Milk provisioning in oviparous caecilian amphibians.

Among vertebrates, the yolk is commonly the only form of nutritional investment offered by the female to the embryo. Some species, however, have developed parental care behaviors associated with specialized food provisioning essential for offspring survival, such as the production of lipidic-rich parental milk in mammals. Here, we show that females of the egg-laying caecilian amphibian Siphonops annulatus provide similarly lipid-rich milk to altricial hatchlings during parental care. We observed that for 2 months, S. annulatus babies ingested milk released through the maternal vent seemingly in response to tactile and acoustic stimulation by the babies. The milk, composed mainly of lipids and carbohydrates, originates from the maternal oviduct epithelium's hypertrophied glands. Our data suggest lactation in this oviparous nonmammalian species and expand the knowledge of parental care and communication in caecilians.

Distribution of major toxins in Rhinella marina parotoid macroglands using Desorption-Electrospray-Ionization mass spectrometry imaging (DESI-MSI).

Amphibian cutaneous glands secrete toxins used in different vital functions including passive defense. Through Desorption Electrospray Ionization-Imaging we analyzed the distribution of the major toxins of the toad Rhinella marina parotoid macroglands. Alkaloids and steroids showed characteristic distribution and intensity within the glands and were also present at lower levels on the skin surface. A comprehensive overview of toxins distribution in toads' skin might help to understand their full biological role within the amphibians.

Structural cutaneous adaptations for defense in toad (Rhinella icterica) parotoid macroglands.

Toads have a pair of glandular accumulations on each side of the dorsal region of the head known as parotoid macroglands. These macroglands consist of secretory units (granular glands), each one capped with an epithelial plug. When threatened, toads point one of the parotoids toward the aggressor, and if the aggressor squeezes the parotoid with its teeth, jets of poison will come out of the secretory units and hit the predator's oral mucosa, thereby causing poisoning. Our study focused on the mechanism of parotoid function by comparing parotoids from toads naturally attacked by dogs with those manually compressed. We verified that the process of glandular emptying in response to dog bites is very similar to that following manual compression. We observed that the structure of the plug plays an essential role in the release of the poison jets. Our results suggest that the parotoids may act as "bulletproof vests," reducing the impact of the force exerted by predator attacks, and thus may function as a passive antipredator mechanism.

Parotoid, radial, and tibial macroglands of the frog Odontophrynus cultripes: Differences and similarities with toads.

Anuran integument is characterized by the presence of glands, some of which are responsible for toxin production. In some species these glands accumulate in parts of the body strategically located against predators, forming structures known as macroglands. This is the case for parotoid macroglands, on the dorsum of the head, tibial macroglands, on the rear limbs, and radial macroglands, on the forelimbs of toads and some other anurans. The toad Rhinella jimi, for example, simultaneously displays all three types of macroglands, which is unusual even among bufonids. Interestingly, considering the phylogenetic distance, the frog Odontophrynus cultripes (Odontophrynidae) also presents these three macroglandular types. In this study we analyze the morphology of O. cultripes macroglands and the chemical composition of their poison using an interdisciplinary approach. In this species, the parotoid, tibial, and radial macroglands consist of aggregates of elongated and juxtaposed poison glands, arranged in a honeycomb style, very similar to that of toads. Comparative analysis of these three macrogland types shows significant differences in both the morphology of secretory granules and biochemical composition. The present work on O. cultripes contributes to the evidence that amphibians, or at least anurans, share a basic design for all cutaneous glandular accumulations. The determinant factor for macroglandular formation may be the selective pressure for defense against predators.

Functional assessment of toad parotoid macroglands: a study based on poison replacement after mechanical compression.

Toads have a pair of parotoid macroglands behind the eyes that secrete poison used in passive defence against predators. These macroglands are composed of juxtaposed alveoli, each one bearing a syncytial gland, all connected to the exterior by ducts. When the parotoids are bitten, the poison is expelled on the predator oral mucosa in the form of jets, causing several pharmacological actions. After poison release, the empty secretory syncytia immediately collapse in the interior of their respective alveoli and gradually start refilling. After parotoid manual compression, simulating a predator's bite, we studied, by means of morphological methods, the replacement of the poison inside the alveoli. The results showed that after compression, a considerable number of alveoli remained intact. In the alveoli that were effectively affected the recovery occurs in different levels, from total to punctual and often restrict to some areas of the syncytia. The severely affected alveoli seem not recover their original functional state. The fact that only a part of the parotoid alveoli is compressed during an attack seems to be crucial for toad survival, since the amphibian, after being bitten by a predator, do not lose all its poison stock, remaining protected in case of new attacks.

Passive and active defense in toads: the parotoid macroglands in Rhinella marina and Rhaebo guttatus.

Amphibians have many skin poison glands used in passive defense, in which the aggressor causes its own poisoning when biting prey. In some amphibians the skin glands accumulate in certain regions forming macroglands, such as the parotoids of toads. We have discovered that the toad Rhaebo guttatus is able to squirt jets of poison towards the aggressor, contradicting the typical amphibian defense. We studied the R. guttatus chemical defense, comparing it with Rhinella marina, a sympatric species showing typical toad passive defense. We found that only in R. guttatus the parotoid is adhered to the scapula and do not have a calcified dermal layer. In addition, in this species, the plugs obstructing the glandular ducts are more fragile when compared to R. marina. As a consequence, the manual pressure necessary to extract the poison from the parotoid is twice as high in R. marina when compared to that used in R. guttatus. Compared to R. marina, the poison of R. guttatus is less lethal, induces edema and provokes nociception four times more intense. We concluded that the ability of R. guttatus to voluntary squirt poison is directly related to its stereotyped defensive behavior, together with the peculiar morphological characteristics of its parotoids. Since R. guttatus poison is practically not lethal, it is possibly directed to predators' learning, causing disturbing effects such as pain and edema. The unique mechanism of defense of R. guttatus may mistakenly justify the popular myth that toads, in general, squirt poison into people's eyes.