Transcriptome analysis of bovine day 16 conceptus derived after transfer of blastocyst from somatic cell nuclear transfer or in vitro production

In vitro embryo production (IVP) and somatic cell nuclear transfer (SCNT) have been used as tools of assisted reproductive technology to produce bovine pre-implantation embryos independent of the maternal environment. However, the embryonic and fetal losses after transfer of SCNT and IVP derived embryos is higher compared to the in vivo (AI) counterparts. This may be associated with the alterations in the molecular signatures and pathways at any stage of embryonic and /or fetal development. Therefore, to identify the molecular changes that could occur at day 16 SCNT and IVP derived embryos, large scale transcriptomic analysis was performed using Affymetrix-Bovine Genome Array. For this, day 7 blastocysts derived from SCNT, IVP and AI were transferred to oestrus synchronized Simmental heifers. Recipients were then slaughtered at day 16 of gestation and conceptuses were retrieved. Following morphological examination, filamentous embryos with visible embryonic disc were subjected to global tanscriptome analysis. The result demonstrated comparable in vivo development rate in SCNT (72.7%), IVP (62.2%) and AI (77.3%) embryo groups. However, considerable reduction in the trophoblast elongation size was observed in SCNT (93.3mm) compared to IVP (186.6mm) and AI (196.3mm) derived embryos. In addition, more than 20% of SCNT (10.7 mm ± 1.08) and IVP (20.1 mm ± 0.15) conceptuses had tubular shape, suggesting a delay in recapitulating filamentous morphology. Gene expression profiling analysis revealed that the transcript levels of 477 genes, which are involved in various pathways including arginine and proline, glycerolipid and fatty acid metabolism, were significantly altered in SCNT embryos compared to AI. Similarly, 365 genes were differentially expressed in IVP embryos compared to AI. Thus, several canonical pathways including TNRF-1 and tight junction signalling pathways were affected in IVP derived conceptuses. To predict whether the altered transcripts were associated with pre-elongation in vitro culture environment or errors in transcriptional reprogramming, unique or commonly differentially expressed genes were analyzed in SCNT and IVP embryos compared to AI or donor cells (fibroblast). Accordingly, 71 transcripts including (FOLR1, MYO1B, RCN2, H2AFJ, HSPB1 and GATM) were found to be not transcriptionally reprogrammed as their expression resembled more the donor cells than AI embryos. The remaining transcripts were either partially or incompletely reprogrammed. In addition, quantitative real time PCR (qPCR) based expression profiling of candidate transcripts in developmentally delayed SCNT or IVP embryos showed low mRNA levels of IFNt, FGFR2, CLDN1 and ARHGEF2 in developmentally lagging IVP and SCNT embryos compared to their respective elongated counterparts. In conclusion, the present study identified deviation in elongation size, gene expression and the corresponding molecular pathways in day 16 SCNT and IVP conceptuses compared to their AI counterparts which may subsequently be associated with fetal development.Transkriptom-analyse von bovinen 16 Tage alten Embryonen, gewonnen durch den Transfer von Blastozysten aus klonierten somatischen Zellen sowie der in vitro Produktion In vitro Embryo Produktion (IVP) und somatischer Kerntransfer (SCNT) sind Werkzeuge der assistierten Reproduktionstechnologien und finden ihren Einsatz um bovine Präimplantations- embryonen unabhängig von der mütterlichen Umwelt zu erzeugen. Allerdings sind embryonale und fetale Verluste nach dem Transfer von SCNT und IVP gewonnenen Embryonen höher im Vergleich zu in vivo (AI) erzeugten Embryonen. Dies kann mit den Veränderungen der molekularen Signaturen sowie Signalwegen in den unterschiedlichen Stadien der embryonalen und/oder fetalen Entwicklung zusammenhängen. Um molekulare Veränderungen zu identifizieren, die am Tag 16 von SCNT und IVP gewonnene Embryonen auftreten können, wurde mit Affymetrix-Bovine Genome Arrays eine Transkriptomanalyse durchgeführt. Hierzu wurden Tag 7 Blastozysten von SCNT, IVP und AI erzeugten Embryonen in Östrus synchronisierte Fleckviehfärsen übertragen. Am Tag 16 der Trächtigkeit wurden die Rezipienten geschlachtet und die Embryonen entnommen. Nach morphologischen Untersuchungen wurden filamentöse Embryonen mit sichtbarer Keimscheibe einer globale Tanskriptomanalyse unterzogen. Das Ergebnis zeigte in den verschiedenen Embryogruppen SCNT (72,7%), IVP (62,2%) und AI (77,3%) eine vergleichbare in vivo Entwicklung. Allerdings konnte eine erhebliche Verringerung in der Größe der Trophoblasten Elongation in SCNT (93,3 mm) im Vergleich zu IVP (186,6 mm) und AI (196,3 mm) Embryonen beobachtet werden. Darüber hinaus wiesen mehr als 20% der SCNT (10,7 mm ± 1,08) und IVP (20,1 mm ± 0,15) Embryonen eine Röhrenform auf, was auf eine verzögerte rekapitulierte filamentöse Morphologie hindeutet. Die Auswertung der Transkriptomanalyse zeigte beim Vergleich von SCNT mit AI 477 signifikant unterschiedlich expremierte Gene, die in verschiedenen Signalwegen beteiligt sind, einschließlich Arginin und Prolin, Glycerolipid und Fettsäure-Metabolismus. Des Weiteren wurden 365 signifikant unterschiedlich exprimierte Gene beim Vergleich von IVP Embryonen mit AI Embryonen identifiziert. Relevante Signalwege dieser Gene waren unter anderem TNRF-1 und Tight-Junction Signalisierung. Um festzustellen, ob die veränderten Transkripte mit der in in vitro Kultur bedingten Präelongation oder mit Fehlern der transkriptionellen Reprogrammierung assoziiert sind, wurden einzigartige oder häufig unterschiedlich exprimierte Gene in SCNT und IVP Embryonen gegenüber AI oder Donorzellen (Fibroblasten) analysiert. Dementsprechend zeigten 71 Transkripte einschließlich FOLR1, MYO1B, RCN2, H2AFJ, HSPB1 und GATM keine transkriptionelle Reprogrammierung, da deren Expressionprofil mehr dem der Donorzellen als dem der AI Embryonen ähnelte. Die restlichen Transkripte waren entweder teilweise oder vollständig reprogrammiert. Zusätzlich, zeigten auf quantitative Real Time PCR (qPCR) basierende Kandidatengenexpressionsprofile in entwicklungsverzögerten SCNT oder IVP Embryonen niedrigere mRNA Spiegel in IFNtau, FGFR2, CLDN1 und ARHGEF2 im Vergleich zu ihren elongierten Gegenstücken. Schlussfolgernd konnten mit dieser Studie Abweichungen in den Elongationsgrößen, den Expressionsprofilen und den entsprechenden molekularen Signalwegen in Tag 16 SCNT und IVP produzierten Embryonen im Vergleich zu AI produzierten Embryonen beobachtet werden. Diese Ergebnisse könnten in Zusammenhang mit den weiteren fötalen Entwicklung gebracht werden

Characterization of the Indigenous Goat Populations of South Gonder Based on their Morphometric Traits and Body Indices

  አህፅሮት በዳሰሳ ላይ የተደገፈ ጥናት በደቡብ ጎንደር ዞን የሞርፎሜትሪክ ባህሪያት እና የአካል መረጃ ጠቋሚዎች መሠረት በማድረግ በአገር በቀል ፍየል የምርት አቅም ላይ የምርምር ሥራ ተካሂዷል፡፡ የፍየሎችን የርባታ ባህሪያትን በተመለከተ ከ153 አባወራዎች በመጠይቅ የተደገፈ መረጃ የተሰበሰበ ሲሆን፤ የሞርፎሜትሪክ ባህሪያት መለኪያዎች ከፋርጣ ወረዳ 152፤ ከፎገራ ወረዳ 154፤ እና ከሊቦ ከምከም ወረዳ 204 በአጠቃላይ ከሁለቱም ፆታዎች በ510 ፍየሎች ላይ መረጃ ተወስዷል፡፡ በዚሁ መሠረት 19 የፍየል አካላት መለኪያ ኢንዴክሶች የሞሮፎሞትሪክ ባህሪያትን መሠረት በማድረግ ተሰልቷል፡፡ በጥናቱ በተገኘው ውጤት መሠረት የሴት ፍየሎች ለመጀሪያ ጊዜ የወለዱበት ዕድሜ 13.6 ወር ሲሆን በሁለት ወሊዶች መካከል ያለው ልዩነት 8.39 ወራት ነበር፡፡ ባንድ ጊዜ የተወለዱት የፍየል ግልገሎች መጠን 1.54 እንደሆነ ታውቋል፡፡ የፍየሎች ፆታ ከጀሮ ርዝመት፤ ከዳሌ ስፋት እና ከእግር አገዳ ቅልጥም ክብ ዙሪያ በሰተቀር በሌሎች ባህሪት ላይ ጉልህ ተፅዕኖ ነበረው (p<0.001)፡፡ በተጨማሪም ከጀሮ ርዝመት በሰተቀር የፍየሎች ዕድሜ በሁሉም የሞርፎሜትሪክ ባህሪያት ላይ የነበረው ተፅዕኖ ከፍተኛ ነበር (p<0.001)፡፡ የፎገራ ወረዳ ፍየሎች በክብደት፤ በደረት ክብ ዙሪያ፤ ከፊት እግር ጀርባ ቁመት፤ በአካል ርዝመት፤ በደርት ስፋት፤ ከኋላ እግር ፊት ቁመት፤ በዳሌ ርዝመት እና በጡት ርዝመት ከሊቦ ከምከም ወረዳ ፍየሎች የበለጡ ነበሩ፡፡ የሊቦ ከምከም ፍየሎች ከፎገራና ፋርጣ ፍየሎች ጋር ሲነፃፀሩ ጥናት በተደረገባቸው በአብዛኛዎቹ የሞርፎሜትሪክ ባህሪያት ዝቀተኛ እንደነበሩ ከጥናቱ የተገኘው ውጤት ያመላክታል፡፡ የደረት ክብ ዙሪያ እና የፊት እግር ጀርባ ቁመት ልኬቶች የሴት እና የወንድ ፍየሎችን የክብደት መጠን ለመተንበይ ቀዳማይ ባህሪያት እንደሆኑ ተለይተው ታውቀዋል፡፡ በተገኘው የፍየል አካላት መለኪያ ኢንዴክሶች መሠረት፤ የደቡብ ጎንደር ፍየሎች አጭር እግር ያላቸው እና ለወይና ደጋ አየር ንብረት የሚስማሙ ሲሆኑ ለስጋ ምርት አቅም እንዳላቸው ታውቋል፡፡ በማጠቃለያም፤ በደቡብ ጎንደር ዞን የሚገኙ አገር በቀል ፍየሎች ለስጋ ምርት እምቅ አቅም ሊኖራቻው እንደሚችል የጥናቱ ውጤት በጉልህ አመላክቷል፡፡   Abstract A survey-based study was undertaken to explore the potentials of indigenous goat populations of the South Gondar zone based on morphometric traits and body indices. Reproductive traits were obtained from 153 households through a questionnaire while morphometric measurements were taken from 510 goats of both sexes drawn from Farta (n=152), Fogera (n=154), and Libokemkem (n=204) districts. Nineteen structural and functional indices were computed from morphometric measurements. In this study, age at first kidding and kidding intervals was 13.6 and 8.39 months, respectively with an average litter size of 1.54. Sex had a significant (p<0.001) effect on all quantitative traits except ear length (EL), rump width, and canon circumference. Except for EL, the age effect was significant (p<0.001) for all morphometric traits. Goats at Fogera district had higher live weight (LW), heart girth (HG), height at withers (HW), body length, chest width, rump height, rump length, and teat length than those at Libokemkem. Goats from Libokemkem district were inferior for most of the studied morphometric traits as compared to those at Fogera and Farta. Both HG and HW were identified as best predictors of LW in both sexes. Based on the values of structural indices, the goat populations could be characterized as meat phenotype with short legs being well adapted to midland altitudes. In conclusion, goats reared in the South Gondar zone could have the genetic potential for meat production with high prolificacy

Assessment of Breeding Practice and Statues of Estrus Synchronization and Mass Insemination of Dairy Cattle in North Shewa Zone, Amhara Region, Ethiopia

The general objective of this study was to assess breeding practice, and status of OSMI of dairy cattle. Data were analyzed using SPSS (version.20) and Ms-Excel (2010). Milk yield, breeding ability, body weight, fertility, and udder size are the major traits perceived by farmers. 92.6% of the respondents reared all cattle categories together except higher Holstein Friesian cross breeds and lactating cows. Conception rate (CR) and number of service per conception (NSPC) were affected (p<0.05) by year, breed and districts. Majority (67.1%) of the respondents were not satisfied with the results of the estrus synchronization and mass insemination (OSMI) program as the conception rate was low. There was better perception of farmers to OSMI technology in Angolelanatera (50%) compared to the two district. There were better opportunities for the application of OSMI technologies in Angolelanatera and Basonaworena districts due to availability of improved breed and milk receiver union. In conclusion the status of OSMI in the study area was increasing starting 2013-2015as shown in table 26. The main reason for farmers’ dissatisfaction on OSMI program   was low conception rate of dairy cows.. In addition procurement of the necessary facilities should be in place before implementing an estrus synchronization program. Keywords: Breeding practice, Estrus synchronization and mass insemination. DOI: 10.7176/JBAH/14-1-01 Publication date: January 31st 202

Morphological descriptors of indigenous chickens: as a selection guide

ABSTRACTThe visual morphologies in animals are associated positively or negatively with production, adaptation, and behavioral characteristics. For rural farmers with limited records, these features are considered reliable phenotypic markers. Thus, this study was intended to explore the unique morphological features of indigenous chickens as a farmer's selection guide. A total of 1,060 adult chickens from four rural districts were investigated for nine morphological traits. The single-comb type, supposed to be of good fertility, was the most prevalent. Yellow and white shanks were predominant, followed by gray, greenish-yellow, green, and black pigmentations. Crested and silky chickens were more frequent in highland areas, whereas naked-neck chickens were observed in lowlands. Chickens of the lowlands have brighter, more uniform plumage colors and yellow shank, beak, and earlobe colors. On the other hand, the highland and mid-altitude chickens are characterized by dark, colorful combinations of various feather colors: laced plumage, white shank, rose, and deformed rose combs. Moreover, multiple correspondence analyses revealed that morphological features have special associations with districts. Thus, the observed morphological heterogeneity of the indigenous chickens can be used as a good indicator of selective breeding. Therefore, the associations of these characteristics with productivity qualities could be confirmed by quantitative and molecular tools

Production, reproduction and some adaptation characteristics of Boran cattle breed under changing climate: A systematic review and meta-analysis.

IntroductionClimate change affects livestock production and productivity, which could threaten livestock-based food security in pastoral and agro-pastoral production systems of the tropics and sub-tropics. Boran cattle breed is one of the hardiest Zebu cattle reared by Borana Oromo pastoralists for milk and meat production. However, there is limited comprensive information on production, reproduction and adaption traits of the Boran cattle in Ethiopia. Thus, this paper aims to compile the main production, reproduction and some adaptation traits of Boran cattle based on systematic review and meta-analysis of peer reviewed published and unpublished literature.MethodologyA combination of systematic review and meta-analysis based on PRISMA guideline was employed. Accordingly, out of 646 recorded articles identified through database searching, 64 were found to be eligible for production, reproduction and adaptation characteristics of the Boran cattle, 28 articles were included in qualitative systematic review while 36 articles were used for quantitative meta-analysis.ResultThe Boran cattle breed has the ability to survive, produce and reproduce under high ambient temperature, utilize low quality forage resources, and resist water shortage or long watering intervals and tick infestations. The review revealed that the breed employs various adaptation responses (morphological, physiological, biochemical, metabolic, cellular and molecular responses) to cope with harsh environmental conditions including climate change, rangeland degradation, seasonal feed and water shortages and high incidences of tick infestations. The meta-analysis using a random-effects model allowed provision of pooled estimates of heritability and genetic correlations for reproduction and production traits, which could be used to solve genetic prediction equations under a population level in purebred Boran cattle. In addition, heritability and genetic-correlation estimates found in the present study suggest that there is high genetic variability for most traits in Boran cattle, and that genetic progress is possible for all studied traits in this breed.ConclusionThe Boran cattle breed has the ability to survive, produce and reproduce under high ambient temperature, utilize low quality forage resources, and resist water shortage or long watering intervals and tick infestations. However, currently there are several challenges such as recurrent droughts, pasture deterioration and lack of systematic selection and breeding programs that play to undermine the realization of the potential of the breed. Thus, we recommend systematic selection for enhancing the reproductive and production performances without compromising the adaptation traits of the breed coupled with improved management of rangelands

Transcriptome profile of bovine elongated conceptus obtained from SCNT and IVP pregnancies

In the present study we analyzed the gene expression changes induced by somatic cell nuclear transfer (SCNT) and in vitro production (IVP) in bovine elongated embryos using Affymetrix bovine genome array. For this, Day-16 bovine embryos from SCNT, IVP, and artificial insemination (AI) were recovered from recipients and used for transcriptome analysis. Despite comparable in vivo development rates, considerable reduction in elongation size was observed in SCNT compared to non-cloned embryos (93.3mm for SCNT vs. 186.6mm and 196.3mm for IVP and AI embryos, respectively). Gene expression analysis revealed that the transcript levels of 477 genes, which are involved in various pathways including arginine and proline or glycerolipid and fatty acid metabolism, were significantly altered in SCNT compared to AI embryos. Similarly, 365 genes were differentially expressed in IVP embryos compared to AI. Thus, several pathways including TNRF-1 signaling and tight junction pathways were affected. To predict whether the altered transcripts were associated with culture condition or errors in transcriptional reprogramming, unique or common differentially expressed genes were analyzed in SCNT and IVP embryos compared to AI or fibroblast donor cells. Accordingly, 71 transcripts were found to be not transcriptionally reprogrammed, as their expression resembled the donor cells more than AI embryos; the remaining transcripts were either partially or incompletely reprogrammed. In conclusion, the present study identified deviations in elongation size, gene expression, and the corresponding molecular pathways in Day-16 SCNT and IVP conceptuses compared to their AI counterparts, which may subsequently be associated with the outcome of fetal development. Mol. Reprod. Dev. 80: 315333, 2013. (c) 2013 Wiley Periodicals, Inc.In the present study we analysed the gene expression changes induced by somatic cell nuclear transfer (SCNT) and in vitro production (IVP) in bovine elongated embryos using the Affymetrix bovine genome array. For this, Day-16 bovine embryos from SCNT, IVP, and artificial insemination (AI) were recovered from recipients and used for transcriptome analysis. Despite comparable in vivo development rates, considerable reduction in elongation size was observed in SCNT compared to non-cloned embryos (93.3 mm for SCNT vs. 186.6 mm and 196.3 mm for IVP and AI embryos, respectively). Gene expression analysis revealed that the transcript levels of 477 genes, which are involved in various pathways including arginine and proline or glycerolipid and fatty acid metabolism, were significantly altered in SCNT compared to AI embryos. Similarly, 365 genes were differentially expressed in IVP embryos compared to AI. Thus, several pathways including TNRF-1 signalling and tight junction pathways were affected. To predict whether the altered transcripts were associated with culture condition or errors in transcriptional reprogramming, unique or common differentially expressed genes were analyzed in SCNT and IVP embryos compared to AI or fibroblast donor cells. Accordingly, 71 transcripts were found to be not transcriptionally reprogrammed as their expression resembled the donor cells more than AI embryos. The remaining transcripts were either partially or incompletely reprogrammed. In conclusion, the present study identified deviations in elongation size, gene expression, and the corresponding molecular pathways in Day 16 SCNT and IVP conceptuses compared to their AI counterparts, which may subsequently be associated with the outcome of fetal development