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Rattlesnakes are born with fully functioning fangs and venom, and are capable of killing prey at birth. Some species of ants in the genus Formica are known to prey upon neonates, and Solenopsis invicta (fire ants) likely do, as well. The Ancient Maya considered the rattlesnake to be a "vision serpent" that acted as a conduit to the "otherworld". Rattlesnakes are predators that live in a wide array of habitats, hunting small animals such as birds and rodents. When they realize a kingsnake is nearby, they begin enacting a set of defensive postures known as "body bridging".

If drinking dew or small puddles, they sip the liquid either by capillary action or by flattening and flooding their lower jaws. Once a receptive female has been located, the male often spends several days following her around (a behavior not common outside of the mating season), frequently touching and rubbing her in an attempt to stimulate her.

The left atrium receives oxygenated blood from the lungs in the pulmonary circuit and pumps it to the ventricle and through the systemic circuit via capillaries and arteries.

Their results demonstrated that the more closely related a species was to rattlesnakes, the more similar that species was to rattlesnakes in both duration and rate of tail vibration. Once the prey has become incapacitated, the rattlesnake locates its head by odors emitted from the mouth. Rattlesnakes travel with their rattles held up to protect them from damage, but in spite of this precaution, their day-to-day activities in the wild still cause them to regularly break off end segments.The resulting serum is purified into a lyophilized powder, which is packaged for distribution and later use by human patients. In more heavily populated and trafficked areas, reports have been increasing of rattlesnakes that do not rattle. These pits have a relatively short effective range of about 1ft, but give the rattlesnake a distinct advantage in hunting for warm-blooded creatures at night. Nevertheless, rattlesnakes superimpose visual images created from information from the eyes with these thermal images from the pit organs to more accurately visualize their surroundings in low levels of light.

Methods of preparation include barbecueing [106] and frying; author Maud Newton, following a recipe by Harry Crews, described the taste, "at least when breaded and fried, like a sinewy, half-starved tilapia. They regularly share their winter burrows with a wide variety of other species (such as turtles, small mammals, invertebrates, and other types of snakes). Optimal digestion occurs when the snake maintains a body temperature between 80 and 85°F (25 and 29°C).Rattlesnake fangs are connected by venom ducts to large venom glands near the outer edge of the upper jaw, towards the rear of the head. The female produces the ova ("eggs") in her ovaries, after which they pass through her body cavity and into one of her two oviducts. Feeding habits play an important ecological role by limiting the size of rodent populations, which prevents crop damage and stabilizes ecosystems.

Rattlesnakes prefer a temperature range between 80 and 90°F (26 and 32°C), but can survive temperatures below freezing, recovering from brief exposure to temperatures as low as 4°F (−16°C), and surviving for several days in temperatures as low as 37°F (3°C). Rapid habitat destruction by humans, mass killings during events such as rattlesnake round-ups, and deliberate extermination campaigns all pose threats to rattlesnake populations in many areas. Aztec paintings, Central American temples, and the great burial mounds in the Southern United States are frequently adorned with depictions of rattlesnakes, often within the symbols and emblems of the most powerful deities.Females secrete small amounts of sex pheromones, which leave a trail the males follow using their tongues and Jacobson's organs as guides. Because antivenom is derived from animal antibodies, people generally display an allergic response during infusion, known as serum sickness. Rattlesnakes are the leading contributor to snakebite injuries in North America, but rarely bite unless provoked or threatened; if treated promptly, the bites are seldom fatal.