ALBERT

Description: Real-time PCR is widely used to study the relative abundance of mRNA due to its specificity, sensitivity, and repeatability quantification. However, relative quantification requires a reference gene, which should be stable in its expression, showing lower variation by experimental conditions or tissues. The aim of this study was to evaluate the stability of the expression of five commonly used reference genes (actb, ywhaz, b2m, sdha, and 18s rRNA) at different physiological stages (alert and emergency) in three different cattle breeds. In this study, five genes (actb, ywhaz, b2m, sdha, and 18s rRNA) were selected as candidate reference genes for expression studies in the whole blood from three cattle breeds (Romosinuano, Gyr, and Brahman) under heat stress conditions. The transcription stability of the candidate reference genes was evaluated using geNorm and NormFinder. The results showed that actb, 18SrRNA, and b2m expression were the most stable reference genes for whole blood of Gyr and Brahman breeds under two states of livestock weather safety (alert and emergency). Meanwhile, actb, b2m, and ywhaz were the most stable reference genes for the Romosinuano breed.

Description: This study aimed to investigate the effects of multi-bacteria solid-state fermented diets with different crude fiber (CF) levels on growth performance, nutrient digestibility, and microbial flora of finishing pigs. The multi-bacteria solid-state fermented diets were made up of Lactobacillus amylovorus, Enterococcus faecalis, Bacillus subtilis, and Candida utilis. According to a 2 (factors) × 2 (levels) design, with the two factors being multi-bacteria solid-state fermentation (fed non-fermented diet or multi-bacteria fermentation) or CF levels (fed a basal diet containing 2.52% CF or 7.00% CF), a total of 36 finishing pigs (70.80 ± 5.75 kg) were divided into 4 treatments with 9 barrows per group: (1) pigs fed a diet containing 7.00% CF (HF), (2) pigs fed a multi-bacteria fermentation diet containing 7.00% CF (HFM), (3) pigs fed a diet containing 2.52% CF (LF), and (4) piglets fed a multi-bacteria fermentation diet containing 2.52% CF (LFM). This experiment lasted 28 days. The multi-bacteria solid-state fermented diet increased the backfat thickness (p 〈 0.05) and apparent total tract nutrient digestibility (ATTD) of CF, neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), 8 amino acids (Trp, Asp, Gly, Cys, Val, Met, Ile, and Leu), total essential amino acids (EAA), total non-essential amino acids (NEEA), and total amino acids (TAA) (p 〈 0.05). Multi-bacteria solid-state fermented diet increased serum concentrations of HDL-c, ABL, TP, and GLU, the serum enzyme activities of GSH-Px, T-AOC, SOD, and CAT (p 〈 0.05), the relative abundance of Lactobacillus, Oscillospira, and Coprococcus (p 〈 0.05), and the abundance of YAMINSYN3-PWY, PWY-7013, GOLPDLCAT-PWY, ARGORNPROST-PWY, and PWY-5022 pathways (p 〈 0.05). The multi-bacteria solid-state fermented diet reduced the digestion amount of CF, NDF, and ADF (p 〈 0.05), the serum concentrations of TC, TG, LDL-c, BUN, and MDA (p 〈 0.05), the relative abundance of Streptococcaceae (p 〈 0.05), and the abundance of PWY-6470, PWY0-862, HSERMETANA-PWY, LACTOSECAT-PWY, MET-SAM-PWY, PWY-6700, PWY-5347, PWY0-1061, and LACTOSECAT-PWY pathways (p 〈 0.05). The high-fiber diet increased average daily feed intake (p 〈 0.05), the serum concentrations of TC, TG, LDL-c, BUN, and MDA (p 〈 0.05), the relative abundance of Clostridiaceae_Clostridium and Coprococcus (p 〈 0.05), and the abundance of TCA-GLYOX-BYPASS, GLYCOLYSIS-TCA-GLYOX-BYPASS, and PWY-6906 pathways (p 〈 0.05). The high-fiber diet reduced chest circumference (p 〈 0.05) and ATTD of ether extract (EE), CF, NDF, ADF, Ca, CP, 18 amino acids (Trp, Thr, Val, Met, Ile, Leu, Phe, Lys, His, Arg Asp, Ser, Glu, Gly, Ala, Cys, Tyr, and Pro), EAA, NEAA, and TAA (p 〈 0.05). The high-fiber diet also reduced the serum concentrations of HDL-c, TP, ABL, and GLU, the serum enzyme activities of T-AOC, GSH-Px, SOD, and CAT (p 〈 0.05), and the relative abundance of Akkermansia and Oscillospira (p 〈 0.05). There was no significant effect of the interaction between multi-bacteria fermentation and dietary CF levels, except on the digestion amount of CF (p 〈 0.05). The 7.00% CF had a negative effect on the digestion of nutrients, but multi-bacteria solid-state fermentation diets could relieve this negative effect and increase backfat thickness. High-fiber diets and multi-bacteria solid-state fermentation improved the diversity and abundance of fecal microorganisms in finishing pigs.

Description: The in-vitro ileal digestibility of dry matter (DM), organic matter (OM), and crude protein (CP) of field beans treated with propionic acid (trFB) and extruded trFB (exFB) was determined in experiment 1. The DE and dCP values of trFB and exFB were determined using the difference method in experiment 2. The effect of replacing SBM with trFB and exFB in grow-finisher diets on growth, carcass quality, apparent ileal digestibility (AiD), and total tract digestibility (ATTD) of DM, OM, gross energy (GE), and CP were investigated in experiment 3. In exp. 1, in-vitro digestibility of exFB compared to trFB was unchanged for DM (p = 0.12), increased for OM (p 〈 0.05), and increased for CP (p 〈 0.05). In exp. 2, the DE value of trFB and exFB was 14.38 and 15.75 MJ/kg respectively; and the dCP value was 21.35% and 21.42% respectively (on DM basis). In exp. 3, ADFI was higher for pigs fed trFB and exFB compared to the control diet (CON; p 〈 0.05), while ADG, FCR and carcass quality parameters of pigs did not differ among treatments (p 〉 0.05).

Description: Ridden horses have been reported to be fearful of cows. We tested whether cows could provoke behavioural and cardiac fear responses in horses, and whether these responses differ in magnitude to those shown to other potential dangers. Twenty horses were exposed to cow, a mobile object or no object. The time spent at different distances from the stimulus was measured. In a separate test, heart rate (HR), root mean square of successive differences between heartbeats (RMSSD) and the horses’ perceived fear were assessed at various distances from the stimuli. The horses avoided the area nearest to all stimuli. During hand-leading, the cow elicited the highest HR and lowest RMSSD. Led horses’ responses to the cow and box were rated as more fearful as the distance to the stimulus decreased. Mares had a higher HR than geldings across all tests. HR positively correlated with the fearfulness rating at the furthest distance from the cow and box, and RMSSD negatively correlated with this rating in cow and control conditions. Our results show that these horses’ avoidance response to cows was similar or higher to that shown towards a novel moving object, demonstrating that potentially, both neophobia and heterospecific communication play a role in this reaction.

Description: The present study was conducted to investigate how raw rapeseed cake (RRC), hydrobarothermally-treated rapeseed cake (HRC) and fermented rapeseed cake (FRC) fed to laying hens over a period of 12 weeks affected their performance, and the quality, fatty acid (FA) profile and oxidative stability of eggs. A total of 304 Hy-Line Brown laying hens at 36 weeks of age were distributed in a completely randomized design to four treatment groups with 38 replicates per treatment and two hens per replicate. The birds had ad libitum access to feed and water throughout the study. During the experiment, the birds were fed isonitrogenous and isocaloric diets in mash form, with various protein sources. In the control group (C), soybean meal (SBM) was the main source of dietary protein, whereas the experimental groups were fed diets containing 20% of RRC, HRC or FRC. Hydrobarothermal treatment and fermentation decreased the glucosinolate (GLS) content of RC, and fermentation reduced the concentration of phytate phosphorus (PP). In comparison with the RRC group, layers from the HRC and FRC groups were characterized by higher laying performance, comparable with that in group C. Irrespective of its physical form, RC added to layer diets adversely affected eggshell quality in all experimental groups, whereas albumen quality was highest in the FRC group. In comparison with group C, diets containing RRC, HRC and HRC led to a significant decrease in the content of saturated fatty acids (SFAs), an increase in the proportion of n-3 and n-6 polyunsaturated fatty acids (PUFAs) in the total FA pool in egg yolks, and a decrease in the n-6/n-3 PUFA ratio. The inclusion of RRC, HRC and FRC in layer diets decreased the activity of superoxide dismutase (SOD) in egg yolks, relative to group C. Group FCR eggs were characterized by the highest activity of catalase (CAT) and the lowest lipid peroxides LOOH concentration, compared with the remaining groups. The addition of RC to layer diets did not compromise the sensory quality of eggs, and eggs produced in group FRC received the highest overall score. It can be concluded that the inclusion of 20% RRC, HRC and FRC in layer diets does not compromise the sensory quality of eggs and has a beneficial influence on the FA profile and antioxidant potential of egg yolks. The use of FRC is recommended because it contributes to the highest laying performance, superior albumen quality and the highest sensory quality of eggs, relative to RRC and HRC.

Description: Severe feather pecking (SFP) is a major animal welfare problem in layers. It results in pain and injuries in the affected animal. It was the aim of this study to gain insight into the actual pecking behavior of laying hens kept on commercial farms with flock sizes common in practice. We observed aggressive pecking and SFP in non-beak-trimmed and beak-trimmed flocks of laying hens and investigated possible influencing factors. The study took place on eight conventional farms in Germany with aviaries, including three farms with a free range and a winter garden, one with a free range and one with a winter garden. Pecking behavior was observed during three observational periods (OPs): OP 1, at the peak of the laying period between the 28th and 33rd week of life; OP 2, in the middle of the laying period between the 42nd and 48th week of life; and OP 3, at the end of the laying period between the 63rd and 68th week of life in one laying period. Videos were analyzed using behavior sampling and continuous recording. We found that SFP occurred in all flocks, but the pecking rate differed significantly between the flocks. SFP correlated positively with the number of hens per square meter of usable area, with statistical significance in the litter area (r = 0.564; p = 0.045). The multivariate analysis revealed that access to a winter garden or free range significantly reduced the SFP rate on perches (p = 0.001). The stocking density (number of birds per usable square meter) had a significant influence on the SPF rate in the nest-box area (p = 0.001). The hybrid line had a significant effect on the SFP rate on perches and in the nest-box area (p = 0.001 each). Lohmann Brown hens in mixed flocks had a higher SFP rate (significant in OP 2) than those in homogeneous flocks, indicating that mixed flocks may be a risk factor for SFP. Lohmann Brown hens pecked significantly less than Dekalb White hens in the litter area (p = 0.010) and in the nest-box area (p = 0.025) and less than Lohmann Selected Leghorn hens in the litter area (p = 0.010). Lohmann Brown and Lohmann Selected Leghorn hens showed increasing SFP rates during the laying period. All hybrid lines had significantly higher SFP rates in the litter area, followed by the nest-box area and perches. These findings emphasize the importance of providing enough litter, litter areas and environmental enrichment. We found a significant positive correlation between aggressive pecking and SFP—in OP 1: rho (Spearman) = 0.580, p 〈 0.001; OP 2: rho = 0.486, p = 0.002; and OP 3: rho = 0.482, p = 0.002 (n = 39) —indicating that SFP may lead to a higher stress level in the flock. Beak trimming reduced pecking rates but did not entirely prevent SFP. Instead of subjecting chicks to this potentially painful procedure, reasons for SFP should be addressed. In conclusion, our data suggest a positive influence of a lower stocking density and the provision of a winter garden or free range for additional space. The hybrid line had a significant influence on the feather-pecking rate on perches and the nest-box area. Aggressive pecking and severe feather pecking correlated positively. We assume that vigorous and painful AP were an additional stress factor, especially in non-beak-trimmed flocks, leading to more SFP in due course. Beak trimming had a reducing effect on SFP. However, our results showed that non-beak-trimmed flocks could be kept without major outbreaks of SFP.

Description: Equine oocyte vitrification would benefit the growing in vitro embryo production programs, but further optimization of the protocol is necessary to reach clinical efficiency. Therefore, we aimed to perform a direct comparison of non-permeating and permeating cryoprotective agents (CPAs) during the vitrification and warming of equine immature oocytes. In the first experiment, cumulus oocytes complexes (COCs) were vitrified comparing sucrose, trehalose, and galactose in combination with ethylene glycol (EG) and dimethyl sulfoxide (DMSO). In the second experiment, the COCs were vitrified using three mixtures of permeating CPAs in a 50:50 volume ratio (ethylene glycol-dimethyl sulfoxide (ED), propylene glycol-ethylene glycol (PE), and propylene glycol-dimethyl sulfoxide (PD)) with galactose and warmed in different galactose concentrations (0.3 or 0.5 mol/L). Overall, all the treatments supported blastocyst formation, but the developmental rates were lower for all the vitrified groups in the first (4.3 to 7.6%) and the second (3.5 to 9.4%) experiment compared to the control (26.5 and 34.2%, respectively; p 〈 0.01). In the first experiment, the maturation was not affected by vitrification. The sucrose exhibited lower cleavage than the control (p = 0.02). Although the galactose tended to have lower maturation than trehalose (p = 0.060) and control (p = 0.069), the highest numerical cleavage and blastocyst rates were obtained with this CPA. In the second experiment, the maturation, cleavage, and blastocyst rates were similar between the treatments. Compared to the control, only the ED reached similar maturation (p = 0.02) and PE similar cleavage (p = 0.1). The galactose concentration during warming did not affect the maturation, cleavage, or blastocyst rates (p 〉 0.1), but the PE-0.3 exhibited the highest blastocyst rate (15.1%) among the treatments, being the only one comparable to the control (34.2%). As such, PE–galactose provides a valuable option for equine immature oocyte vitrification and should be considered for the future optimization of the protocol.

Description: Responses to stress are unavoidable, adaptive mechanisms in humans and non-human animals. However, in humans, chronic stress has been linked to poor health outcomes and early mortality. Allostatic load, the physiologic dysregulation that occurs when an organism is exposed to chronic stressors, has been used to assess stress in humans; less work has been done using non-human primates. Our aim was to determine the relationship between allostatic load in ring-tailed lemurs (Lemur catta) under human care and potentially stressful individual, social, medical and husbandry factors, as well a sex and age. An allostatic load index (ALI) was calculated for 38 lemurs using six biomarkers measured in serum (albumin, cortisol, dehydroepiandrosterone-sulfate, DNA damage, glucose and prostaglandin E2). Potentially stressful factors were recorded over the lifetime of each lemur using medical and husbandry records. Animals with a higher percentage of time spent indoors, those kept in smaller average group sizes, and those with fewer minor group composition changes had, or tended to have, higher ALI. There was no relationship between ALI and sex or age. Some social and husbandry factors were associated with allostatic load in lemurs, indicating that this index may be a useful tool in assessing and determining factors contributing to stress of lemurs and other animals under human care.

Description: This study was designed to compare the fatty acid (FA) profiles of four types of fat depots from crossbred bulls (Polish Holstein-Friesian × Limousin, PHF × LM) fattened semi-intensively (SI) and intensively (I). Intramuscular fat was most abundant in total polyunsaturated fatty acids (PUFAs), n-3 and n-6 PUFAs, and functional fatty acids C 18:2, C 18:3, C 20:4 and C 20:5 in comparison with the remaining fat types. Furthermore, note that external fat was more abundant in conjugated linoleic acid (CLA) than other fat types. Grass silage fed to cattle during fattening had a beneficial influence on the FA profile of beef, and an increase in the amount of concentrate in the ration did not decrease beef quality.

Description: The objective of this study was to evaluate the effects of Bacillus subtillis PB6, chromium propionate or a combination of the two on the performance, egg and eggshell quality, nutrient metabolizability and serum biochemistry of layer breeders. White Plymouth Rock and Red Rhodes Island breeder hens at 55 weeks of age were allocated in individual cages using a completely randomized block design with 16 replicates. Hens were fed control, control + probiotic (500 g/ton of Bacillus subtilis PB6), control + CrProp (50 g/ton of chromium propionate) and control + probiotic + CrProp diets from 55 to 70 weeks of age. Productive parameters and eggshell quality as well as cortisol and blood biochemistry were grouped each 28 d as well as for the overall period. The metabolizability of nutrients and energy was determined at 70 weeks of age. In the overall period, hens fed the control + probiotic or control + probiotic + CrProp diets had significantly higher egg production, egg mass, shell percentage, thickness and shell strength. The metabolizability of dry matter, nitrogen and energy increased in hens that were fed the control + probiotic + CrProp diet. In conclusion, diets supplemented with Bacillus subtillis PB6 and chromium propionate resulted in improved productive performance, eggshell quality and nutrient metabolizability of layer breeders, without modifying serum cortisol, albumin and triglycerides.