Due to their widespread use in consumer products and industrial applications and their inherent chemical stability, perfluoroalkyl and polyfluoroalkyl substances (PFAS) are now recognised as persistent environmental contaminants. Recent epidemiological data indicates that PFAS pose a risk to human health, prompting us to explore their effects on male reproductive function via a chronic in vivo exposure model wherein adult male mice received a cocktail of nine PFAS for 12-weeks via their drinking water at doses that mimic those found in the Australian environment. Thereafter, we assessed effects on overall body and reproductive organ weights, sperm functional parameters and in vitro fertilization. Whilst whole body, testis and epididymal weights were not affected by PFAS-treatment, both seminal vesicle (p<0.05) and seminal fluid weights (p<0.01) were significantly reduced in PFAS-treated males. Additionally, daily sperm production was also significantly reduced among PFAS treated mice (p<0.05). Interrogation of the impact of PFAS on sperm function revealed a significant reduction in the ability of sperm to bind the zona pellucida surrounding ovulated oocytes (p<0.05). More detailed assessment of the functional competence of PFAS exposed sperm in terms of their ability to support in vivo fertilization and embryonic development following natural mating demonstrated a significant reduction in the size of litters sired from PFAS treated males (p<0.01, average 10.25 vs. 12.11 pups). Notably, the pups born from PFAS treated fathers were also significantly heavier than then untreated pups at day of birth (p<0.01). At 7-days of age, pups sired from PFAS treated males also displayed significantly larger anogenital distances (p<0.01). Collectively, these studies provide new insight into the effect of PFAS on male reproductive health and have prompted us to begin to explore the mechanistic basis of such effects as well to elucidate whether they are perpetuated across generations.