A cloacal opening in a non-avian dinosaur.

The Frankfurt specimen of Psittacosaurus sp. (SMF R 4970) from the Early Cretaceous Jehol deposits of Liaoning (Figure S1) exhibits exceptional preservation of scale-clad integument1. Preservation of colour patterns and countershading allowed a detailed reconstruction of this individual's physical appearance. It was previously noted that the cloacal region was preserved2, but its detailed anatomy was incorrectly reconstructed. We show here that the fine anatomy of the vent is remarkably well preserved and can be retrodeformed to illustrate its three-dimensional nature. The vent's scale anatomy and pigmentation are distinct from adjacent body regions, and although its anatomy does not reveal much information about the ecology, or sex, of this dinosaur, it suggests possible roles for visual and olfactory signalling.

Revalidation of the Argentinian pouched lamprey Geotria macrostoma (Burmeister, 1868) with molecular and morphological evidence.

BACKGROUND: The Argentinian pouched lamprey, classified as Petromyzon macrostomus Burmeister, 1868 was first described in 1867 in De La Plata River, in Buenos Aires, Argentina, and subsequently recorded in several rivers from Patagonia. Since its original description, the validity of P. macrostomus was questioned by several ichthyologists and 36 years after its original discovery it was considered a junior synonym of Geotria australis Gray, 1851. For a long time, the taxonomic status of G. australis has been uncertain, largely due to the misinterpretations of the morphological alterations that occur during sexual maturation, including the arrangement of teeth, size and position of fins and cloaca, and the development of an exceptionally large gular pouch in males. In this study, the taxonomic status of Geotria from across the "species" range was evaluated using both molecular analysis and examination of morphological characteristics. METHODOLOGY/PRINCIPAL FINDINGS: Phylogenetic and species delimitation analyses based on mitochondrial DNA sequences of Cytochrome b (Cyt b) and Cytochrome C Oxidase Subunit 1 (COI) genes, along with morphological analysis of diagnostic characters reported in the original descriptions of the species were used to assess genetic and morphological variation within Geotria and to determine the specific status of the Argentinian lamprey. These analyses revealed that Geotria from Argentina constitutes a well differentiated lineage from Chilean and Australasian populations. The position of the cloaca and the distance between the second dorsal and caudal fins in sub-adult individuals, and at previous life stages, can be used to distinguish between the two species. In addition, the genetic distance between G. macrostoma and G. australis for the COI and Cyt b mitochondrial genes is higher than both intra- and inter-specific distances reported for other Petromyzontiformes. CONCLUSIONS/SIGNIFICANCE: Our results indicate that the Argentinian pouched lamprey, found along a broad latitudinal gradient on the south-west Atlantic coast of South America, should be named as Geotria macrostoma (Burmeister, 1868) and not as G. australis Gray 1851, returning to its earliest valid designation in Argentina. Geotria macrostoma can now be considered as the single lamprey species inhabiting Argentinian Patagonia, with distinct local adaptations and evolutionary potential. It is essential that this distinctiveness is recognized in order to guide future conservation and management actions against imminent threats posed by human actions in the major basins of Patagonia.

A teleost structural analogue to the avian bursa of Fabricius.

The bursa of Fabricius is a primary and secondary lymphoid organ considered exclusively present in birds, and studies of this structure have been vital to our current understanding of the adaptive immune system of vertebrates. In this study, we reveal substantial lymphoepithelial tissue in a previously undescribed bursa in Atlantic salmon (Salmo salar), situated caudal to the urogenital papilla of the cloaca and thus analogous to the anatomical placement of the bursa of Fabricius. We investigated three groups of Atlantic salmon at different maturational stages and characterized the structure by applying dissection, radiology, scanning electron microscopy and histological techniques, including immunohistochemistry and in situ hybridization. We found that the epithelial anlage of the salmon cloacal bursa developed into substantial lymphoepithelial tissue and subsequently regressed following sexual maturation. Such a dynamic development is also a key characteristic of the avian bursa. The presence of intraepithelial lymphocytes was concomitant with expression of the leukocyte-attracting chemokine CCL19, indicative of lymphoid organ functions. We did not observe recombination or gene conversion in salmon bursal lymphocytes at any developmental stage, indicating the absence of primary lymphoid organ functions in contrast to the bursa of Fabricius. However, the possibility of the bursa to trap both enteric and environmental antigens, combined with the presence of several antigen-presenting cells residing within the lymphoepithelium, suggest the structure has secondary lymphoid organ functions. We present the discovery of a lymphoid organ in Atlantic salmon with striking topographical similarities to that of the bursa of Fabricius in birds. In addition, the age-dependent dynamics of its lymphoepithelium suggest functions related to the maturation processes of lymphocytes.

Male age and its association with reproductive traits in captive and wild house sparrows.

Evolutionary theory predicts that females seek extra-pair fertilizations from high-quality males. In socially monogamous bird species, it is often old males that are most successful in extra-pair fertilizations. Adaptive models of female extra-pair mate choice suggest that old males may produce offspring of higher genetic quality than young males because they have proven their survivability. However, old males are also more likely to show signs of reproductive senescence, such as reduced sperm quality. To better understand why old males account for a disproportionally large number of extra-pair offspring and what the consequences of mating with old males are, we compared several sperm traits of both captive and wild house sparrows, Passer domesticus. Sperm morphological traits and cloacal protuberance volume (a proxy for sperm load) of old and young males did not differ substantially. However, old males delivered almost three times more sperm to the female's egg than young males. We discuss the possibility of a post-copulatory advantage for old over young males and the consequences for females mated with old males.

Transcending Dimensions: a Comparative Analysis of Cloaca Imaging in Advancing the Surgeon's Understanding of Complex Anatomy.

Surgeons have a steep learning capacity to understand 2-D images provided by conventional cloacagrams. Imaging advances now allow for 3-D reconstruction and 3-D models; but no evaluation of the value of these techniques exists in the literature. Therefore, we sought to determine if advances in 3-D imaging would benefit surgeons, lead to accelerated learning, and improve understanding for operative planning of a cloaca reconstruction. Questionnaires were used to assess the understanding of 2-D and 3-D images by pediatric surgical faculty and trainees. For the same case of a cloacal malformation, a 2D contrast study cloacagram, a 3D model rotatable CT scan reconstruction, a software enhanced 3D video animation (which allowed the observer to manipulate the structure in any orientation), and a printed physical 3D cloaca model that could be held in the observer's hand were employed. Logistic mixed effect models assessed whether the proportion of questions about the case that were answered correctly differed by imaging modality, and whether the proportion answered correctly differed between trainee and attending surgeons for any particular modality. Twenty-nine pediatric surgery trainees (27 pediatric general surgery and 2 pediatric urology surgery trainees) and 30 pediatric surgery and urology faculty participated. For trainees, the percentage of questions answered correctly was: 2-D 10.5%, 3-D PACS 46.7%, 3-D Enhanced 67.1%, and 3-D Printed 73.8%. For faculty, the total percentage of questions answered correctly was: 2-D 22.2%, 3-D PACS 54.8%, 3D Enhanced 66.2%, and 3-D printed 74.0%. The differences in rates of correctness across all four modalities were significant in both fellows and attendings (p < 0.001), with performance being lowest for the 2-D modality, and with increasing percentage of correct answers with each subsequent modality. The difference between trainees and attendings in correctness rate was significant only for the 2-D modality, with attendings answering correctly more often. The 2-D cloacagram, as the least complex model, was the most difficult to interpret. The more complex the modality, the more correct were the responses obtained from both groups. Trainees and attendings had similar levels of correct answers and understanding of the cloacagram for the more advanced modalities. Mental visualization skills of anatomy and complex 3-D spatial arrangements traditionally have taken years of experience to master. Now with novel surgical education resources of a 3-D cloacagram, a more quickly advancing skill is possible.

Morfologia da bolsa cloacal de emas, Rhea americana americana Linnaeus, 1758 / Morphology of rhea's cloacal bursa, Rhea americana americana Linnaeus, 1758

A bolsa cloacal é o órgão das aves responsável pela maturação e transferência de linfócitos para outros tecidos. Apesar da importância deste órgão nos mecanismos imunológicos desses animais, são escassas as informações a respeito de sua morfologia em emas. Neste estudo, objetivou-se descrever o desenvolvimento morfológico da bolsa cloacal de emas jovens. Utilizou-se 12 animais de ambos os sexos (6 machos e 6 fêmeas) para a microscopia de luz, eletrônica de transmissão e varredura. Microscopicamente, a bolsa cloacal da ema apresentou, em todas as idades a mucosa interna pregueada composta por lóbulos linfoides de diversos tamanhos, organizados como estrutura alveolar. Em cada prega verificou-se quatro componentes histológicos: as camadas mucosa, submucosa, muscular e adventícia. Esses lóbulos eram compostos de uma zona cortical, uma zona corticomedular e uma zona medular. Verificou-se a existência de linfócitos de tamanhos variados, linfoblastos, capilares sanguíneos, células reticulares epiteliais e macrófagos. Pela microscopia eletrônica de varredura, verificou-se que a superfície da mucosa dos lóbulos bursais apresentaram projeções poligonais, com a presença de curtas microvilosidades em toda a superfície. A comparação nas idades de 0 e 15 semanas de vida demostrou o desenvolvimento dos lóbulos bursais. O padrão morfológico da bolsa cloacal de emas difere do padrão comumente reportado para outras aves tais como pato selvagem, galinha da angola, ganso nativo, peru, codorna japonesa e falcão.(AU)

The cloacal bursa is the bird's organ responsible for maturation and transfer of lymphocytes to other tissues. Despite the importance of this organ in the immunological mechanisms of these animals, information about their morphology in rhea are scarce. We used 12 animals (6 males and 6 females) for light, transmission electron, and scanning microscopy. Microscopically, the cloacal bursa presented the inner mucosa consists of pleated lymphoid lobes of various sizes, organized as alveolar structure, in all ages. In each nail was found four histological components: mucosa, submucosa, muscular and adventitia layers. These lobes were composed of a cortical zone, a corticomedular zone and a medular area. It was verified the existence of varying sizes lymphocytes, lymphoblasts, blood capillaries, epithelial reticular cells and macrophages. By scanning electron microscopy, it was found that the mucous membrane surface of the bursal lobes showed polygonal projections, with the presence of short microvilli membranes throughout the surface. The comparison between 0 and 15 weeks demonstrated the development of the bursal lobes. The morphological pattern of the rhea cloacal bursa differs from standard commonly reported for other birds such as wild duck, Angola's chicken, native goose, turkey, Japanese quail, and Hawk.(AU)

Comb, cloaca and feet scores and testis morphometry in male broiler breeders at two different ages / Escores de crista, cloaca e de pé e morfometria testicular em galos de matriz pesada em duas diferentes idades

In broiler breeder roosters, the weight of the testes is positively associated with daily sperm production and fertility. In birds, the testes are located in the coelomic cavity, not being accessible to reproductive evaluation as mammalian testes. The reproductive evaluation of roosters is based on phenotypic traits. Any changes on testes will be reflected on fertility levels. The aim of this paper was to evaluate the potential of association of phenotypic traits as body weight (BW), comb scores (COS), cloaca scores (CLS) and feet scores (FS) with testicular morphometric parameters in adult Cobb® Mx roosters in two ages (25 and 45 weeks of age). The low BW roosters had lower testes weight (TW), smaller seminiferous tubule diameter (STD) and lower comb and cloaca scores than the medium and high BW roosters (P≤0.05). Heavy roosters had the highest COS and bigger seminiferous epithelium height (SEH). There was no statistical difference between the BW categories in FS and volumetric proportion of seminiferous tubule (ST) and interstitial tissue (IT). Considering the age effect, roosters with 45 weeks of age had lower TW and ST than 25-week roosters. Positive, moderate and significant correlation was found between testes weight and phenotypic characteristics as BW, COS and CLS in 45week roosters. A positive, moderate and significant correlation was found between body weight and histologic testicular characteristics as STD and SHE in both evaluated ages. In conclusion, there is a regression in the testes weight with age and it is more intense in Cobb® Mx roosters with lower BW. Furthermore, it is concluded that the BW positively influences the testes weight and histological quality of the testes. COS and CLS can be used with moderate potential prediction to identify roosters with low testes weight at 45 weeks of age.(AU)

Em galos de matriz pesada sabe-se que o peso dos testículos pode ser associado positivamente com a produção diária de espermatozoides e que alterações nos testículos irão refletir na fertilidade. Nas aves, os testículos estão localizados dentro da cavidade celomática, e, portanto, não são acessíveis para exame andrológico direto, como em mamíferos. A avaliação reprodutiva de galos se baseia principalmente em características fenotípicas. Buscou-se avaliar o potencial de associação de alguns parâmetros fenotípicos, como peso corporal (PC), escores de crista (ECR), escores de cloaca (ECL) e escores de pés (EP), com parâmetros morfológicos de testículos em galos Cobb® Mx em duas idades (25 e 45 semanas de idade). Galos leves apresentaram menor peso testicular (PT), menor diâmetro de túbulo seminífero (DT), e menores ECR e ECL que galos médios e pesados (P≤0.05). Galos pesados apresentaram os maiores ECR e maiores valores para altura do epitélio seminífero (AE), e DT. Não houve diferença estatisticamente significativa entre as categorias de peso para escores de pé e proporções de túbulo seminífero (TS) e interstício (INT). Considerando-se o efeito da idade, galos com 45 semanas de idade apresentaram menores PT e TS que galos com 25 semanas de idade. Correlação positiva, moderada e significativa foi encontrada entre peso de testículo e características fenotípicas como ECR, ECL e PC em galos de 45 semanas de idade. Foi encontrada correlação positiva, moderada e significativa entre PC e características testiculares como DT e AE nas duas idades avaliadas. Conclui-se que em galos Cobb® Mx ocorre regressão do testículo com o avanço da idade, e ela é mais intensa em galos com menor peso corporal. Conclui-se que o PC tem influência positiva no peso e índices histomorfométricos dos testículos, e que escores de crista e de cloaca podem ser utilizados com potencial moderado de predição para identificar galos com baixo peso de testículos em galos com 45 semanas de idade.(AU)

Functional morphology of the gonoduct of the viviparous teleost Poeciliopsis gracilis (Heckel, 1848) (Poeciliidae).

Female teleosts do not develop Müllerian ducts; consequently, the ovary of teleosts contains two zones: germinal and gonoduct. The gonoduct lacks germinal cells, but has relevant functions in the reproductive process. We describe the functional morphology of the gonoduct in the viviparous teleost Poeciliopsis gracilis during nongestation and gestation stages. This study tests the hypothesis that the gonoduct functions as a barrier between the germinal zone and the exterior. By providing information about morphology and function of the gonoduct we show that this part of the ovary has an essential role in the reproduction of teleosts. The ovaries were processed by histological technique and stained with hematoxylin-eosin (H-E), Masson's trichrome, toluidine blue and periodic acid-Schiff (PAS). The gonoduct is divided into three regions: cephalic, middle, and caudal. In the cephalic and middle regions there are mucosal folds that extend into the gonoductal lumen, forming structures similar to a cervix. The caudal region has two portions: the anterior contains a dorsal invagination and exocrine glands among columnar cells; the posterior has a ventral flexion and stratified epithelium with apical secretory cells. The morphology of this epithelium indicates two functions: (a) secretory by the apical columnar cells, and (b) protection through the stratification. Another peculiarity of the caudal region is that both ducts, reproductive and digestive, converge in a common cavity at their caudal ends, forming a cloacal region. The histology of the gonoduct indicates relevant functions including: (1) the control of the luminal diameter by the muscle and the presence of mucosal folds, like a cervix; (2) the relationship with the spermatozoa during insemination and storing them in mucosal folds; (3) the support of immunological processes; (4) secretory activities; (5) forming the duct during birth; and (6) possibly, acts as a barrier against parasite infestations.

Reorganization of mammalian body wall patterning with cloacal septation.

Septation of the cloaca is a unique mammalian adaptation that required a novel reorganization of the perineum-the caudal portion of the trunk body wall not associated with the hindlimb. Fish, the basal vertebrates, separate ventrolateral body wall musculature of the trunk into two discrete layers, while most tetrapods expand this pattern in the thorax and abdomen into four. Mammals, the only vertebrate group to divide the cloaca into urogenital and anorectal portions, exhibit complex muscle morphology in the perineum. Here we describe how perineal morphology in a broad sample of mammals fits into patterning of trunk musculature as an extension of the four-layer ventrolateral muscular patterning of the thorax and abdomen. We show that each perineal muscle layer has a specific function related to structures formed by cloacal septation. From superficial to deep, there is the subcutaneous layer, which regulates orifice closure, the external layer, which supplements both erectile and micturition function, the internal layer, which provides primary micturition and defecation regulation, and the transversus layer, which provides structural support for pelvic organs. We elucidate how the four-layer body wall pattern, restricted to the non-mammal tetrapod thorax and abdomen, is observed in the mammalian perineum to regulate function of unique perineal structures derived from cloacal septation.

Prenatal sonographic diagnosis of urorectal septum malformation sequence and chromosomal microarray analysis: A case report and review of the literature.

INTRODUCTION: Urorectal septum malformation sequence (URSMS) is a rare congenital abnormal syndrome that is caused by the incomplete division of the cloaca. Based on whether the cloaca membrane breaks down or not, the URSMS are classified as full and partial forms. The prenatal diagnosis of URSMS remains challenging because of poor recognition to this malformation and the relatively non-specific sonographic features. We report a prenatally sonographic diagnosed case of the partial URSMS, and review the literature to summarize the prenatal features. CASE REPORT AND REVIEW: A 37-year old woman was referred at 24 weeks of gestation for fetal abdominal cyst. Detailed sonographic examination was done and revealed the vesicocolic fistula, distended colon, absence of perianal hypoechoic ring, pyelectasis, and small stomach bubble. The URSMS was suspected.Amniocentesis was done and karyotyping revealed 46,XY. Furthermore, chromosomal microarray analysis (CMA) was performed for the first time in URSMS and an alteration of 111.8Kb deletion was detected in 16p13.3 which was located inside the RBFOX1 gene. Parental studies showed that the deletion was inherited from the father who has nomal clinical phenotype.The woman elected to terminate the pregnancy at 25 weeks gestation and postmortem examination confirmed the diagnosis of partial URSMS.The published studies were reviewed and 28 cases of URSMS with conducted prenatal ultrasonography were collected in this report. The most common sonographic description, as suspicious signs of URSMS, were severe oligohydramnios or anhydramnios, urinary tract anomalies, fetal intra-abdominal cysts, and dilated bowel. Also, enterolithiasis and vesicocolic fistula were relatively infrequent but highly specific feature of URSMS. CONCLUSIONS: URSMS is difficult to be diagnosed prenatally. However, it has characteristic features that can be detected by fetal ultrasonography, and a precise prenatal sonographic examination is crucial for diagnosing URSMS. Besides, more genomic profiling studies are needed to elucidate the causality.

Historical considerations and comments on the type series of Cyrtodactylus marmoratus Gray, 1831, with an updated comparative table for the bent-toed geckos of the Sunda Islands and Sulawesi.

Cyrtodactylus marmoratus Gray, 1831, a species of bent-toed gecko exhibiting a precloacal groove in males, was described on the basis of specimens collected by Heinrich Kuhl and Johan Conrad van Hasselt in Java, Greater Sunda Islands, Indonesia. Kluge (1985) subsequently designated a lectotype for C. marmoratus from a series of these specimens (i.e., syntypes), now housed in the herpetological collection at Naturalis (formerly the Rijksmuseum van Natuurlijke Historie; RMNH), Leiden, the Netherlands. Our work at Naturalis shows that the type series of C. marmoratus at RMNH actually comprises two sets of specimens, and that examination of specimens from one set or the other by different authors, including Kluge (1985), is responsible for some confusion surrounding the type series of this species. As a consequence, we present relevant morphological data for all 14 specimens constituting the type series of C. marmoratus at RMNH for the first time. The type status of two specimens of C. marmoratus in the collection at the Muséum National d'Histoire Naturelle, Paris, France, remains unresolved at present. Owing to the inconsistent naming and application of terms for some key characters (e.g., groove, sulcus, pit, hollow, depression) used in the diagnoses of Cyrtodactylus species, we here propose a set of novel and useful definitions that are supported by photographs. We also illustrate the sexually dimorphic expression of this character in C. marmoratus. Finally, we present a revised comparative table for the bent-toad geckos of the Sunda Islands and Sulawesi.

Clarification of mammalian cloacal morphogenesis using high-resolution episcopic microscopy.

The developmental process through which the cloaca transforms from one hollow structure to two separated urinary and digestive outlets remains controversial and speculative. Here, we use high-resolution episcopic microscopy to examine a comprehensive series of normal and mutant mouse cloaca in which the detailed 3-dimensional (3-D) morphological features are illuminated throughout the development. We provide evidence that the dorsal peri-cloacal mesenchyme (dPCM) remains stationary while other surrounding tissues grow towards it. This causes dramatic changes of spatial relationship among caudal structures and morphological transformation of the cloaca. The 3-D characterizations of Dkk1 mutants reveal a hyperplastic defect of dPCM, which leads to a significant anterior shift of the caudal boundary of the cloaca, premature occlusion of the cloaca and, imperforate anus phenotype. Conversely, Shh knockout causes a severe hypoplastic defect of cloaca mesenchyme including dPCM and persistent cloaca. Collectively, these findings suggest that formation of the dPCM is critical for cloacal morphogenesis and furthermore, growth and movement of the mesenchymal tissues towards the dPCM lead to the cloaca occlusion and separation of the urinary and digestive outlets.

The phylogenetic distribution, anatomy and histology of the post-cloacal bones and adnexa of geckos.

Post-cloacal bones of gekkotans may be present as a single (medial) pair, two pairs (medial and lateral), or may be lacking. We, herein, demonstrate that the presence of a single medial pair is the ancestral condition for the Gekkota, that the lateral pair is of sporadic occurrence within and between families, except for the Eublepharidae where it is universal, and that absence is also of sporadic occurrence except for the Sphaerodactylidae where it is the ancestral condition. Adult male Tokay geckos (Gekko gecko) possess only the medial pair of bones, and these exhibit a regionally-specific expression of woven, fibrolamellar, and lamellar bone, and an enclosed medullary cavity. Females and small juvenile males lack bony elements but exhibit a conspicuous band of dense connective tissue located about the anterior and lateral margins of the cloacal sacs. As males grow and attain sexual maturity, the medial post-cloacal bones condense in this band of dense connective tissue, and are thus shown to be dermal ossifications, similar to osteoderms but with muscular associations (although this is also known for crocodylians). Based upon ontogenetic data we set forth a scenario to explain the loss of the medial post-cloacal bones in various lineages. Differential staining of the cloacal sacs failed to reveal any specialized glandular structures. Investigation of the post-cloacal spurs shows them to be associated with cellular connective tissue of a type similar to that found in the vicinity of the medial post-cloacal bones. This suggests that the lateral post-cloacal bones may also be dermal bones, but histological evidence is needed to corroborate this.

Systematic stereoscopic analyses for cloacal development: The origin of anorectal malformations.

The division of the embryonic cloaca is the most essential event for the formation of digestive and urinary tracts. The defective development of the cloaca results in anorectal malformations (ARMs; 2-5 per 10,000 live births). However, the developmental and pathogenic mechanisms of ARMs are unclear. In the current study, we visualized the epithelia in the developing cloaca and nephric ducts (NDs). Systemic stereoscopic analyses revealed that the ND-cloaca connection sites shifted from the lateral-middle to dorsal-anterior part of the cloaca during cloacal division from E10.5 to E11.5 in mouse embryos. Genetic cell labeling analyses revealed that the cells in the ventral cloacal epithelium in the early stages rarely contributed to the dorsal part. Moreover, we revealed the possible morphogenetic movement of endodermal cells within the anterior part of the urogenital sinus and hindgut. These results provide the basis for understanding both cloacal development and the ARM pathogenesis.

Gender Identification by Cloacoscopy and Cystoscopy in Juvenile Chelonians.

Cloacoscopy and cystoscopy are simple, noninvasive to minimally invasive techniques that provide excellent visualization, and result in fast recovery. General or intrathecal anesthesia is sufficient. They can be performed in free-ranging turtles under field conditions. Cloacoscopic gender identification of external genitalia is not reliable because of the high degree of misinterpretation between phallus and clitoris, especially in juveniles. However, saline-infusion or air insufflation cystoscopy through the urinary bladder (or accessory vesicles/bladders) is often effective for the visualization of gonads and to identify the sex. Visualization of gonads is feasible through the urinary bladder or accessory vesicle wall in many species.

A relative shift in cloacal location repositions external genitalia in amniote evolution.

The move of vertebrates to a terrestrial lifestyle required major adaptations in their locomotory apparatus and reproductive organs. While the fin-to-limb transition has received considerable attention, little is known about the developmental and evolutionary origins of external genitalia. Similarities in gene expression have been interpreted as a potential evolutionary link between the limb and genitals; however, no underlying developmental mechanism has been identified. We re-examined this question using micro-computed tomography, lineage tracing in three amniote clades, and RNA-sequencing-based transcriptional profiling. Here we show that the developmental origin of external genitalia has shifted through evolution, and in some taxa limbs and genitals share a common primordium. In squamates, the genitalia develop directly from the budding hindlimbs, or the remnants thereof, whereas in mice the genital tubercle originates from the ventral and tail bud mesenchyme. The recruitment of different cell populations for genital outgrowth follows a change in the relative position of the cloaca, the genitalia organizing centre. Ectopic grafting of the cloaca demonstrates the conserved ability of different mesenchymal cells to respond to these genitalia-inducing signals. Our results support a limb-like developmental origin of external genitalia as the ancestral condition. Moreover, they suggest that a change in the relative position of the cloacal signalling centre during evolution has led to an altered developmental route for external genitalia in mammals, while preserving parts of the ancestral limb molecular circuitry owing to a common evolutionary origin.

Escores de cloaca e de crista e morfometria testicular em galos de matriz pesada com 71 semanas de idade e três categorias de peso corporal / Cloaca and comb scores and testicular morphometry in broiler breeder roosters at 71 weeks of age and three body weight categories

Em galos de matriz pesada, o peso dos testículos se associa positivamente com a produção diária de espermatozoides e com a fertilidade. Contudo, os testículos de aves se localizam na cavidade abdominal, não sendo acessíveis para exame andrológico como os testículos de mamíferos. A avaliação reprodutiva de galos de matriz pesada é subjetiva e se baseia em características como peso corporal (PC), qualidade de pés e pernas e morfologia de cloaca e crista. Estudos que validem o potencial de associação dessas características fenotípicas com parâmetros da morfometria testicular são raros, principalmente quando se consideram galos de matriz pesada manejados em lotes comerciais. Escores de cloaca e de crista e parâmetros da morfometria testicular foram avaliados em galos Cobb 500 Slow adultos das categorias de PC leve (<4,300kg), médio (entre 4,300 e 5,250kg) e pesado (≥5,250kg). O coeficiente de variação para PC foi de 12%. Os galos da categoria de PC pesado apresentaram testículos mais pesados e maiores escores de cloaca e de crista do que os galos da categoria de PC leve. Não houve diferença estatística entre as categorias de PC, considerando-se índice gonadossomático, porcentagens de túbulo seminífero e de tecido intersticial, diâmetro de túbulo seminífero e altura de epitélio seminífero. Correlação positiva, moderada e significativa foi observada entre peso de testículo e características de PC, escores de cloaca e de crista, diâmetro de túbulo seminífero e altura de epitélio seminífero. Um dos seis galos da categoria de PC leve avaliados quanto à histologia de testículo apresentou atrofia testicular. Em galos Cobb 500 Slow, conclui-se que o PC influencia o peso e a qualidade histológica de testículos e que os escores de cloaca e de crista podem ser utilizados com potencial moderado de predição para identificar galos com baixo peso de testículos...

In broiler breeder roosters the weight of the testes is positively associated with daily sperm production and fertility. However, the avian testes are located in the abdominal cavity, not being accessible for reproductive evaluation as mammalian testes. The reproductive evaluation of broiler breeder roosters is subjective and is based on characteristics such as body weight (BW), quality of feet and legs and morphology of cloaca and comb. Studies that validate the potential of associating these phenotype characteristics with testicular morphometric parameters are rare, especially when broiler breeder roosters of commercial flocks are considered. Cloaca and comb scores and testicular morphometric parameters were evaluated in adult Cobb 500 Slow roosters with low (<4.300kg), medium (between 4.300 and 5.250kg) and high (≥5.250kg) BW categories. The coefficient of variation for BW was 12%. The high BW roosters have heavier testes and higher cloaca and comb scores than the low BW roosters. There was no statistical difference between the BW categories in gonadosomatic index, percentage of seminiferous tubule and interstitial tissue, diameter of the seminiferous tubule and height of the seminiferous epithelium. Testis weight is positive, moderate and significantly correlated with BW, cloaca and comb scores, diameter of seminiferous tubule and height of seminiferous epithelium. One of six low BW roosters evaluated for histology of the testis had testicular atrophy. We conclude that in Cobb 500 Slow roosters BW influences the weight and histological quality of the testes. Besides this, cloaca and comb scores can be used with moderate prediction potential to identify roosters with underweight testes...

Cloacal anatomy of the male Carpathian newt, Lissotriton montandoni (Amphibia, Salamandridae), in the breeding season.

This study presents the first light microscopy-based description of the cloacal anatomy of male Carpathian newts (Lissotriton montandoni). This European newt species hybridizes with its sister species, the smooth newt (L. vulgaris), despite a high level of prezygotic isolation. The goal of the study was to ascertain possible anatomical differences in cloacal anatomy, especially pheromone and spermatophore producing glands, which might potentially affect reproductive isolation between L. montandoni and L. vulgaris. The cloaca of L. montandoni males consists of the cloacal tube, cloacal chamber, pseudopenis, and aggregations of cloacal glands. Four main types of cloacal glands were recognized. Pheromone-producing dorsal glands are of two types due to differences in their secretory epithelium: cuboidal and high prismatic. The remaining glands: ventral, pelvic, and Kingsbury's glands, are most likely involved in the synthesis of spermatophore components. Two distinct groups of ventral glands were identified: posterior and anterior ventral glands. In comparison with L. vulgaris, no evident differences were found that could potentially affect courtship pheromone synthesis by dorsal glands or the base and the cap of the spermatophore produced by the remaining cloacal glands.

Gross anatomical features of the gastrointestinal tract (GIT) of blue-and-yellow macaws (Ara ararauna) - oesophagus to cloaca.

Morphological studies of the gastrointestinal tract of blue-and-yellow macaws (Ara ararauna) are scarce. In view of the paucity of information regarding the digestive tract of macaws, this study aims to describe the gross anatomical features (oesophagus to cloaca) as part of a broad study of the gastrointestinal tract (GIT) of these birds. Three animals (two males and one female) adult macaws were anatomically dissected from the oropharynx to the cloaca to expose the GIT. The oesophagus was identified as a muscle-membranous tube continuous with the crop, which was intimately attached to the skin. The internal longitudinal folds of the cervical oesophagus were sparser cranial to the crop and less evident compared to the portion caudal to the crop. The duodenum began in the pylorus and was grey-coloured exhibiting a large lumen. The jejunum was formed by loops in a spiral-fashion model supported by mesojejunum. The ileum was also composed by small loops and was continuous with the colo-rectum forming the large intestine, because the caeca were absent. The large intestine was short, median in position, suspended in the dorsal wall of the abdominal cavity by mesentery and ended in the cloaca. The GIT was similar to the basic patterns in birds, in general, and also presented new unreported morphological data that might be important when studying nutrition and health of the macaws.

Efficiency of the cloacal sexing technique in greater rhea chicks (Rhea americana).

1. The feasibility and accuracy of the cloacal sexing technique in greater rhea chicks was assessed using chicks of two captive populations of greater rhea in Córdoba, Argentina. 2. A total of 46 greater rhea chicks of 2 to 3 months of age were randomly arranged into three groups and the members of each group were sexed by a different operator. 3. A feather of each chick was plucked for sexing through a molecular method and results were used as controls. 4. Sex was correctly assigned by cloacal inspection in 98% of the cases. Chick manipulation was easily performed and no infections or traumatic lesions were observed a posteriori. 5. Cloacal sexing of rhea chicks up to 3 months of age does not affect animal welfare and should be considered an efficient alternative to molecular methods.