THE ANATOMICAL RECORD 298:1836–1847 (2015)

Development of the Superaltricial Monk

Parakeet (Aves, Psittaciformes): Embryo
Staging, Growth, and Heterochronies
JULIETA CARRIL1,2* AND CLAUDIA P. TAMBUSSI2,3
1
C
atedra De Histologıa Y Embriologıa Animal, Facultad De Ciencias Naturales Y Museo,
Universidad Nacional De La Plata, La Plata, Argentina
2
Consejo Nacional De Investigaciones Cientıficas Y Tecnicas (CONICET), Argentina
3
Centro De Investigaciones En Ciencias De La Tierra (CICTERRA), CONICET-UNC,
Cordoba, Argentina

ABSTRACT
Knowledge about the embryonic stages of birds is important in
answering many questions about development and evolution. We give the
first description of 41 embryological stages of the monk parakeet (Myiop-
sitta monachus) on the basis of external morphology and comparison with
the chicken. We also provide measurements of some external morphologi-
cal characters (i.e. body mass, crown-rump, beak, forelimb, and third toe
lengths) and perform comparisons with other precocial and altricial birds
with the aim of identifying heterochronous developmental features. The
following differences in the development of characters in the monk para-
keet when compared with other birds were found: (1) delay of the feath-
ers primordia, (2) wing buds initially greater than leg buds, (3) forelimbs
and hindlimbs with similar relative size, (4) retroversion of the toe IV, (5)
ventral curvature of the upper jaw, (6) positive regressions between
stages and beak length with acceleration and higher values and III toe
lengths with deceleration and lower values in the monk parakeet com-
pared to the chicken. The growth pattern of the monk paraket Myiopsitta
monachus could be influenced by some heterochronic processes like post-
displacement, acceleration and/or deceleration. Results of this research
allow the standard identification of stages in different species of parrots,
recognize similarities and differences between precocial (the chicken) and
altricial species (Myiopsitta), and provide planning data for future stud-
ies. Anat Rec, 298:1836–1847, 2015. V C 2015 Wiley Periodicals, Inc.

Key words: Evo-Devo; morphogenesis; Myiopsitta monachu;

neotropical parrots

Knowledge about the embryonic stages of birds is extensively studied. Most of the literature is based on
important in answering many questions in development, Gould (1977) and Alberch et al. (1979) approaches, but
evolution, and conservation. Biologists have long sought the models proposed by the authors led to ambiguity in
to understand the relationship between ontogeny and
phylogeny, particularly after the Haeckel’s and von
Baers drawings of embryos in the late nineteenth cen- *Correspondence to: Julieta Carril, Calle 64 N 3, La Plata
tury (Gould, 1977; Hopwood, 2007). The interest in avian B1900BVA, Buenos Aires, Argentina.
development had a significant relaunching a decade ago E-mail: [email protected]
with the Evo-Devo that aims to understand the mecha- Received 27 January 2015; Revised 29 May 2015; Accepted 12
nisms governing the evolution of development. Hetero- June 2015.
chronies or evolutionary changes in the timing and/or DOI 10.1002/ar.23256
rates of processes underlying the ontogenetic formation Published online 12 August 2015 in Wiley Online Library
of morphological traits (Reilly et al., 1997) has been (wileyonlinelibrary.com).

C 2015 WILEY PERIODICALS, INC.

V
 EMBRYONIC DEVELOPMENT OF THE MONK PARAKEET 1837
the application of certain concepts (Reilly et al., 1997). availability of specimens. Also is an interesting parrot to
In his classic review, Reilly et al. (1997) proposed six study because of all Psittaciformes, it is the only one
processes of heterochronic change that result from shifts that builds a stick nest instead of using a hole in a tree
in rate, offset, or onset of the developmental trajectories. and breeds in a single large nest with separate entran-
They include: acceleration and deceleration (in replace- ces for each couple (Collar, 1997 and bibliography cited
ment of neoteny) resulting from an increase or decrease therein). Monk parakeet eggs are small, white, and mea-
of growth rate; hypermorphosis and hypomorphosis (in sure 3 cm in length approximately. Clutch size is 7 on
replacement of enanism) as a result of an extended or average (one deposited every two days), and the incuba-
reduced offset time; and predisplacement and postdis- tion period is 24 days. The superaltricial chicks hatch
placement when the onset time of a structure begins with eyes closed, nearly without feathers and stay in the
before or after respectively. These processes can produce nest until 40 days after hatching (Navarro and Bucher,
truncated (paedomorphosis), extended (peramorphosis) 1992; Navarro et al., 1995; Salvador and Arambur u,
or the same (isomorphosis) traits when comparing two 1995).
ontogenetic trajectories. The study of comparative devel- To the best of our knowledge, this is the first thorough
opment gives the empirical basis for advances on this investigation dedicated to the development of parrots
matter. giving the first description of embryological development
Developmental biology is unimaginable in contempo- of a Neotropical parrot, the monk parakeet Myiopsitta
rary science without standard definition of normal monachus, and the first complete staging of any Psittaci-
stages (Hopwood, 2007). Widely used as model is the formes embryo. We provide measurements of some exter-
chick embryos development, extensively studied by nal morphological characters and performs comparisons
many researchers and discretized in 46 chronological with other precocial (e.g. chicken) and altricial (e.g. cock-
stages by Hamburger and Hamilton (1951). Since then, atiel) birds.
the normal stages of the embryo development of Gallus Our objectives were to identify heterochronus develop-
gallus domesticus have been applied as a model to solve mental features, to provide normative information that
several questions such as migration of germ cells, cellu- would enable standard identification of stages in differ-
lar and molecular interactions during development, and ent species of parrots, to recognize similarities and dif-
production of transgenic birds and chimeras (Ainsworth ferences between the chicken (precocial) and Myiopsitta
et al., 2010; Le Douarin and Dieterlen-Lièvre, 2013). (altricial) species, and to provide planning data for
Nevertheless, knowledge about prenatal development in future studies.
birds grew very slowly. Complete or partial embryonic
development is known of few precocial or semiprecocial MATERIALS AND METHODS
birds, particularly poultry and/or species of commercial
importance like chickens, quail and other Phasianidae, A total of 200 specimens of monk parakeet Myiopsitta
and ducks (e.g. Fant, 1957; Rempel and Eastlick, 1957; monachus were used in the development staging series.
Koecke, 1958; Mun and Kosin, 1960; Phillips and Wil- Eggs were obtained from nests in Dean Funes locality,
liams, 1964; Hendrickx and Hanzlik, 1965; Kaltofen, Cordoba province (Argentina) during October and
1971; Dupuy et al., 2002; Ainsworth et al., 2010; Ram- November of the breeding periods of 2012 and 2013. The
teke et al., 2013). embryos were removed with forceps from the extraem-
A few works have described all or some of the develop- bryonic membranes. Each embryo was weighed with a
mental stages of wild birds like Society finch Lonchura digital scale (0.001 g precision) and fixed by immersion
striata (Yamasaki and Tonosaki, 1988), Brandt cormo- in a 4% neutral buffered formaldehyde solution. The
rant Phalacrocorax penicillatus (Price, 1938), Barn owl external morphology of early stages embryos was studied
Tyto alba (K€oppl et al., 2005), emu Dromaius novaehol- under a stereomicroscope Leica S6D. To enhance struc-
landiae (Nagai et al., 2011), lapwing Vanellus cristatus ture visibility (e.g. somites, pharyngeal arches, limb
(Grosser and Tandler, 1909), Adelie penguin Pygoscelis buds, eyelids, feather germs) a 0.5% neutral red in PBS
adeliae (Herbert, 1967), black stilt Himantopus novaeze- (phosphate buffered saline solution) staining solution
landiae (Browne, 2006), and rock pigeon Columba livia was used (technique based on that used by Ainsworth
(Olea and Sandoval, 2012). et al., 2010). Due to the great height of nests and their
Works regarding embryological development of Psitta- composition of interlaced sticks, access to eggs was diffi-
ciformes are scarce, incomplete, and restricted to Austra- cult and it was not possible to keep track of incubation
lian and African species. Braun (1879), Keibel (1901), time in the field, so stage designation was made only on
and Abraham (1901) were the first to study embryos of the basis of external morphology and comparison with
the Psittacidae budgerigar Melopsittacus undulatus. Hamburger and Hamilton (1951) stages.
Also Mebes (1984) made observations on the embryogen- The assignment of stages was performed based on a
esis of the Rosy-faced lovebird Agapornis roseicollis. set of “events” taking into account some morphological
Tokita (2004) was the only researcher to perform a features. These are morphological states of one or more
description of the last normal developmental stages embryological structures (arbitrarily selected by Ham-
(from stage 17 onwards) of a member of the Cacatuidae burger and Hamilton, 1951) that define a stage and dis-
family, the cockatiel Nymphicus hollandicus, although tinguish a previous stage from the next (Bininda-
brief, superficial and focused on the morphogenesis of Emonds et al., 2002). When the size and degree of
jaw muscles. embryo development allowed it, crown-rump length,
The monk parakeet (Myiopsitta monachus, Boddaert beak length (from the base to the tip), forelimb autopo-
1783) is one of the few Neotropical species whose cap- dium length, and third toe length including claw were
ture has no restrictions because its pest status (Cana- measured using a digital calliper to the nearest
velli et al. 2013), providing benefits for the abundant 0.01 mm. All measurements were log10-transformed and
1838 CARRIL AND TAMBUSSI

descriptive statistics, growth curves, correlation and lin- Stage 24. Maxillary process anterior to lens and
ear regression analysis were performed using the Past longer than the mandibular process. Distal tip of the
3.02a software (Hammer et al., 2001). The results were mandibular process at the mid-line of the eye. Limbs
compared with those available for other species in the greater in length than width (Fig. 2D).
literature (i.e. Nymphicus, Tokita, 2004; and Gallus, Stage 25. Presence of protuberances delimiting the
Hamburger and Hamilton, 1951). Because this study external auditory opening and forming a collar dorsally
presents data from relatively few individuals per stage, to the pharyngeal arches. Optic fissure evident. Limbs
non-parametric statistics were applied. Finally, photos with elbow and knee distinct and with rounded distal
were taken with a Nikon D-40 digital camera (macro digital plate (Fig. 3A).
128 mm). Stage 27. The region of the beak can be identified.
Maxillary and mandibular processes anterior to the eye.
RESULTS External nares evident. Elongated limbs, with enlargement
of the distal plate. Urogenital papilla evident (Fig. 3B).
Developmental Stages Description Stages 29. Eyelids begin to grow. Beak defined with
Stage 1. After laying. The peripheral area opaca egg tooth. The collar disappeared. Elongated neck. Dis-
and the central area pellucida are evident. tinguishable auditory meatus. Limbs with three seg-
Stage 2. The primitive streak has triangular shape. ments delimited (stylopod, zeugopod, and autopod). All
It is observed in the outer margin of the area pellucida fingers distinct by grooves and with interdigital webs.
and it extends toward its center (Fig. 1A). Stages 31. Some scleral papillae are distinguishable
Stage 4. The area pellucida becomes pear-shaped. in the caudo-ventral margin of the eye. Fingers elon-
The primitive streak reaches its maximum length. The gated with thin and concave interdigital webs. First
primitive groove, primitive pit and Hensen’s node are feather primordia in two rows in dorsal region, limbs
observed (Fig. 1B). base and tail (Fig. 3C).
Stage 8. The neural folds meet at the anterior Stage 34. Eyelids start to grow and feather primor-
region. Four pairs of somites and blood islands are dia are present around them. The nictitating membrane
present. begins to grow in the anterior region of the eye. Up to
Stage 9. Primary optic vesicles begin to form. Six 14 scleral papillae in eye. Beak elongated and more
pairs of somites are distinguishable (Fig. 1C). defined. Maxillary process anterior to the telencephalon
Stage 11. All primary brain vesicles are evident: and longer than mandibular process, the latter with ser-
telencephalon, diencephalon, mesencephalon, metence- rated edges at the anterior end. Egg tooth cornified.
phalon and myelencephalon. Slight cranial flexion. The Elongated limbs with interdigital webs reduced. In the
anterior neuropore begins to close. From a ventral view forelimbs, the alula is differentiated. The digit II of the
the triangular heart with a slight bent to right can be forelimb is elongated. Feathers primordia on the cervical
region, the ventral edge of the forelimbs, base of the
observed. Thirteen pairs of somites are distinguishable
humerus, dorsal and ventral region of the trunk (except
(Fig. 1D).
central rows), hindlimbs, and tail (Fig. 3D).
Stage 15. Cranial and cervical flexures. Hindbrain
Stage 35. Brain projections disappear. The nictitat-
nearby the heart, oriented ventrally and forming a 908
ing membrane approaches to the sclerotic papillae. Ser-
angle with the midbrain. Pineal gland visible. Three
rated edges at the anterior end of both maxillary and
pharyngeal arches evident. Otic and optic vesicles pres-
mandibular processes. Interdigital webs absent. In the
ent. Heart with tubular torsion. Tail bud begins to form. forelimbs, the digit II is more elongated than in previous
Somites hard to count. No wing or leg buds are evident stage. In the hindlimbs, the toe phalanges are evident,
(Fig. 2A). toe IV starts to rotate laterally and the hallux medially.
Stage 19. Defined body with head, lateral and tail The urogenital papilla is globular. The old feather pri-
folds complete. Head with brain parts delimited. Hind- mordia have grown in length. Feathers germs also on
brain contacts the heart. Pineal gland present in the frontals and nasals, eyelids, auditory meatus, cheeks,
diencephalon. Eyes without pigmentation. Otic vesicles and neck (Fig. 4A).
dorsal to the pharyngeal arches. Nasal pits located in Stages 36. Ellipsoidal and translucent eyelids. Cere
the ventro-lateral part of the head. Four pharyngeal surrounding external nares. Toe rotation complete. Claws
arches evident, the second arch is the largest. The man- of hindlimbs toes slightly curved. Feather primordia
dibular process is longer than the maxillary process. lengthened in the dorsal region and pygostyle (Fig. 4B).
Limb buds present and greater in width than length. Stages 38. Lower eyelid covers half of the eye. Egg
Forelimbs larger than hindlimbs. Somites extends to tail tooth and surrounding areas cornified. Primordia of
that begins to curve. Amnion closed and small allantois scales are evident in hindlimbs (Fig. 4C).
present. Stage 40. Eyelids are translucent and close the
Stage 20. Head and tail folds converge in the umbil- eyes. Beak almost fully cornified and with fleshy com-
ical area. Eye pigmentation faint. Pharyngeal arches missures. Cornified and curved claws are evident. Feath-
conspicuous. Limb buds greater in width than length. ers yellow colored and lengthened, especially in the
Forelimbs larger than hindlimbs. Allantois with vesicu- dorsal region, hindlimbs, and pygostyle.
lar shape and similar size that the telencephalon (Fig. Stage 401 (hatchling). Eyelids not translucent.
2B). Beak fully cornified. Ventrally curved maxilla with lat-
Stages 23. Telencephalic vesicles are distinguished. eral tomial teeth. Curved toes with fully cornified claws.
Eye pigmentation evident. Maxillary and mandibular Cloaca defined. Swollen crop and abdomen (Fig. 4D).
processes similar in length. Limb buds equal in width As a result of our sampling, there is a restricted avail-
and length. Forelimbs larger than hindlimbs (Fig. 2C). ability of a set of embryos of continuous stages.
 EMBRYONIC DEVELOPMENT OF THE MONK PARAKEET 1839

Fig. 1. Developmental stages of Myiopsitta monachus. A: stage 2; Hensen s node; lm, lateral plate mesoderm; mi, mielencephalon; ms,
B: stage 4; C: stage 9; and D: stage 11 in dorsal (left) and ventral mesencephalon; mt, metencephalon, n, notochord; nf, neural folds;
(right) views. Abbreviations: an, anterior neuropore; ao, area opaca; opv, optic vesicles; pg, primitive groove; pp, primitive pit; ps, primitive
ap, area pellucida; av, area vasculosa; d, diencephalon; h, heart; hn, streak; s, somites; t, telencephalon. Scale bar: 1 mm.
1840 CARRIL AND TAMBUSSI

Fig. 2. Developmental stages of Myiopsitta monachus. A: stage 15; B: stage 20; C: stage 23; and D:
stage 24 in lateral views. Abbreviations: a, alantoids; an, anterior neuropore; e, eye; fb, forelimb bud; h,
heart; hb, hindlimb bud; md, mandibular process; mx, maxillary process; np, nasal pit; ov, otic vesicle;
pa, pharyngeal arches; pg, pineal gland; s, somites; tb, tail bud. Scale bar: 1 mm. Detail of limbs not in
scale.

Morphological Measurements and Growth terization as a baseline. Average embryo body masses,
crown-rump length (CRL), beak length, forelimb autopo-
Quantitative measurements were taken for monk par- dium length, and third toe length including claw are
akeet embryos in order to provide developmental charac- presented in Table 1. Body mass and CRL were
 EMBRYONIC DEVELOPMENT OF THE MONK PARAKEET 1841

Fig. 3. Developmental stages of Myiopsitta monachus. A: stages 25; B: stage 27; C: stage 31; and D:
stage 34 in lateral views. Abbreviations: am, auditory meatus, b, beak; c, collar; dp, digital plate; el,
elbow; en, external nares; et, egg tooth; fl, forelimb; fp, feathers primordia; hl, hindlimb; iw, interdigital
webs; k, knee; nm, nictitating membrane; np, nasal pit; of, optic fissure; up, urogenital papilla. Scale bar:
2 mm. Detail of limbs not in scale.
1842 CARRIL AND TAMBUSSI

Fig. 4. Developmental stages of Myiopsitta monachus. A: stage 35; B: stage 36; C:, stage 38; and D:
stage 401 in lateral views. Abbreviations: fl, forelimb; hl, hindlimb. Scale bar: 5 mm. Detail of limbs not in
scale.

measured from stage 24 while the other measurements tively constant in stages 36 to 40 and increased again in
from stage 34 onwards. the final stages.
Correlation analysis showed a positive relation
between body mass and CRL (r2 5 0.96, P < 0.05; Fig.
5A). The growth curves of external morphological char- DISCUSSION
acters (beak length, forelimb and third toe length) as a How the Development of the Monk Parakeet
function of embryonic mass are given in Fig. 5B–D. Fits in the Chick Embryo Model?
Besides shape change, the growth of the beak showed an
increase in size faster in the early stages (34 to 36) with Hamburger and Hamilton (1951, HH) described a
a slow decrease in the growth rate towards the end of series of 46 stages of the Gallus gallus domesticus chick
the embryonic stages. Third toe length followed a similar development, a species model widely used in the embryo-
pattern while forelimb length showed a faster increase logical studies of birds (Bellairs and Osmond, 2005;
in the early stages (34 to 36), and then remained rela- Davey and Tickle, 2007; Vergara and Canto-Soler, 2012).
 EMBRYONIC DEVELOPMENT OF THE MONK PARAKEET 1843
TABLE 1. Developmental stages of Myiopsitta monachus embryos and its average measurements (with SE,
standard error; df: n 2 1) from stage 24 onward
Stage N M CRL Beak FL HL dIII
24 3 0.103 6 0.049 5.147 6 0.689 – – –
25 14 0.131 6 0.028 7.041 6 1.827 – – –
27 16 0.234 6 0.033 9.718 6 2.106 – – –
29–30 7 0.344 6 0.044 10.236 6 1.743 – – –
31–33 16 0.412 6 0.052 13.007 6 2.282 – – –
35 11 0.780 6 0.080 18.008 6 1.024 3.458 6 0.285 4.736 6 0.531 2.363 6 0.221
36–37 28 1.095 6 0.124 20.888 6 1.928 4.420 6 0.502 5.483 6 0.620 3.159 6 0.396
38–39 24 1.561 6 0.281 23.950 6 3.502 5.160 6 0.354 5.838 6 0.388 3.773 6 0.375
40 28 2.509 6 0.431 29.079 6 4.021 6.022 6 0.543 5.965 6 0.600 4.368 6 0.479
41 13 3.810 6 0.385 30.033 6 3.962 6.808 6 0.328 7.117 6 0.620 4.893 6 0.339

Abbreviations: n, number of specimens; M, body mass (in grams); CRL, crown-rump length (in mm); Beak, beak length (in
mm); FL, forelimb autopodium length (in mm); and HL dIII, third toe length including claw (in mm).

Developmental stages are an appropriate tool for the indicate a post-displacement and afterwards acceleration
comparison of development within Aves since they processes in Myiopsitta because isomorphic adults are
standardize homologous structural events independently produced (complete plumage in both species). Similar
of time and body size, and changes from the normal processes might occur in the semialtricial Barn owl that
stages could indicate heterochronic events (Starck, 1993; hatches covered with plumage and which feathers germs
Rice, 1997). This approach is useful to compare not only appear also in the stage 32 of development (K€oppl et al.,
embryo growth of the same avian species but also in an 2005).
interspecific comparative frame. However, HH character- Wing buds are initially greater than leg buds in the
izes their last stages only based on the length of beak monk parakeet, similar condition to that described for
and third toe and, to some extent, this makes it inappli- the cockatiel (Tokita, 2004) and different from the
cable in a comparative context. Ricklefs and Starck chicken (Hamburger and Hamilton, 1951). Also, we
(1998) in a shorter model consider that the stage 40 is found that forelimbs and hindlimbs have similar relative
the sum of 40-44 HH stages and stages 41 and 42 are size in the monk parakeet, contrary to the chicken that
characterized by behaviour of the embryo, pipping, and shows a greater size of the hindlimbs (Hamburger and
hatching respectively. Our description follows this sec- Hamilton, 1951). This differential development degree of
ond scheme (401 stages). the limbs could be due to changes in the rate of develop-
ment in Myiopsitta when compared with Gallus: initially
Comparison Between Development of the Monk there is an acceleration of the wing buds and a decelera-
Parakeet and Other Aves tion of the leg buds followed by a deceleration of hin-
dlimbs. These features are associated with different
The embryonic development of birds in general is mor- locomotor habits between the flying monk parakeet and
phologically conserved, although variable in growth the more terrestrial chicken, a fact that is evident from
rates (Nagai et al., 2011). We found differences in devel- the moment of hatch. Differences in the growth rate of
opment of characters in the monk parakeet when com- limbs have been demonstrated in other species, for
pared with those of the chicken and other birds, which example, the precocial and cursorial ostrich and emu
evidence some heterochronic events mainly in the latest which have a noticeable reduced growth rate in the fore-
stages. limbs (Blom and Lilja, 2005; Nagai et al., 2011). As well,
Parrots and galliforms were long thought to be distant in the altricial and flying fieldfare the forelimbs are sig-
relatives. In contrast to parrots, galliforms are precocial nificantly larger than the hindlimbs (Blom and Lilja,
birds whose offspring follow their parents and feed on 2005).
their own (Ricklefs and Starck, 1998). Altricial offspring During the stages 35 and following, substantial
remain relatively helpless and dependent on their changes occur in the foot. Since the chicken have aniso-
parents after hatchling. This is in line with the delayed dactyl foot and the monk parakeet zygodatil one, it is
maturation of the legs, plumage, vision, and also of the not surprising that the major differences between both
brain growth as indicated by Charvet and Striedter species happen at these last stages (see below). The
(2008) and Charvet et al. (2011). In most phylogentic peculiar ventral curvature of the upper jaw occurs in the
schemes (e.g. Jarvis et al., 2014), Galliformes are consid- later stages of the intra egg development at stage 401
ered basal while parrots are derived; also, precociality is in the monk parakeet. Therefore, the chicks have their
the ancestral condition regarding altriciality (Starck, characteristic beak at hatch but begin to feed themselves
1993). In this context, here direction of changes is deter- 20 days after that. Images available of the cockatiel
mined in Myiopsitta compared with Gallus. The appear- (Tokita, 2004) allow inferring that the same happens in
ance of the feathers primordia occurs later in the monk this species. Of course, this does not occur in the chicken
parakeet (stage 31) and in the cockatiel (stage 32; embryo.
Tokita, 2004) when compared with the chicken (stage 30; In sum, prominent differences exist between precocial
Hamburger and Hamilton, 1951). As consequence, the and altricial embryos at least in the later stages. In
newborn chick has virtually no external feathers, a fea- addition, certain characters present in the same stage,
ture shared with other altricial species. This difference may have significant qualitative differences that are par-
in the time of appearance of feathers primordia could ticularly manifest after hatchling. For example, the
1844 CARRIL AND TAMBUSSI

Fig. 5. Scatterplot of log10 measurements versus log10 body mass in Myiopsitta monachus (A–D). Scat-
terplot of correlation of log10 CRL versus stages in Myiopsitta monachus compared with Nymphicus hol-
landicus (E), and linear regressions of beak length and III toe length (dotted line and empty circles)
compared with Gallus gallus domesticus (F).
 EMBRYONIC DEVELOPMENT OF THE MONK PARAKEET 1845
density of feathers (high in the chicken and low in par- interdigital webs that binds all fingers from stage 29 to
rots), and the ability to open the eyes (the chickens are 34 in both parrots: monk parakeet (this paper) and the
born with their eyes open while parrots with eyes budgerigar (Botelho et al., 2014). Comparing with other
closed). birds, between stages 29 and 35 the deeper changes in
the foot occur. In the chicken, webs between digits and
Morphological Measurements and Growth toes disappear during stage 35 and they become sepa-
Comparisons rated (Sanz-Ezquerro and Tickle, 2003; Bellairs and
Osmond, 2005). During the stage 29 of the Barn owl
Quantitative measurements of external morphological development, the four toes have webs; toes with slightly
characters for the monk parakeet embryos proved to be concave webbing in between are visible during stage 32
useful in developmental characterization and to make and have nearly disappeared during stage 35 (K€oppl
comparisons with other birds.
et al., 2005).
A positive correlation between stages and CRL was
Another important process during digit morphogenesis
found in both the monk parakeet (r2 5 0.98, P < 0.05)
is the reduction (in parrots) or not (in the chicks’
and the cockatiel (r2 5 0.98, P < 0.05). In the latter, CRL
embryo) of the musculus extensor brevis digiti IV during
values are higher throughout the whole development
the stage 35 (Botelho et al., 2014). These authors states
(Fig. 5E), probably due to its larger size (adult N. hol-
that the decrease may be related to the retroversion of
landicus 33 cm; Rowley, 1997; and adult M. monachus
28–29 cm; Collar, 1997). Also, cockatiels have a growth the 4th digit in the budgerigar. The musculus extensor
spurt of CRL at later stages of development whereas the brevis digiti IV is also completely absent in the monk
monk parakeet could have a decrease of growth rate parakeet (Carril et al., 2014).
(deceleration). The digit formation and growth can provide important
In comparison with the available data on the precocial information about evolutionary morphological diversity.
chicken (Hamburger and Hamilton, 1951), the positive Particularly, the development of the zygodactyl foot
regressions between stages and beak length showed could supplement morphological systematic and evolu-
acceleration (higher slope) and higher values for the tionary studies of Psittaciformes, relationships within
monk parakeet (Fig. 5F; G. gallus, r2 5 0.98, p<0.05, Telluraves (the clade including most arboreal birds in
slope 0.04; M. monachus, r2 5 0.88, P < 0.05, slope 0.06), the sense of Yuri et al., 2013) and other Neornithes.
which results in larger beaks with higher growing rates.
Conversely, positive regressions between stages and III
toes lengths showed deceleration (lower slope) and lower
CONCLUSIONS
values for the monk parakeet in comparison with the Comparative embriology is the empirical basis for dis-
chicken, which results in shorter III toes with lower cover how developmental processes have evolved and is
growing rates (Fig. 5F, dotted lines; G. gallus, r2 5 0.98, the motor of contemporary field of Evo-Devo. At the
P < 0.05, slope 0.08; M. monachus, r2 5 0.89, P < 0.05, dawn of the studies on avian development, chick embryo
slope 0.06). Even though the correlation coefficients for was used as a useful model and has become one of the
the chicken are higher than they are for the monk para- most versatile systems in developmental biology (Stern,
keets, “r” values are close to one in both species and 2004), even in the incredibly fast-growing molecular sci-
there are no significant differences between them. ence. Considering that birds are the most diverse conti-
nental vertebrates, knowledge about their development
Acquisition of Zygodactil Arrangement of Toes is scarce. Our work partially fills that gap. Most develop-
Study of digits position is interesting for several rea- mental stages proposed Hamburger and Hamilton (1951)
sons: implies a deep transformation or repositioning, for the chicken can be recognized in the development of
because the type of foot is usually employed for estab- the monk parakeet and the main differences between
lishing relationships within Aves, and because one of the species were found in the later stages. Standardizing the
ongoing discussions in the field of ornithology concerns stages of development is an important tool in compara-
homologies between fingers. A zygodactyl or yoke-toed tive studies of embryology. Our study is the first dedi-
foot (digits II and III directed forwards, digits I and IV cated to the development of a Neotropical parrot and is
pointing backwards) is a specialization of the avian leg among the few dedicated to Psittaciformes in general.
identified with perching, climbing or manipulation hab- Most of the morphological variation within groups is a
its (Carril et al., 2014 and bibliography cited therein). It consequence of changes in developmental timing and
is considered a modification of the anisodactyl foot (dig- rates (heterochrony). The growth pattern of the monk
its II, III and IV oriented forward) that evolved inde- paraket Myiopsitta monachus may be the product of a
pendently at least in two other clades other than combination of heterochronic processes like post-
Psittaciformes (Cuculidae and Piciformes; Mayr, 2014). displacement, acceleration and/or deceleration.
In the monk parakeet, the retroversion of digit IV starts Since ontogenetic studies of avian development in rep-
during the stage 35 (Fig. 4A) and is complete at stage resentative orders are essential for addressing micro and
36. These observations are completely consistent with macroevolution questions (Nagai et al., 2011), results of
that seen in the budgerigar (Botelho et al., 2014). To our this research allow the standard identification of stages
knowledge, there is no more information in the litera- in different species of parrots, recognize similarities and
ture about the lateral rotation of digit IV in any other differences between precocial (the chicken) and altricial
Psittaciformes. Tokita (2004) makes no reference to this species (Myiopsitta), and provide planning data for
aspect about the cockatiel development. Finger IV rota- future studies, such as ossification sequences, myogene-
tion occurs after the reduction (by apoptosis) of the sis of the mandibular muscles, and brain development.
1846 CARRIL AND TAMBUSSI

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