19 LIFETIME MORTALITY OF CLONED PIGLETS

2014 ◽  
Vol 26 (1) ◽  
pp. 124
Author(s):  
M. Schmidt ◽  
K. D. Winther ◽  
H. Callesen

Perinatal mortality of cloned animals is a well-recognised problem, but it is not clear if and how the problems continue throughout the life of cloned animals. Here we report occurrence and causes of lifetime mortality of cloned piglets from their perinatal period and onwards. From 2006 until 2013, 178 Large White (LW) sows received Day 5–6 handmade cloned morulae or blastocysts where donor cells were from LW, Yucatan, or Göttingen, and were either transgenic or not. Pregnancy rate on Day 30 was 73%, but 19% terminated before term, resulting in 104/178 deliveries. For each litter, each piglet was characterised as either alive or stillborn (i.e. dead without signs of autolysis or dying during the first minutes after birth). Data were analysed by Fisher's Exact test with a significance level of P < 0.05. In the 104 litters, stillborn piglets were found in 54 (from 1 to 7 stillborn piglets in a litter), and the highest number of stillborns was found in Göttingen piglets (P < 0.05; see Table 1). Alive piglets were followed and their mortality registered, and they either died spontaneously (arthritis, pericarditis, enteritis, septicemia, lethal malformations) or were killed if they had other malformations, no weight gain, anorexia, bristly hair coat or signs of compromised well-being. Piglet mortality was highest in the perinatal period but remained high until weaning at Day 28 (see Table 1), so almost half of the alive piglets died during the first month; this preweaning mortality was highest in Yucatan piglets (P < 0.05), which also had the highest cumulative mortality. Further analyses are needed to describe differences observed between donor cell breeds. After weaning, where the pigs were kept under standard conditions, the mortality was at the normal level for the farm. Few died of infections with no breed differences, so most were killed over the subsequent months and years for various experimental purposes. The oldest pig lived until 6 years of age. During their life, no differences compared with noncloned pigs were observed in behaviour, growth, occurrence of diseases, or viability. This study confirms the well-known high perinatal mortality of cloned piglets and shows also high mortality until weaning. However, after weaning, mortality was not different from that of noncloned piglets. Table 1.Lifetime mortality rates in cloned piglets of different breeds and at different time periods after birth

2010 ◽  
Vol 22 (1) ◽  
pp. 197 ◽  
Author(s):  
M. Schmidt ◽  
P. M. Kragh ◽  
J. Li ◽  
L. Lin ◽  
Y. Liu ◽  
...  

Production of cloned piglets, using normal or transgenic donor cells, has been of limited efficiency. One reason could be breed differences between recipient and embryos. The aim of this study was to compare pregnancies after transfer to Large White (LW) sow recipients of cloned LW embryos or cloned, transgenic minipig embryos. Large White donor cells were from LYxD, and minipig cells were transgenic with 1 of 5 genes related to different human diseases and from either Göttingen or Yucatan. The cells were used on Day 0 for handmade cloning (Du et al. 2005 Cloning Stem Cells 7, 199-205). The reconstructed embryos were cultured in vitro until transfer on Day 5 to 6. As recipients, 70 LW sows were weaned and anesthetized 4 days after natural heat. Through an abdominal incision the ovaries were controlled (CL formation, absence of cysts) and the embryos slowly introduced into the uterus via a catheter inserted 5 to 6 cm into the tip of the upper horn. To each of 33 recipients, 40 to 60 LW embryos were transferred, and 37 recipients each received 50 to 100 transgenic minipig embryos. Pregnancies were examined by ultrasound scanning every second week. Abortions were defined as absence of earlier confirmed scanning or delivery of aborted fetuses. Caesarean sections were performed on Day 114 (minipig) or Day 116 (LW) 24 h after injection of a prostaglandin analogue. At delivery, placental gross morphology was recorded with samples taken for later histology. The piglets were fed every 3 h with colostrum for the first 24 h and then by the recipient LW sow. Data were analyzed by Fisher’s Exact test with a significance level of P < 0.05. The overall pregnancy rate was 49% (34/70) with an abortion rate of 29% (8 aborted + 2 resorbed/34) from Day 30 to 45 giving 24/70 deliveries (34% of the transfers). There was no significant difference between minipig (54%, 20/37) and LW pregnancy results (42%, 14/33), although there tended to be more abortions with minipig pregnancies (8/20 v. 2/14; P = 0.14) resulting in 12 minipig and 12 LW litters of which 4 and 9, respectively, have grown up to adulthood. In almost every recipient the placenta and fetal membranes showed abnormal thick and edematous morphology. The total litter sizes ranged from 1 to 10 piglets (mean 4.4 ± 0.6), and in 13 of 24 litters there were 1 to 5 stillborn piglets. Except for one litter of 9 transgenic Yucatan piglets that all died within their first 2 weeks, the postnatal mortality of both LW- and mini-piglets seemed similar to that of farm piglets of the same age, and the piglets appeared normal with respect to weight gain, gross morphology, and behavior. These results demonstrate that, in spite of a rather high abortion rate and some fetal mortality, an acceptable birth rate can be achieved after transfer to LW recipients of cloned LW embryos (36%) as well as cloned, transgenic minipig embryos (32%). Therefore, a breed difference between the embryos and their recipient seems not to influence the pregnancy results. The authors thank B. Synnestvedt, H. Kristiansen, S. Starsig, A. Pedersen, J. Adamsen, R. Kristiansen, and K. Villemoes for invaluable technical assistance.


2015 ◽  
Vol 27 (1) ◽  
pp. 107 ◽  
Author(s):  
M. Schmidt ◽  
K. D. Winther ◽  
H. Callesen

Around 50% of cloned and transgenic piglets are lost during the first month after birth (Reprod. Fertil. Develop. 26, 124), and one reason is malformations of vital organs. The aim of the present study was to describe the distribution of malformations in piglets (until weaning, i.e. age <Day 28) born after transfer to Large White (LW) recipients of cloned embryos. Donor cells were fibroblasts either from LW (non-transgenic) or from Yucatan or Göttingen (made transgenic with 1 of 7 genes related to different human diseases). Handmade cloning was used to produce embryos that, after 5–6 days in vitro culture, were transferred to 202 LW sows 4 days after natural heat. Abortion occurred in 29 sows, and 6.5 ± 0.4 piglets per litter (from 1 to 22) were delivered from 116 sows (46 litters with LW piglets, 40 with Göttingen, 30 with Yucatan). In 78 of these litters (67%), autopsies were performed on 55 ± 4% of piglets stillborn or dead before weaning. Data were analysed by Fisher's Exact test with P < 0.05 as significance level. Malformations were found in 1 to 12 piglets per litter, with a higher malformation rate in transgenic Göttingen and Yucatan piglets (35% and 46% of all born, respectively) than in non-transgenic LW (17%). Many piglets showed 2 (24%) or more (6%) malformations. Some malformations seemed to be related to breed and/or transgene (see Table 1 for most frequent malformations); for example, heart malformations were most frequent in Yucatan litters independent of the transgene, whereas gall bladder and limb malformations were more frequent in Göttingen and in various litters with the same transgene. These results show that pig cloning results in a considerable loss of piglets, and that a majority of these can be related to various malformations. Use of transgenic cells for cloning only adds to this problem. Some malformations are related to the specific breed in use, but the general finding is that the problem is related to the cloning technique as such. However, because approximately half of the cloned piglets still survive, perhaps with unknown minor malformations, use of pigs as a model for human diseases is still realistic. But choice of breed and possible improvements of the transgenic technologies for this kind of work should be considered carefully. Table 1.Malformations in transgenic and nontransgenic cloned piglets dead before weaning


2012 ◽  
Vol 24 (1) ◽  
pp. 123 ◽  
Author(s):  
M. Schmidt ◽  
K. D. Winter ◽  
J. Li ◽  
P. M. Kragh ◽  
Y. Du ◽  
...  

Viability of cloned and transgenic piglets is seriously compromised and one obvious reason could be malformations. The aim of the present study was therefore to describe gross pathological conditions in dead pre-weaned piglets born after transfer to Large White (LW) recipients of cloned (LW donor cells) or transgenic (Yucatan or Göttingen donor cells) embryos. Donor cells were fibroblasts and the Göttingen and Yucatan cells were made transgenic with 1 of 5 genes known to dispose for different human diseases. Handmade cloning was used to produce embryos that after 5 to 6 days of in vitro culture were transferred surgically to 108 LW sows 4 days after their natural heat. Of these, 21 sows delivered cloned LW piglets, whereas 17 and 16 sows, respectively, delivered transgenic Göttingen and Yucatan piglets. Stillborn and dead pre-weaned piglets were necropsied and malformations registered. Data were analysed by Fisher's exact test with a significance level of P < 0.05. In the 54 litters, total litter size ranged from 1 to 22 piglets (mean 5.4 ± 0.5) and the overall mortality rate until weaning on day 28 was 59%. Malformations were found in piglets from 38 litters where an average of 35% of the piglets showed malformations (between 8 and 100%). In those litters, 1 to 7 piglets had 1, 2, or several malformations (Table 1). The malformation rate in the autopsied transgenic Göttingen was 58% and in Yucatan 46%; these were significantly higher than in the autopsied cloned LW piglets with 18%. Some of the malformations seemed to be related to breed and/or transgene; for instance, heart malformations were most frequent in Yucatan litters (70%) independent of the transgene, whereas gallbladder and gonad malformations were more frequent in various litters with the same transgene. These results show that the use of cloning in pigs results in a considerable loss of piglets due to malformations and transgenic transformation of the cells used for cloning superimpose on this problem. In combination, these elements could seriously compromise the use of pigs as a model for human diseases and the choice of breeds and also transgenes for this kind of work should be considered carefully. However, further improvements in production of cloned/transgenic embryos may ultimately reduce the incidence of malformations. Table 1.Number of malformations in 54 litters of cloned or transgenic piglets


2011 ◽  
Vol 23 (5) ◽  
pp. 645 ◽  
Author(s):  
M. Schmidt ◽  
K. D. Winter ◽  
V. Dantzer ◽  
J. Li ◽  
P. M. Kragh ◽  
...  

The perinatal mortality of cloned animals is a well-known problem. In the present retrospective study, we report on mortality of cloned transgenic or non-transgenic piglets produced as part of several investigations. Large White (LW) sows (n = 105) received hand-made cloned LW or minipig blastocysts and delivered either spontaneously or after prostaglandin induction followed by either Caesarean section or vaginal birth. The overall pregnancy rate was 62%, with 26% of pregnancies terminating before term. This resulted in 48 deliveries. The terminated pregnancies consisted of 12 abortions that occurred at 35 ± 2 days gestation and five sows that went to term without returning to heat and then by surgery showed the uterus without fetal content. The gestation length was for sows with LW piglets that delivered by Caesarean section or vaginally was 115.7 ± 0.3 and 117.6 ± 0.4 days, respectively. In sows with minipiglets, the gestation length for those delivered by Caesarean section or vaginally 114.4 ± 0.2 and 115.5 ± 0.3 days, respectively. Of the 34 sows that delivered vaginally, 28 gave birth after induction, whereas 6 farrowed spontaneously. Of the 14 sows that delivered after Caesarean section and in the five empty sows, the endometrium and placenta showed severe oedema. Piglet mortality following vaginal delivery was higher than after Caesarean section (31% v. 10%, respectively; P < 0.001). When vaginal delivery occurred spontaneously, the stillborn rate was greater than after induced delivery (56% v. 24%, respectively; P < 0.0001). Internal organ weights were recorded for seven cloned LW piglets and six normal piglets. The relative weight of the heart, liver, kidneys and small intestine was found to be reduced in the cloned piglets (P < 0.05). The present study demonstrates extensive endometrial oedema in sows pregnant with cloned and transgenic piglets, as well as in empty recipients, at term. The growth of certain organs in some of the cloned piglets was reduced and the rate of stillborn piglets was greater in cloned and transgenic piglets delivered vaginally, possibly because of oedema of the fetal–maternal interface.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Georgina Navoly ◽  
Conor J. McCann

AbstractEnteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. After in vivo transplantation, ENSC and their derivatives have been shown to engraft within colonic tissue, migrate and populate endogenous ganglia, and functionally integrate with the enteric nervous system. However, the mechanisms underlying the integration of donor ENSC, in recipient tissues, remain unclear. Therefore, we aimed to examine ENSC integration using an adapted ex vivo organotypic culture system. Donor ENSC were obtained from Wnt1cre/+;R26RYFP/YFP mice allowing specific labelling, selection and fate-mapping of cells. YFP+ neurospheres were transplanted to C57BL6/J (6–8-week-old) colonic tissue and maintained in organotypic culture for up to 21 days. We analysed and quantified donor cell integration within recipient tissues at 7, 14 and 21 days, along with assessing the structural and molecular consequences of ENSC integration. We found that organotypically cultured tissues were well preserved up to 21-days in ex vivo culture, which allowed for assessment of donor cell integration after transplantation. Donor ENSC-derived cells integrated across the colonic wall in a dynamic fashion, across a three-week period. Following transplantation, donor cells displayed two integrative patterns; longitudinal migration and medial invasion which allowed donor cells to populate colonic tissue. Moreover, significant remodelling of the intestinal ECM and musculature occurred upon transplantation, to facilitate donor cell integration within endogenous enteric ganglia. These results provide critical evidence on the timescale and mechanisms, which regulate donor ENSC integration, within recipient gut tissue, which are important considerations in the future clinical translation of stem cell therapies for enteric disease.


2021 ◽  
pp. jech-2020-216030
Author(s):  
Benjamin J Gray ◽  
Richard G Kyle ◽  
Jiao Song ◽  
Alisha R Davies

BackgroundThe public health response to the SARS-CoV-2 (COVID-19) pandemic has had a detrimental impact on employment and there are concerns the impact may be greatest among the most vulnerable. We examined the characteristics of those who experienced changes in employment status during the early months of the pandemic.MethodsData were collected from a cross-sectional, nationally representative household survey of the working age population (18–64 years) in Wales in May/June 2020 (n=1379). We looked at changes in employment and being placed on furlough since February 2020 across demographics, contract type, job skill level, health status and household factors. χ2 or Fisher’s exact test and multinomial logistic regression models examined associations between demographics, subgroups and employment outcomes.ResultsOf our respondents, 91.0% remained in the same job in May/June 2020 as they were in February 2020, 5.7% were now in a new job and 3.3% experienced unemployment. In addition, 24% of our respondents reported being placed on furlough. Non-permanent contract types, individuals who reported low mental well-being and household financial difficulties were all significant factors in experiencing unemployment. Being placed on ‘furlough’ was more likely in younger (18–29 years) and older (60–64 years) workers, those in lower skilled jobs and from households with less financial security.ConclusionA number of vulnerable population groups were observed to experience detrimental employment outcomes during the initial stage of the COVID-19 pandemic. Targeted support is needed to mitigate against both the direct impacts on employment, and indirect impacts on financial insecurity and health.


2019 ◽  
Vol 165 (5) ◽  
pp. 363-370 ◽  
Author(s):  
Lauren Rose Godier-McBard ◽  
L Ibbitson ◽  
C Hooks ◽  
M Fossey

BackgroundPoor mental health in the perinatal period is associated with a number of adverse outcomes for the individual and the wider family. The unique circumstances in which military spouses/partners live may leave them particularly vulnerable to developing perinatal mental health (PMH) problems.MethodsA scoping review was carried out to review the literature pertaining to PMH in military spouses/partners using the methodology outlined by Arksey and O’Malley (2005). Databases searched included EBSCO, Gale Cengage Academic OneFile, ProQuest and SAGE.ResultsThirteen papers fulfilled the inclusion criteria, all from the USA, which looked a PMH or well-being in military spouses. There was a strong focus on spousal deployment as a risk factor for depressive symptoms and psychological stress during the perinatal period. Other risk factors included a lack of social/emotional support and increased family-related stressors. Interventions for pregnant military spouses included those that help them develop internal coping strategies and external social support.ConclusionsUS literature suggests that military spouses are particularly at risk of PMH problems during deployment of their serving partner and highlights the protective nature of social support during this time. Further consideration needs to be made to apply the findings to UK military spouses/partners due to differences in the structure and nature of the UK and US military and healthcare models. Further UK research is needed, which would provide military and healthcare providers with an understanding of the needs of this population allowing effective planning and strategies to be commissioned and implemented.


Reproduction ◽  
2007 ◽  
Vol 133 (1) ◽  
pp. 219-230 ◽  
Author(s):  
Feikun Yang ◽  
Ru Hao ◽  
Barbara Kessler ◽  
Gottfried Brem ◽  
Eckhard Wolf ◽  
...  

The epigenetic status of a donor nucleus has an important effect on the developmental potential of embryos produced by somatic cell nuclear transfer (SCNT). In this study, we transferred cultured rabbit cumulus cells (RCC) and fetal fibroblasts (RFF) from genetically marked rabbits (Alicia/Basilea) into metaphase II oocytes and analyzed the levels of histone H3-lysine 9-lysine 14 acetylation (acH3K9/14) in donor cells and cloned embryos. We also assessed the correlation between the histone acetylation status of donor cells and cloned embryos and their developmental potential. To test whether alteration of the histone acetylation status affects development of cloned embryos, we treated donor cells with sodium butyrate (NaBu), a histone deacetylase inhibitor. Further, we tried to improve cloning efficiency by chimeric complementation of cloned embryos with blastomeres fromin vivofertilized or parthenogenetic embryos. The levels of acH3K9/14 were higher in RCCs than in RFFs (P<0.05). Although the type of donor cells did not affect development to blastocyst, after transfer into recipients, RCC cloned embryos induced a higher initial pregnancy rate as compared to RFF cloned embryos (40 vs 20%). However, almost all pregnancies with either type of cloned embryos were lost by the middle of gestation and only one fully developed, live RCC-derived rabbit was obtained. Treatment of RFFs with NaBu significantly increased the level of acH3K9/14 and the proportion of nuclear transfer embryos developing to blastocyst (49 vs 33% with non-treated RFF,P<0.05). The distribution of acH3K9/14 in either group of cloned embryos did not resemble that inin vivofertilized embryos suggesting that reprogramming of this epigenetic mark is aberrant in cloned rabbit embryos and cannot be corrected by treatment of donor cells with NaBu. Aggregation of embryos cloned from NaBu-treated RFFs with blastomeres fromin vivoderived embryos improved development to blastocyst, but no cloned offspring were obtained. Two live cloned rabbits were produced from this donor cell type only after aggregation of cloned embryos with a parthenogenetic blastomere. Our study demonstrates that the levels of histone acetylation in donor cells and cloned embryos correlate with their developmental potential and may be a useful epigenetic mark to predict efficiency of SCNT in rabbits.


Development ◽  
1990 ◽  
Vol 108 (1) ◽  
pp. 185-189 ◽  
Author(s):  
J.N. Petitte ◽  
M.E. Clark ◽  
G. Liu ◽  
A.M. Verrinder Gibbins ◽  
R.J. Etches

Cells were isolated from stage X embryos of a line of Barred Plymouth Rock chickens (that have black pigment in their feathers due to the recessive allele at the I locus) and injected into the subgerminal cavity of embryos from an inbred line of Dwarf White Leghorns (that have white feathers due to the dominant allele at the I locus). Of 53 Dwarf White Leghorn embryos that were injected with Barred Plymouth Rock blastodermal cells, 6 (11.3%) were phenotypically chimeric with respect to feather colour and one (a male) survived to hatching. The distribution of black feathers in the recipients was variable and not limited to a particular region although, in all but one case, the donor cell lineage was evident in the head. The male somatic chimera was mated to several Barred Plymouth Rock hens to determine the extent to which donor cells had been incorporated into his testes. Of 719 chicks hatched from these matings, 2 were phenotypically Barred Plymouth Rocks demonstrating that cells capable of incorporation into the germline had been transferred. Fingerprints of the blood and sperm DNA from the germline chimera indicated that both of these tissues were different from those of the inbred line of Dwarf White Leghorns. Bands that were present in fingerprints of blood DNA from the chimera and not present in those of the Dwarf White Leghorns were observed in those of the Barred Plymouth Rocks.(ABSTRACT TRUNCATED AT 250 WORDS)


2016 ◽  
Vol 50 (2) ◽  
pp. 263-271 ◽  
Author(s):  
Sandra Maria Sampaio Enes ◽  
Simone Perufo Opitz ◽  
André Ricardo Maia da Costa de Faro ◽  
Mavilde de Luz Gonçalves Pedreira

Abstract OBJECTIVE To identify the presence of phlebitis and the factors that influence the development of this complication in adult patients admitted to hospital in the western Brazilian Amazon. METHOD Exploratory study with a sample of 122 peripheral intravenous catheters inserted in 122 patients in a medical unit. Variables related to the patient and intravenous therapy were analyzed. For the analysis, we used chi-square tests of Pearson and Fisher exact test, with 5% significance level. RESULTS Complication was the main reason for catheter removal (67.2%), phlebitis was the most frequent complication (31.1%). The mean duration of intravenous therapy use was 8.81 days in continuous and intermittent infusion (61.5%), in 20G catheter (39.3%), inserted in the dorsal hand vein arc (36.9 %), with mean time of usage of 68.4 hours. The type of infusion (p=0.044) and the presence of chronic disease (p=0.005) and infection (p=0.007) affected the development of phlebitis. CONCLUSION There was a high frequency of phlebitis in the sample, being influenced by concomitant use of continuous and intermittent infusion of drugs and solutions, and more frequent in patients with chronic diseases and infection.


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