196 NONINVASIVE HORMONE MEASUREMENTS FOR GENDER IDENTIFICATION IN MONOMORPHIC ANURANS

Noninvasive techniques have been successful in distinguishing gender in dimorphic amphibian models (Szymanski DC et al. 2006 Zoo Biol. 25, 35-46). The aim of this study was to assess gender-specific fecal hormone metabolite concentrations in 2 dimorphic and 2 monomorphic, critically endangered species of amphibians to assist in ex situ breeding efforts. Fecal samples from 2 dimorphic anurans, the Puerto Rican Crested Toad (Bufo lemur) and the Wyoming Toad (Bufo baxteri) and 2 monomorphic species, the Mountain Chicken Frog (Leptodactylus fallax) and the Dusky Gopher Frog (Lithobates sevosa) were measured by enzyme immunoassay for sex-specific hormone differences (Student’s t-test; P < 0.05) among 6 variables (concentrations of estradiol (E2), progesterone (P), and testosterone (T), and ratios of P:E2, T:E2, and P:T). Values are presented as means ± SEM. Mountain Chicken Frog samples were obtained from individuals of known sex. Dusky Gopher Frog samples were from confirmed females but males were presumed based on subtle physical characteristics. The E2 concentrations (ng g-1) were significantly higher in females than males (P < 0.05) in all species except for the Dusky Gopher Frog. However, males in all 4 species, including the presumptive Dusky Gopher Frog males, exhibited significantly higher P:E2 and T:E2 ratios than females. Additionally, Wyoming Toad and Mountain Chicken Frog females had significantly higher P:T ratios than males (0.19 ± 0.05 v. 0.06 ± 0.00 and 0.34 ± 0.03 v. 0.20 ± 0.02, respectively). Similar to the American Toad (Szymanski et al. 2006), presumptive Dusky Gopher Frog males measured significantly higher T concentrations compared with females (376.4 ± 118.5 ng g-1 v. 174.0 ± 23.1 ngg-1, respectively). Although species-specific differences are apparent, fecal hormone measurements can be used to identify sex in monomorphic species and offer a potential tool for the captive breeding of endangered anurans. Table 1.

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Reproductive parameters in some captive-bred cockatoo species (genus Cacatua and Eolophus)

Veterinary Record Open ◽

10.1136/vetreco-2020-000405 ◽

2020 ◽

Vol 7 (1) ◽

pp. e000405

Author(s):

Stefano Paolo Marelli ◽

Ahmad Abdel Sayed ◽

Michele Magni ◽

Lorenzo Crosta ◽

Petra Schnitzer ◽

...

Keyword(s):

Endangered Species ◽

Reproductive Performance ◽

Captive Breeding ◽

Scientific Data ◽

Ex Situ ◽

Reproductive Parameters ◽

In Captivity ◽

Species Effect ◽

Adaptive Ability ◽

Species Specific

BackgroundCockatoo reproduction in captivity supplies a powerful tool to limit the economic motivation to capture endangered species from the wild; nevertheless, scientific data about reproductive parameters in cockatoos are very rare. The aim of the present work was to investigate the reproductive performance of different species of the Cacatua genus reared in the same facility to evaluate adaptability to captive breeding and to identify the main problems in ex situ conservation of some cockatoo species.MethodsData of 28 eggs from 19 reproductive pairs from 9 cockatoo species were analysed. Statistical analysis was carried out by SAS NPAR1WAY procedure: species was considered source of variation.ResultsSpecies effect does not significantly influence reproductive variables; differences were recorded in eggs fertility and embryo liveability. Bird adaptive ability to captive breeding has been described through reproductive parameters.ConclusionOur results show the importance and the maintenance of natural species-specific behaviours and habits, and they underline the relevance of data collection about reproductive performance in endangered species kept in captivity to improve breeding management in conservation programmes.

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Isolation and Characterization of 27 Novel Microsatellite Loci in Critically Endangered Wyoming Toad

Journal of Fish and Wildlife Management ◽

10.3996/042019-jfwm-029 ◽

2019 ◽

Vol 10 (2) ◽

pp. 563-566

Author(s):

Renee M. Martin ◽

Heidi Meador ◽

Lee Bender ◽

Lacey Hopper

Keyword(s):

Captive Breeding ◽

Captive Breeding Program ◽

Isolation And Characterization ◽

The Family ◽

Hardy Weinberg Equilibrium ◽

Laramie Basin ◽

Species Specific ◽

Wyoming Toad ◽

Paired End Sequencing

Abstract Wyoming toad Anaxyrus baxteri is a federally endangered amphibian endemic to the Laramie basin in southwestern Wyoming, USA. A captive breeding program propagates A. baxteri, and the monitoring of genetic diversity in the captive stock can assist in guiding conservation measures of this species. Illumina paired-end sequencing lead to 27 species-specific polymorphic microsatellite genetic markers being developed. Across 24 samples, A. baxteri exhibited two to eight alleles per locus, and observed and expected heterozygosities per locus ranged from 0.292 to 0.958 and from 0.344 to 0.787, respectively. Tests for Hardy–Weinberg equilibrium were nonsignificant except for Abax_13 and Abax_39. These microsatellite markers will be useful for genetic monitoring to aid recovery efforts of A. baxteri captive and wild populations as well as other amphibians in the family Bufonidae.

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Reproductive Anatomy of Chondrichthyans: Notes on Specimen Handling and Sperm Extraction. II. Sharks and Chimaeras

Animals ◽

10.3390/ani11082191 ◽

2021 ◽

Vol 11 (8) ◽

pp. 2191

Author(s):

Pablo García-Salinas ◽

Victor Gallego ◽

Juan F. Asturiano

Keyword(s):

Artificial Insemination ◽

Captive Breeding ◽

Ex Situ ◽

Specimen Handling ◽

Viable Sperm ◽

Abdominal Massage ◽

Reproductive Anatomy ◽

Conservation Projects ◽

Breeding Techniques

The chondrichthyan fishes, which comprise sharks, rays, and chimaeras, are one of the most threatened groups of vertebrates on the planet. Given this situation, an additional strategy for the protection of these species could be the ex situ conservation projects developed in public aquaria and research centers. Nevertheless, to increase sustainability and to develop properly in situ reintroduction strategies, captive breeding techniques, such as sperm extraction and artificial insemination, should be developed. These techniques are commonly used in other threatened species and could be also used in chondrichthyans. However, the different reproductive morphologies found in this group can complicate both processes. Therefore, a comparison of the reproductive anatomy of eight distinct chondrichthyans, with an emphasis on those important differences when performing sperm extraction or artificial insemination, is carried out herein. Sharks and chimaeras belonging to the Scyliorhinidae, Carcharhinidae, Centrophoridae, Etmopteridae, Hexanchidae, and Chimaeridae families were obtained from commercial fisheries, public aquaria, and stranding events. In addition, the process of obtaining viable sperm samples through cannulation, abdominal massage, and oviducal gland extraction is described in detail for both living and dead animals.

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The Role of Gardens in Integrated Conservation Practice

Sibbaldia: the International Journal of Botanic Garden Horticulture ◽

10.24823/sibbaldia.2019.273 ◽

2019 ◽

pp. 177-188

Author(s):

Audrey Denvir ◽

Jeannine Cavender-Bares ◽

Antonio González-Rodríguez

Keyword(s):

Plant Conservation ◽

Ex Situ ◽

Conservation Practice ◽

Integrated Research ◽

Academic Researchers ◽

Conservation Plan ◽

Species Specific ◽

Integrated Conservation

Gardens and horticulturists play an increasingly important role in plant conservation, both in situ and ex situ. Integrated research and conservation of species intends to work across fields to connect science to conservation practice by engaging actors from different sectors, including gardens. The case of integrated conservation of Quercus brandegeei, a microendemic oak species in Baja California Sur, Mexico, is presented as an example of a collaboration between gardens and academic researchers to create a species-specific conservation plan that incorporates horticultural knowledge.

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First Captive Breeding Program for the Endangered Pyrenean Sculpin (Cottus hispaniolensis Bacescu-Master, 1964)

Water ◽

10.3390/w12112986 ◽

2020 ◽

Vol 12 (11) ◽

pp. 2986

Author(s):

Joan Manubens ◽

Oriol Comas ◽

Núria Valls ◽

Lluís Benejam

Keyword(s):

Captive Breeding ◽

Ex Situ Conservation ◽

Breeding Program ◽

Ex Situ ◽

Major Influence ◽

Freshwater Species ◽

Non Profit ◽

Captive Breeding Program ◽

Critical Phases ◽

Conservation Plans

The strong decline of freshwater fish species in Europe implies that further ex-situ conservation plans should be implemented in the near future. The present study reflects our experience with the Pyrenean sculpin (Cottus hispaniolensis Bacescu-Mester, 1964)—a small cottid endemic to the Hispano-French Garona River basin. In recent years, the Spanish Pyrenean sculpin population has reached a limit situation. Because of that, the non-profit association ADEFFA—with support from the public administration—started the first captive breeding program for this species in 2006. Fourteen years later, this study presents the results and evaluates the different steps of the program, with the aim of discussing and improving the ex-situ conservation plans for this and other cold freshwater species. There is a description and a comparison between six consecutive phases during the captive breeding process: nesting behaviour, courtship, egg fixation, parental care (incubation), hatching and survival during juvenile development. The purposes of this project are to: (1) identify the most determining phases for a successful captive breeding; (2) identify the factors that had a major influence to the success of the critical phases; and (3) increase the number of the offspring. This study is based on thirty-three wild individuals collected from Garona River (Val d’Aran, Spanish Pyrenees). During the program, twelve couples spawned in captive conditions, with around 2300 eggs laid. Eight couples bred successfully, with 751 hatched individuals and 608 juveniles reared. The analysis of each step of the captive breeding does not reveal significant differences between phases, so it can be concluded that they are all critical at the same level. In the literature, similar study-cases of captive breeding programs identify incubation and survival phases as the most critical. Consequently, the management made for this project has probably allowed to overcome in part the main impediments described in other similar programs.

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Characterization of the spermiation response, luteinizing hormone release and sperm quality in the American toad (Bufo americanus) and the endangered Wyoming toad (Bufo baxteri)

Reproduction Fertility and Development ◽

10.1071/rd00056 ◽

2000 ◽

Vol 12 (2) ◽

pp. 51 ◽

Cited By ~ 36

Author(s):

Amy R. Obringer ◽

Justine K. O'Brien ◽

Robin L. Saunders ◽

Kazutoshi Yamamoto ◽

Sakae Kikuyama ◽

...

Keyword(s):

Sperm Quality ◽

Sampling Interval ◽

Sperm Production ◽

Bufo Americanus ◽

American Toad ◽

Non Invasive ◽

Lh Release ◽

Wyoming Toad ◽

Toad Bufo

Spermiation and LH release in response to several methods of LHRH administration were assessed in the American toad (Bufo americanus), and the most successful method was tested in the endangered Wyoming toad (Bufo baxteri). Specific objectives were to: (1) compare spermiation responses and plasma LH concentration after invasive and non-invasive LHRH treatments; (2) evaluate sperm production in response to different LHRH dosages; (3) characterize the timing of sperm release post LHRH treatment; and (4) assess sperm quality (motility, viability, morphology and acrosomal status). Male American toads were administered 4 g LHRH by one of four routes: (1) intraperitoneal injection (IP); (2) subcutaneous injection (SQ); (3) dorsal dermis absorption (DDA); and (4) ventral dermis absorption (VDA). Aspermic urine only was collected from saline-treated controls and DDA animals. Several VDA animals released spermic urine; however, all LHRH-injected toads released spermatozoa. IP animals produced higher sperm and LH concentrations than SQ animals. The spermiation response in animals treated IP with 1 g LHRH was similar to that in animals treated with 4 g, but lower LHRH dosages tested produced inferior responses. Sperm production in responsive animals increased over time during the 12-h sampling interval. Regardless of treatment, most American toad spermatozoa were motile, viable, and acrosome-intact. Endangered Wyoming toads were treated IP with 4 g LHRH, and spermic urine was collected. Although most spermatozoa were viable and acrosome-intact, a considerable percentage possessed structurally abnormal heads. A single IP injection of LHRH appears to be a reliable and safe method for controlling spermiation in toads and may be useful for assisting endangered amphibian propagation.

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GIS-Facilitated Effective Propagation Protocols of the Endangered Local Endemic of Crete Carlina diae (Rech. f.) Meusel and A. Kástner (Asteraceae): Serving Ex Situ Conservation Needs and Its Future Sustainable Utilization as an Ornamental

Plants ◽

10.3390/plants9111465 ◽

2020 ◽

Vol 9 (11) ◽

pp. 1465

Author(s):

Katerina Grigoriadou ◽

Virginia Sarropoulou ◽

Nikos Krigas ◽

Eleni Maloupa ◽

Georgios Tsoktouridis

Keyword(s):

Added Value ◽

Ex Situ ◽

Sustainable Utilization ◽

Ms Medium ◽

Sustainable Exploitation ◽

Local Endemic ◽

Species Specific ◽

Shoot Propagation

Conservation and sustainable exploitation of threatened endemic plants with medicinal and/or horticultural/ornamental value can be achieved through the development of effective propagation protocols. After unveiling the bioclimatic preferences of Carlina diae (Asteraceae) with geographic information systems (GIS), four propagation trials were conducted using seeds of this endangered local Cretan endemic for in vivo and in vitro germination, as well as seasonal vegetative propagation trials (softwood cuttings) and micropropagation (nodal explants). Seed germination was accomplished at a level of 77–90% in vivo (30 days) and 96% in vitro (10 days) using an MS medium with 2.9 μM gibberellic acid (GA3). The optimum treatments for cuttings’ rooting were 1000 and 2000 ppm indole-3-butyric acid (IBA) (11–16 roots, 2–3 cm long, 100% rooting) within 40 days in mist. In vitro shoot propagation exhibited a 2.8 proliferation rate after six successive subcultures on an MS medium with 2.9 μM GA3. Both ex vitro rooting and acclimatization were successful in 40 days, with 96% microshoot rooting and an equal survival rate. The GIS-facilitated effective species-specific propagation protocols developed in this study can consolidate the perspective of successful re-introduction of ex situ-raised material of C. diae into wild habitats and may serve its sustainable exploitation for high-added value ornamental products.

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Serratiopeptidase reduces the invasion of osteoblasts by Staphylococcus aureus

International Journal of Immunopathology and Pharmacology ◽

10.1177/0394632017745762 ◽

2017 ◽

Vol 30 (4) ◽

pp. 423-428 ◽

Cited By ~ 6

Author(s):

Laura Selan ◽

Rosanna Papa ◽

Angela Ermocida ◽

Andrea Cellini ◽

Evaristo Ettorre ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Periprosthetic Infection ◽

Bone Cells ◽

Etiologic Agent ◽

Osteoblast Cells ◽

Main Target ◽

Student’S T ◽

Potential Tool ◽

New Strategies ◽

Student’S T Test

Finding new strategies to counteract periprosthetic infection and implant failure is a main target in orthopedics. Staphylococcus aureus, the leading etiologic agent of orthopedic implant infections, is able to enter and kill osteoblasts, to stimulate pro-inflammatory chemokine secretion, to recruit osteoclasts, and to cause inflammatory osteolysis. Moreover, by entering eukaryotic cells, staphylococci hide from the host immune defenses and shelter from the extracellular antibiotics. Thus, infection persists, inflammation thrives, and a highly destructive osteomyelitis occurs around the implant. The ability of serratiopeptidase (SPEP), a metalloprotease by Serratia marcescens, to control S. aureus invasion of osteoblastic MG-63 cells and pro-inflammatory chemokine MCP-1 secretion was evaluated. Human osteoblast cells were infected with staphylococcal strains in the presence and in the absence of SPEP. Cell proliferation and cell viability were also evaluated. The release of pro-inflammatory chemokine MCP-1 was evaluated after the exposure of the osteoblast cells to staphylococcal strains. The significance of the differences in the results of each test and the relative control values was determined with Student’s t-test. SPEP impairs their invasiveness into osteoblasts, without affecting the viability and proliferation of bone cells, and tones down their production of MCP-1. We recognize SPEP as a potential tool against S. aureus bone infection and destruction.

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