Fetal Programming of Semen Quality (FEPOS) Cohort – A DNBC Male-Offspring Cohort

Figures & data

Figure 1 Flowchart of the sampling strategy and recruitment process of the FEPOS cohort.

Figure 2 Overview of the cumulative number of invited and participants in the FEPOS cohort during the study period from March 2017 to December 2019.

Figure 3 Map of the residence of participants in the FEPOS cohort.

Table 1 Overview of Data Available on the FEPOS Cohort

Category	Data Source	Variables
Prenatal exposure data*
Lifestyle	DNBC telephone interview at gestational weeks 16 and 31	Alcohol Caffeine Smoking Physical activity Pre-pregnancy body mass index Stress Dietary supplements
Employment	DNBC telephone interview at gestational weeks 16 and 31	Work Workload (only first interview)
Health and medicine use	DNBC telephone interview at gestational weeks 16 and 31	Diseases Over the counter pain medication Prescribed medication Nonsteroidal anti-inflammatory drugs (NSAIDs)
Reproduction	DNBC telephone interview at gestational weeks 16 and 31	Age of onset of puberty Menstrual cycle characteristics Birth control Infertility treatment Weight gain during pregnancy
Vitamins^†	Maternal plasma from gestational week 7 or 26	Vitamin D3
Biomarkers of perfluoroalkyl acid exposure (PFFA)^†	Maternal plasma from gestational week 7 or 26	Perfluorohexane sulfonic acid (PFHxS) Perfluorooctanoic acid (PFOA) Perfluorooctane sulfonic acid (PFOS) Perfluoroheptanoic acid (PFHpA) Perfluorononanoic acid (PFNA) Perfluorodecanoic acid (PFDA) Perfluoroundecanoic acid (PFUnDA) Perfluorododecanoic acid (PFDoDA)
Biomarkers of phthalate exposure^†	Maternal plasma from gestational week 7 or 26	Mono-(2-ethyl-5-oxohexyl) phthalate (5-oxo-MEHP) Mono-(2-ethyl-5-hydroxyhexyl) phthalate (5-OH-MEHP) Mono-(2-ethyl-5-carboxypentyl) phthalate (5-cx-MEPP) Mono-(4-methyl-7-carboxyheptyl) phthalate (cx-MiNP)
Biomarkers of additional xenobiotics^†	Maternal plasma from gestational week 7 or 26	Triclosan^§ Cotinine^§ Acetaminophen (APAP)
Data on sons’ own exposures
Lifestyle	FEPOS questionnaire	Diet Alcohol Caffeine Smoking Physical activity Narcotic drugs
Education and work	FEPOS questionnaire	Educational level Work
Health	FEPOS questionnaire	Diseases Medication
Puberty and sexual experience	FEPOS questionnaire	Puberty Sexual experience
Physical tests	Clinical examination	Blood pressure Fat and muscle distribution Lung function Testicle size
Semen sample	Semen sample delivered at the clinical examination	Sperm concentration Semen volume Total sperm count Morphology Motility DNA fragmentation index (DFI)^ǂ
Vitamins^†	Blood sample taken at the clinical examination	Vitamin D3
Hormones^‡	Blood sample taken at the clinical examination	Hemoglobin A1c (HbA1c) Follitropin (FSH) Insulin-like Growth Factor I (IGF-1) Insulin Lutropin (LH) Thyrotropine (TSH) Thyroxine (T4) Sexual Hormone Binding Globulin (SHBG) Testosterone Thyroxine (Free T4) Oestradiol Inhibin B
Biomarkers of perfluoroalkyl acid exposure (PFFA)^†	Blood sample taken at the clinical examination	Perfluorohexane sulfonic acid (PFHxS) Perfluorooctane sulfonic acid (PFOS) Perfluoroheptanoic acid (PFHpA) Perfluorooctanoic acid (PFOA) Perfluorononanoic acid (PFNA) Perfluorodecanoic acid (PFDA) Perfluoroundecanoic acid (PFUnDA) Perfluorododecanoic acid (PFDoDA)
Biomarkers of phthalate exposure^†§	Urine sample taken at the clinical examination	Monoethyl phthalate (MEP) Monobutyl phthalate (MBP) Monobenzyl phthalate (MBzP) Mono-(2-ethylhexyl) phthalate (MEHP) Mono-[2-(carboxymethyl)hexyl] Phthalate (2cx-MEHP) Mono-(4-methyl-7-hydroxyloctyl) phthalate (OH-MiNP) Mono-(4-methyl-7oxo octyl) phthalate (oxo-MiNP) Monocarboxyisonoyl Phthalate (cx-MiDP) Mono-(propyl-6-hydroxyheptyl) phthalate (OH-MPHP)
Biomarkers of phthalate exposure^†§	Blood and urine taken at the clinical examination	Mono-(2-ethyl-5-oxohexyl) phthalate (5-oxo-MEHP) Mono-(2-ethyl-5-hydroxyhexyl) phthalate (5-OH-MEHP) Mono-(2-ethyl-5-carboxypentyl) phthalate (5-cx-MEPP) Mono-(4-methyl-7-carboxyheptyl) phthalate (cx-MiNP)
Biomarkers of bisphenol exposure^†§	Urine sample taken at the clinical examination	Bisphenol A (BPA) Bisphenol S (BPS) Bisphenol F (BPF)
Biomarker of pesticide exposure^†§	Urine sample taken at the clinical examination	3-Phenoxybenzoic acid (3PBA) Trichloropyridinol (TCP)
Biomarker of exposure from urban pollution to polycyclic aromatic hydrocarbons^†§	Urine sample taken at the clinical examination	1-Hydroxypyrene (1-HP)
Biomarker of smoking^†§	Blood and urine taken at the clinical examination	Cotinine
Additional xenobiotic chemicals^†§	Urine sample taken at the clinical examination	Di-phenylphosphate (DPP)
Additional xenobiotic chemicals^†§	Blood and urine taken at the clinical examination	Triclosan

Notes: *See DNBC website (https://www.dnbc.dk/) for all data available on mothers. ^†Analyses are ongoing. ^‡Analyses await funding. ^§Information on dilution with density or creatinine is available on all analyses in urine.

Table 2 Selected Maternal Characteristics of the 1057 Mothers to the 1058 Included Sons in the FEPOS Cohort*

Characteristics	All
	N=1057
Age in years, mean [SD]	30.5 [4.2]
Pre-pregnancy body mass index (kg/m^2), mean [SD]	22.8 [3.6]
Infertility treated, N (%)	67 (6)
Smokers, N (%)	245 (23)
Cotinine plasma levels (ng/mL)^†, median [p10-p90]	0.4 [0.1–50.4]
Number of samples <LOD of 0.2 ng/mL, N (%)	146 (27)
Alcohol consumption (units/week), mean [SD]	0.7 [1.1]
Family occupational status, N (%)
High grade professional	357 (34)
Low grade professional	350 (33)
Skilled worker	203 (19)
Unskilled worker	98 (9)
Student	45 (4)
Unclassified	≤5
Plasma sample available^†, N (%)
1st trimester blood sample	489 (92)
2nd trimester blood sample	42 (8)
3rd trimester blood sample	≤5
Biomarkers of perfluoroalkyl acid exposure (PFFA)^†, median [p10-p90]
Perfluorooctanoic acid (PFOA) (ng/mL)	4.6 [2.5–7.2]
Number of samples <LOD of 0.02 ng/mL, N (%)	0 (0)
Perfluorooctane sulfonic acid (PFOS) (ng/mL)	26.3 [17.0–40.1]
Number of samples <LOD of 0.02 ng/mL, N (%)	0 (0)
Biomarkers of phthalate exposure^†, median [p10-p90]
Mono-(2-ethyl-5-oxohexyl) phthalate (5-oxo-MEHP) (ng/mL)	0.1 [<LOD-0.3]
Number of samples <LOD of 0.1 ng/mL, N (%)	271 (51)
Mono-(2-ethyl-5-hydroxyhexyl) phthalate (5-OH-MEHP) (ng/mL)	0.2 [0.1–0.9]
Number of samples <LOD of 0.05 ng/mL, N (%)	52 (10)
Mono-(2-ethyl-5-carboxypentyl) phthalate (5-cx-MEPP) (ng/mL)	0.7 [0.4–1.5]
Number of samples <LOD of 0.05 ng/mL, N (%)	0 (0)
Mono-(4-methyl-7-carboxyheptyl) phthalate (cx-MiNP) (ng/mL)	0.3 [0.2–0.8]
Number of samples <LOD of 0.02 ng/mL, N (%)	0 (0)
Biomarkers of additional xenobiotics^†, median [p10-p90]
Triclosan (ng/mL)	1.3 [<LOD-15.1]
Number of samples <LOD of 0.1 ng/mL, N (%)	107 (20)
Acetaminophen (APAP) (ng/mL)	<LOD [<LOD-1.4]
Number of samples <LOD of 0.5 ng/mL, N (%)	424 (80)

Notes: *One mother had twins. ^†Analysis is ongoing, so far 532 samples have been analysed. p10-p90: 10th-90th percentile.

Abbreviation: LOD, Limit of detection.

Table 3 Selected Characteristics of 1058 Sons Included in the FEPOS Cohort*

Characteristics	All	Copenhagen Clinic	Aarhus Clinic
	N=1058	N=830	N=228
Body Mass Index (kg/m^2), mean [SD]	22.5 [3.4]	22.5 [3.3]	22.7 [3.6]
Body Composition analysed, N (%)	1055 (100)	829 (100)	226 (99)
Body fat mass (kg), mean [SD]	11.5 [6.7]	11.3 [6.6]	12.3 [6.8]
Muscle mass (kg), mean [SD]	60.8 [7.1]	61.0 [7.1]	60.0 [7.1]
Smoking habits, N (%)
Daily smoking	250 (24)	198 (24)	52 (23)
Occasional/former smoker	296 (28)	241 (29)	55 (24)
Never smoker	508 (480)	389 (47)	119 (52)
Cotinine plasma levels (ng/mL)^†§, median [p10-p90]	5.8 [0.3–2951.9]	11.5 [0.4–2540.1]	1.5 [0.3–3124.4]
Number of samples <LOD of 0.3 ng/mL, N (%)	46 (8)	24 (7)	22 (10)
Alcohol drinker, N (%)	984 (93)	769 (93)	215 (94)
Physical activity, N (%)
No physical activity	184 (18)	149 (18)	35 (16)
1 time a week	139 (13)	107 (13)	32 (14)
2 times a week	201 (19)	165 (20)	36 (16)
3 times a week	220 (21)	164 (20)	56 (25)
4 times a week	155 (15)	121 (15)	34 (15)
≥5 times a week	155 (15)	122 (15)	33 (15)
Biospecimens collected, N (%)
Semen	1054 (100)	829 (100)	225 (99)
Blood	1047 (99)	821 (99)	226 (99)
Urine	1042 (99)	819 (99)	223 (98)
Hair	690 (65)	484 (58)	206 (90)
Lung function measured, N (%)	1052 (100)	826 (100)	226 (99)
Forced Expiratory Volume (FVC), mean [SD]	5.6 [0.8]	5.6 [0.8]	5.3 [1.0]
Forced Expired Volume in the first second (FEV1), mean [SD]	4.6 [0.7]	4.7 [0.6]	4.4 [0.9]
Ejaculation at the clinic, N (%)	910 (87)	720 (87)	190 (86)
Spillage of semen sample, N (%)	182 (17)	146 (18)	36 (16)
Season of ejaculation, N (%)
Winter	199 (19)	146 (18)	53 (23)
Spring	205 (19)	142 (17)	63 (28)
Summer	258 (24)	213 (26)	45 (20)
Autumn	396 (37)	329 (40)	67 (29)
Biomarkers of perfluoroalkyl acid exposure (PFFA)^†‡, median [p10-p90]
Perfluorooctanoic acid (PFOA)^‡ (ng/mL)	1.3 [0.9–2.0]	1.4 [0.9–2.1]	1.3 [0.9–2.0]
Number of samples <LOD of 0.02 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
Perfluorooctane sulfonic acid (PFOS)^‡ (ng/mL)	4.3 [2.6–7.2]	4.1 [2.6–6.5]	4.4 [2.8–7.7]
Number of samples <LOD of 0.02 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
Biomarkers of pesticide exposure^§, median [p10-p90]
Trichloropyridinol (TCP)^§ (ng/mL)	1.0 [0.3–3.5]	1.0 [0.4–3.7]	0.9 [0.3–3.2]
Number of samples <LOD of 0.1 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
3-Phenoxybencoic acid (3PBA)^§ (ng/mL)	0.3 [0.1–1.1]	0.3 [0.1–1.1]	0.3 [0.1–1.3]
Number of samples <LOD of 0.05 ng/mL, N (%)	31 (6)	20 (6)	11 (5)
Bisphenol A (BPA)^†§, median [p10-p90]	1.3 [0.3–5.5]	1.4 [0.4–6.6]	1.2 [0.3–4.7]
Number of samples <LOD of 0.2 ng/mL, N (%)	19 (3)	7 (2)	12 (5)
Biomarkers of phthalate exposure measured in urine^†§, median [p10-p90]
Mono-(4-methyl-7-carboxyheptyl) phthalate (cx-MiNP)^§ (ng/mL)	3.6 [1.0–12.6]	3.7 [1.0–13.4]	3.5 [1.0–10.5]
Number of samples <LOD of 0.05 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
Mono-(2-ethyl-5-oxohexyl) phthalate (5-oxo-MEHP)^§ (ng/mL)	2.6 [0.8–8.1]	2.7 [0.8–8.2]	2.4 [0.7–7.5]
Number of samples <LOD of 0.2 ng/mL, N (%)	≤5	≤5	0 (0)
Mono-(2-ethyl-5-hydroxyhexyl) phthalate (5-OH-MEHP)^§ (ng/mL)	4.5 [1.5–14.3]	4.7 [1.6–15.2]	4.3 [1.3–13.0]
Number of samples <LOD of 0.1 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
Mono-(2-ethyl-5-carboxypentyl) phthalate (5-cx-MEPP)^§ (ng/mL)	3.7 [1.1–11.9]	3.9 [1.3–12.4]	3.5 [1.0–10.1]
Number of samples <LOD of 0.07 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
Biomarkers of phthalate exposure measured in blood^†‡, median [p10-p90]
Mono-(4-methyl-7-carboxyheptyl) phthalate (cx-MiNP)^‡	0.5 [0.2–1.5]	0.5 [0.2–1.5]	0.5 [0.2–1.5]
Number of samples <LOD of 0.02 ng/mL, N (%)	0 (0)	0 (0)	0 (0)
Mono-(2-ethyl-5-oxohexyl) phthalate (5-oxo-MEHP)^‡	<LOD [<LOD-0.1]	<LOD [<LOD-0.1]	<LOD [<LOD-0.1]
Number of samples <LOD of 0.1 ng/mL, N (%)	532 (96)	314 (95)	218 (96)
Mono-(2-ethyl-5-hydroxyhexyl) phthalate (5-OH-MEHP)^‡	<LOD [<LOD-0.1]	<LOD [<LOD-0.1]	0.1 [<LOD-0.1]
Number of samples <LOD of 0.05 ng/mL, N (%)	305 (55)	193 (59)	112 (50)
Mono-(2-ethyl-5-carboxypentyl) phthalate (5-cx-MEPP)^‡	0.2 [0.1–0.4]	0.2 [0.1–0.4]	0.2 [0.1–0.3]
Number of samples <LOD of 0.05 ng/mL, N (%)	12 (2)	7 (2)	≤5
Triclosan^†‡, median [p10-p90]	<LOD [<LOD-0.2]	<LOD [<LOD-0.2]	<LOD [<LOD-0.2]
Number of samples <LOD of 0.1 ng/mL, N (%)	451 (81)	262 (80)	189 (84)

Notes: *One mother had twins. ^†Analysis is ongoing, so far 559 urine samples and 555 blood samples have been analysed. ^‡Measured in blood. ^§Measured in urine. p10-p90: 10th-90th percentile.

Abbreviation: LOD, limit of detection.

Table 4 Semen Characteristics of 1054 Sons Who Delivered a Semen Sample in the FEPOS Cohort

Characteristics	All	Copenhagen Clinic	Aarhus Clinic
	N=1054	N=829	N=225
	Median (5–95 Percentile)	Median (5–95 Percentile)	Median (5–95 Percentile)
Abstinence time (days)	2.0 (0.5–4.5)	2.0 (0.5–4.5)	2.5 (0.5–4.5)
Semen volume (mL)*	2.7 (1.0–5.4)	2.7 (1.0–5.4)	2.6 (1.0–5.2)
Sperm concentration (million/mL)	38.7 (2.7–138.0)	38.7 (2.8–138.8)	38.6 (2.4–134.8)
Total sperm count (million)*	102.6 (8.1–409.2)	104.9 (7.2–407.2)	95.9 (8.4–417.1)
Motile sperm (%)^†	63 (30–83)	64 (31–84)	63 (28–80)
Morphologically normal sperm (%)^†	6 (0–15)	6 (0–15)	6 (1–15)
Testicular size (mL)	15 (8–25)	15 (8–25)	15 (8–25)
Minutes until motility analysis	45 (30–90)	45 (35–90)	35 (25–90)

Notes: *182 samples excluded due to spillage. ^†Motility and morphology not available for 17 sons with azoospermia.