A genotyping array for the globally invasive vector mosquito, Aedes albopictus.

BACKGROUND: Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito (Aedes albopictus), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. METHODS: We designed a SNP chip for Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. RESULTS: Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth < 20, while there was near complete agreement with WGS read depths > 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. CONCLUSIONS: The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS.

The impact of thawing duration on leptin and adiponectin levels in frozen human milk samples.

BACKGROUND: Measurements of hormone concentrations in human milk from understudied populations across the world is of growing scientific interest. Due to limited access to suitable laboratory facilities at many research sites, human milk samples are often shipped frozen to distant laboratories for analysis. Shipping and handling exposes samples to the risk of thawing and degradation as the results of delays, mishandling, or other unforeseen circumstances. Similar degradation risk factors are well known in serum samples. However, the vulnerability of hormone degradation in human milk to thawing during transportation is largely unexplored. Leptin and adiponectin are of particular importance due to their roles in feeding behavior and metabolism in infants, hence our focus on these two hormones. In addition, leptin and adiponectin have been shown to be vulnerable to pasteurization temperature degradation in human milk samples. AIMS: We tested the degradation of human milk metabolic hormones - leptin and adiponectin - in a controlled environment in response to a variety of freeze/thaw conditions that are reflective of potential temperature fluctuations during transportation. MATERIALS & METHODS: Human milk samples were experimentally subjected to thaw durations ranging from 4 to 16 h and assayed for hormonal levels to observe changes from baseline. Samples were also subjected to two additional experimental conditions, re-freezing after an extended period outside freezers or remaining thawed with additional ice packs, to investigate the impact of common transportation conditions. RESULTS: We found the assayed levels of leptin were not significantly impacted by an extended thaw cycle of 16 h. However, leptin levels were impacted by the extended period of 40 h outside of the freezer. Adiponectin showed a decrease in concentration percentage after the initial 16-h thaw period, but the increased degradation between 0-16 h and 16-40 h was not as severe as that seen in the leptin samples. DISCUSSION & CONCLUSION: The results of this experiment can be used to inform hormone measurement consistency given different thawing lengths or freezing conditions during transportation. Additionally, this research informs decisions regarding transportation, storage/handling, and data analysis in human milk research.

C-reactive protein in adult Samoans: Population variation and physiological correlates.

OBJECTIVES: C-reactive protein (CRP) has been associated with adiposity and cardiometabolic disease risk in many populations but remains remarkably understudied in Pacific Islander populations. Here, we provide the first examination of correlates of CRP in adult Samoans (n = 108, ages 35-55 years) to test the hypotheses that CRP exhibits sex-dependent associations with measures of BMI, adiposity, and cardiometabolic disease risks. METHODS: We analyzed associations between measures of adiposity (total fat mass, visceral fat mass, percent total body fat), body mass index (BMI), cardiometabolic risks, behaviors, demographics, and CRP. Unadjusted analyses of CRP were undertaken using Pearson's pairwise, and Spearman's rank correlations; one-way analysis of variance and Kruskal-Wallis tests assessed variables by CRP quartiles. Adjusted analyses of CRP correlates were examined using generalized linear regression. RESULTS: Serum CRP ranged from 0.08 to 13.3 mg/L (median 1.4 mg/L) and varied significantly by sex t (108) = -2.47, p = .015. CRP was weakly to moderately associated with measures of adiposity and BMI (r and ρ ranged between 0.25 and 0.50, p < .05) and some cardiometabolic markers (including HbA1c, fasting insulin, and insulin resistance). CRP was significantly associated with percent body fat in women and men, adjusting for other variables. CONCLUSIONS: These data are among the first to demonstrate CRP correlates in a sample of adult Samoans. CRP differed by sex and was associated with BMI, adiposity, and some cardiometabolic risk markers. These data align with findings in other populations.

Associations between fecal cortisol and biparental care in a pair-living primate.

OBJECTIVES: We quantified variation in fecal cortisol across reproductive periods in Azara's owl monkeys (Aotus azarae) to examine physiological mechanisms that may facilitate biparental care. Specifically, we evaluated evidence for the explanation that owl monkeys have hormonal mechanisms to mobilize energy during periods when each sex is investing heavily in reproduction, that is, the gestation period for females and the infant care period for males. MATERIALS AND METHODS: Between 2011 and 2015, we monitored 10 groups of Azara's owl monkeys from a wild population in Formosa, Argentina and collected fecal samples from 26 adults (13 males, 13 females). Using enzyme-linked immunosorbent assays, we quantified fecal cortisol as a proxy for evaluating stress responses, including energetic demands, on both sexes during periods of reproduction and parental care. RESULTS: Male cortisol was lowest during periods when they were caring for young infants (<3 months) compared with periods with older infants or no infant. Female cortisol was elevated during gestation compared with other periods. Mean fecal cortisol in both males and females was lower when an infant was present compared with when females were gestating. DISCUSSION: Our results do not support the hypothesis that owl monkey males have elevated fecal cortisol during periods when they need to mobilize energy to provide intensive infant care. Our findings are also inconsistent with the Maternal Relief hypothesis. However, results from studies measuring fecal cortisol must be interpreted with care and alternative explanations, such as seasonal fluctuations in diet and thermoenergic demands, should be considered when drawing conclusions.

Comparison of two human infant urine collection methods for measuring estrone-3-glucuronide.

OBJECTIVES: Current human infant urine collection methods for the field are problematic for the researcher and potentially uncomfortable for the infant. In this study, we compared two minimally invasive methods for collecting infant urine: organic cotton balls and filter paper. MATERIALS AND METHODS: We first collected urine from infants using the clean catch method. We then used those samples to compare the performance of filter paper and cotton ball collection protocols. We analyzed the clean catch and cotton samples using commercial estrone-3-glucuronide (E1G) kits and tried two different extraction methods for the filter paper. Using a paired t-test (n = 10), we compared clean catch and cotton samples. We also compared effect sizes within and between methods. RESULTS: We were unable to extract enough urine from the filter paper to successfully assay the samples for E1G. The paired t-test revealed a statistically significant difference between the clean catch and cotton methods (t = 2.63, p-value = 0.03). However, the effect size was small (5.91 µg/ml, n = 10, 95% CI = 3.80, 8.02) and similar to or larger than the difference seen between duplicate wells for clean catch and cotton values. DISCUSSION: While this study is limited by sample size, our results indicate that filter paper is not a field-friendly method for collecting infant urine. However, we found that organic cotton balls showed similar values to the clean catch method, and we propose this method as an alternative, minimally invasive method for study of E1G in human infant urine.

The role of intragroup agonism in parent-offspring relationships and natal dispersal in monogamous owl monkeys (Aotus azarae) of Argentina.

Agonistic behaviors are common in many group-living taxa and may serve a variety of functions, ranging from regulating conflicts over reproduction to defending food resources. However, high rates of agonism are not expected to occur among close relatives or individuals in established mating relationships, which are characteristics of monogamous groups. To contribute to our understanding of agonism within socially monogamous groups, we collected behavioral and demographic data from Azara's owl monkeys (Aotus azarae) in the Gran Chaco of Argentina over 14 years. We examined factors related to age, sex, kinship, and behavioral context to evaluate predictions of the hypotheses that agonism functions to regulate dispersal and that it mediates competition for food and/or mates. Intragroup agonism was relatively rare: the group rate was approximately one event every three and a half hours. Rates of agonism were generally similar for both sexes, but there were marked differences among age categories. Agonism performed by adults was more frequently directed at subadults than at younger offspring. In contrast, agonistic interactions involving infants were very rare. Among interactions between adults and subadults, adults were much more frequently the actors than the recipients, suggesting that agonism from adults may influence natal dispersal of subadults. Agonistic events were most frequent during foraging, but also occurred more frequently than expected during bouts of social behavior. Overall, our results suggest that agonism in owl monkeys serves as a mechanism for regulating dispersal, and also likely plays a role in mediating mating and feeding competition.

Hormonal correlates of development and natal dispersal in wild female owl monkeys (Aotus azarae) of Argentina.

Pair-living and socially monogamous primates typically do not reproduce before dispersing. It is currently unclear whether this reproductive suppression is due to endocrine or behavioral mechanisms. Cooperatively breeding taxa, like callitrichids, may forego reproduction in natal groups because they reap inclusive fitness benefits and/or they are avoiding inbreeding. However, neither of these benefits of delayed reproduction appear to adequately explain the lack of reproduction prior to leaving the natal group in pair-living monogamous species. In this study, we determined whether wild Azara's owl monkeys (Aotus azarae) in the Argentinean Chaco establish reproductive maturity prior to dispersing. We utilized 635 fecal extracts to characterize reproductive hormone profiles of 11 wild juvenile and subadult females using enzyme immunoassays. Subadult females showed hormone profiles indicative of ovulatory cycling and had mean PdG and E1G concentrations approximately five times higher than juveniles. Contrary to expectations from the inbreeding avoidance hypothesis, female owl monkeys do not delay puberty, but rather commence ovarian cycling while residing in their natal group. Still, subadults appear to have a period during which they experience irregular, non-conceptive cycles prior to reproducing. Commencing these irregular cycles in the natal group may allow them to develop a state of suspended readiness, which could be essential to securing a mate, while avoiding costs of ranging solitarily. Our results indicate that reproductive suppression in female owl monkeys is not due to endocrine suppression. We suggest that adults likely use behavioral mechanisms to prevent subadults from reproducing with unrelated adult males in their natal group.