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      Sex Determination in Two Species of Anuran Amphibians by Magnetic Resonance Imaging and Ultrasound Techniques

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          Abstract

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          Amphibians are of critical importance among vertebrates. They play a critical role in their ecosystems, are commonly used as environmental health indicators and are also considered as exotic pets throughout the world. Among amphibians, many anuran species are included in active conservation programs as they are listed as endangered species. Thus, it is important for veterinarians and biologists to examine their sanitary status and to find a non-invasive tool to evaluate the health status of these individuals, particularly the state of their reproductive system and to be able to carry out a sex determination in case of no sexual dimorphism. For the first time, we demonstrate that benchtop magnetic resonance imaging and high-resolution ultrasound are suitable non-invasive imaging techniques for an accurate sex determination of two anuran species. Both techniques allowed the identification of ovaries and testes. Therefore, our data constitute an important contribution for clinical diagnostic and conservation purposes in amphibians, as it is possible to distinguish males and females in a quick, safe and relatively easy way.

          Abstract

          The objective of the present study was to evaluate whether gender determination in two amphibian species ( Kaloula pulchra and Xenopus laevis) can be reliably carried out by means of magnetic resonance imaging (benchtop magnetic resonance imaging; BT-MRI) or ultrasound (high-resolution ultrasound; HR-US) techniques. Two species of healthy, sexually mature anurans have been used in the present study. Eight Kaloula (blind study) and six Xenopus were used as controls. Magnetic resonance imaging experiments were carried out on a low-field (1 Tesla) benchtop-MRI (BT-MRI) system. HR-US examination was performed with high-resolution equipment. Low-field BT-MRI images provided a clear and quantifiable identification of all the sexual organs present in both genders and species. The HR-US also allowed the identification of testes and ovaries in both species. Results indicate that BT-MRI allowed a very precise sex identification in both anuran species, although its use is limited by the cost of the equipment and the need for anesthesia. HR-US allowed an accurate identification of ovaries of both species whereas a precise identification of testes is limited by the ultrasonographer experience. The main advantages of this technique are the possibility of performing it without anesthesia and the higher availability of equipment in veterinary and zoo institutions.

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          Status and trends of amphibian declines and extinctions worldwide.

          The first global assessment of amphibians provides new context for the well-publicized phenomenon of amphibian declines. Amphibians are more threatened and are declining more rapidly than either birds or mammals. Although many declines are due to habitat loss and overutilization, other, unidentified processes threaten 48% of rapidly declining species and are driving species most quickly to extinction. Declines are nonrandom in terms of species' ecological preferences, geographic ranges, and taxonomic associations and are most prevalent among Neotropical montane, stream-associated species. The lack of conservation remedies for these poorly understood declines means that hundreds of amphibian species now face extinction.
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            Applied reproductive technologies and genetic resource banking for amphibian conservation.

            As amphibian populations continue to decline, both government and non-government organisations are establishing captive assurance colonies to secure populations deemed at risk of extinction if left in the wild. For the most part, little is known about the nutritional ecology, reproductive biology or husbandry needs of the animals placed into captive breeding programs. Because of this lack of knowledge, conservation biologists are currently facing the difficult task of maintaining and reproducing these species. Academic and zoo scientists are beginning to examine different technologies for maintaining the genetic diversity of founder populations brought out of the wild before the animals become extinct from rapidly spreading epizootic diseases. One such technology is genetic resource banking and applied reproductive technologies for species that are difficult to reproduce reliably in captivity. Significant advances have been made in the last decade for amphibian assisted reproduction including the use of exogenous hormones for induction of spermiation and ovulation, in vitro fertilisation, short-term cold storage of gametes and long-term cryopreservation of spermatozoa. These scientific breakthroughs for a select few species will no doubt serve as models for future assisted breeding protocols and the increasing number of amphibians requiring conservation intervention. However, the development of specialised assisted breeding protocols that can be applied to many different families of amphibians will likely require species-specific modifications considering their wide range of reproductive modes. The purpose of this review is to summarise the current state of knowledge in the area of assisted reproduction technologies and gene banking for the conservation of amphibians.
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              Amphibian conservation, land-use changes and protected areas: A global overview

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                Author and article information

                Journal
                Animals (Basel)
                Animals (Basel)
                animals
                Animals : an Open Access Journal from MDPI
                MDPI
                2076-2615
                18 November 2020
                November 2020
                : 10
                : 11
                : 2142
                Affiliations
                [1 ]Departamento de Medicina y Cirugía, Facultad de Veterinaria, Universidad Complutense, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain; mariajoseruizfernandez@ 123456gmail.com (M.J.R.-F.); isagreal@ 123456vet.ucm.es (M.I.G.-R.)
                [2 ]Departamento de Biología Celular, Facultad de Biología, Universidad Complutense, Avenida José Antonio Novais 12, 28040 Madrid, Spain; sajime01@ 123456ucm.es (S.J.); nerea@ 123456bio.ucm.es (N.M.)
                [3 ]Unidad de RMN—CAI Bioimagen Complutense, Universidad Complutense, Paseo de Juan XXIII 1, 28040 Madrid, Spain; evalle@ 123456ucm.es (E.F.-V.); dcastejon@ 123456ucm.es (D.C.)
                [4 ]Centro Veterinario Los Sauces, Calle de Santa Engracia 63, 28010 Madrid, Spain; ardiaca.m@ 123456outlook.com (M.A.); amontesinosbarcelo@ 123456gmail.com (A.M.); salvadorariza@ 123456msn.com (S.A.)
                [5 ]Departamento de Anatomía y Embriología, Sección Departamental de Anatomía y Embriología (Veterinaria), Facultad de Veterinaria, Universidad Complutense, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain
                Author notes
                [* ]Correspondence: juncalgs@ 123456vet.ucm.es
                Author information
                https://orcid.org/0000-0002-7433-5976
                https://orcid.org/0000-0002-5578-192X
                https://orcid.org/0000-0002-6375-2459
                Article
                animals-10-02142
                10.3390/ani10112142
                7698606
                33217993
                c0d1d5e1-72a6-4564-af1b-0d5a60a4c12d
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 18 October 2020
                : 16 November 2020
                Categories
                Article

                amphibians,anurans,sex determination,magnetic resonance,ultrasonography

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