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: 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.

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: 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 pharmacokinetic of enrofloxacin was investigated in brown trout (Salmo trutta) following oral administration of 10, 20, and 40 mg/kg doses at 11 ± 1.5 °C. Furthermore, MICs of enrofloxacin against Aeromonas hydrophila and A. sobria were determined. The plasma concentrations of enrofloxacin and ciprofloxacin were determined using HPLC–UV and analyzed by non-compartmental method. Following oral administration at dose of 10 mg/kg, total clearance (CL/F), area under the concentration–time curve (AUC0−∞) and peak plasma concentrations (Cmax) were 41.32 mL/h/kg, 242.02 h*μg/mL and 4.63 μg/mL, respectively. When compared to 10 mg/kg dose, the dose-normalized AUC0–∞ and Cmax were increased by 56.30% and 30.08%, respectively, while CL/F decreased by 38.4% at 40 mg/kg dose, suggesting the non-linearity. Ciprofloxacin was not detected in the all of plasma samples. The MIC values of enrofloxacin were ranged 0.0625–4 μg/mL for A. hydrophila and 0.0625–2 μg/mL for A. sobria. The oral administration of enrofloxacin at 10 (for 192 h) and 20 (for 240 h) mg/kg doses provided the AUC of enrofloxacin equal to 1.23 and 1.96-fold MICs, respectively, for A. hydrophila and A. sobria with the MIC90 values of 1 µg/mL. However, further researches are needed on the PK/PD study of enrofloxacin for the successful treatment of infections caused by A. hydrophila and A. sobria in brown trout.

Description: Insects can play an important role to upgrade waste streams into high-grade proteins and fats as food and feed ingredients or non-food products. The aim of this research was to assess the feasibility to use waste streams with a low value for direct application as animal feed as substrates to grow BSF larvae in terms of larval growth rate, waste reduction index, and efficiency of conversion of ingested feed. The growth of black soldier fly (BSF), Hermetia illucens larvae and conversion of biowaste was assessed in triplicate in biowaste substrates: chicken feed (CF; reference diet), pig manure solid (PMS), Betafert® solid (BTFS), swill (SW), olive pulp (OP), pig manure liquid mixed with chicken feed (PMLCF), and silage grass (SG). Per kilogram fresh substrate 2500 starter (8-days-old, second instar) larvae were incubated in 21 plastic containers (75 × 47 × 15 cm). The BSF larvae were fed according to a batch feeding system. Highest growth rate was found in larvae reared on SW (13.4 mg/d). Larval growth rate was even higher than in larvae reared on the reference substrate CF (7.2 mg/d). Growth rate in larvae reared on PMLCF (7.3 mg/d) did not differ from CF, whereas growth rate of larvae reared on PMS (3.2 mg/d) was lower than on CF. Growth rate of larvae reared on BTFS, OP and SG was very low (0.6, 0.2 and 0.7 mg/d, respectively). Waste Reduction Index (WRI) was highest on SW (11.3), followed by PMLCF (9.3), and both were higher than WRI on CF (8.5). Waste Reduction Index further decreased in descending order from PMS, SG, BTFS to OP (7.6, 4.0, 2.9 and 1.7, respectively). The Efficiency of Conversion of Ingested substrate (ECI) was highest on SW (0.31), followed in descending order by PMLCF, CF and PMS (0.25, 0.21 and 0.18, respectively). The substrates OP, BTFS and SG (0.16, 0.15 and 0.14, respectively) resulted in a lower ECI than other substrates. Highest CO2 and lowest NH3 concentrations were found above substrates with the highest larval growth performances. This study showed that BSF larvae can be reared on different biowaste substrates; the growth rate of the larvae was extremely high on SW. The effects of chemical composition and physical properties of the substrates on larval growth and gas emissions should be further considered.

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: Fibropapillomatosis (FP), a debilitating, infectious neoplastic disease, is rarely reported in endangered Kemp’s ridley sea turtles (Lepidochelys kempii). With this study, we describe FP and the associated chelonid alphaherpesvirus 5 (ChHV5) in Kemp’s ridley turtles encountered in the United States during 2006–2020. Analysis of 22 case reports of Kemp’s ridley turtles with FP revealed that while the disease was mild in most cases, 54.5% were adult turtles, a reproductively valuable age class whose survival is a priority for population recovery. Of 51 blood samples from tumor-free turtles and 12 tumor samples from turtles with FP, 7.8% and 91.7%, respectively, tested positive for ChHV5 DNA via quantitative polymerase chain reaction (qPCR). Viral genome shotgun sequencing and phylogenetic analysis of six tumor samples show that ChHV5 sequences in Kemp’s ridley turtles encountered in the Gulf of Mexico and northwestern Atlantic cluster with ChHV5 sequences identified in green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtles from Hawaii, the southwestern Atlantic Ocean, and the Caribbean. Results suggest an interspecific, spatiotemporal spread of FP among Kemp’s ridley turtles in regions where the disease is enzootic. Although FP is currently uncommon in this species, it remains a health concern due to its uncertain pathogenesis and potential relationship with habitat degradation.

Description: The purpose of this study was to evaluate the effect of full-fat almonds (FFA) as an alternative protein and fat source for broiler feed on broiler productivity, nutrient digestibility, blood characteristics, cecal-fecal microflora, and foot-pad dermatitis (FPD). A total of 96, one-day-old broiler chickens (Arbor Acres) with initial body weight 41.61 ± 0.36 g were placed in 16 cages. In each trial, four treatments were set up: a basal diet partially replacing animal fat with FFA 0% (Control, CON), a basal diet partially replacing animal fat with FFA 1% (T1), a basal diet partial replacing animal fat with FFA 2% (T2), a basal diet partially replacing animal fat with FFA 4% (T3). The experiment was conducted for a total of 4 weeks. Feed conversion ratio (FCR) was higher (p 〈 0.05) in the T3 group of broilers at weeks 0 to 1 than in the CON group of broilers. From weeks 3 to 4, and for the entire experimental period, FCR was lower (p 〈 0.05) in the T3 group of broilers than in the CON and T1 groups of broilers. The apparent ileal digestibility (AID) of the ether extract (EE) was higher (p 〈 0.05) in the T3 group than in the other treatment groups, and AID of crude protein (CP) was higher (p 〈 0.05) in the T3 group than in the CON group. The apparent total tract digestibility (ATTD) of EE was lower (p 〈 0.05) in the CON group than in the other treatment groups, and the ATTD of CP and energy was higher (p 〈 0.05) in the T3 group of broilers than in the CON group of broilers. The AID and ATTD of total amino acids were higher (p 〈 0.05) in the T3 group than in the other treatment groups. Blood cholesterol levels were lower (p 〈 0.05) in the T3 group of broilers than in the CON and T1 groups of broilers, and higher (p 〈 0.05) in the CON group of broilers than in the T2 and T3 groups of broilers. The amount of E. coli in the cecal and fecal was lower (p 〈 0.05) in the T3 group than in CON and T1 groups. FPD score was higher (p 〈 0.05) in the T3 group of broilers than in the CON group of broilers. In conclusion, replacing a partial of animal fat with at least 4% FFA in broiler diets can increase growth performance and nutrient digestibility in broiler nutrition.

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.