Numerous studies have assessed single nutrient and multiple nutrient supplements in the treatment of psychiatric symptoms. Recently there has been growing evidence suggesting that treatment with broad-spectrum micronutrient supplements (Rucklidge et al., 2012), B vitamin combinations (Stough et al., 2011) and vitamin C (Hughes et al., 2011) may have benefits for emotional and cognitive functioning. Whether a combination of nutrients acting synergistically is more effective than the administration of a B vitamin complex or vitamin C alone is yet to be determined. The present study was designed to address this question. To investigate this, 40 male and 40 female rats were administered a broad-spectrum micronutrient supplement (Berocca), a B vitamin complex supplement, or vitamin C alone in their drinking water, for seven weeks. A control group was provided unadulterated water. Animals were then examined through several behavioural measures of anxiety and one of short-term memory. The results revealed a number of significant behavioural differences between treatment groups, with the Y maze showing the greatest number of significant treatment effects. The Berocca group had a significantly higher percentage of entries into and occupancy of the changed arm, as well as total entries of both arms (ambulation) than all other treatment groups. There were also significant sex x treatment interactions in change arm entries and occupancy, with both measures increasing with Berocca supplementation in males, and increasing with both Berocca and B vitamin supplementation in females. This suggests that different treatments affected each sex differently. Overall rats receiving broad-spectrum supplementation (Berocca) showed significantly less emotionality and significantly improved short-term spatial memory when compared to other treatment groups.