Genomic Signatures of Local Adaptation under High Gene Flow in Lumpfish-Implications for Broodstock Provenance Sourcing and Larval Production.

Aquaculture of the lumpfish (Cyclopterus lumpus L.) has become a large, lucrative industry owing to the escalating demand for "cleaner fish" to minimise sea lice infestations in Atlantic salmon mariculture farms. We used over 10K genome-wide single nucleotide polymorphisms (SNPs) to investigate the spatial patterns of genomic variation in the lumpfish along the coast of Norway and across the North Atlantic. Moreover, we applied three genome scans for outliers and two genotype-environment association tests to assess the signatures and patterns of local adaptation under extensive gene flow. With our 'global' sampling regime, we found two major genetic groups of lumpfish, i.e., the western and eastern Atlantic. Regionally in Norway, we found marginal evidence of population structure, where the population genomic analysis revealed a small portion of individuals with a different genetic ancestry. Nevertheless, we found strong support for local adaption under high gene flow in the Norwegian lumpfish and identified over 380 high-confidence environment-associated loci linked to gene sets with a key role in biological processes associated with environmental pressures and embryonic development. Our results bridge population genetic/genomics studies with seascape genomics studies and will facilitate genome-enabled monitoring of the genetic impacts of escapees and allow for genetic-informed broodstock selection and management in Norway.

Mitogenomics of the suborder Cottoidei (Teleostei: Perciformes): Improved assemblies, mitogenome features, phylogeny, and ecological implications.

We determined the mitogenome of Cyclopterus lumpus using a hybrid sequencing approach, and another four closely related species in the Liparidae based on available next-generation sequence data. We found that the mitogenome of C. lumpus was 17,266 bp in length, where the length and organisation were comparable to those reported for cottoids. However, we found a GC-homopolymer region in the intergenic space between tRNALeu2 and ND1 in liparids and cyclopterids. Phylogenetic reconstruction confirmed the monophyly of infraorders and firmly supported a sister-group relationship between Cyclopteridae and Liparidae. Purifying selection was the predominant force in the evolution of cottoid mitogenomes. There was significant evidence of relaxed selective pressures along the lineage of deep-sea fish, while selection was intensified in the freshwater lineage. Overall, our analysis provides a necessary expansion in the availability of mitogenomic sequences and sheds light on mitogenomic adaptation in Cottoidei fish inhabiting different aquatic environments.

Genome- and transcriptome-derived microsatellite loci in lumpfish Cyclopterus lumpus: molecular tools for aquaculture, conservation and fisheries management.

The lumpfish Cyclopterus lumpus is commercially exploited in numerous areas of its range in the North Atlantic Ocean, and is important in salmonid aquaculture as a biological agent for controlling sea lice. Despite the economic importance, few genetic resources for downstream applications, such as linkage mapping, parentage analysis, marker-assisted selection (MAS), quantitative trait loci (QTL) analysis, and assessing adaptive genetic diversity are currently available for the species. Here, we identify both genome- and transcriptome-derived microsatellites loci from C. lumpus to facilitate such applications. Across 2,346 genomic contigs, we detected a total of 3,067 microsatellite loci, of which 723 were the most suitable ones for primer design. From 116,555 transcriptomic unigenes, we identified a total of 231,556 microsatellite loci, which may indicate a high coverage of the available STRs. Out of these, primer pairs could only be designed for 6,203 loci. Dinucleotide repeats accounted for 89 percent and 52 percent of the genome- and transcriptome-derived microsatellites, respectively. The genetic composition of the dominant repeat motif types showed differences from other investigated fish species. In the genome-derived microsatellites AC/GT (67.8 percent), followed by AG/CT (15.1 percent) and AT/AT (5.6 percent) were the major motifs. Transcriptome-derived microsatellites showed also most dominantly the AC/GT repeat motif (33 percent), followed by A/T (26.6 percent) and AG/CT (11 percent). Functional annotation of microsatellite-containing transcriptomic sequences showed that the majority of the expressed sequence tags encode proteins involved in cellular and metabolic processes, binding activity and catalytic reactions. Importantly, STRs linked to genes involved in immune system process, growth, locomotion and reproduction were discovered in the present study. The extensive genomic marker information reported here will facilitate molecular ecology studies, conservation initiatives and will benefit many aspects of the breeding programmes of C. lumpus.

Initiation of complementary feeding and duration of total breastfeeding: unlimited access to lactation consultants versus routine care at the well-baby clinics.

INTRODUCTION: Breastfeeding has several advantages for both mother and child. Lactation consultants may promote prolonged breastfeeding, but little is known about their impact on the initiation of complementary feeding. SUBJECTS AND METHODS: Dietary intake during the initial complementary feeding period from 5 to 6 months was collected on mother-infant pairs who had unlimited access to lactation consultants along with those mother-infant pairs who received routine care at the well-baby clinics. The total duration of breastfeeding in each study population was also recorded, and total breastfeeding durations of infants receiving complementary foods from 4 months and those exclusively breastfed for 6 months in each of the two study populations were compared. RESULTS: Higher proportion of infants of mothers with unlimited access to lactation consultants were fed vegetable and vegetable purées (p=0.05) and more than one food type (p=0.05) at 5 months. Furthermore, a lower percentage of them had three meals per day at 6 months (p=0.001) compared with those receiving routine care at the well-baby clinics. Infants exclusively breastfed for 6 months all had similar duration of total breastfeeding. CONCLUSIONS: Mother-infant pairs with unlimited access to lactation consultant had slower introduction of complementary foods at the initial complementary feeding period, according to number of infant's meals at 6 months of age. Furthermore, those exclusively breastfed for 6 months had more prolonged breastfeeding compared with mothers who began complementary feeding at 4 months regardless of exposure to lactation consultants.

Exclusive breastfeeding for 4 versus 6 months and growth in early childhood.

AIM: To investigate the growth and the prevalence of overweight in early childhood among infants exclusively breastfed for 6 months (EBF) compared with those receiving complementary foods from 4 months of age in addition to breast milk (CF). METHODS: A total of 119 mother-infant pairs were randomised either in the CF or in the EBF group. Weight, length and head circumference of the infants were measured at birth, 6 weeks, and 3-6 months of age. In the follow-up, the children's weight, length and head circumference were measured at 8, 10, 12 and 18 months and weight and height at 29-38 months. RESULTS: There were no differences between groups in the anthropometric outcome measures of weight-for-age (p = 0.78), length-for-age (p = 0.59), head-circumference-for-age (p = 0.82) and BMI-for-age (p = 0.61), using repeated measurements ANOVA. Furthermore, no difference was seen in the prevalence between groups in risk of being overweight or in those who were overweight at 18 months and 29-38 months of age. CONCLUSION: Exclusive breastfeeding for the first 4 or 6 months of life does not seem to affect the risk of being overweight or the prevalence of those who were overweight in early childhood.

Exclusive breastfeeding and developmental and behavioral status in early childhood.

Breastfeeding during infancy may have beneficial effects on various developmental outcomes in childhood. In this study, exclusively breastfed infants were randomly assigned to receive complementary foods from the age of 4 months in addition to breast milk (CF, n = 60), or to exclusively breastfeed to 6 months (EBF, n = 59). At 18 months and again at 30-35 months of age, the children were evaluated with the Parent's Evaluation of Developmental Status questionnaire (PEDS) and the Brigance Screens-II. The parents completed the PEDS questionnaire at both time intervals and the children underwent the Brigance Screens-II at 30-35 months. At 30-35 months, no significant differences were seen in developmental scores from the Brigance screening test (p = 0.82). However, at 30-35 months a smaller percentage of parents in group CF (2%) had concerns about their children's gross motor development compared to those in group EBF (19%; p = 0.01), which remained significant when adjusted for differences in pre-randomization characteristics (p = 0.03). No sustained effect of a longer duration of exclusive breastfeeding was seen on selected measures of developmental and behavioral status at 18 months, although at 30-35 months, a smaller percentage of parents of children introduced to complementary foods at four months of age expressed concerns about their gross motor development.

Timing of the introduction of complementary foods in infancy: a randomized controlled trial.

OBJECTIVE: To increase knowledge on iron status and growth during the first 6 months of life. We hypothesized that iron status would be better in infants who received complementary foods in addition to breast milk compared with those exclusively breastfed. METHODS: One hundred nineteen healthy term (≥37 weeks) singleton infants were randomly assigned to receive either complementary foods in addition to breast milk from age 4 months (CF) or to exclusive breastfeeding for 6 months (EBF). Dietary data were collected by 3-day weighed food records, and data on iron status and growth were also collected. RESULTS: One hundred infants (84%) completed the trial. Infants in the CF group had higher mean serum ferritin levels at 6 months (P = .02), which remained significant when adjusted for baseline characteristics. No difference was seen between groups in iron deficiency anemia, iron deficiency, or iron depletion. The average daily energy intake from complementary foods of 5-month-olds in the CF group was 36.8 kJ per kg body weight. Infants in both groups grew at the same rate between 4 and 6 months of age. CONCLUSIONS: In a high-income country, adding a small amount of complementary food in addition to breast milk to infants' diets from 4 months of age does not affect growth rate between 4 and 6 months, but has a small and positive effect on iron status at 6 months. The biological importance of this finding remains to be determined.

Randomized controlled trial of 4 compared with 6 mo of exclusive breastfeeding in Iceland: differences in breast-milk intake by stable-isotope probe.

BACKGROUND: The WHO recommends exclusive breastfeeding (EBF) for 6 mo after birth. However, the time at which breast milk ceases to provide adequate energy and nutrition, requiring the introduction of complementary foods, remains unclear. Most studies that investigated this issue were observational and potentially confounded by variability in social circumstances or infant growth. OBJECTIVE: We hypothesized that EBF infants would consume more breast milk at age 6 mo than infants receiving breast milk and complementary foods. DESIGN: We measured anthropometric outcomes, body composition, and breast-milk intake at age 6 mo in infants who were randomly assigned at age 4 mo either to 6-mo EBF or to the introduction of complementary foods with continued breastfeeding. We recruited 119 infants from health centers in Reykjavik and neighboring municipalities in Iceland. In 100 infants who completed the protocol (50/group), breast-milk intake was measured by using stable isotopes, and complementary food intakes were weighed over 3 d in the complementary feeding (CF) group. RESULTS: Breast-milk intake was 83 g/d (95% CI: 19, 148 g/d) greater in EBF (mean ± SD: 901 ± 158 g/d) than in CF (818 ± 166 g/d) infants and was equivalent to 56 kcal/d; CF infants obtained 63 ± 52 kcal/d from complementary foods. Estimated total energy intakes were similar (EBF: 560 ± 98 kcal/d; CF: 571 ± 97 kcal/d). Secondary outcomes (anthropometric outcomes, body composition) did not differ significantly between groups. CONCLUSIONS: On a group basis, EBF to age 6 mo did not compromise infant growth or body composition, and energy intake at age 6 mo was comparable to that in CF infants whose energy intake was not constrained by maternal breast-milk output.

Safety and immunogenicity of CRM197-conjugated pneumococcal-meningococcal C combination vaccine (9vPnC-MnCC) whether given in two or three primary doses.

This randomized trial compares safety and immunogenicity when vaccinating infants with a pneumococcal-meningococcal conjugate vaccine in two doses vs. three doses. Infants (N=223) received 9vPnC-MnCC (CRM197-conjugated pneumococcal serotypes 1, 4, 5, 6B, 9V, 14, 18C, 19F, 23F and meningococcal C polysaccharides) either at 3 and 5 or 3, 4 and 5 months and a booster with either 9vPnC-MnCC or 23-valent pneumococcal-polysaccharide vaccine (23vPPS) and CRM197-MnCC, at 12 months. Safety was monitored and IgG measured at 3, 6, 12 and 13 months in all subjects and serum bactericidal activity (SBA) in half. The 9vPnC-MnCC vaccine was safe and induced significant IgG to all components. Three doses induced higher antibody GMCs (geometric mean concentrations) at 6 months to seven of nine pneumococcal serotypes. This was most significant for 6B and 23F (p<0.001), that also showed lower rate of responders>0.35 (6B, 23F) and >0.5 microg/mL (6B). Antibody GMCs remained lower following 9vPnC-MnCC booster in subjects primed with two doses although only significant for serotype 18C. Significant memory responses were observed 1 week after the 23vPPS toddler dose. MnCC-IgG GMC was lower after two doses, however with comparable SBA. This study shows that the 9vPnC-MnCC vaccine is safe and induces successful immunological memory, whether given in two or three primary doses.