ObjectiveThe aim of our study was to determine the associations of heifer reproductive performance with survival up to the first calving, first-lactation milk yield, and the probability of being culled within 50 days after first calving.MethodsData from 33 large Holstein-Friesian commercial dairy herds were gathered from the official milk recording database in Hungary. The data of heifers first inseminated between January 1, 2011 and December 31, 2014 were analyzed retrospectively, using Cox proportional hazards models, competing risks models, multivariate linear and logistic mixed-effects models.ResultsHeifers (n = 35,128) with younger age at conception were more likely to remain in the herd until calving, and each additional month in age at conception increased culling risk by 5.1%. Season of birth was related to first-lactation milk yield (MY1; n = 19,931), with cows born in autumn having the highest milk production (p<0.001). The highest MY1 was achieved by heifers that first calved between 22.00 and 25.99 months of age. Heifers that calved in autumn had the highest MY1, whereas calving in summer was related to the lowest milk production (p<0.001). The risk of culling within 50 days in milk in first lactation (n = 21,225) increased along with first calving age, e.g. heifers that first calved after 30 months of age were 5.52-times more likely to be culled compared to heifers that calved before 22 months of age (p<0.001). Calving difficulty was related to higher culling risk in early lactation (p<0.001). Heifers that required caesarean section were 24.01-times more likely to leave the herd within 50 days after first calving compared to heifers that needed no assistance (p<0.001).ConclusionReproductive performance of replacement heifers is closely linked to longevity and milk production in dairy herds.

ObjectiveWe examined the localization and expression of H+ pumping vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) in the epididymis of pigs, expressed in clear and basal cells, respectively, during postnatal development.MethodsEpididymides were obtained from pigs at 1, 7, 21, 60, 120, and 180 days of age; we observed the localization and expression patterns of V-ATPase and KRT5 in the different regions of these organs, namely, the caput, corpus, and cauda. The differentiation of epididymal epithelial cells was determined by immunofluorescence labeling using cell-type-specific markers and observed using confocal microscopy.ResultsAt postnatal day 5 (PND5), the localization of clear cells commenced migration from the cauda toward the caput. Although at PND120, goblet-shaped clear cells were detected along the entire length of the epididymis, those labeled for V-ATPase had disappeared from the corpus to cauda and were maintained only in the caput epididymis in adult pigs. In contrast, whereas basal cells labeled for KRT5 were only present in the vas deferens at birth, they were detected in all regions of the epididymis at PND60. These cells were localized at the base of the epithelium; however, no basal cells characterized by luminally extending cell projections were observed in any of the adult epididymides examined.ConclusionThe differentiation of clear and basal cells progressively initiates in a retrograde manner from the cauda to the caput epididymis. The cell-type-specific distribution and localization of the epithelial cells play important roles in establishing a unique luminal environment for sperm maturation and storage in the pig epididymis.

ObjectiveThis study assessed the effects of probiotics on cecal microbiota, gene expression of intestinal tight junction proteins, and immune response in the cecal tonsil of broiler chickens challenged with Salmonella enterica subsp. enterica.MethodsOne-day-old broiler chickens (n = 240) were randomly allocated to four treatments: negative control (Cont), multi-strain probiotic-treated group (Pro), Salmonella-infected group (Sal), and multi-strain probiotic-treated and Salmonella-infected group (ProSal). All chickens except those in the Cont and Pro groups were gavaged with 1×108 cfu/mL of S. enterica subsp. enterica 4 days after hatching.ResultsOur results indicated that body weight, weight gain, and feed conversion ratio of birds were significantly reduced (p<0.05) by Salmonella challenge. Chickens challenged with Salmonella decreased cecal microbial diversity. Chickens in the Sal group exhibited abundant Proteobacteria than those in the Cont, Pro, and ProSal groups. Salmonella infection downregulated gene expression of Occludin, zonula occludens-1 (ZO1), and Mucin 2 in the jejunum and Occludin and Claudin in the ileum. Moreover, the Sal group increased gene expression of interferon-γ (IFN-γ), interleukin-6 (IL-6), IL-1β, and lipopolysaccharide-induced tumor necrosis factor-alpha factor (LITAF) and reduced levels of transforming growth factor-β4 and IL-10 compared with the other groups (p<0.05). However, chickens receiving probiotic diets increased Lactobacillaceae abundance and reduced Enterobacteriaceae abundance in the ceca. Moreover, supplementation with probiotics increased the mRNA expression of Occludin, ZO1, and Mucin 2 in the ileum (p<0.05). In addition, probiotic supplementation downregulated the mRNA levels of IFN-γ (p<0.05) and LITAF (p = 0.075) and upregulated IL-10 (p = 0.084) expression in the cecal tonsil.ConclusionThe administration of multi-strain probiotics modulated intestinal microbiota, gene expression of tight junction proteins, and immunomodulatory activity in broiler chickens.

ObjectiveThe efficiency of the knock-in process is very important to successful gene editing in domestic animals. Recently, it was reported that transient loosening of the nucleosomal folding of transcriptionally inactive chromatin might have the potential to enhance homologous recombination efficiency. The objective of this study was to determine whether histone deacetylases (HDAC) inhibitor and RAD51 recombinase (RAD51) expression were associated with increased knock-in efficiency on the β-casein (bCSN2) gene locus in mammary alveolar-large T antigen (MAC-T) cells using the transcription activator-like effector nucleases (TALEN) system.MethodsMAC-T cells were treated with HDAC inhibitors, valproic acid, trichostatin A, or sodium butyrate for 24 h, then transfected with a knock-in vector, RAD51 expression vector and TALEN to target the bCSN2 gene. After 3 days of transfection, the knock-in efficiency was confirmed by polymerase chain reaction and DNA sequencing of the target site.ResultsThe level of HDAC 2 protein in MAC-T cells was decreased by treatment with HDAC inhibitors. The knock-in efficiency in MAC-T cells treated with HDAC inhibitors was higher than in cells not treated with inhibitors. However, the length of the homologous arm of the knock-in vector made no difference in the knock-in efficiency. Furthermore, DNA sequencing confirmed that the precision of the knock-in was more efficient in MAC-T cells treated with sodium butyrate.ConclusionThese results indicate that chromatin modification by HDAC inhibition and RAD51 expression enhanced the homologous recombination efficiency on the bCSN2 gene locus in MAC-T cells.

ObjectiveMeat quality characteristics and sensory attributes were evaluated in three age groups (12, 18, and 24 mo) of one-humped camels of the Saudi Arabian Najdi breed.MethodsThirty-six male camels (12 for each age group) were used. The Longissimus dorsi muscle from each carcass was divided into three parts and subjected to three ageing periods (1, 5, or 10 d) and evaluated for shear force, myofibril fragmentation index (MFI), expressed juice, cooking loss, and sensory attributes.ResultsAge had a significant effect on shear force, MFI, expressed juice quantity, and organoleptic properties. Camels slaughtered at 12 mo exhibited lower shear force and MFI, and higher expressed juice quantity, tenderness, juiciness, and overall acceptability than those slaughtered at 24 mo. Ageing had a significant influence on shear force, MFI, expressed juice quantity, but not on cooking loss. Camel meat aged for 10 d exhibited significantly lower shear force values and expressed juice quantity, and significantly higher MFI compared to that aged for 1 d. However, ageing did not significantly affect sensory attributes, except for tenderness, in camels slaughtered at 18 mo.ConclusionBoth instrumental and sensory evaluations showed that young camel meat has desirable quality characteristics, with superior tenderness and juiciness.

ObjectiveThis study was conducted to determine the composition and diversity of the fungal flora at various control points in cheese ripening rooms of 10 dairy farms from six different provinces in the Republic of Korea.MethodsFloor, wall, cheese board, room air, cheese rind and core were sampled from cheese ripening rooms of ten different dairy farms. The molds were enumerated using YM petrifilm, while isolation was done on yeast extract glucose chloramphenicol agar plates. Morphologically distinct isolates were identified using sequencing of internal transcribed spacer region.ResultsThe fungal counts in 8 out of 10 dairy farms were out of acceptable range, as per hazard analysis critical control point regulation. A total of 986 fungal isolates identified and assigned to the phyla Ascomycota (14 genera) and Basidiomycota (3 genera). Of these Penicillium, Aspergillus, and Cladosporium were the most diverse and predominant. The cheese ripening rooms was overrepresented in 9 farms by Penicillium (76%), while Aspergillus in a single farm. Among 39 species, the prominent members were Penicillium commune, P. oxalicum, P. echinulatum, and Aspergillus versicolor. Most of the mold species detected on surfaces were the same found in the indoor air of cheese ripening rooms.ConclusionThe environment of cheese ripening rooms persuades a favourable niche for mold growth. The fungal diversity in the dairy farms were greatly influenced by several factors (exterior atmosphere, working personnel etc.,) and their proportion varied from one to another. Proper management of hygienic and production practices and air filtration system would be effective to eradicate contamination in cheese processing industries.

ObjectiveHuman mesenchymal stromal cells (MSCs) exhibit variable differentiation potential and can be divided accordingly into distinct subpopulations whose ratios vary with donor age. However, it is unknown whether the same is true in pigs. This study investigated MSC subpopulations in miniature pig and compared their characteristics in young (2 to 3 months) and adult (27 to 35 months) pigs.MethodsOsteogenic, chondrogenic, and adipogenic capacity of isolated MSCs was evaluated by von Kossa, Alcian blue, and oil red O staining, respectively. Cell surface antigen expression was determined by flow cytometry. Proliferative capacity was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Expression of marker genes was detected by quantitative real-time polymerase chain reaction.ResultsPorcine MSCs comprised cells with trilineage and bilineage differentiation potential (tMSCs and bMSCs, respectively) and non-differentiating stromal cells (NDSCs). The tMSC and bMSC fractions were smaller in adult than in young pigs (63.0% vs 71.2% and 11.6% vs 24.0%, respectively, p<0.05); NDSCs showed the opposite trend (25.4% vs 4.8%; p<0.05). Subpopulations showed no differences in morphology, cell surface antigen expression, or proliferative capacity, but octamer-binding transcription factor 4 (OCT4) expression was higher in tMSCs than in bMSCs and NDSCs (p<0.05), whereas sex determining region Y-box 2 (SOX2) expression was higher in tMSCs and bMSCs than in NDSCs (p<0.05). Aging had no effect on these trends.ConclusionPorcine MSCs comprise distinct subpopulations that differ in their differentiation potential and OCT4 and SOX2 expression. Aging does not affect the characteristics of each subpopulation but alters their ratios.

ObjectiveEggshells with a uniform color and intensity are important for egg production because many consumers assess the quality of an egg according to the shell color. In the present study, we evaluated the influence of dominant effects on the variations in eggshell color after 32 weeks in a crossbred population.MethodsThis study was conducted using 7,878 eggshell records from 2,626 hens. Heritability was estimated using a univariate animal model, which included inbreeding coefficients as a fixed effect and animal additive genetic, dominant genetic, and residuals as random effects. Genetic correlations were obtained using a bivariate animal model. The optimal diagnostic criteria identified in this study were: L* value (lightness) using a dominance model, and a* (redness), and b* (yellowness) value using an additive model.ResultsThe estimated heritabilities were 0.65 for shell lightness, 0.42 for redness, and 0.60 for yellowness. The dominance heritability was 0.23 for lightness. The estimated genetic correlations were 0.61 between lightness and redness, −0.84 between lightness and yellowness, and −0.39 between redness and yellowness.ConclusionThese results indicate that dominant genetic effects could help to explain the phenotypic variance in eggshell color, especially based on data from blue-shelled chickens. Considering the dominant genetic variation identified for shell color, this variation should be employed to produce blue eggs for commercial purposes using a planned mating system.

ObjectiveAn experiment was conducted to determine the effect of partial replacement of soybean meal (SBM) by white lupine seeds (WLS) on milk yield and quality, feed efficiency and rumen fermentation of high-yielding dairy cows.MethodsThirty multiparous cows of two breeds (20 Holstein and 10 Czech Pied cows) in early mid-lactation received three diets (treatments) in a 3×3 Latin square design with a 28-d period. The dietary treatments were as follows: CON (control total mixed ration with SBM, no WLS), WLS30 (30% of the SBM was replaced, on a dry matter basis, by WLS), and WLS50 (50% of the SBM was replaced by WLS).ResultsFeed intake by the cows was not affected (p = 0.331) by the diets. Milk production decreased with increasing proportions of WLS in the diet. Cows fed WLS50 yielded approximately 1 kg/d (p<0.001) less milk than cows fed the CON diet. The proportions of milk fat (p = 0.640), protein (p = 0.507), and lactose (p = 0.709) were not altered by the diet. For milk fat, feeding with WLS50 reduced the proportion of total saturated fatty acids (p<0.001) and increased the proportion of total monounsaturated fatty acids (p<0.001), mainly through oleic acid (p<0.001). No differences were found in feed efficiency, body weight, and blood plasma metabolites between groups. Rumen ammonia-N levels tended (p = 0.087) to increase with increasing proportions of WLS in the diet, whereas no effect of diet on rumen pH was found (p = 0.558).ConclusionWe did not identify the safe range within which raw WLS can efficiently replace SBM in the diet of high-producing dairy cows. In contrast, even partial replacement of SBM by WLS favorably changed the milk fatty acid profile.

ObjectiveTo evaluate the effects of the carmine cochineal-resistant spineless cactus genotypes cv. Orelha de Elefante Mexicana (Opuntia) and Miúda (Nopalea) on the intake and digestibility of nutrients, ingestive behavior, performance, and ultrasound measurements of growing lambs.MethodsThirty-six male (non-castrated) Santa Inês lambs were used, with an average age of 6 months and an initial average weight of 22.0±2.9 kg. They were distributed in a completely randomized design with 3 treatments (Tifton hay, Nopalea and Opuntia) and 12 replications, using initial weight as a covariate. The experimental period was 86 days, with the first 30 days used for the adaptation of the animals to the facilities, diets and management, and the remaining 56 days used for evaluation and data collection.ResultsThe intake and apparent digestibility of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), total carbohydrates (TC), non-fibrous carbohydrates (NFC), and total digestible nutrients (TDN) showed a significant difference (p<0.05) as a function of the diets, with the Nopalea treatment (p<0.05) increasing DM intake (g/kg and % body weight [BW]), CP, TDN, and TC digestibility, whereas the Tifton hay diet led to a high (p<0.001) neutral detergent fiber corrected for ash and protein (NDFap) g/d intake, NDFap (BW %) and digestibility of said nutrient. There was no effect of treatments (p>0.05) on feeding time, however, rumination time and total chewing time were higher (p<0.05) for animals fed Tifton hay. The performance of the animals was similar (p>0.05). For the ultrasound measurements, Nopalea promoted an increase in the final loin eye area, compared to Tifton hay.ConclusionThe use of spineless cactus variety Miúda leads to the greater intake and digestibility of nutrients. The evaluated carmine cochineal-resistant spineless cactus genotypes are alternatives for semi-arid regions as they do not negatively affect the performance of growing lambs.