Glutamate, which is one of the most important contributors to oxidative metabolism in the intestinal mucosa, is mainly transported by the excitatory amino acids transporters (EAATs) that are expressed in enterocytes. The objective of this study was to evaluate the effects of in ovo administration of l-trans pyrrolidine-2,4-dicarboxylic acid (l-trans-PDC), a potent competitive inhibitor of glutamate uptake by EAATs, on the growth of the small intestine in chicks. Two series of experiments were conducted with hatching eggs; 100 μl of various l-trans-PDC solutions (0, 0.075 or 0.225 mg/egg for the Control group, low-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (L-PDC) or high-dose l-trans pyrrolidine 2,4-dicarboxylic acid group (H-PDC), respectively) was injected into the albumen sac of these hatching eggs before incubation. Hatchlings were sacrificed by cervical dislocation to determine the embryonic development in Experiment I, whereas the birds in Experiment II were raised or sampled at hatching, days 7 and 14 (D7 and D14) for further study. Gene expression in the small intestines was determined by real-time RT-PCR; and serum concentration of free amino acids was determined by an amino acid analyzer. The results showed that the hatchability was decreased by in ovo administration of l-trans-PDC. The small intestinal weights of the H-PDC group were decreased (P<0.05) at hatching and increased (P<0.05) on D7 and D14 compared with those in the Control group. In addition, the gene expression of EAAT2 in the completed or segmental small intestines was not changed (P>0.05); EAAT3 gene expression in the duodenum (P<0.05), jejunum (P=0.084) and ileum (P=0.060) on D14 was lower in the H-PDC group than in the Control group. Furthermore, the serum concentrations of free proline, threonine and phenylalanine but not glutamate or aspartate were increased (P<0.06) in H-PDC group. In conclusion, this paper is the first to report that in ovo administration of l-trans-PDC induces small intestinal growth retardation during the embryonic period and catch-up growth after hatching.

If loose-housed farrowing systems are to be an alternative to traditional farrowing crates, it is important that they can deliver the same production results as can be achieved in farrowing crates under commercial conditions. The aim of this study was to compare preweaning mortality in farrowing crates and free farrowing pens (FF-pens) within herds that had both systems. The study was conducted over 2 years in three commercial Danish herds that had FF-pens as well as traditional farrowing crates in their farrowing unit. Piglet mortality was analysed in two periods: before litter equalisation and after litter equalisation. Linear models were used to analyse effects of housing (crate or pen), herd (Herd A, B or C), parity (parities 1, 2, 3 to 4 or 5 to 8) as well as the effect of number of total born piglets on mortality before litter equalisation, and the effect of equalised litter size on piglet mortality after litter equalisation. All corresponding interactions were included in the models. Before litter equalisation piglet mortality was higher (P<0.001) in pens (13.7%) than in crates (11.8%). Similarly, piglet mortality after litter equalisation was higher in pens than in crates in all three herds, but the difference between pens and crates were dissimilar (P<0.05) in the different herds. In addition, piglet mortality, both before (P<0.001) and after litter equalisation (P<0.001), grew with increasing parity of the sows. Mortality before litter equalisation moreover increased with increasing number of total born piglets per litter (P<0.001), and mortality after equalisation increased when equalised litter size increased (P<0.001). No significant interactions were detected between housing and parity or housing and litter size for any of the analysed variables. In conclusion, there is knowledge how to design pens for free farrowing; but this study showed a higher preweaning mortality in the FF-pen. Nonetheless a noteworthy proportion of the sows in the FF-pens delivered results comparable to those farrowing in crates. This indicates that FF-pens are not yet a robust type of housing for farrowing sows.

Rapid rebreeding of winter- and spring-lambing ewes is essential if ewes are to lamb more than once per year, but fertility of lactating ewes is often low and early weaning of lambs may be undesirable in forage-based production systems. Selection to improve fertility in spring matings has been successful and led to development of ewes with a reduced seasonal anestrus. Potential for rapid rebreeding of lactating out-of-season breeding (OOS) ewes was tested in three studies. In Experiment 1, effects of short-term lamb removal on rebreeding was evaluated over 2 years using 71 January-lambing OOS ewes. At an average of 63 days postpartum, 36 ewes had lambs removed for 72 h, and all ewes were joined with rams. Circulating progesterone levels indicated that 74% of ewes ovulated before lamb separation; 91% of ewes mated within 5 weeks of ram exposure, 85% were diagnosed as pregnant and 75% lambed. The average interval between lambings was 225 days. In contrast to results observed in cattle, none of the measured variables was affected by lamb separation (P>0.20). Experiment 2 compared rebreeding performance of 24 OOS and 23 St. Croix ewes that lambed in January and averaged 60 days postpartum at ram introduction. More OOS ewes ovulated, mated and became pregnant during the first 21 days of ram exposure (83.3%, 58.3%, and 41.7%, respectively; P<0.001) compared with St. Croix ewes (26.1%, 0%, and 0%, respectively). After 39 days of ram exposure, pregnancy rates still favored OOS ewes (66.7% v. 39.1%; P=0.06), but the percentage of ewes that lambed did not differ (P>0.20) between OOS (47.8%) and St. Croix ewes (34.8%). In the third study, 34 March-lambing OOS ewes were exposed to rams on May 3 at an average of 40 days postpartum to characterize their reproductive performance. After 39 days of ram exposure, 52.9±8.7% of the ewes had mated, and 38.2±8.5% were diagnosed as pregnant. However, only 20.6±7.0% of the ewes produced viable lambs, suggesting a high level of uterine insufficiency. Spring fertility of lactating OOS ewes in these studies was one of the highest reported in the literature and indicated that selection for fertility in spring mating would improve reproductive performance in accelerated lambing programs. However, exposure of lactating OOS ewes to rams at 30 to 50 days postpartum was associated with high prenatal lamb mortality.

Despite the widespread adoption of hormonal synchronization protocols that allow for timed artificial insemination (AI), detection of estrus plays an important role in the reproductive management program on most dairies in the United States. Increased physical activity is a secondary sign of estrus in dairy cattle, and a new generation of electronic systems that continuously monitor physical activity to predict timing of AI have been developed and marketed to the dairy industry. A variety of management and physiologic challenges inhibit detection of behavioral estrus on farms, but the prevalence of anouvular cows near the end of the voluntary waiting period is particularly problematic. Only 70% of lactating Holstein cows were detected in estrus when using an activity monitoring system, with the remaining 20% of cows classified as anovular and 10% ovulating without showing signs of activity. Mean time of AI in relation to ovulation based on the activity monitoring system was acceptable for most of the cows with increased activity, however, variability in the duration of estrus and timing of AI in relation to ovulation could result in poor pregnancy outcomes in some cows. Use of a Presynch–Ovsynch protocol for submission of cows for first AI has been widely adopted by dairies in the United States, and a combined approach in which AI based on activity is followed by submission of cows not detected with activity to timed AI after synchronization of ovulation may be an effective strategy for submission of cows to first AI. Based on a field trial on a large commercial dairy in the United States, the activity monitoring system detected 70% of cows with increased activity after the second PGF2α injection of a Presynch–Ovsynch protocol, however, cows inseminated to increased activity had fewer pregnancies per AI (P/AI) compared with cows with increased activity after the second PGF2α injection that received timed AI after completing the Presynch–Ovsynch protocol. Based on an economic model comparing reproductive management programs with varying levels of AI to estrus v. timed AI, the rate of estrus detection and the P/AI to inseminations based on AI to detected estrus v. timed AI affected the decision to inseminate based on activity v. timed AI. In conclusion, an activity monitoring system detected increased activity in about 70% of lactating Holstein cows on a large commercial dairy in the United States, however, synchronization of ovulation and timed AI was beneficial to inseminate cows not detected with increased activity by the activity monitoring system.

The objective of this field study with an automatic milking system was to evaluate the effects of omitting the dry period on health and productivity during the subsequent lactation in dairy cows. A total of 98 German Simmental cows of six Southern German farms were assigned randomly to two experimental groups: The first group was dried-off 56 days before calving (D for dried-off, n=49), and the second group was milked continuously during this period until calving (CM for continuous milking, n=49). From the latter a third group emerged, including cows that dried-off themselves spontaneously (DS for dried-off spontaneously, n=14). Blood serum values of glucose, β-hydroxybutyrate (BHBA), non-esterified fatty acids (NEFA) and IGF-1 showed most pronounced fluctuations in D cows. Over the entire study period, the concentrations of BHBA and NEFA were markedly lower in the CM and DS groups. Furthermore, IGF-1 concentration was lowest for D cows and also decrease in back fat thickness was more pronounced. Mean concentration of milk protein was markedly higher in CM and DS cows (3.70% and 3.71%) compared with D cows (3.38%). Owing to the lower 305-day milk yield (−15.6%) and the lower total milk yield (−3.1%), the total amount of produced protein in the subsequent lactation was 2.5% (6.8 kg) lower, although the additional protein amount in CM cows from week −8 to calving was 35.7 kg. The greatest benefit resulted from positive effects on fertility and the lower incidence of diseases: CM cows had their first oestrus 1 week earlier compared with D cows, they also conceived earlier and showed a significantly lower risk of developing hypocalcaemia, ketosis and puerperal disorders. The present study showed that the costs of medical treatment and milk losses were twice as high in D cows, compared with CM and DS cows, and thus the reduced costs because of the more stable health outweighed the financial losses of milk yield by +18.49 € per cow and lactation.

Many traits associated with domestication are suggested to have developed as correlated responses to reduced fear of humans. Tameness may have reduced the stress of living in human proximity and improved welfare in captivity. We selected Red Junglefowl (ancestors of all domestic chickens) for four generations on high or low fear towards humans, mimicking an important aspect of the earliest period of domestication, and tested birds from the third and fourth generation in three different social tests. Growth and plumage condition, as well as size of eggs and offspring were also recorded, as indicators of some aspects of welfare. Birds selected for low fear had higher weight, laid larger eggs and generated larger offspring, and had a better plumage condition. In a social dominance test they also performed more aggressive behaviour and received less of the same, regardless of whether the restricted resource was feed or not. Hence, dominance appeared to increase as a consequence of reduced fear of humans. Furthermore, egg size and the weight of the offspring were larger in the less fearful birds, and plumage condition better, which could be interpreted as the less fearful animals being better adapted to the environment in which they were selected.

This study examined the effect of long-term selection of a maternal rabbit line, solely for a reproductive criterion, on the ability of female rabbits to deal with constrained environmental conditions. Female rabbits from generations 16 and 36 (n=72 and 79, respectively) of a line founded and selected to increase litter size at weaning were compared simultaneously. Female rabbits were subjected to normal (NC), nutritional (NF) or heat (HC) challenging conditions from 1st to 3rd parturition. Animals in NC and NF were housed at normal room temperatures (18°C to 25°C) and respectively fed with control (11.6 MJ digestible energy (DE)/kg dry matter (DM), 126 g digestible protein (DP)/kg DM, and 168 g of ADF/kg DM) or low-energy fibrous diets (9.1 MJ DE/kg DM, 104 g DP/kg DM and 266 g ADF/kg DM), whereas those housed in HC were subjected to high room temperatures (25°C to 35°C) and the control diet. The litter size was lower for female rabbits housed in both NF and HC environments, but the extent and timing where this reduction took place differed between generations. In challenging conditions (NF and HC), the average reduction in the reproductive performance of female rabbits from generation 16, compared with NC, was −2.26 (P<0.05) and −0.51 kits born alive at 2nd and 3rd parturition, respectively. However, under these challenging conditions, the reproductive performance of female rabbits from generation 36 was less affected at 2nd parturition (−1.25 kits born alive), but showed a greater reduction at the 3rd parturition (−3.53 kits born alive; P<0.05) compared with NC. The results also showed differences between generations in digestible energy intake, milk yield and accretion, and use of body reserves throughout lactation in NC, HC and NF, which together indicate that there were different resource allocation strategies in the animals from the different generations. Selection to increase litter size at weaning led to increased reproductive robustness at the onset of an environmental constraint, but failure to sustain the reproductive liability when the challenge was maintained in the long term. This response could be directly related to the short-term environmental fluctuations (less severe) that frequently occur in the environment where this line has been selected.

This study aimed to investigate the effects and possible interactions of birth weight and n-3 polyunsaturated fatty acid (PUFA) supplementation of the maternal diet on the fatty acid status of different tissues of newborn piglets. These effects are of interest as both parameters have been associated with pre-weaning mortality. Sows were fed a palm oil diet or a diet containing 1% linseed, echium or fish oil from day 73 of gestation. As fish oil becomes a scarce resource, linseed and echium oil were supplemented as sustainable alternatives, adding precursor fatty acids for DHA to the diet. At birth, the lightest and heaviest male piglet per litter were killed and samples from liver, brain and muscle were taken for fatty acid analysis. Piglets that died pre-weaning had lower birth weights than piglets surviving lactation (1.27±0.04 v. 1.55±0.02 kg; P<0.001), but no effect of diet on mortality was found. Lower DHA concentrations were observed in the brain of the lighter piglets compared with their heavier littermates (9.46±0.05 v. 9.63±0.04 g DHA/100 g fatty acids; P=0.008), suggesting that the higher incidence of pre-weaning mortality in low birth weight piglets may be related to their lower brain DHA status. Adding n-3 PUFA to the sow diet could not significantly reduce this difference in DHA status, although numerically the difference in the brain DHA concentration between the piglet weight groups was smaller when fish oil was included in the sow diet. Independent of birth weight, echium or linseed oil in the sow diet increased the DHA concentration of the piglet tissues to the same extent, but the concentrations were not as high as when fish oil was fed.

Thymol is the most common molecule in thyme and has been proposed as an oral alternative to antibiotics in the feed of pigs and broilers. The knowledge of the in vivo physiological effects of thymol on tissues is limited, particularly its impact on the gastric mucosa, where it is primarily absorbed when it is orally supplied. In this study, thymol (TH, 50 mg/ kg BW) or a placebo (CO) was introduced directly into the stomach of 8 weaned pigs that were slaughtered 12 h later and sampled for gastric oxyntic and pyloric mucosa. The analysis of whole transcript expression was performed using Affymetrix© Porcine Gene 1.1 ST array strips. Affymetrix Transcripts IDs were associated with 13 406 human gene names based on Sus scrofa Ensemble. Gene Set Enrichment Analysis was performed, comparing TH and CO pigs. For each gene set, the normalized enrichment score (NES) was defined as significant when the false discovery rate % was <25 and the P-value of NES was <0.05. In response to TH, 72 and 19 gene sets were significantly enriched in the oxyntic and pyloric mucosa, respectively. Several gene sets involved in mitosis and its regulation ranked near the top, primarily in the oxyntic mucosa; the gene set DIGESTION ranked first and ninth in the pyloric and oxyntic mucosa, respectively. Within this group, somatostatin (SST), SST receptors, peptide transporter 1 (SLC15A1) and calpain 9 (gastrointestinal tract-specific calpain) were the most strongly upregulated genes. Thymol reduced the enrichment of 120 and 59 gene sets in the oxyntic and pyloric mucosa, respectively. Several gene sets related to ion transport and channeling and aqueous pores across membranes, including short transient receptor potential (TRP) channel 4, potassium voltage-gated channel members 1 and 2, and ryanodine receptors 2 and 3, were less enriched. The downregulation of these genes sensitive to thymol in vitro could depend on the thymol dose and contact with the gastric tissues that causes an adaptive response with their reduced activation. Conversely, the activation of the TRPA1 gene (ranked 1072 and 128 among all the genes in the oxyntic and pyloric mucosa, respectively) indicates the involvement of another TRP-regulating cellular calcium storage. In conclusion, the stimulation of gastric proliferative activity and the control of digestive activity by thymol can influence positively gastric maturation and function in the weaned pigs. These properties should be considered in addition to thymol’s antimicrobial properties when supplementation of this molecule in feed is evaluated.

The metabolic state of pregnant mammals influences the offspring’s development and risk of metabolic disease in postnatal life. The metabolic state in a lactating dairy cow differs immensely from that in a non-lactating heifer around the time of conception, but consequences for their calves are poorly understood. The hypothesis of this study was that differences in metabolic state between non-lactating heifers and lactating cows during early pregnancy would affect insulin-dependent glucose metabolism and development in their neonatal calves. Using a mixed linear model, concentrations of glucose, IGF-I and non-esterified fatty acids (NEFAs) were compared between 13 non-lactating heifers and 16 high-yielding dairy cows in repeated blood samples obtained during the 1st month after successful insemination. Calves born from these dams were weighed and measured at birth, and subjected to intravenous glucose and insulin challenges between 7 and 14 days of age. Eight estimators of insulin-dependent glucose metabolism were determined: glucose and insulin peak concentration, area under the curve and elimination rate after glucose challenge, glucose reduction rate after insulin challenge, and quantitative insulin sensitivity check index. Effects of dam parity and calf sex on the metabolic and developmental traits were analysed in a two-way ANOVA. Compared with heifers, cows displayed lower glucose and IGF-I and higher NEFA concentrations during the 1st month after conception. However, these differences did not affect developmental traits and glucose homeostasis in their calves: birth weight, withers height, heart girth, and responses to glucose and insulin challenges in the calves were unaffected by their dam’s parity. In conclusion, differences in the metabolic state of heifers and cows during early gestation under field conditions could not be related to their offspring’s development and glucose homeostasis.

The production of pork with high amounts of intramuscular fat (IMF) without an increase in subcutaneous fat is highly desirable for the pig industry and consumers. Herein, we question the impact of dietary protein reduction (18% v. 13%) on fat deposition in the subcutaneous adipose tissue (SAT) and longissimus lumborum (LL) muscle using genetically diverse pigs for body fatness (lean v. fat). A clear effect of genotype was observed on plasma insulin (P=0.004) and leptin (P<0.001), as well as on backfat thickness (P<0.001), with the fat pigs having higher values. Accordingly, IMF was higher in the fat pigs, when compared with their lean counterparts (P=0.003), which was supported by enlarged adipocytes (P<0.001). The area of lipid droplets within the LL fibres (P=0.039) and extramyocellular lipids number (P=0.017) were increased in pigs fed reduced protein diets, regardless of genotype, which is consistent with higher levels of plasma triacylglycerols (P=0.002). The gene-expression pattern of lipogenic factors in the SAT was distinct from the LL muscle. In the SAT, PPARG expression was similar among genotypes (P>0.05), whereas in the LL muscle it was higher in the lean pigs (P=0.023), especially when fed on low protein diet (P=0.057). The CEBPA and FABP4 mRNA levels were increased in the SAT of fat pigs (P<0.001), without changes in the LL muscle (P>0.05). The influence of diet on FABP4 expression in the SAT was dependent on pig’s genetic background (P=0.005). In conclusion, fat deposition was clearly influenced by genotype and, to a lesser extent, by dietary protein level, the SAT being more sensitive than the LL muscle. One can speculate that the pathways involved in lipid metabolism are downregulated in intramuscular adipocytes when compared with SAT fat cells. This result might be a direct consequence of the relatively low proportion of adipocytes found in the LL muscle.

Lameness in sows is an animal welfare problem which also presents an economic challenge to pig producers. Information about the prevalence of herd lameness in organic sows is relatively scarce. The first objective of this study was to establish the prevalence of lameness and to identify risk factors associated with sow lameness in Danish outdoor organic sow herds by analysing the association between risk factors at both sow and herd level using clinical records of lameness. A total of 1850 sows from nine organic herds were included in the study. Second, the study examined differences in the prevalence of sow lameness between outdoor organic and indoor conventional herds. An additional aim here was to identify risk factors associated with clinical records of sow lameness in Danish sow herds by analysing the association between risk factors with lameness at sow and herd level. One thousand and fifty four gestation sows from 44 indoor conventional and nine organic sow herds were included in this study. The nine organic herds were visited twice: once in summer/autumn 2011, and once in winter/spring 2012. In winter/spring 2011, a total of 44 indoor conventional herds were visited. Risk factors included in the study were clinical parameters and factors related to the production system. Sows were examined visually by one of four trained observers. The organic sows were assigned scores for lameness, body condition, hoof length, bursitis, abscesses and leg wounds, while the conventional sows were assigned scores for lameness, body condition and bursitis. A multivariable analysis was carried out by logistic regression with the herd and observer as random effects. The average herd lameness prevalence in gestation and lactation sows in organic herds was 11% in summer/autumn and 4.6% in winter/spring. ‘Wounds, bursitis and abscess’ on legs (OR=4.7, P<0.001) and body condition score >3 (OR=1.79, P=0.008) were associated with increased risk of lameness in Danish organic sow herds. Season (winter/spring v. summer/autumn) lowered the risk of lameness (OR=0.37, P<0.001). Average prevalence of lameness in gestation sow herds in winter/spring in conventional herds was 24.4%, and in organic herds it was 5.4%. An organic sow had a decreased risk of lameness (OR=0.28, P<0.001) as compared with a conventional sow. Bursitis was associated with increased risk of lameness (OR=2.08, P=0.002) regardless of the production system (i.e. whether the herd was organic or conventional).

The objective of the present study was to investigate the effects of dietary-induced insulin enhancement during the late luteal phase on subsequent fertility of gilts. Fifty-two littermate cyclic gilts were subjected to dietary treatments where two energy sources were tested: corn starch (T1) and soybean oil (T2). The experimental diets were supposed to provide similar amounts of dietary energy, but from different sources. Gilts were fed ad libitum, starting day 8 of the estrous cycle, until the next standing heat. Blood sampling was performed in a subgroup of 20 gilts on days 14 and 21 of the cycle for analyses of glucose and insulin, and after ovulation detection until 18 h after ovulation for progesterone. All gilts were slaughtered on day 28 of pregnancy and the reproductive tracts recovered for further analysis. T1 gilts showed higher postprandial insulin peak on days 14 and 21 and lower glucose levels 4 h after feeding on day 14 (P<0.05), however, there were no treatment effects on plasma progesterone concentrations. Dietary energy sources did not affect average daily feed intake, body weight and backfat on day 28 of pregnancy. Estrous cycle length, estrus duration and time of ovulation were not affected by previous nutritional treatments either. T1 gilts showed higher ovulation rates, number of embryos, embryo weight and placental weight (P<0.05). There were no treatment effects on pregnancy rate, embryo survival rate and volume of amniotic fluid. A positive correlation between progesterone concentration 18 h after ovulation and ovulation rate was observed (r=0.75; P<0.01). These results suggest that it is possible to manipulate dietary insulin response in cyclic gilts and, thus, improve reproductive efficiency when feeding starch as the main energy source during the late luteal and follicular phases of the cycle.

Consistency over time of (on-farm) animal welfare assessment systems forms part of reliability, meaning that results of the assessment should be representative of the longer-term welfare state of the farm as long as the housing and management conditions have not changed considerably. This is especially important if assessments are to be used for certification purposes. It was the aim of the present study to investigate consistency over time of the Welfare Quality® (WQ®) assessment system for fattening cattle at single measure level, aggregated criterion and principle scores, and overall classification across short-term (1 month) and longer-term periods (6 months). We hypothesized that consistency over time of aggregated criterion and principle scores is higher than that of single measures. Consistency was also expected to be lower with longer intervals between assessments. Data were obtained using the WQ® protocol for fattening cattle during three visits (months 0, 1 and 7) on 63 beef farms in Austria, Germany and Italy. Only data from farms where no major changes in housing and management had taken place were considered for analysis. At the single measure level, Spearman rank correlations between visits were >0.7 and variance was lower within farms than between farms for six and two of 19 measures after 1 month and 6 months, respectively. After aggregation of single measures into criterion and principle scores, five and two of 10 criteria and three and one of four principles were found reliable after 1 and 6 months, respectively. At the WQ® principle level, this was the case for three and one of four principles. Seventy-nine per cent and 75% of the farms were allocated to the same overall welfare category after 1 month and 6 months. Possible reasons for a lack of consistency are seasonal effects or short-term fluctuations that occur under normal farm conditions, low prevalence of clinical measures and probably insufficient sample size, whereas poor inter-observer agreement leading to inflation of correlation can be ruled out. At the criterion and principle level, aggregation of information into scores appears to partly smoothen undirected variation at the single measure level without losing sensitivity in terms of welfare evaluation. Reliable on-farm animal welfare assessments should therefore be based on repeated assessments. Further long-term studies are recommended to better understand the factors influencing consistency over time.

The world’s first cloned swamp buffalo (Bubalus bubalis) derived from adult ear skin fibroblast has been reported. Donor fibroblast cells were produced from biopsies taken from adult male ear skin and in vitro matured oocytes obtained from a slaughterhouse were used as cytoplasts. A total of 39 blastocysts and 19 morulae fresh embryos were transferred into 12 recipient buffaloes. Progesterone assays indicated establishment of pregnancy in 10 of the 12 buffaloes (83.3%) after 45 days, with six animals still pregnant at 3 months. One recipient maintained pregnancy to term and naturally delivered a 40 kg male calf after 326 days of gestation. DNA analysis showed that the cloned calf was genetically identical to the donor cells. Genotype analyses, using 12 buffalo microsatellite markers, confirmed that the cloned calf was derived from the donor cell lines. In conclusion, the present study reports, for the first time, the establishment of pregnancy and birth of the first cloned Thai swamp buffalo derived from adult ear skin fibroblast cells.

MicroRNAs are endogenous ~22nt RNAs that negatively regulate gene expression at the posttranscriptional level via binding to the 3′-untranslated region (3′UTR) of target mRNAs. The microRNA miR-27a was reported to depress the expression of myostatin, a critical inhibitor of skeletal myogenesis, by binding to its 3′UTR in mouse. In this study, we cloned the full-length 3′UTR of porcine myostatin by rapid amplification of 3′-cDNA ends (3′-RACE) and demonstrated that the 3′UTR of porcine myostatin is targeted by miR-27a. The phenomenon that the level of myostatin inversely correlated with miR-27a was observed in fat and heart of pigs and also in proliferating porcine myoblasts. Besides, overexpression of miR-27a in porcine myoblasts promoted cell proliferation by reducing the expression of myostatin. Our data suggest that miR-27a positive regulates porcine myoblast proliferation via targeting myostatin.

The main objective of the implementation of artificial insemination (AI) in cattle is to produce a sustained genetic progress in the herd. Although AI is an old reproductive biotechnology, its widespread implementation is very recent and is mainly because of the use of protocols that allows AI without oestrus detection, commonly called fixed-time artificial insemination (FTAI). The development of FTAI protocols also allowed the application of AI in larger, extensively managed herds and especially in suckled cows instead of restricting the breeding programmes to the heifers. Fixed-time AI treatments are widely used in South America, with about 3 000 000 cows inseminated in the last season in Argentina and about 8 000 000 in Brazil. The objective of this review is to present and describe the various treatments available and some of the factors that may affect pregnancy rates in beef cattle.

The characterization of blood metabolite concentrations over the circadian period and across physiological stages is important for understanding the biological basis of feed efficiency, and may culminate in indirect methods for assessing feed efficiency. Hematological analyses for albumin, urea, creatine kinase, glutamate dehydrogenase, aspartate aminotransferase, carbon dioxide, and acetate were carried out in growing and gestating heifers. These measures were carried out in a sample of 36 Bos taurus crossed beef heifers held under the same husbandry conditions. Hourly blood samples were collected over a 24-h period on three separate sampling occasions, corresponding approximately to the yearling (and open), early-gestation and late-gestation stages. This design was used to determine variation throughout the day, effects due to physiological status and any associations with feed efficiency, as measured by residual feed intake. Blood analyte levels varied with time of day, with the most variation occurring between 0800 and 1600 h. There were also considerable differences in analyte levels across the three physiological stages; for example, creatine kinase was higher (P<0.05) in open heifers, followed by early- and late-gestation heifers. Feed efficiency was also associated with analyte abundance. In more feed-efficient open heifers, there were higher activities of creatine kinase (P<0.05) and aspartate aminotransferase (P<0.05), and lower concentrations of carbon dioxide (P<0.05). Furthermore, in late gestation, more efficient heifers had lower urea concentrations (P<0.05) and lower creatine kinase levels (P<0.05). Over the whole experimental period, carbon dioxide concentrations were numerically lower in more feed efficient heifers (P=0.079). Differences were also observed across physiological stages. For instance, open heifers had increased levels (P<0.05) of creatine kinase, aspartate aminotransferase, carbon dioxide than early and late pregnancy heifers. In essence, this study revealed relevant information about the metabolic profile in the context of feed efficiency and physiological stages. Further optimization of our approach, along with the evaluation of complementary analytes, will aid in the development of robust, indirect assessments of feed efficiency.

In pasture-based automatic milking systems (AMS), feed is the main incentive that can be managed to encourage reliable and consistent voluntary and distributed cow traffic. Modifying timing, placement and size of feed allocations is expected to have an effect on cow behavior that could avoid the occurrence of extended milking intervals, which have a negative effect on milk yield. Therefore, behavioral studies provide information on how cows modify their actions under different management regimes and can help explain the impact of those regimes. Behavioral observations were conducted in spring 2011 at the FutureDairy AMS research farm, as part of a study where a herd of 175 cows was split into two groups that received supplementary feed either before (PRE), or immediately after (POST) milking. In addition, all cows were offered access to two daily pasture allocations. Observations were conducted in the pasture allocation on 15 focal cows from each treatment group during four periods of 24 h to detect presence and behavior (grazing, ruminating, idling and other) every 15 min. In addition, bite rate and pasture biomass were measured every hour. Overall, despite the finding that more POST cows than PRE cows entered the pasture allocation during the first 8 h of active access, there was no difference in the total proportion of cows that had gained access by the end of the active access period (average 68% for both treatments). Cows in the PRE treatment started exiting the pasture allocation just 6 h after entering, compared with 8 h for POST cows, although their behaviors in the pasture allocation did not differ. Behaviors and bite rate were more dependent on pasture biomass than on supplementary feeding management.

A prototype of an integrated closed system for fish-plankton aquaculture was developed in Iquitos (Peruvian Amazonia) in order to cultivate the Tiger Catfish, Pseudoplatystoma punctifer (Castelnau, 1855). This freshwater recirculating system consisted of two linked sewage tanks with an intensive rearing unit (a cage) for P. punctifer placed in the first, and with a fish-plankton trophic chain replacing the filters commonly used in clear water closed systems. Detritivorous and zooplanktivorous fishes (Loricariidae and Cichlidae), maintained without external feeding in the sewage volume, mineralized organic matter and permitted the stabilization of the phytoplankton biomass. Water exchange and organic waste discharge were not necessary. In this paper we describe the processes undertaken to equilibrate this ecosystem: first the elimination of an un-adapted spiny alga, Golenkinia sp., whose proliferation was favored by the presence of a small rotifer, Trichocerca sp., and second the control of this rotifer proliferation via the introduction of two cichlid species, Acaronia nassa Heckel, 1840 and Satanoperca jurupari Heckel, 1840, in the sewage part. This favored some development of the green algae Nannochloris sp. and Chlorella sp. At that time we took the opportunity to begin a 3-month rearing test of P. punctifer. The mean specific growth rate and feed conversion ratio (FCR) of P. punctifer were 1.43 and 1.27, respectively, and the global FCR, including fish in the sewage part, was 1.08. This system has proven to be suitable for growing P. punctifer juveniles out to adult, and provides several practical advantages compared with traditional recirculating clear water systems, which use a combination of mechanical and biological filters and require periodic waste removal, leading to water and organic matter losses.