two apparent exclusions of paternity were obtained (at the loci FESFPS and TPOX), from a total of nineteen loci examined (short tandem repeats, AMFLPs, DQA1, Polymarkers). The combined paternity index of the other seventeen loci, and of two multilocus probes (33.15 and 33.6) would exceed twenty-eight trillion to one. In the absence of any alternative putative father (including close relatives of the man tested), we concluded that the most likely explanation for these results was that the putative father was indeed the biological father, but that two independent mutations had occurred in either the ovum and/or spermatozoon. We have altered our internal laboratory criterion for exclusion of paternity to require exclusions in three independent loci.

Assessing the evidential value of DNA profiles matching without using the assumption of independent loci.

DNA profiling allows determination of the alleles at multiple loci on an individual's genome. The frequencies of these alleles are then estimated from a sample drawn from the population. If the occurrences of alleles at different loci are independent, the frequencies may be multiplied together to give an estimate of the probability of DNA from a randomly-chosen member of the population matching the DNA in question. However, there is doubt as to whether the assumption of independence can be justified. This paper discusses a method of calculating the probability of a match that does not require the assumption of independence. A suitable set of criteria is also derived that offer an objective approach to the determination of a match from two DNA samples.

Maternal reports of behavior of siblings of children with Down syndrome.

Mothers of children, ages 6 to 13 years, representing two target groups were interviewed: siblings of children with Down syndrome and a comparison group of siblings of children without Down syndrome. Mothers were interviewed regarding their expectations of typical child behavior, their perceptions of conduct problems of their child, and the extent to which that child shared household responsibilities. There were no significant differences between the groups regarding expectations of "typical" children, but the mothers of girls who had a sibling with Down syndrome reported significantly more conduct problems than did either mothers of children without Down syndrome or mothers of boys with a sibling who had Down syndrome. Groups did not differ with regard to the number of household tasks, but reports of conduct problems in the girls who had a sibling with Down syndrome were inversely related to the number of tasks undertaken.

Human population genetic studies using hypervariable loci. I. Analysis of Assamese, Australian, Cambodian, Caucasian, Chinese and Melanesian populations.

Population genetic studies, in Australian, Assamese, Cambodian, Chinese, Caucasian and Melanesian populations, were performed with several highly polymorphic DNA loci. Results showed that the Caucasian and Chinese had the highest level of heterozygosity. The size range of the majority of the polymorphic DNA fragments of a locus was the same in the different populations. The distinguishing feature of each ethnic group was the relative frequency of a particular set or group of alleles. For example, alleles greater than 9.0 kb in size, in D14S13, or from 4.5 to 4.7 kb, in D18S27, were less than half as frequent in Caucasians than in the other populations. Overall, there were groups of alleles, at one or more loci, whose frequencies were different among some of the ethnic groups and therefore could be used to differentiate one group from the other.

Seasonality of births in nineteenth-century urban Tasmania.

The monthly distribution of births for Hobart Town, Van Diemen's Land, during the period 1839-1859 is examined. Prior research on two rural registration districts in nineteenth-century Tasmania revealed patterns of birth seasonality. The pattern was responsive to both the distinctive seasonal rhythm of regional economic activities and the birth interval and differed from the pattern for all of Tasmania in the twentieth century. Here, I argue that the aggregate monthly pattern of births in a mid-nineteenth-century urban registration district was, by contrast, not seasonal. Some seasonality was found among farmers, seamen, and dealers in foodstuffs but not in other industry groups. Class differences were not apparent. The research establishes that the seasonal distribution of mid-nineteenth-century urban births corresponds neither to nineteenth-century rural patterns nor to the patterns evident in the twentieth century.

PIP: Monthly data on 16,677 births in the Hobart registration district in the English colony of Van Diemen's Land (Tasmania) between 1839-59 were applied to the circular goodness of fit test and the Maag's U2 to prove that births were not seasonal as often believed. Also examined were 19th century data from the rural districts of Oatlands and Bothwell which demonstrated patterns of birth seasonality. The monthly birth distributions for the Hobart registration district were not seasonal. Even in the years when there was an indication of seasonality 91840-44 and 1955-59), no consistent pattern of monthly distribution if births emerged, which supported the general interpretation that the births were not seasonally distributed. The observed pattern of births in Hobart differed from that of the 2 rural districts. Both urban and rural data indicated apparent seasonality of births for farming families but not for families of farm laborers. Occupation had more importance in seasonality than the direct effects of climate. For example, the only industrial groups that exhibited any seasonality at all in the urban district were those that depended on the demands of the climate--farming, the maritime trade (but not from coastal whaling and fishing), and dealing in foodstuffs (most were butchers). Yet seasonality was marginal. No significant differences in seasonality of births existed between the classes. Seasonal distribution of urban births did not parallel the seasonality patterns observed in 20th century Hobart. The 20th century seasonality patterns occurred because women had contraceptive technology available to time births. The mid-19th century distribution of births pattern in Hobart did not match either that of mid-10th century rural districts or that of the 20th century.

Counting by children with Down syndrome.

Children's understanding of one-to-one, stable order, and cardinal principles was tested through error-detection and self-performance counting tasks. Gelman's modified counting task was used to test understanding of the order irrelevance principle. There were no significant differences between the mean task scores of 15 children with Down syndrome and 15 preschoolers with similar revised Peabody Picture Vocabulary test (PPVT-R) scores (mean age equivalent, 4 years, 7 months). Partial correlations showed several significant associations between counting principles and PPVT-R that were independent of chronological age. The results support the view that counting by children with Down syndrome can be guided by counting principles and that developmental level rather than the syndrome is associated with counting behavior.