Gene Banks and AI: Minitube´s Contribution to Biodiversity

It is estimated that every year, up to 100 animal species disappear from the earth forever, and thousands of animals are so severely endangered that they may soon face the same fate. Therefore, some experts are already talking about the sixth great mass extinction in the history of the earth.

The Red List shows that around a quarter of all mammal species are threatened with extinction (IUCN 2023). In addition to the obvious exotic wild animal species, (such as the white rhinoceros, the Amur leopard, the Orangutan, or dolphin) a number of domestic and farm animal breeds are now so rare that measures to preserve them have become necessary (among them cattle, sheep, goats, pigs, horses, or dogs). Politicians, scientists, and conservationists are therefore working to find ways of halting species extinction and saving biodiversity on our planet. So-called “bio banks”, or “gene banks” are seen as one possible solution to this problem. In this modern “Noah's Ark” of the 21st century, oocytes, sperm, or embryos from animals are frozen and stored in liquid nitrogen, theoretically for a thousand years, waiting for science to catch up.

As a specialist in artificial reproduction of animals, Minitube has exactly the equipment needed to set up and operate gene banks. Deploying products and services for obtaining sperm and oocytes and for preserving them through cryopreservation is the core area of our work, as well as IVF, embryo transfer, and related biotechnologies.

Minitube equipment is used and appreciated in some of the already existing gene banks and laboratories for the reproduction of rare animal species. Examples are shown below. Other projects, such as for big cats and polar bears in the US, or for dolphins in the UAE, involve Minitube equipment and expertise too. What the experts from the "Food and Agriculture Organization of the United Nations" recommend as required equipment, for the cryoconservation of animal genetic resources is summarized in the product list on this page. All parts can be supplied/ordered directly from Minitube.

Panda Center Chengdu,
China Yak Research Center,
India Bovine AI Centre Preska,
Slovenia Wageningen University &
Research, The Netherlands Animal Reproduction
Center A.R.C., Slovakia Institute of Farm Animal
Genetics (ING), Germany Leibniz-IZW and BioRescue
Project, Germany

PRODUCT LIST

Panda Center Chengdu, China

The Chengdu Research Base for Giant Panda Breeding is home to 42 male pandas, from which sperm was frozen as part of the creation of a gene database. This is important to maintain the diversity of the species and prevent inbreeding-induced decline in viability. For this reason, sperm is also exchanged with centers and zoos in other Chinese provinces. Since 2017, sperm has been regularly collected by electroejaculation from anesthetized animals. A total of 6,391 straws and 1,484 pellets are stored in nitrogen tanks. The ejaculates are tested for quality using AndroVision®.

The breeding season of the Giant Panda is February to May. Only within these months it is possible to obtain sperm of good quality. Female pandas are also inseminated endoscopically under anesthesia. Fresh semen is preferred if it is available. All these efforts lead to 10 to 15 newborn pandas each year.

Yak Research Center, India

In India, a Yak Research Centre known as The ICAR-National Research Centre on Yak is a premier research institute proactively engaged in research and development of yak (Peophagus grunniens L.), and is the sole authority on research strategy formulation for sustainable development of yak husbandry in the country of India.

The institute has been mandated to identify, conserve, characterize, and evaluate yak germplasm and improvement of yak for draught and milk purposes.

They have been using Minitube MPP Uno for filling and sealing yak semen. Minitube´s cold handling cabinet and EasyCoder are additionally in use in the genetic laboratory, for ensuring the optimal workflow.

Bovine AI Centre Preska, Slovenia

The Insemination Center Preska is operating within the Chamber of Agriculture and Forestry of Slovenia, and has been preparing bull semen for artificial insemination for more than 60 years.

The Centre is involved in cryopreservation of semen from bovine autochthonous, such as CIKA and traditional breeds such as Brown Swiss, Limousin, and Charolais. Their bank also comprises traditional breeds that are in Slovenia longer than 50 years.

For semen analysis, an AndroVision® CASA system from Minitube is in effective use.

Wageningen University & Research, The Netherlands

The Centre for Genetic Resources, the Netherlands (CGN) manages gene bank collections of Dutch livestock species which are important for the (future) production of milk, meat, or eggs, as well as other species closely related to agriculture.

The gene bank is open to all livestock breeds, but CGN has a focus on Dutch (rare) breeds with a significant cultural-historic value. CGN conserves over 350,000 doses of sperm, of about 150 breeds of twelve livestock species in liquid nitrogen at -196° Celsius. CGN also stores embryos, oocytes, and DNA. The collection is for the long-term conservation, as a safe card in case of extinction or loss of genetic diversity and for research purposes. CGN develops cryoconservation protocols, collects material in the field and freezes most of the collection itself.

For its daily work, the CGN uses Minitube equipment such as AndroVision®, EasyCoder, or IceCube.

Animal Reproduction Center A.R.C., Slovakia

The company Animal Reproduction Center A.R.C. Slovakia, specifically Dr. Jaroslav Pokorádi and Dr. Zuzana Krchniková, founded a cryobank of protected animal species in 2007. In addition to this, their primary business is domestic and farm animals and their andrological examinations. Today, they have optimized methods and successfully frozen several species of protected animals, mainly the wolf, brown bear, serval, High Tatras chamois, red deer, fallow deer, mouflon, maral deer, American bison, European pronghorn, and other endangered ruminant species. Recently, sperm of raptors, such as falcons, hawks, and golden eagles, have also been successfully archived.

All insemination doses are archived in 0.25 or 0.5 ml vials in liquid nitrogen at -196° Celsius. If it is a matter of protected species, then these allowances are the property of the Slovak Republic and the A.R.C. workplace performs their processing and archiving without the right to remuneration, that is, free of charge.

All biotechniques are solved in cooperation with Minitube. Among other products, an AndroVision® provides optimum support for workflow processes at A.R.C., extracting any information needed through precise, standardized, and interactive semen analysis.

Institute of Farm Animal Genetics (ING), Germany

The Institute of Farm Animal Genetics (ING) at the Friedrich-Loeffler-Institute, the Federal Research Institute for Animal Health in Mariensee, Germany, has assumed responsibility for building up a national cryoreserve for maintaining the genetic diversity of economically important farm animal. For that purpose, the year 2010 brought the construction of liquid nitrogen tanks and backup systems for each species and establishment of an office for the national cryoreserve.
Source: https://www.fli.de/en/institutes/institute-of-farm-animal-genetics-ing/german-gene-bank/

Numerous Minitube products are in use at the institute and help to advance the work.

Source: IZW Berlin

Leibniz-IZW and BioRescue Project, Germany

The Leibniz Institute for Zoo and Wildlife Research (Leibniz-IZW) is an internationally renowned German research institute. The goal is to understand the adaptability of wildlife in the context of global change and to contribute to the enhancement of the survival of viable wildlife populations.

Based on a comprehensive understanding of the evolutionary equipment of wildlife and the challenges they face, the institute develops novel concepts and methods for conservation. They aim to improve population viability on different levels: The IZW develops the scientific basis for conservation decisions and transfers recommendations to conservationists, wildlife professionals, zoological gardens, and policy makers. Science-based veterinary interventions are performed directly by Leibniz-IZW staff, e.g., in assisted reproduction.

In the IZW Department of Reproduction Management, the ultimate goal is to enhance reproduction performance in endangered and critically endangered species, improving population viability and species survival. Research activities focus on understanding specific reproductive traits and reproduction morphology and physiology in wildlife species to develop new reproductive management programs.

One of the main research goals is the development of rescue strategies by the use of advanced Assisted Reproduction Technologies and Stem Cell Associated Techniques for endangered or even critically endangered species such as the Sumatran or Northern white rhinoceros. Specifically, the department pioneers artificial insemination, gamete collection, in vitro embryo production, and embryo transfer for endangered and critically endangered wildlife species.

For long-term preservation of biodiversity and basic research, biobanking became increasingly important. The Department of Reproduction Management collects and manages frozen sperm samples from more than 45 endangered, or even extinct species and a collection of cryopreserved tissues and fibroblast cultures from more than 150 endangered species as a contribution to the IZW and other European cryobanks. As a special achievement, it holds a collection of induced pluripotent and embryonic stem cell lines from 9 endangered species

The BioRescue project under the leadership of the Leibniz-IZW is contributing scientific and veterinary expertise to the Northern white rhino rescue mission. It is an international project funded by the German Federal Ministry of Education and Research and aims to make the seemingly impossible a reality and is developing methods of assisted reproduction and stem cell research.

Minitube is proud to be part of the project, by supplying equipment such as the video endoscope CowScope, the mobile CASA system AndroScope, or simple consumables like ReproJelly.

In the IZW Department of Reproduction Biology, the research focus is to understand basic reproductive characteristics and strategies of wildlife in order to assess the effects of environmental factors on fertility. The group investigates the generation and maturation of germ cells, their functional interactions in the male and female genital tract as well as the endocrine regulation of reproductive processes. The implementation of cell-based techniques bridges the gap between the genetic and organismic level of research.

The expertise in gamete biology is consequently used for the development and refinement of assisted reproduction techniques, in particular cryopreservation of sperm cells, oocytes, gonadal tissue and embryos as well as in vitro maturation and fertilization of oocytes.

In 2007, a Felid-Gamete-Rescue-Project was established, specifically dedicated to the conservation of genetic diversity of felid ex situ populations. Of the 39 cat species that exist in the world, 18 are listed on the "Red List" of the International Union for the Conservation of Nature (IUCN) as threatened or endangered. The associated cryobank includes semen samples from 18 felid species, in 9 of which in vitro embryos were generated and frozen.

Minitube has been a recurring cooperation partner in basic research projects investigating the multiple effects of sperm extender components on sperm function, the results of which have been published in a number of papers: