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In alpacas (Vicugna pacos), the high cost of in vitro embryo production is also a consequence of the use of several substances in the culture medium. In addition, embryo production rates in this species are still considered low. Thus, in attempt to reduce the cost and to improve the in vitro embryo production rates, this study evaluates the effect of adding follicular fluid (FF) in the in vitro maturation medium on oocyte maturation and subsequent embryo production. After ovary collection at the local slaughterhouse, the oocytes were recovered, selected, and allocated in experimental groups: standard maturation medium (G1) and simplified medium added by 10% FF (G2). The FF was acquired from follicles between 7- and 12-mm diameter. The cumulus cell expansion and the embryo production rates were analyzed by chi-square with p < 0.05. No differences (p > 0.05) were observed in maturation rate between G1 (66.36%) and G2 (63.12%) groups. Likewise, no significant difference (p > 0.05) was verified between G1 and G2 for morula (40.85 vs 38.45%), blastocyst (7.01 vs 6.93%), and total number of embryos (47.87 vs 45.38%). In conclusion, it was possible to simplify the medium used for in vitro maturation of alpaca oocytes resulting in embryo production rates similar to the standard medium.

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Under natural and well-managed conditions, the buffalo has good reproductive and productive indices. However, in vitro embryo production (IVEP) has been used commercially to maximise the number of elite animals. In this species, several factors (donor management, in vitro culture medium, semen, in vitro conditions, embryo transfer) still affect the IVEP results. In addition, the cost of this technique is very high for this purpose. Therefore, more studies, as well as adequate plans, are needed to achieve this objective efficiently. In this review, we discussed the current commercial status, influencing factors (in vivo and in vitro), and the progress and future challenges of IVEP in buffalo. A total of 81 references were used from 1979 to 2022. The relevant data or literature were searched using the following databases: Google, ResearchGate, Science Alert, Science Direct and PubMed, using the following keywords: buffalo oocytes/COCs, buffalo embryos, pregnancy and calving or live birth rate after embryo transfer. The best maturation, cleavage and blastocyst rates in the in vitro production of buffalo embryos were 95.8, 75.2 and 33.4%, respectively. The pregnancy and live birth rates ranged from 22.2% to 43.5% and from 15.3% to 36.5%, respectively, after the transfer of fresh embryos produced in vitro to the recipients. This review will help to contextualise IVEP in buffaloes, as well as create an adequate plan for implementing IVEP in buffaloes.

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Mesenchymal stem cells (MSC) from the umbilical cord (UC) have several attractive properties for clinical use. This study aimed to verify the impact of a lipid-rich diet during late gestation of donor goats on the growth and differentiation of MSCs from UC. From the 100th day of pregnancy to delivery, 22 goats were grouped based on their diet into the donor-lipid (DLD; n = 11) and donor-baseline (DBD; n = 11) diet groups. Diets were isonitrogenous and isoenergetic, differing in fat content (2.8% vs. 6.3% on a dry matter basis). Wharton's jelly (WJ) fragments were cultured. After primary culture, samples of WJ-MSCs were characterized by the expression of CD90, CD73, CD34, CD45, CD105, and Fas genes, mitochondrial activity using MitoTracker (MT) fluorescence probe, and growth kinetics. Population doubling time (PDT) was also determined. WJ-MSCs were differentiated into chondrocytes, adipocytes and osteocytes, and the mineralized area and adipocytes were determined. The lipid diet significantly increased triglyceride and cholesterol levels during pregnancy. The DLD group showed sub-expression of the CD90 gene, a high MT intensity, and a low proliferation rate at the end of the subculture. The mean PDT was 83.9 ± 1.3 h. Mineralized area and lipid droplet stain intensity from osteogenic and adipogenic differentiations, respectively, were greater in DLD. We conclude that in donor goats, dietary dyslipidemia during late pregnancy affects the ability of UC-derived MSCs to express their developmental potential in vitro, thus limiting their possible use for therapeutic purposes.

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BACKGROUND: Crotalicidin (Ctn), a snake venom cathelicidin-related antimicrobial peptide, is a 34-residue-long linear lysine-rich vipericidin obtained from the South American rattlesnake, Crotalus durissus terrificus. Ctn contains tandem repeats of nine amino acid residues (1KRFKKFFKK9 and 16KRLKKIFKK24; consensus: 1KRhKKhFKK9, h = hydrophobic amino acid) as an integral part of its structure. OBJECTIVES: The aim of this study was to evaluate the antimicrobial activity of the encrypted vipericidin nonapeptide KRFKKFFKK, designated as Ctn[1-9], and its structural analogue, rhodamine- B‒conjugated Ctn[1-9], designated as RhoB-Ctn[1-9]. METHODS: The susceptibility of representative pathogenic bacteria and yeasts to antimicrobial agents was determined using the broth microdilution minimum inhibitory concentration (MIC) method. Cytotoxicity was estimated using a hemolytic assay. The accumulation of RhoB-Ctn[1-9] in microbial cells was observed by fluorescence microscopy. The antimicrobial synergism of RhoB-Ctn[1-9] with antimicrobials was evaluated using a checkerboard analysis. RESULTS: RhoB-conjugated Ctn[1-9] displayed selective antimicrobial activity against infectious gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and pathogenic species of Candida with low hemolytic effects on human erythrocytes which were not observed with unconjugated Ctn[1-9]. RhoB-Ctn[1-9] could permeate cell membranes and accumulate intracellularly in microbial cells. RhoB-Ctn[1-9] exhibits synergistic effects when used with antibiotics or antifungal agents and reduced the MICs of the peptide and antimicrobials. CONCLUSION: These findings indicate the potential of crotalicidin-related short peptides as structural motifs for the diversification of biological functionalities. Further, they set the stage to investigate the molecular mechanisms by which chemically modified vipericidin repeats modulate cell fate.

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One of the most significant challenges in deer is the ability to maintain genetic diversity, avoiding inbreeding and sustaining population health and reproduction. Although our general knowledge of reproductive physiology is improving, it appears that the application of assisted reproductive technology (ART) will more efficiently advance wildlife conservation efforts and preserve genetic diversity. The purpose of this review is to present the most important results obtained with the use of ART in Neotropical deer. Thus, the state-of-the-art for estrus synchronization, semen technology, artificial insemination, and in vivo embryo production will be presented. In vitro embryo production (IVP) is also a biotechnology that is taking initial steps in deer. In this aspect, the approach with the proteomics of ovarian follicular fluid is being used as a tool for a better understanding of oocyte maturation. Finally, cell banks and the use of interspecific somatic cell nuclear transfer (iSCNT) as well as the use of stem cells for gametes differentiation are promising techniques.

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Ovinos e caprinos são espécies que devido ao seu tamanho adequado, curto período gestacional e produção leiteira, tornaram-se importantes modelos na pesquisa pecuária, farmacêutica e biomédica. Assim, a engenharia genética tem sido amplamente aplicada à pesquisa com pequenos ruminantes. Caprinos e ovinos modificados geneticamente fornecem modelos valiosos para pesquisa sobre as funções dos genes, melhorando a exploração, obtendo produtos farmacêuticos no leite e aumentando a resistência às doenças. Além disso, o uso conjunto da clonagem e transgênese já foi responsável por marcos importantes na biotecnologia animal. Esta revisão destaca os avanços da clonagem e da engenharia genética nessas espécies obtidos nas últimas quatro décadas, com ênfase nas pesquisas realizadas no Brasil.

Sheep and goats are valuable species that due to their suitable size, short pregnancy period, and secretion of milk, have become important model in agricultural, pharmaceutical, and biomedical research. Thus, genome engineering has been widely applied to sheep and goat research. Gene-edited sheep and goats provide valuable models for investigations on gene functions, improving animal breeding, producing pharmaceuticals in milk and improving disease resistance. Additionally, the joint use of cloning and transgenesis has already been responsible for major milestones in animal biotechnology. This review highlights the advances of cloning and genome engineering in these species over the past four decades with particular emphasis on research in Brazil.

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A espécie caprina tornou-se um excelente modelo animal objetivando a produção de proteínas recombinantes no leite. A obtenção de machos fundadores transgênicos é importante, pois os mesmos permitem aumentar o número de crias transgênicas pelo simples uso da inseminação artificial. Este trabalho teve por objetivo, comparar parâmetros seminais de machos caprinos transgênicos (TG) e não transgênicos (NTG) para o Fator Estimulante de Colônia de Granulócitos humano (hG-CSF). Para tanto, semanalmente,foram colhidas amostras de sêmen de todos os animais através de vagina artificial. Foram avaliados os seguintes parâmetros: volume, motilidade massal, concentração espermática, motilidade individual progressiva (MIP), vigor e patologias espermáticas. Foram observadas diferenças estatísticas somente para volume e MIP, os quais tiveram valores superiores (p0,05). Em conclusão, a presença do transgene não afetou a qualidade do sêmen favorecendo e contribuindo para a formação de um rebanho transgênico.

Goats became an excellent animal model aiming the production of recombinant proteins in the milk. Transgenic founder males are important to increase the number of transgenic kids by artificial insemination. This work aimed to compare seminal parameters of transgenic (TG) and non-transgenic goats (NTG) for the human Granulocyte Colony Stimulating Factor (hG-CSF). Thus, weekly, semen samples were collected from all transgenic males by artificial vagina and evaluated: volume, mass motility, sperm concentration, individual progressive motility (IPM), vigor and sperm pathologies. Statistical differences were observed only for volume and IPM, which had higher values (p0.05). In conclusion, the presence of the transgene did not affect the semen quality and these animals can be used to obtain kids and enable to create of a transgenic herd.

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Neste estudo, a avaliação do sêmen foi utilizada para investigar prováveis efeitos da presença do transgene sobre os parâmetros seminais de caprinos. Para tanto, amostras de sêmen foram colhidas semanalmente de três caprinos, sendo um não transgênico (NTG) e dois transgênicos (TG) para o Fator Estimulante de Colônia de Granulócitos humano (hG-CSF). Observou-se que, com exceção do volume ejaculado, não foram verificadas diferenças estatísticas para os dados relativos aos demais parâmetros: motilidade massal, concentração espermática, motilidade individual progressiva e patologias espermáticas. Concluiu-se que a presença do transgene hG-CSF não teve influência sobre os principais parâmetros seminais dos animais avaliados.

In this study, semen evaluation was used to investigate likely effects of transgene presence on seminal parameters of goats. For this purpose, semen samples were collected weekly from three goats, one non-transgenic (NTG) and two transgenic (TG) for the Human Granulocyte Colony Stimulating Factor (hG-CSF). It was observed that, with the exception of the ejaculated volume, no statistical differences were verified for the data related to the other parameters: mass motility, sperm concentration, individual progressive motility and spermatic pathologies. It was concluded that the presence of the hG-CSF transgene had no influence on the main seminal parameters of the animals evaluated.

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A produção in vitro de embriões (PIVE) bovinos no Brasil é dependente de transporte de oócitos das fazendas para laboratórios, sendo útil um sistema de incubação que mantenha a competência ao desenvolvimento. Assim, comparou-se a dois sistemas de incubação (mini vs portátil) quanto à qualidade dos embriões produzidos. Oócitos bovinos foram maturados, fecundados e cultivados in vitro. Além disso, foi verificada a produção de espécies reativas de oxigênio (EROs). As taxas de clivagem e blastocisto foram similares (p>0,05) para ambos os sistemas. Os níveis de EROs foram significativamente maiores (p<0,05) em embriões de dia 2 produzidos na incubadora mini. Em conclusão, o sistema de incubação portátil mostrou ser uma alternativa viável para PIVE bovina.

The in vitro production of bovine embryos (PIVE) in Brazil, is dependent on oocyte transport from farms to laboratories, and an incubation system that maintains developmental competence is useful. Thus, it was compared to two incubation systems (mini vs portable) regarding the quality of the embryos produced. Bovine oocytes were matured, fertilized and cultured in vitro. In addition, the production of reactive oxygen species (ROS) was verified. The cleavage and blastocyst rates were similar (p>0.05) for both systems. ERO levels were significantly higher (p<0.05) in day 2 embryos produced in the mini incubator. In conclusion, the portable incubation system proved to be a viable alternative for bovine PIVE.

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A resposta ovariana de catetos tratados com eCG/hCG foi analisada pelo desenvolvimento folicular e pela expressão gênica. O tratamento DS (dose utilizada em suínos) e DA (dose ajustada alometricamente) foram comparados. Todas as fêmeas apresentaram corpos hemorrágicos (CHs) e as taxas de ovulação foram semelhantes entre DS (4,00±1,17) e DA (2,50±0,43). O número de folículos por animal e as quantidades de folículos P (≤3mm), M (3-5mm) e G (≥5mm) foram similares entre os grupos. Os folículos ovarianos do mesmo tamanho expressaram FSHR e LHCGR em níveis similares entre os grupos. FSHR foi mais expresso na fase inicial, e LHCGR, na fase tardia da foliculogênese ovariana de cateto. A expressão de LHCGR nos CHs foi igual entre os tratamentos e entre os ovários direito e esquerdo.

The ovarian response of catches treated with eCG/hCG was analyzed by follicular development and gene expression. The treatment DS (dose used in pigs) and AD (dose adjusted alometrically) were compared. All females showed hemorrhagic bodies (CHs) and ovulation rates were similar between DS (4.00±1.17) and AD (2.50±0.43). The number of follicles per animal and the numbers of follicles P (≤3mm), M (3-5mm) and G (≥5mm) were similar between the groups. Ovarian follicles of the same size expressed FSHR and LHCGR at similar levels between groups. FSHR was more expressed in the early phase, and LHCGR, in the late phase of ovarian catheter folliculogenesis. LHCGR expression in CHs was equal between treatments and between right and left ovary.

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