<b><i>Background:</i></b> Genes involved in the serotonin pathway may determine the susceptibility to alcohol dependence and its severity. The present study explored whether specific polymorphisms in the serotonin pathway could be associated with alcohol dependence or alcohol-related psychopathological symptoms. <b><i>Methods:</i></b> The cohort comprised 101 currently and 100 formerly alcohol-dependent males, as well as 97 male healthy blood donors. The following questionnaires were employed: the Alcohol Use Disorders Identification Test, the Zung Depression and Anxiety Scale, the Brief Social Phobia Scale, the Yale-Brown Obsessive Compulsive Scale and Obsessive Compulsive Drinking Scale, and the Buss-Durkee Hostility Inventory. Subjects were genotyped for bi- and triallelic <i>SLC6A4 5-HTTLPR,</i><i>HTR1A</i> rs6295, and <i>HTR1B</i> rs13212041. <b><i>Results:</i></b> Statistical differences in bi- and triallelic <i>SLC6A4 5-HTTLPR</i> genotype distribution were observed between the 3 groups investigated (p = 0.008 and p = 0.023, respectively); however, no gene-dose effect was observed. The severity of the alcohol problems was higher in currently alcohol-dependent subjects with the <i>5-HTTLPR</i> LL (p = 0.039) and L′L′ genotypes (p = 0.027). Formerly dependent subjects with the <i>5-HTTLPR</i> S′S′ genotype showed more social anxiety, depressive, and anxiety traits (p = 0.009, p = 0.006, and p = 0.036, respectively). Healthy controls with the <i>5-HTTLPR</i> SS genotype showed more traits of social anxiety (p = 0.020). <b><i>Conclusions:</i></b> Our findings suggest that bi- and triallelic <i>SLC6A4 5-HTTLPR</i> has some effects on the severity of alcohol dependence. Triallelic <i>5-HTTLPR</i> was associated with social anxiety, anxiety, and depressive traits in alcohol-dependent subjects.