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

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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Reproductive Anatomy of Chondrichthyans: Notes on Specimen Handling and Sperm Extraction. I. Rays and Skates

Animals ◽

10.3390/ani11071888 ◽

2021 ◽

Vol 11 (7) ◽

pp. 1888

Author(s):

Pablo García-Salinas ◽

Victor Gallego ◽

Juan F. Asturiano

Keyword(s):

Threatened Species ◽

Artificial Insemination ◽

Captive Breeding ◽

Ex Situ ◽

Breeding Programs ◽

Specimen Handling ◽

Abdominal Massage ◽

Reproductive Anatomy ◽

Captive Breeding Programs

The superorder Batoidea (rays, skates, and relatives), constitutes one of the most threatened group of vertebrates. Strengthening ex situ conservation programs developed in research centers and public aquaria could be a way of addressing this situation. However, captive breeding programs must be improved to prevent the capture of wild animals and to develop proper in situ reintroduction strategies. Sperm extraction and artificial insemination are two techniques commonly used in other threatened species, which could also be used in rays and the like. However, the different reproductive morphologies present within this group of animals may hamper both processes. Here, we present a comparison of the reproductive anatomies of 11 distinct batoid species, emphasizing the important differences between the species when performing sperm extraction or artificial insemination. Both male and female animals, belonging to the Rajidae, Dasyatidae, Torpedinidae and Myliobatidae families, from the Mediterranean Sea were studied. In addition, we describe the procedure to extract sperm using both cannulation and abdominal massage, either from live or dead batoids Finally, the obtention of motile sperm recovered from the oviducal gland of females is described. These techniques generate a new range of possibilities for the conservation of these threatened species.

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Conservation of wild silkworm genetic resources through cryopreservation: Standardization of sperm processing

Journal of Applied and Natural Science ◽

10.31018/jans.v10i2.1733 ◽

2018 ◽

Vol 10 (2) ◽

pp. 544-547

Author(s):

G. Lokesh ◽

Geetha N Murthy ◽

Veeranna Gowda ◽

Alok Sahay ◽

Gargi Gargi

Keyword(s):

Genetic Resources ◽

Seminal Vesicle ◽

Artificial Insemination ◽

Biologically Active ◽

Ex Situ ◽

Female Moth ◽

Male Moth ◽

Semen Collection ◽

Wild Silkworm ◽

Viable Sperm

Conservation of the invaluable sericigenous genetic resources is of prime importance with respect to their utilization and improvement for wider exploitation. Conservation of wild silkworms and its applicability in hybridization have limitations due to incompatibility, less amenability, change of behaviour under ex situ conditions, non-synchronization of moth eclosion and difficulties in mating between variables. In view of this, the newer technologies such as cryopreservation and artificial insemination are offering better strategies for preservation of biologically active samples like semen at sub-zero temperature (-196º C) conditions for longer duration. In this context, under standardization of sperms preservation from wild silkworms, two methods of semen collection were scrutinized for obtaining active and viable sperm for cryopreservation and further artificial insemination. Semen collection from the seminal vesicle of freshly emerged male moth and the other from the bursa copulatrix (BC) and spermatheca of the female moth after mating. The sperms in the semen collected from seminal vesicle are in the form of bundles known as eupyrene sperm bundles and apyrene sperms. The morphology and behaviour of these sperm bundles were recorded through microscopic examination. To study the density and motility behaviour of the sperms, sperm bundles were treated with proteolytic enzyme (~2-3µg/ml) to digest the membrane and release the sperms. The density and motility behaviour of sperms in the semen recovered from the BC and spermatheca of female moth after mating were higher compared to those released after digestion of sperm bundles from seminal vesicle of the male moth.

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Ark or park: the need to predict relative effectiveness of ex situ and in situ conservation before attempting captive breeding

Journal of Applied Ecology ◽

10.1111/1365-2664.12449 ◽

2015 ◽

Vol 52 (4) ◽

pp. 841-850 ◽

Cited By ~ 22

Author(s):

Paul M. Dolman ◽

Nigel J. Collar ◽

Keith M. Scotland ◽

Robert. J. Burnside

Keyword(s):

Captive Breeding ◽

Relative Effectiveness ◽

In Situ Conservation ◽

Ex Situ

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Conservation and behavior of Africa’s “Big Five”

Current Zoology ◽

10.1093/czoolo/60.4.486 ◽

2014 ◽

Vol 60 (4) ◽

pp. 486-499 ◽

Cited By ~ 8

Author(s):

Tim Caro ◽

Jason Riggio

Keyword(s):

Big Five ◽

Captive Breeding ◽

Conservation Status ◽

In Situ Conservation ◽

Animal Husbandry ◽

Ex Situ ◽

Flagship Species ◽

African Species ◽

In The Wild

Abstract We examine the conservation status of Africa’s “Big Five”: lion, leopard, buffalo, black and white rhinoceros and elephant, and the role of behavioral knowledge in their conservation. Efforts to conserve these flagship species consist of in situ conservation, captive breeding and reintroductions. With a few exceptions, we find limited evidence that knowledge of behavior informs conservation programs targeted at these species. For management in the wild, knowledge of infanticide and ranging can provide guidelines for realistic hunting quotas and corridors between protected areas, respectively. For ex situ and reintroduction programs, behavioral knowledge is chiefly focused on improved animal husbandry. Despite a formidable understanding of these species’ behavior, the practicalities of using such knowledge may be diminished because exploitation of these species is so forceful and the bulk of efforts aimed at conserving these species (and indeed most other African species) are primarily in situ where behaviorally driven interventions are limited. Our comparative findings suggest that behavior has been of rather narrow use in the conservation of these flagship species [Current Zoology 60 (4): 486–499, 2014].

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Elephants on the Move: Spatial Formations of Wildlife Exchange

Environment and Planning D Society and Space ◽

10.1068/d210t ◽

2000 ◽

Vol 18 (2) ◽

pp. 185-203 ◽

Cited By ~ 57

Author(s):

Sarah Whatmore ◽

Lorraine Thorne

Keyword(s):

Wildlife Conservation ◽

Captive Breeding ◽

Science Studies ◽

In Situ Conservation ◽

Global Networks ◽

Feminist Science ◽

The Status ◽

Conservation Projects ◽

Spatial Formations

In this paper we explore tensions between the notions and spaces of social agency mobilised in actant network theory and feminist science studies by focusing on their implications for the status and treatment of nonhuman animals, in this case the African elephant. The notion of a spatial formation of wildlife exchange (SFWE) is deployed to trace the diverse modalities and spatialities of social networks in which such creatures are caught up and the ways in which these practical orderings work through the bodies of elephants, both in the sense of their energies being variously transduced and of their experiences being reconfigured in the process. These themes are pursued through two contemporary global networks of wildlife conservation/science. The first, characterised as a mode of ordering of foresight, is a network concerned with ‘captive breeding’ and configured through the coding and exchange of computerised information on the lineages and breeding properties of animals held in zoological collections worldwide. The second, characterised as a mode of ordering of authenticity, is a network concerned with ‘in-situ’ conservation projects and configured through the recruitment of paying volunteers, corporate donors, and field scientists to a global programme of research expeditions. Our account traces three simultaneous moments in the patterning of elephants in each network—as virtual bodies, as bodies in place, and as living spaces.

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In-Situ and Ex-Situ status of the Javan Gibbon and the role of zoos in conservation of the species

Contributions to Zoology ◽

10.1163/18759866-0750304005 ◽

2006 ◽

Vol 75 (03-04) ◽

pp. 161-168 ◽

Cited By ~ 16

Author(s):

Vincent Nijman

Keyword(s):

Captive Breeding ◽

Conservation Status ◽

Ex Situ ◽

Captive Breeding Programme ◽

In Captivity ◽

Hylobates Moloch ◽

In The Wild ◽

Set Up ◽

Javan Gibbon

The Javan gibbon Hylobates moloch is confined to the last remnants of rainforest on the island of Java, Indonesia. As of 2002, the species has been recorded in 29 forest areas, and the wild population is conservatively estimated at 4100-4500 individuals. Over 95% of the gibbons are in populations of >100 individuals, and the four largest areas support populations of >500 individuals each. In 2003, 56 Javan gibbons were maintained at eight Indonesian zoos, 15 at four Indonesian wildlife rescue centres, with five potential breeding pairs. There is no evidence that the species has bred successfully in captivity in Indonesia. Outside the range country, 48 Javan gibbons were maintained at ten institutions in nine countries, with six breeding pairs. The total ex-situ population is some 120 individuals, the majority of which is wild-caught. At present most initiatives relating to the conservation of the Javan gibbon have targeted small isolated forest areas and the ex-situ population, whereas in-situ protection of the largest populations in the wild has been largely ignored. Significant populations are currently found in unprotected forests. The large captive population of Javan gibbons within Indonesia allows, with improved co-operation, to set up an integrated captive- breeding programme. This should, not be seen, however, as a means to improve the conservation status of the wild Javan gibbons, which needs to be achieved through protection of remaining habitat, but could be used for improving the prevailing low levels of conservation awareness in Java.

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An in situ and ex situ TEM study of the formation of thin cobalt silicide films by molecular beam epitaxy on the silicon (100) surface

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010010648x ◽

1988 ◽

Vol 46 ◽

pp. 884-885

Author(s):

D. Loretto ◽

J. M. Gibson ◽

S. M. Yalisove ◽

R. T. Tung

Keyword(s):

Molecular Beam Epitaxy ◽

Molecular Beam ◽

Defect Density ◽

High Vacuum ◽

Ultra High Vacuum ◽

Ex Situ ◽

Metal Base ◽

In Situ Experiments ◽

Potential Applications

The cobalt disilicide/silicon system has potential applications as a metal-base and as a permeable-base transistor. Although thin, low defect density, films of CoSi2 on Si(111) have been successfully grown, there are reasons to believe that Si(100)/CoSi2 may be better suited to the transmission of electrons at the silicon/silicide interface than Si(111)/CoSi2. A TEM study of the formation of CoSi2 on Si(100) is therefore being conducted. We have previously reported TEM observations on Si(111)/CoSi2 grown both in situ, in an ultra high vacuum (UHV) TEM and ex situ, in a conventional Molecular Beam Epitaxy system.The procedures used for the MBE growth have been described elsewhere. In situ experiments were performed in a JEOL 200CX electron microscope, extensively modified to give a vacuum of better than 10-9 T in the specimen region and the capacity to do in situ sample heating and deposition. Cobalt was deposited onto clean Si(100) samples by thermal evaporation from cobalt-coated Ta filaments.

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The use of transmission and analytical electron microscopy in studying reactions and phase transformations in thin film

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100150046 ◽

1993 ◽

Vol 51 ◽

pp. 842-843

Author(s):

K. Barmak

Keyword(s):

Thin Film ◽

Thin Films ◽

Phase Transformations ◽

Analytical Electron Microscopy ◽

Ex Situ ◽

Multilayer Thin Films ◽

Absolute Concentration ◽

Analytical Electron ◽

Deposition Step

Generally, processing of thin films involves several annealing steps in addition to the deposition step. During the annealing steps, diffusion, transformations and reactions take place. In this paper, examples of the use of TEM and AEM for ex situ and in situ studies of reactions and phase transformations in thin films will be presented.The ex situ studies were carried out on Nb/Al multilayer thin films annealed to different stages of reaction. Figure 1 shows a multilayer with dNb = 383 and dAl = 117 nm annealed at 750°C for 4 hours. As can be seen in the micrograph, there are four phases, Nb/Nb3-xAl/Nb2-xAl/NbAl3, present in the film at this stage of the reaction. The composition of each of the four regions marked 1-4 was obtained by EDX analysis. The absolute concentration in each region could not be determined due to the lack of thickness and geometry parameters that were required to make the necessary absorption and fluorescence corrections.

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Transmission Electron Microscopy of Epitaxial silicides grown by ultra high vacuum deposition

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100154287 ◽

1989 ◽

Vol 47 ◽

pp. 462-463

Author(s):

D. Loretto ◽

J. M. Gibson ◽

S. M. Yalisove

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Electron Diffraction ◽

High Vacuum ◽

High Energy ◽

Energy Electron ◽

Ultra High Vacuum ◽

Ex Situ ◽

Transmission Electron

The silicides CoSi2 and NiSi2 are both metallic with the fee flourite structure and lattice constants which are close to silicon (1.2% and 0.6% smaller at room temperature respectively) Consequently epitaxial cobalt and nickel disilicide can be grown on silicon. If these layers are formed by ultra high vacuum (UHV) deposition (also known as molecular beam epitaxy or MBE) their thickness can be controlled to within a few monolayers. Such ultrathin metal/silicon systems have many potential applications: for example electronic devices based on ballistic transport. They also provide a model system to study the properties of heterointerfaces. In this work we will discuss results obtained using in situ and ex situ transmission electron microscopy (TEM).In situ TEM is suited to the study of MBE growth for several reasons. It offers high spatial resolution and the ability to penetrate many monolayers of material. This is in contrast to the techniques which are usually employed for in situ measurements in MBE, for example low energy electron diffraction (LEED) and reflection high energy electron diffraction (RHEED), which are both sensitive to only a few monolayers at the surface.

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Atomic Layer Deposition of LiF and Lithium Ion Conducting (AlF3)(LiF)x Alloys Using Trimethylaluminum, Lithium Hexamethyldisilazide and Hydrogen Fluoride

10.26434/chemrxiv.5459653 ◽

2017 ◽

Author(s):

Younghee Lee ◽

Daniela M. Piper ◽

Andrew S. Cavanagh ◽

Matthias J. Young ◽

Se-Hee Lee ◽

...

Keyword(s):

Lithium Ion ◽

Atomic Layer ◽

Mass Gain ◽

Alloy Film ◽

Ex Situ ◽

X Ray ◽

Layer Deposition ◽

Ion Conducting ◽

Good Agreement

<div>Atomic layer deposition (ALD) of LiF and lithium ion conducting (AlF3)(LiF)x alloys was developed using trimethylaluminum, lithium hexamethyldisilazide (LiHMDS) and hydrogen fluoride derived from HF-pyridine solution. ALD of LiF was studied using in situ quartz crystal microbalance (QCM) and in situ quadrupole mass spectrometer (QMS) at reaction temperatures between 125°C and 250°C. A mass gain per cycle of 12 ng/(cm2 cycle) was obtained from QCM measurements at 150°C and decreased at higher temperatures. QMS detected FSi(CH3)3 as a reaction byproduct instead of HMDS at 150°C. LiF ALD showed self-limiting behavior. Ex situ measurements using X-ray reflectivity (XRR) and spectroscopic ellipsometry (SE) showed a growth rate of 0.5-0.6 Å/cycle, in good agreement with the in situ QCM measurements.</div><div>ALD of lithium ion conducting (AlF3)(LiF)x alloys was also demonstrated using in situ QCM and in situ QMS at reaction temperatures at 150°C A mass gain per sequence of 22 ng/(cm2 cycle) was obtained from QCM measurements at 150°C. Ex situ measurements using XRR and SE showed a linear growth rate of 0.9 Å/sequence, in good agreement with the in situ QCM measurements. Stoichiometry between AlF3 and LiF by QCM experiment was calculated to 1:2.8. XPS showed LiF film consist of lithium and fluorine. XPS also showed (AlF3)(LiF)x alloy consists of aluminum, lithium and fluorine. Carbon, oxygen, and nitrogen impurities were both below the detection limit of XPS. Grazing incidence X-ray diffraction (GIXRD) observed that LiF and (AlF3)(LiF)x alloy film have crystalline structures. Inductively coupled plasma mass spectrometry (ICP-MS) and ionic chromatography revealed atomic ratio of Li:F=1:1.1 and Al:Li:F=1:2.7: 5.4 for (AlF3)(LiF)x alloy film. These atomic ratios were consistent with the calculation from QCM experiments. Finally, lithium ion conductivity (AlF3)(LiF)x alloy film was measured as σ = 7.5 × 10-6 S/cm.</div>

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