Browsing by Author "Fermin LM"

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    Controlled Cytoplast Arrest and Morula Aggregation Enhance Development, Cryoresilience, and In Vivo Survival of Cloned Sheep Embryos
    (Mary Ann Liebert, Inc, 2021-02-02) McLean ZL; Appleby SJ; Fermin LM; Henderson HV; Wei J; Wells DN; Oback B
    Zona-free somatic cell transfer (SCT) and embryo aggregation increase throughput and efficiency of cloned embryo and offspring production, respectively, but both approaches have not been widely adopted. Cloning efficiency is further improved by cell cycle coordination between the interphase donor cell and metaphase-arrested recipient cytoplast. This commonly involves inclusion of caffeine and omission of calcium to maintain high mitotic cyclin-dependent kinase activity and low calcium levels, respectively, in the nonactivated cytoplast. The aim of our study was to integrate these various methodological improvements into a single work stream that increases sheep cloning success. We show that omitting calcium during zona-free SCT improved blastocyst development from 6% to 13%, while caffeine treatment reduced spontaneous oocyte activation from 17% to 8%. In a retrospective analysis, morula aggregation produced high morphological quality blastocysts with better in vivo survival to term than nonaggregated controls (15% vs. 9%), particularly after vitrification (14% vs. 0%). By combining cytoplast cell cycle control with zona-free embryo reconstruction and aggregation, this novel SCT protocol maximizes the benefits of vitrification by producing more cryoresilient blastocysts. The presented cloning methodology is relatively easy to operate and further increases throughput and efficiency of cloned embryo and offspring production. Integration of additional reprogramming steps or alternate donor cells is straightforward, providing a flexible workflow that can be adapted to changing experimental requirements.
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    Smooth muscle hamartoma in a castrated male red deer (Cervus elaphus) in New Zealand.
    (Taylor and Francis Group, 2023-07-01) Johnson SG; Fermin LM; Aberdein D; Lawrence KE
    Reports of neoplasia in deer remain rare (Hill and Staples Citation1999), despite the conviction that as deer farming became more common, a greater number of pathological processes, including tumours, would be recognised in deer (Pérez et al. Citation1998). Skin tumours are among the most common neoplasms reported in red deer (Cervus elaphus) and are usually papillomavirus-associated dermal fibropapillomas and papillomas (Erdélyi et al. Citation2009; Vaatstra et al. Citation2014; Garcês et al. Citation2020). Additional reports of cutaneous and subcutaneous tumours in red deer include malignant schwannoma and dermal malignant melanoma (Pérez et al. Citation1998; Scandrett and Wobeser Citation2004). In related deer species, subcutaneous dermoid cysts have been described in caribou (Rangifer tarandus) (Wobeser et al. Citation2009) and cutaneous fibromas in predominantly male white-tailed deer (Odocoileus virginianus) (Berry Citation1925; Friend Citation1967; Sundberg and Nielsen Citation1982).
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    Theileria orientalis Ikeda infection detected in red deer but not dogs or horses in New Zealand.
    (2024-09-02) Lawrence KE; Gedye K; Carvalho L; Wang B; Fermin LM; Pomroy WE
    AIMS: To determine whether evidence for infection with Theileria orientalis (Ikeda) could be identified in samples of commercial red deer (Cervus elaphus), horses, and working farm dogs in New Zealand. METHODS: Blood samples were collected during October and November 2019 from a convenience sample of red deer (n = 57) at slaughter. Equine blood samples (n = 50) were convenience-sampled from those submitted to a veterinary pathology laboratory for routine testing in January 2020. Blood samples, collected for a previous study from a convenience sample of Huntaway dogs (n = 115) from rural regions throughout the North and South Islands of New Zealand between August 2018 and December 2020, were also tested. DNA was extracted and quantitative PCR was used to detect the T. orientalis Ikeda major piroplasm surface protein (MPSP) gene. A standard curve of five serial 10-fold dilutions of a plasmid carrying a fragment of the T. orientalis MPSP gene was used to quantify the number of T. orientalis organisms in the samples. MPSP amplicons obtained by end-point PCR on positive samples were isolated and subjected to DNA sequencing. The resulting sequences were compared to previously published T. orientalis sequences. RESULTS: There were 6/57 (10%) samples positive for T. orientalis Ikeda from the deer and no samples positive for T. orientalis Ikeda from the working dogs or horses. The mean infection intensity for the six PCR-positive deer was 5.1 (min 2.2, max 12.4) T. orientalis Ikeda organisms/µL. CONCLUSIONS AND CLINICAL RELEVANCE: Red deer can potentially sustain low infection intensities of T. orientalis Ikeda and could act as reservoirs of infected ticks. Further studies are needed to determine whether naïve ticks feeding on infected red deer can themselves become infected. ABBREVIATIONS: Cq: Quantification cycle; LOQ: Limits of quantification; MPSP: Major piroplasm surface protein; qPCR: Quantitative polymerase chain reaction.