Although our knowledge regarding oocyte quality and development has improved significantly, the molecular mechanisms that regulate and determine oocyte developmental competence are still unclear. Therefore, the objective of this study was to identify and analyze the transcriptome profiles of porcine oocytes derived from large or small follicles using RNA high-throughput sequencing technology. RNA libraries were constructed from oocytes of large (LO; 3-6 mm) or small (SO; 1.5-1.9 mm) ovarian follicles and then sequenced in an Illumina HiSeq4000. Transcriptome analysis showed a total of 14,557 genes were commonly detected in both oocyte groups. Genes related to the cell cycle, oocyte meiosis, and quality were among the top highly expressed genes in both groups. Differential expression analysis revealed 60 up- and 262 downregulated genes in the LO compared with the SO group. BRCA2, GPLD1, ZP3, ND3, and ND4L were among the highly abundant and highly significant differentially expressed genes (DEGs). The ontological classification of DEGs indicated that protein processing in endoplasmic reticulum was the top enriched pathway. In addition, biological processes related to cell growth and signaling, gene expression regulations, cytoskeleton, and extracellular matrix organization were among the highly enriched processes. In conclusion, this study provides new insights into the global transcriptome changes and the abundance of specific transcripts in porcine oocytes in correlation with follicle size.
- MeSH
- genové regulační sítě fyziologie MeSH
- oocyty metabolismus MeSH
- oogeneze genetika MeSH
- ovariální folikul cytologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- prasata genetika růst a vývoj MeSH
- signální transdukce genetika MeSH
- stanovení celkové genové exprese MeSH
- transkriptom * MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- vývojová regulace genové exprese fyziologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We evaluated the prevalence and epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates in pigs during production cycle on a Czech farm with the history of previous use of ceftiofur. ESBL-producing E. coli isolates were obtained from rectal swabs from pigs of different age groups (suckling piglets, weaned piglets, growers and sows). Collected samples were directly cultivated on MacConkey agar with cefotaxime (2 mg l-1 ), whereas intestinal swabs of slaughtered pigs and surface swabs from pig carcasses were also pre-enriched in buffered peptone water without antimicrobials before the cultivation. Clonal relationship of selected isolates was determined by XbaI pulse-field gel electrophoresis and multi-locus sequence typing. The transferability of plasmids carrying blaCTX-M genes was tested by conjugation experiments. From all examined samples, 141 (43·7%, n = 323) were positive for ESBL-producing E. coli. All ESBL-producing isolates showed resistance to multiple antimicrobials and were positive for blaCTX-M genes. The blaCTX-M-1 was carried by conjugative IncN/ST1 plasmids (c. 40-45 kb) while the blaCTX-M-15 was located on conjugative F plasmids with F:18:A5:B1 formula (c. 165 kb). This study demonstrated the persistence of CTX-M-positive E. coli isolates 2 months after banner of ceftiofur usage and indicated possible risk of transmission of these isolates to humans via the food chain.
- MeSH
- beta-laktamasy genetika metabolismus MeSH
- Escherichia coli enzymologie genetika izolace a purifikace MeSH
- farmy MeSH
- infekce vyvolané Escherichia coli mikrobiologie patofyziologie veterinární MeSH
- lidé MeSH
- multilokusová sekvenční typizace MeSH
- nemoci prasat mikrobiologie patofyziologie MeSH
- plazmidy genetika metabolismus MeSH
- prasata růst a vývoj mikrobiologie MeSH
- proteiny z Escherichia coli genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Porcine thymus contains three independent populations of cells that have rearranged immunoglobulin heavy chain VDJH genes. The first population can be found exclusively in medulla and it consists of existing mature B cells and plasma cells. The second consists of developing B cells characterized by the presence of selected VDJH rearrangement, similar to B cell lymphogenesis in the bone marrow. The third population is entirely unaffected by selection mechanism for productive VDJH rearrangement and represents T lineage cells that rearrange immunoglobulin genes. Transcription of unselected VDJH repertoire is not allowed in T cells. Sequence analysis of unselected VDJH repertoire from T cells also revealed important consequences for B cell lymphogenesis and selection of B cell repertoire. As far as we know, this is the first evidence that some species completely rearrange VDJH genes in T cells. Our results also support the finding that B cells actively develop in the thymus.
- MeSH
- B-lymfocyty imunologie MeSH
- druhová specificita MeSH
- geny pro těžké řetězce imunoglobulinů genetika MeSH
- lidé MeSH
- plod imunologie MeSH
- podskupiny lymfocytů imunologie MeSH
- prasata genetika růst a vývoj imunologie MeSH
- T-lymfocyty imunologie MeSH
- thymus růst a vývoj imunologie MeSH
- V(D)J rekombinace genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Farrowing pens with temporary crating have been developed as a compromise between conventional farrowing crates and pens to better accommodate the welfare of both sow and piglets during lactation. However, not much is known about the behavioral and physiological consequences of early removal of confinement on the sow and piglets during lactation. The aim of this study was to assess the effects on sow and piglet performance of temporary crating until 3-d postpartum at 2 times points, immediately after confinement removal and 25 d into lactation. Sows were crated from 5-d prepartum either to weaning (permanently crated-PC group; N =14) or to D3 (83.0 ± 1.3 h) postpartum (Temporarily crated - TC group N = 13). Sow postural changes, activity, cortisol and IgA concentrations, and piglet body weight gain and behavior were assessed on D4 and on D25 postpartum, whereas piglet mortality was assessed throughout lactation. Data were analyzed using PROC GLM and PROC GENMOD of SAS. On D4 postfarrowing, TC sows were more active (10.9% vs. 7.1%; SEM: 0.8; P = 0.002), rolled more frequently (21.3% vs. 14.4%; SEM: 1.6; P = 0.008), and had lower IgA concentrations (139.7 vs. 75.2 µg/mL; SEM: 20.3; P = 0.040) than PC sows. No effects of housing were found (P > 0.05) on standing-to-lying movement or cortisol concentrations. No differences for any variables were found (P > 0.05) on D25. Mortality, body weights, and activity levels at the udder or in the pen of pigs born to PC sows did not differ (P > 0.05) from those of piglets born to TC sows on D4 nor on D25. This study indicates that removal of confinement on the 4th-d postpartum may have had small short-term positive effects on sow behavior and stress levels (as measured by IgA), and that it did not impair piglets' behavior and performance during lactation. Therefore, this work suggests that temporary crating limited to the first 3-d postpartum might be a feasible alternative to improve welfare under intensive production conditions.
- MeSH
- bydlení zvířat * MeSH
- chování zvířat * MeSH
- hmotnostní přírůstek MeSH
- hydrokortison analýza MeSH
- imunoglobulin G analýza MeSH
- laktace MeSH
- mléčné žlázy zvířat fyziologie MeSH
- náhodné rozdělení MeSH
- novorozená zvířata MeSH
- odstavení MeSH
- poporodní období MeSH
- prasata růst a vývoj fyziologie psychologie MeSH
- těhotenství MeSH
- tělesná hmotnost MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Mammalian cumulus-oocyte complexes (COCs) reach full developmental capability during folliculogenesis and oogenesis. It is well recognized that only gametes achieving MII stage after in vivo or in vitro maturation (IVM) are successfully fertilized by a single spermatozoon. Although the process of oocyte nuclear and/or cytoplasmic maturation in pigs is well determined, there exist many differences that promote these processes in vivo and in vitro. Therefore, this study aimed to investigate the differences in RNA expression profiles between porcine oocytes before and after IVM using microarray and real-time quantitative polymerase chain reaction (RT-qPCR) assays. Experiments were performed on oocytes isolated from 55 pubertal crossbred Landrace gilts. The oocytes were analyzed both before and after IVM and only Brilliant Cresyl Blue (BCB)-positive gametes were used for subsequent microarray analysis (Affymetrix) and RT-qPCR analysis. The microarray assay, which measures expression of 12,258 transcripts, revealed 419 differentially expressed transcripts in porcine oocytes, from which 379 were downregulated and 40 were upregulated before IVM compared to those analyzed after IVM. After DAVID analysis, we found eight different transcripts, including IHH, BMP1, WWTR1, CHRDL1, KLF10, EIF2AK3, MMP14, and STC1. Their expression is related to the "bone development" ontology group and was further subjected to hierarchical clusterization. Using RT-qPCR analysis, we confirmed the results of the microarray assay, showing increased expression of the eight genes in oocytes before IVM compared to oocytes after maturation in vitro. It has been suggested that "bone development" belongs to one ontological group involving genes substantially upregulated in porcine oocytes before IVM. We suggest that the gamete mRNA expression profile before IVM may comprise stored transcripts, which are templates for protein biosynthesis following fertilization. We also hypothesize that these mRNAs may be a specific "fingerprint" of folliculogenesis and oogenesis in pigs.
- MeSH
- aktivace transkripce MeSH
- down regulace MeSH
- exprese genu fyziologie MeSH
- genová ontologie MeSH
- IVM techniky MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- messenger RNA biosyntéza genetika MeSH
- oocyty cytologie metabolismus MeSH
- oogeneze genetika MeSH
- prasata genetika růst a vývoj MeSH
- prekurzory RNA MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody MeSH
- stanovení celkové genové exprese MeSH
- transkriptom fyziologie MeSH
- upregulace MeSH
- vývoj kostí genetika fyziologie MeSH
- vývojová regulace genové exprese * MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Hydrogen sulfide, one of three known gasotransmitters, is involved in physiological processes, including reproductive functions. Oocyte maturation and surrounding cumulus cell expansion play an essential role in female reproduction and subsequent embryonic development. Although the positive effects of exogenous hydrogen sulfide on maturing oocytes are well known, the role of endogenous hydrogen sulfide, which is physiologically released by enzymes, has not yet been described in oocytes. In this study, we observed the presence of Cystathionine β-Synthase (CBS), Cystathionine γ-Lyase (CTH) and 3-Mercaptopyruvate Sulfurtransferase (3-MPST), hydrogen sulfide-releasing enzymes, in porcine oocytes. Endogenous hydrogen sulfide production was detected in immature and matured oocytes as well as its requirement for meiotic maturation. Individual hydrogen sulfide-releasing enzymes seem to be capable of substituting for each other in hydrogen sulfide production. However, meiosis suppression by inhibition of all hydrogen sulfide-releasing enzymes is not irreversible and this effect is a result of M-Phase/Maturation Promoting Factor (MPF) and Mitogen-Activated Protein Kinase (MAPK) activity inhibition. Futhermore, cumulus expansion expressed by hyaluronic acid (HA) production is affected by the inhibition of hydrogen sulfide production. Moreover, quality changes of the expanded cumuli are indicated. These results demonstrate hydrogen sulfide involvement in oocyte maturation as well as cumulus expansion. As such, hydrogen sulfide appears to be an important cell messenger during mammalian oocyte meiosis and adequate cumulus expansion.
- MeSH
- imunohistochemie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- kyselina hyaluronová chemie MeSH
- oocyty enzymologie růst a vývoj MeSH
- prasata růst a vývoj fyziologie MeSH
- sulfan metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Free farrowing pens (pens) improve the welfare of sows but may increase sow activity and negatively influence piglet production. The aim of this study was to assess the effect of pens and crates on sow postural changes, piglet trapping, sow responses to piglet screams, piglet mortality, and piglet BW gain. It was predicted that provision of greater space (in pens) would increase not only the frequency of sow postural changes and the probability of trappisng but also sow responses to the screams of piglets; thus, the outcome would be no differences in fatal piglet crushing or overall mortality between the housing systems. Sows were randomly moved to either a farrowing pen (n = 20) or farrowing crate (n = 18). Sow behavior was recorded and analyzed for 72 h from the birth of the first piglet (BFP). Sow postural changes included rolling from a ventral to lateral position and vice versa and going from standing to sitting, standing to lying, and sitting to lying. Occurrences of piglet trapping and sow responsiveness to real crushing situations were analyzed. Sow responsiveness was assessed in response to audio playbacks (PB) of piglet screams on d 3 postpartum (48 to 72 h after BFP; PB crush calls) and real piglet crushing during the first 72 h after BFP (real crush calls). Piglet BW gain was estimated 24 h after BFP, piglet BW was recorded at weaning, and piglet crushing and piglet mortality were recorded during the 72 h after BFP. Data were analyzed using PROC MIXED and PROC GENMOD of SAS. Sows in pens showed more postural changes (P = 0.04) and tended to have greater incidences of piglet trapping (P = 0.07) than those in crates. Sow response to PB crush calls was greater in pens (P = 0.04) but did not differ for real crush calls between pens and crates (P = 0.62). There was no effect on the probability of piglet crushing (P = 0.38) and mortality (P = 0.41) during the 72 h after BFP nor in piglet mortality at weaning (P = 0.81) between pens and crates. Piglet BW gain at 24 h after BFP (P = 0.01) and piglet BW at weaning (P = 0.02) were greater in pens. Sows in pens showed more postural changes and tended to trap more piglets; however, the response to real crush calls did not differ between the two housing systems. Despite this, there was no increase in piglet crushing or mortality in pens, which might be influenced by the better piglet body condition observed in pens, which in turn could influence their ability to avoid crushing by the sow.
- MeSH
- bydlení zvířat MeSH
- chování zvířat MeSH
- mortalita MeSH
- novorozená zvířata růst a vývoj psychologie MeSH
- postura těla MeSH
- prasata růst a vývoj fyziologie psychologie MeSH
- vokalizace zvířat * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The effects of live yeast Saccharomyces cerevisiae (strain CNCM I-4407, 10(10) cfu/g; Actisaf; Lesaffre Feed Additives, Marcq-en-Baroeul, France) on the severity of diarrhea, immune response, and growth performance in weaned piglets orally challenged with enterotoxigenic Escherichia coli (ETEC) strain O149:K88 were investigated. Live yeast was fed to sows and their piglets in the late gestation, suckling, and postweaning periods. Sows were fed a basal diet without (Control; n = 2) or with (Supplemented; n = 2) 1 g/kg of live yeast from d 94 of gestation and during lactation until weaning of the piglets (d 28). Suckling piglets of the supplemented sows were orally treated with 1 g of live yeast in porridge carrier 3 times a week until weaning. Weaned piglets were fed a basal starter diet without (Control; n = 19) or with (Supplemented; n = 15) 5 g of live yeast/kg feed for 2 wk. Significantly lower daily diarrhea scores (P < 0.05), duration of diarrhea (P < 0.01), and shedding of pathogenic ETEC bacteria (P < 0.05) in feces was detected in the supplemented piglets. Administration of live yeast significantly increased (P < 0.05) IgA levels in the serum of piglets. Evidence indicates that decreased infection-related stress and severity of diarrhea in yeast-fed weaned piglets positively affected their growth capacity in the postweaning period (P < 0.05). The results suggest that dietary supplementation with live yeast S. cerevisiae to sows and piglets in the late gestation, suckling, and postweaning periods can be useful in the reduction of the duration and severity of postweaning diarrhea caused by ETEC.
- MeSH
- dieta veterinární MeSH
- enterotoxigenní Escherichia coli klasifikace MeSH
- fyziologie výživy v mateřství MeSH
- fyziologie výživy zvířat MeSH
- imunoglobulin A krev MeSH
- infekce vyvolané Escherichia coli prevence a kontrola veterinární MeSH
- krmivo pro zvířata MeSH
- nemoci prasat etiologie prevence a kontrola MeSH
- prasata růst a vývoj imunologie MeSH
- průjem etiologie prevence a kontrola veterinární MeSH
- Saccharomyces cerevisiae * MeSH
- těhotenství MeSH
- zvířata MeSH
- Check Tag
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky kontrolované MeSH
- práce podpořená grantem MeSH
Pharmacological dietary zinc (Zn) concentrations of 1,000 to 3,000 mg/kg diet from Zn oxide have been found to increase growth in post-weaning pigs. However, results were inconsistent among studies. A frequentist meta-analysis, in which effects were numerically described with standardized effect sizes (Hedges's g), was conducted in order to identify and quantify the responses in average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G/F) in post-weaning pigs upon dietary Zn supplementation from Zn oxide. The inability of independent continuous variables to explain significant heterogeneity obtained with fixed effect models necessitated the use of random effects models to calculate summary statistics. Dietary Zn supplementation increased (P < 0.05) ADG (mean effect size = 1.086, 95 % confidence intervals = 0.905-1.266, 26 studies, 72 comparisons), ADFI (mean effect size = 0.794, 95 % confidence intervals = 0.616-0.971, 25 studies, 71 comparisons), and G/F (mean effect size = 0.566, 95 % confidence intervals = 0.422-0.710, 24 studies, 70 comparisons). Zinc oxide provided a feasible alternative to in-feed antibiotics to improve growth in post-weaning pigs, and its reduction in diets due to potential environmental pollution will have to be negated by alternative feed additives and management strategies in order to prevent economic losses.
- MeSH
- fyziologie výživy zvířat * MeSH
- hmotnostní přírůstek účinky léků MeSH
- krmivo pro zvířata * analýza MeSH
- odstavení MeSH
- oxid zinečnatý aplikace a dávkování farmakologie MeSH
- potravní doplňky * MeSH
- prasata růst a vývoj MeSH
- přijímání potravy fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- metaanalýza MeSH
- práce podpořená grantem MeSH
A general transformation formula is presented for interval traits in pigs, such as weaning-to-first-service interval or farrowing interval. In the logarithmic transformation, only observations that are greater than the median of the trait as originally recorded are transformed. The transformation considerably reduces the skewness and kurtosis of the trait so that the distribution of transformed data approaches normality. In analysis of >12,000 records for weaning-to-first-service interval and >9,000 records for farrowing interval, heritability estimates for the transformed data were greater than for the data as recorded. Therefore, selection on the transformed trait should be more effective than selection on the original trait. The transformation formula is likely to be appropriate for other livestock species and traits.
