Means of Livelihood, Clean Environment to Women Empowerment: The Multi-Faceted Role of Donkeys.

Despite the substantial contribution donkeys make to the livelihood of the world's poorest populations, the existence of donkeys has received little notice worldwide. This article reviews the value of donkeys in a variety of sectors, including agriculture, construction industry, and mining, as well as their role in empowering women and achieving sustainable development goals. However, donkeys and mules are not given enough credit or attention in terms of developing strategies regarding their role in reducing poverty. There is a dearth of information and statistics on their impact across industries, the factors contributing to the donkey population dropping, the socioeconomic status of the dependent communities, and related animal and human welfare issues.

An Overview of Transboundary Animal Diseases of Viral Origin in South Asia: What Needs to Be Done?

Transboundary animal diseases (TADs) pose a serious threat to the sustainability and economic viability of the existing animal agriculture ecosystem in south Asia. The rapid spread of African swine fever and lumpy skin diseases in south Asia must be considered a wake-up call to prevent the entry, spread, and establishment of new exotic TADs, as south Asia has the highest density of livestock populations, and it will have a huge socioeconomic impact. Regional cooperation for the prevention and control of TADs is necessary, but rational decisions should be made to initiate even sub-regional cooperation in the present geopolitical situation. Cross-border collaboration for surveillance, early warnings, and animal movement control should be encouraged on a bilateral or multilateral basis as many countries share a porous border. Foot-and-mouth disease (FMD), peste des petits ruminants (PPR), and avian influenza (AI) have been identified as regional priority TADs, and many regional and country initiatives have been undertaken in the last two decades that need to be translated into action. The incursion of exotic TADs into south Asia has compelled us to rethink overall policies and strategies for prevention and control of TADs. This paper took into consideration six emerging and endemic TADs of viral origin to suggest a future course of action.

Impact of COVID-19 and associated lockdown on livestock and poultry sectors in India.

The COVID-19 pandemic and the associated lockdown for a long period have created a significant adverse impact on different sectors, including that of the agriculture and other allied sub-sectors in India and several other countries. The present review aimed to depict the impact of this pandemic and the lockdown on the livestock and poultry sectors in the country, which has been one of the fastest-growing sectors in recent years. Inadequacy of country-wide information has been a major bottleneck for having a thorough understanding of the impact of the prolonged lockdown on different sub-sectors of livestock and poultry. In the present case, an in-depth analysis of the subject has been made through the collation of available published materials and information collected through public contacts. The pandemic and the associated lockdown has not only caused enormous distress to the millions of poor and marginal farmers for saving their crops and/or livestock and thereby assuring their livelihoods but also impacted the overall poultry, dairy, and other livestock production systems and associated value chains, nutrition and health care, and labor availability. The paper highlights various dimensions of the impacts, namely, reduction in demand of different commodities, wastage of the produce due to the closure of transport and market chains, distress sale of the produce, and labor shortage and revival strategies taken by the government and associated enterprises. The present impact study although gives a picture about the overall present scenario, a systematic study through the collection of primary data from all over the country is suggested, which will provide a holistic view of the impact on each of the sub-sectors and the associated value chains.

Semen parameters and fertility potency of a cloned water buffalo (Bubalus bubalis) bull produced from a semen-derived epithelial cell.

Semen contains epithelial cells that can be cultured in vitro. For somatic cell nuclear transfer applications, it is essential to know whether clone(s) produced from semen-derived epithelial cells (SedECs) are healthy and reproductively competent. In this study, the semen and fertility profile of a cloned bull (C1) that was produced from a SedEC were compared with its donor (D1) and with two cloned bulls (C2, C3) that were produced from commonly used skin-derived fibroblast cells (SkdFCs). We observed variations in some fresh semen parameters (ejaculated volume and mass motility), frozen-thawed sperm parameters (plasma membrane integrity, and computer-assisted semen analysis (CASA) indices), but values are within the normal expected range. There was no difference in sperm concentration of ejaculated semen and frozen-thawed semen parameters which include sperm motility, percentage of live and normal morphology sperm, and distance traveled through oestrus mucus. Following in vitro fertilization (IVF) experiments, zygotes from C1 had higher (P < 0.05) cleavage rates (81%) than C2, C3, and D1 (71%, 67%, and 75%, respectively); however, blastocyst development per cleaved embryo and quality of produced blastocysts did not differ. The conception rate of C1 was 46% (7/15) and C2 was 50% (8/15) following artificial insemination with frozen-thawed semen. Established pregnancies resulted in births of 7 and 6 progenies sired by C1 and C2, respectively, and all calves show no signs of phenotypical abnormalities. These results showed that semen from a cloned bull derived from SedECs is equivalent to semen from its donor bull and bulls cloned from SkdFCs.

Transposon mediated reprogramming of buffalo fetal fibroblasts to induced pluripotent stem cells in feeder free culture conditions.

Commonly, induced pluripotent stem (iPS) cells are generated by viral transduction of four core reprogramming genes, but recent evidences suggest that slightly different combination of transcription factors improve the efficiency and quality of generated iPS cells. However, vectors like retro- and lentiviral may cause insertional mutagenesis due to its integrating ability. Hence, alternate methods with safety concerns are needed to be investigated. Therefore, the present study was undertaken to reprogram buffalo fibroblasts using non-viral piggyBac (PB) transposon mediated transfer of six transcription factors. To generate buffalo iPS cells, fibroblasts were isolated from buffalo fetus at passage 2. The cells were co-electroporated with a PB transposon having CAGGS promoter driven cassette of Oct4, Sox2, Klf4, cMyc, Nanog, and Lin28 transcription factors separated by self-cleaving 2A peptide and a helper plasmid pCMV-PB transposase. After 12-14 days post electroporation, fibroblast cells morphology was observed to change to round structures which formed loose aggregates of cells on day 18. Putative iPS cell colonies were propagated in feeder free system and characterized through expression of pluripotency markers such as alkaline phosphatase, SSEA-1, SSEA-4, SSEA-5, TRA-1-81, Oct4, Nanog and Sox2 and endogenous genes supported the stemness property of the generated cells. These cells differentiated in vitro to form embryoid bodies and were found to express three germ layers markers. In conclusion, generation of buffalo iPS cells using transposon system provides insights into viral-free iPS technology which will facilitate genetic modification of the buffalo genome and help in the production of transgenic animals using genetically modified iPS cells.

Establishment of a Somatic Cell Bank for Indian Buffalo Breeds and Assessing the Suitability of the Cryopreserved Cells for Somatic Cell Nuclear Transfer.

Biobanks of cryopreserved gametes and embryos of domestic animals have been utilized to spread desired genotypes and to conserve the animal germplasm of endangered breeds. In principle, somatic cells can be used for the same purposes, and for reviving of animals, the somatic cells must be suitable for animal cloning techniques, such as somatic cell nuclear transfer. In the present study, we derived and cryopreserved somatic cells from three breeds of riverine and swamp-like type buffaloes and established a somatic cell bank. In total, 350 cryovials of 14 different individual animals (25 cryovials per animal) were cryopreserved and informative data such as breed value, origin, and others were documented. Immunostaining of the established cells against vimentin and cytokeratin suggested a commitment to the fibroblast lineage. In addition, microsatellite analysis was performed and documented for unambiguous parentage verification of clones in the future. Subsequently, the cryopreserved cells were tested for their suitability as nuclear donors (n = 7) using handmade cloning, and the reconstructed embryos were cultured in vitro. The cleavage rates (95.99% ± 2.17% vs. 82.18% ± 2.50%) and blastocyst rates (37.73% ± 1.54% vs. 24.31% ± 1.78%) were higher (p < 0.05) for riverine buffalo cells than that of swamp-like buffalo cells, whereas the total cell numbers of blastocysts (258.16 ± 36.25 vs. 198.16 ± 36.25, respectively) were similar. In conclusion, we demonstrated the feasibility of biobanking of buffalo somatic cells, and that the cryopreserved cells can be used to produce cloned embryos. This study encourages the development of somatic cell biobanks of domestic livestock, including endangered breeds of buffalo, to preserve valuable genotypes for future revitalization by animal cloning techniques.

Generation of Venus fluorochrome expressing transgenic handmade cloned buffalo embryos using Sleeping Beauty transposon.

The objective of this study was to optimise the electroporation conditions for efficient integration of Venus construct in buffalo fetal fibroblasts using Sleeping Beauty (SB) based transposition and to produce Venus expressing transgenic cloned embryos through handmade cloning (HMC) approach. Primary culture of buffalo fetal fibroblast cells was established and subsequently cultured cells were co-transfected with Venus and helper plasmid at different combinations of electroporation condition. In different combinations of voltage, time and plasmid dose, we observed that 300 V, single pulse for 10 ms in 2 mm cuvette and 1.5-2.0 µg transposons with 200-300 ng transposase dose was optimum for expressing Venus fluorescence in cells via electroporation. After electroporation, the cells were cultured for 2-3 days and then Venus expressing cells were picked with the help of a Pasteur pipette under the fluorescence microscope to enrich them through single cell culture method before using as donor cells for HMC. In vitro matured oocytes were reconstructed with either transfected or non-transfected buffalo somatic cells by electric fusion followed by activation. The reconstructed, activated embryos were cultured in 400 µL of Research Vitro Cleave medium supplemented with 1% fatty acid-free BSA in 4-well dish, covered with mineral oil and incubated in an incubator (5% CO2 in air) at 38.5 °C for 8 days and the developmental competence was observed. The percentage of cleaved, 4-8 and 8-16 cells stage embryos generated through Venus expressing cells were comparable with control, whereas, the morula (21.0 vs 53.0%) and blastocysts (10.5 vs 30.6%) produced through Venus expressing cells was found low as compared to control. These results indicate that fetal fibroblasts transfected with Venus could be used as donor cells for buffalo cloning and that Venus gene can be safely used as a marker of foreign gene in buffalo transgenesis.