Coastal Gulf of Mexico habitats are susceptible to saltwater intrusion from storm surges, drought, and human development altering freshwater flow. Alligators living in these habitats have low tolerance to saltwater due to their inability to excrete high sodium (Na+) concentrations. Previous research has demonstrated that long- (5 weeks) and short-term (1 week) 12‰ saltwater exposure significantly alter regulation of salt/water balance and cause disruptions of sex steroid hormones in juvenile alligators. This study investigates whether similar physiological effects occur in juvenile and sub-adult wild alligators caught in varying salinities (0.4 to 22.2‰). To this end, blood plasma samples obtained from alligators sampled in Grand Chenier, Louisiana were analyzed for concentrations of 15 hormones (renin-angiotensin-aldosterone system hormones, glucocorticoids, progestogens, androgens, estrogens) via liquid chromatography and tandem mass spectrometry (LC-MS/MS). Plasma samples were further analyzed for levels of various blood plasma biochemistry parameters (electrolytes, liver enzymes, proteins, glucose, cholesterol). The multivariate analyses of all blood chemistry parameters showed seasonal variations in select parameters and significant positive correlations for selected steroid hormones and blood sodium and chloride ion concentrations in juvenile wild alligators. Further understanding seasonal variation of various biomarkers and salinity’s effects on alligator physiology will better inform management decisions regarding wild alligator populations.