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Saponin possessed antiprotozoal activity; therefore, it is generally used to mitigate enteric methane (CH4) emission in ruminant nutrition. The objective of this study was to evaluate the effects of Aloe saponaria (AS) supplementation on rumen fermentation, CH4, and microbial diversity in an in vitro experiment over 48 h incubation. Five treatments were used: (1) no additives (CON), 1% and 2% dosage of heat-dried AS (HAS), and freeze-dried AS (FAS), and the experiments were performed three times. After the incubation, volatile fatty acids (VFAs), CH4 proportion in total gas, and microbial diversity were evaluated. Gas production was assessed at 3, 6, 9, 12, 24, 36, and 48 h incubation. FAS addition significantly increased gas production compared to CON except for 6 and 48 h, whereas HAS addition did not significantly affect the gas production at all observed time points. AS addition significantly increased total VFA and the absolute abundances of fungi and Ruminococcus albus regardless of drying method. There was no significant change in the CH4 proportion at 48 h, although the reduction in ciliate protozoa was detected in FAS-treated groups. In conclusion, FAS may be an effective feed additive to improve ruminal fermentation via enhancing feed utilization by rumen microbes.

Objective The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p

The aim of this study was to investigate the effects of heat stress on milk traits in South Korea using comprehensive data (dairy production and climate). The dataset for this study comprised 1,498,232 test-day records for milk yield, fat- and protein-corrected milk, fat yield, protein yield, milk urea nitrogen (MUN), and somatic cell score (SCS) from 215,276 Holstein cows (primiparous: n = 122,087; multiparous: n = 93,189) in 2,419 South Korean dairy herds. Data were collected from July 2017 to April 2020 through the Dairy Cattle Improvement Program, and merged with meteorological data from 600 automatic weather stations through the Korea Meteorological Administration. The segmented regression model was used to estimate the effects of the temperature-humidity index (THI) on milk traits and elucidate the break point (BP) of the THI. To acquire the least-squares mean of milk traits, the generalized linear model was applied using fixed effects (region, calving year, calving month, parity, days in milk, and THI). For all parameters, the BP of THI was observed; in particular, milk production parameters dramatically decreased after a specific BP of THI (p < 0.05). In contrast, MUN and SCS drastically increased when THI exceeded BP in all cows (p < 0.05) and primiparous cows (p < 0.05), respectively. Dairy cows in South Korea exhibited negative effects on milk traits (decrease in milk performance, increase in MUN, and SCS) when the THI exceeded 70; therefore, detailed feeding management is required to prevent heat stress in dairy cows.

Abstract Canine obesity is a major health concern that predisposes dogs to various disorders. The fecal microbiome has been attracting attention because of their impact on energy efficiency and metabolic disorders of host. However, little is known about specific microbial interactions, and how these may be affected by obesity in dogs. The objective of this study was to investigate the differences in fecal microbiome and specific microbial networks between obese and normal dogs. A total of 20 beagle dogs (males = 12, body weight [BW]: 10.5 ± 1.08 kg; females = 8, BW: 11.3 ± 1.71 kg; all 2-year-old) were fed to meet the maintenance energy requirements for 18 weeks. Then, 12 beagle dogs were selected based on body condition score (BCS) and divided into two groups: high BCS group (HBCS; BCS range: 7–9, males = 4, females = 2) and normal BCS group (NBCS; BCS range: 4–6, males = 4, females = 2). In the final week of the experiment, fecal samples were collected directly from the rectum, before breakfast, for analyzing the fecal microbiome using 16S rRNA gene amplicon sequencing. The HBCS group had a significantly higher final BW than the NBCS group (P

This study aimed to investigate Pinus koraiensis cone essential oil (PEO) as a methane (CH4) inhibitor and determine its impact on the taxonomic and functional characteristics of the rumen microbiota in goats. A total of 10 growing Korean native goats (Capra hircus coreanae, 29.9 ± 1.58 kg, male) were assigned to different dietary treatments: control (CON; basal diet without additive) and PEO (basal diet +1 g/d of PEO) by a 2 × 2 crossover design. Methane measurements were conducted every 4 consecutive days for 17–20 days using a laser CH4 detector. Samples of rumen fluid and feces were collected during each experimental period to evaluate the biological effects and dry matter (DM) digestibility after PEO oral administration. The rumen microbiota was analyzed via 16S rRNA gene amplicon sequencing. The PEO oral administration resulted in reduced CH4 emission (eructation CH4/body weight0.75, p = 0.079) without affecting DM intake; however, it lowered the total volatile fatty acids (p = 0.041), molar proportion of propionate (p = 0.075), and ammonia nitrogen (p = 0.087) in the rumen. Blood metabolites (i.e., albumin, alanine transaminase/serum glutamic pyruvate transaminase, creatinine, and triglyceride) were significantly affected (p

This study investigated the effect of different dietary fat levels in concentrate mixes on the growth performance, rumen characteristics, digestibility, blood metabolites, and methane emissions in growing Hanwoo steers. Thirty steers (386 ± 24.6 kg of body weight [BW]; 12 months old), blocked by BW, were randomly assigned to three dietary treatments with varying fat concentrations in concentrate mix (48, 74, and 99 g of ether extract per kg dry matte [DM]). The fat intake of the low-fat treatment represented 4.15% of the total dry matter intake (DMI), while the medium- and high-fat treatments accounted for 5.77% and 7.23% of total DMI, respectively. Concentrate mix DMI decreased with increasing fat level (p < 0.01). The growth rate and digestibility did not significantly differ based on the fat level (p > 0.05). As the fat level increased, propionate in the total ruminal volatile fatty acids increased, and butyrate and acetate-to-propionate decreased (p < 0.01). Cholesterol in blood serum increased significantly with increasing dietary fat levels (p < 0.01). Methane emissions exhibited a linear decrease with increasing fat level (p < 0.05). In conclusion, elevating fat content in the concentrates up to 100 g/kg DM reduced methane emissions without compromising the growth performance of growing Hanwoo steers.

This study aimed to assess the influence of rice grain in the total mixed ration (TMR) on the growth performance, blood metabolites, rumen fermentation, and rumen microbial community of fattening Hanwoo steers. Two experimental diets were prepared: (i) a TMR containing 33% dry matter (DM) corn grains (Corn TMR) and (ii) a TMR containing 33% DM rice grains (Rice TMR). Twenty-two Hanwoo steers (body weight [BW], 498 ± 32 kg; months, 17 ± 0.5) were distributed into two treatment groups in a completely randomized block design according to BW. The Rice TMR group had a higher final BW and DM intake (DMI) compared to those in the Corn TMR group (p < 0.01). However, no difference was observed in the average daily gain (ADG) and feed conversion ratio (FCR) between the two treatments. For the rumen fermentation parameters, the molar portion of butyrate in the Rice TMR was higher than in the Corn TMR (p < 0.01). Streptococcus bovis tended to be higher in the Rice TMR (p = 0.09). The results of this study suggest that using rice grain as the primary starch source in TMRs may be an alternative option for fattening Hanwoo steers.

Abstract Several seaweed extracts have been reported to have potential antimethanogenic effects in ruminants. In this study, the effect of three brown seaweed species (Undaria pinnatifida, UPIN; Sargassum fusiforme, SFUS; and Sargassum fulvellum, SFUL) on rumen fermentation characteristics, total gas, methane (CH4), carbon dioxide (CO2) production, and microbial populations were investigated using an in vitro batch culture system. Seaweed extract and its metabolites, total flavonoid and polyphenol contents were identified and compared. For the in vitro batch, 0.25 mg∙mL−1 of each seaweed extract were used in 6, 12, 24, 36 and 48 h of incubation. Seaweed extract supplementation decreased CH4 yield and its proportion to total gas production after 12, 24, and 48 h of incubation, while total gas production were not significantly different. Total volatile fatty acid and molar proportion of propionate increased with SFUS and SFUL supplementation after 24 h of incubation, whereas UPIN was not affected. Additionally, SFUS increased the absolute abundance of total bacteria, ciliate protozoa, fungi, methanogenic archaea, and Fibrobacter succinogenes. The relative proportions of Butyrivibrio fibrisolvens, Butyrivibrio proteoclasticus, and Prevotella ruminicola were lower with seaweed extract supplementation, whereas Anaerovibrio lipolytica increased. Thus, seaweed extracts can decrease CH4 production, and alter the abundance of rumen microbial populations.

A series of in vitro batch culture incubations were carried out to investigate changes in rumen fermentation characteristics, methane (CH4) production, and microbial composition in response to supplementation with five different red seaweed species (Amphiroa anceps, AANC; Asparagopsis taxiformis, ATAX; Chondracanthus tenellus, CTEN; Grateloupia elliptica, GELL; and Gracilaria parvispora, GPAR). Prior to the incubations, the total flavonoid and polyphenol content of the red seaweed extracts was quantified. The incubated substrate consisted of timothy hay and corn grain [60:40 dry matter (DM) basis]. Treatments were substrate mixtures without seaweed extract (CON) or substrate mixtures supplemented with 0.25 mg/mL of red seaweed extract. Samples were incubated for 6, 12, 24, 36, and 48 h. Each sample was incubated in triplicates in three separate runs. In vitro DM degradability, fermentation parameters (i.e., pH, volatile fatty acids, and ammonia nitrogen), total gas production, and CH4 production were analyzed for all time points. Microbial composition was analyzed using 16S rRNA amplicon sequencing after 24 h of incubation. The highest CH4 reduction (mL/g DM, mL/g digested DM, and % of total gas production) was observed in ATAX (51.3, 50.1, and 51.5%, respectively, compared to CON; P < 0.001) after 12 h of incubation. The other red seaweed extracts reduced the CH4 production (mL/g DM; P < 0.001) in the range of 4.6–35.0% compared to CON after 24 h of incubation. After 24 h of incubation, supplementation with red seaweed extracts tended to increase the molar proportion of propionate (P = 0.057) and decreased the acetate to propionate ratio (P = 0.033) compared to the CON. Abundances of the genus Methanobrevibacter and total methanogens were reduced (P = 0.050 and P = 0.016) by red seaweed extract supplementation. The linear discriminant analysis effect size (P < 0.05, LDA ≥ 2.0) showed that UG Succinivibrionaceae, Anaeroplasma, and UG Ruminococcaceae, which are associated with higher propionate production, starch degradation, and amylase activity were relatively more abundant in red seaweed extracts than in the CON. Our results suggest that supplementation with red seaweed extracts altered the microbiota, leading to the acceleration of propionate production and reduction in CH4 production.

Recently, a high level of dietary crude protein (CP) has become of interest as a possible practice to improve the carcass quality of beef cattle, and its level has been increasing in the field. However, there is little scientific evidence that supports this. This study was conducted to test whether a high dietary CP level would improve growth performance, body metabolism, and carcass traits in Hanwoo beef cattle. A total of 32 Hanwoo finishing beef cattle (18 multiparous cows, six heifers, and eight steers) participated in a 12-weeks feeding trial. Two kinds of total mixed rations were prepared to contain two different CP; 156 g/kg for the control (CON) and 173 g/kg of CP for the treatment (HCP), while maintaining a similar level of metabolizable energy. The experiment was ended when more than half of the steers reached the target body weight (730 kg). Blood was collected at the end of the experiment. After harvesting, the carcass trait was evaluated at the slaughterhouse according to Korean standards. The carcass yield score and grade were also calculated based on revised criteria. Overall, dry matter intake, average daily gain, blood metabolites concentration, and the carcass traits, except for backfat thickness and the yield score, did not differ between the treatments. The HCP had lower backfat thickness than those of CON. There was no difference in the carcass yield grade, but the yield score was higher in the HCP treatment. According to the newly revised carcass grading criteria, both yield score and grade were higher in HCP than in CON. Increasing CP supply decreased the carcass’s backfat thickness without altering growth performance and body metabolism, resulting in improved yield score and grade. Therefore, feeding a high CP diet may be beneficial in the farm income, although it may also increase feed cost and nitrogen excretion to the environment.

Abstract Background Endolysins, the bacteriophage-originated peptidoglycan hydrolases, are a promising replacement for antibiotics due to immediate lytic activity and no antibiotic resistance. The objectives of this study were to investigate the lytic activity of endolysin LyJH307 against S. bovis and to explore changes in rumen fermentation and microbiota in an in vitro system. Two treatments were used: 1) control, corn grain without LyJH307; and 2) LyJH307, corn grain with LyJH307 (4 U/mL). An in vitro fermentation experiment was performed using mixture of rumen fluid collected from two cannulated Holstein steers (450 ± 30 kg) and artificial saliva buffer mixed as 1:3 ratio for 12 h incubation time. In vitro dry matter digestibility, pH, volatile fatty acids, and lactate concentration were estimated at 12 h, and the gas production was measured at 6, 9, and 12 h. The rumen bacterial community was analyzed using 16S rRNA amplicon sequencing. Results LyJH307 supplementation at 6 h incubation markedly decreased the absolute abundance of S. bovis (approximately 70% compared to control, P = 0.0289) and increased ruminal pH (P = 0.0335) at the 12 h incubation. The acetate proportion (P = 0.0362) was significantly increased after LyJH307 addition, whereas propionate (P = 0.0379) was decreased. LyJH307 supplementation increased D-lactate (P = 0.0340) without any change in L-lactate concentration (P > 0.10). There were no significant differences in Shannon’s index, Simpson’s index, Chao1 estimates, and evenness (P > 0.10). Based on Bray-Curtis dissimilarity matrices, the LyJH307 affected the overall shift in microbiota (P = 0.097). LyJH307 supplementation induced an increase of 11 genera containing Lachnoclostridium, WCHB1–41, unclassified genus Selenomonadaceae, Paraprevotella, vadinBE97, Ruminococcus gauvreauii group, Lactobacillus, Anaerorhabdus furcosa group, Victivallaceae, Desulfuromonadaceae, and Sediminispirochaeta. The predicted functional features represented by the Kyoto Encyclopedia of Genes and Genomes pathways were changed by LyJH307 toward a decrease of carbohydrate metabolism. Conclusions LyJH307 caused a reduction of S. bovis and an increase of pH with shifts in minor microbiota and its metabolic pathways related to carbohydrate metabolism. This study provides the first insight into the availability of endolysin as a specific modulator for rumen and shows the possibility of endolysin degradation by rumen microbiota.

The increasing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in the dairy industry has become a fundamental concern. Endolysins are bacteriophage-derived peptidoglycan hydrolases that induce the rapid lysis of host bacteria. Herein, we evaluated the lytic activity of endolysin candidates against S. aureus and MRSA. To identify endolysins, we used a bioinformatical strategy with the following steps: (1) retrieval of genetic information, (2) annotation, (3) selection of MRSA, (4) selection of endolysin candidates, and (5) evaluation of protein solubility. We then characterized the endolysin candidates under various conditions. Approximately 67% of S. aureus was detected as MRSA, and 114 putative endolysins were found. These 114 putative endolysins were divided into three groups based on their combinations of conserved domains. Considering protein solubility, we selected putative endolysins 117 and 177. Putative endolysin 117 was the only successfully overexpressed endolysin, and it was renamed LyJH1892. LyJH1892 showed potent lytic activity against both methicillin-susceptible S. aureus and MRSA and showed broad lytic activity against coagulase-negative staphylococci. In conclusion, this study demonstrates a rapid strategy for the development of endolysin against MRSA. This strategy could also be used to combat other antibiotic-resistant bacteria.

The main objective of this in vitro study was to evaluate red ginseng byproduct (RGP) as a protein resource and its effects on rumen fermentation characteristics, microflora, CO2, and CH4 production in ruminants. Four treatments for in vitro fermentation using buffered rumen fluid over a 48 h incubation period were used: 1, RGP; 2, corn gluten feed (CGF); 3, wheat gluten (WG); and 4, corn germ meal. In vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), in vitro crude protein digestibility (IVCPD), volatile fatty acids, pH, and ammonia nitrogen (NH3-N) were estimated after 48 h incubation. Gas production was investigated after 3, 6, 12, 24, 36 and 48 h. The CO2 and CH4 were evaluated after 12, 24, 36, and 48 h. A significant difference in total gas production and CO2 emissions was observed (p < 0.01) at all incubation times. CH4 production in RGP were higher (p < 0.05) than that in other treatments but a higher CH4 portion in the total gas production was observed in WG (p < 0.05) at 48 h incubation. The IVDMD, IVNDFD, and IVCPD of RGP was lower than those of other conventional ingredients (p < 0.01). The RGP had the lowest NH3-N value among the treatments (p < 0.01). The RGP also had the lowest total VFA concentration (p < 0.01), but presented the highest acetate proportion and acetate to propionate ratio among the treatments (both, p < 0.01). The abundance of Prevotella ruminicola was higher in RGP than in WG (p < 0.01), whereas RGP has lower methanogenic archaea (p < 0.01). In conclusion, based on the nutritive value, IVDMD, low NH3-N, and decreased methanogenic archaea, RGP inclusion as a protein source in ruminant diets can be an option in replacing conventional feed sources.

Objective The objective of this study was to evaluate the effects of lysophospholipids (LPL) supplementation on rumen fermentation, degradability, and microbial diversity in forage with high oil diet in an in vitro system. Methods Four experimental treatments were used: i) annual ryegrass (CON), ii) 93% annual ryegrass +7% corn oil on a dry matter (DM) basis (OiL), iii) OiL with a low level (0.08% of dietary DM) of LPL (LLPL), and iv) OiL with a high level (0.16% of dietary DM) of LPL (HLPL). An in vitro fermentation experiment was performed using strained rumen fluid for 48 h incubations. In vitro DM degradability (IVDMD), in vitro neutral detergent fiber degradability, pH, ammonia nitrogen (NH3-N), volatile fatty acid (VFA), and microbial diversity were estimated. Results There was no significant change in IVDMD, pH, NH3-N, and total VFA production among treatments. The LPL supplementation significantly increased the proportion of butyrate and valerate (Linear effect [Lin], p = 0.004 and

This study determined the substitution effects of rice for corn as the main grain source in a total mixed ration (TMR). In vitro rumen fermentation characteristics and microbes were assessed using two experimental diets. Diets included 33% dry matter (DM) of either corn (Corn TMR) or rice grains (Rice TMR). In a 48-h in vitro incubation, DM digestibility (IVDMD), neutral detergent fiber degradability (IVNDFD), crude protein digestibility (IVCPD), volatile fatty acids (VFAs), pH and ammonia nitrogen (NH3-N) were estimated. Gas production has been calculated at 3, 6, 12, 24 and 48 h. Our results indicate that the gas production, VFAs, IVDMD, and IVNDFD of Rice TMR were higher than those of Corn TMR (p < 0.05). Ruminal pH and total fungi were significantly higher in Corn TMR (p < 0.05) than in Rice TMR; however, NH3-N and IVCPD were not affected by treatment type. In conclusion, substituting rice for corn at 33% DM in TMR appears to have no negative effects on in vitro rumen fermentation characteristics. Therefore, rice grains are an appropriate alternative energy source in early fattening stage diets of beef cattle.

This study investigated the effect of variation in the number of somaticcell-cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100- 150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.

Objective This study was realized to evaluate the nutritional value of rice grains as a replacement for corn grains in the diet of growing Hanwoo steers. Methods Two experimental diets were prepared: i) Corn total mixed ration (TMR) consisting of 20% corn grains and ii) Rice TMR consisting of 20% rice grains, in a dry matter (DM) basis. These treatments were used for in vitro rumen fermentation and in vivo growth trials. In the rumen fermentation experiment, the in vitro DM digestibility (IVDMD), in vitro crude protein digestibility (IVCPD), in vitro neutral detergent fiber digestibility, pH, ammonia nitrogen, and volatile fatty acids (VFA) were estimated at 48 h, and the gas production was measured at 3, 6, 12, 24, and 48 h. Twenty four growing Hanwoo steers (9 months old; body weight [BW]: 259±13 kg) were randomly divided into two treatment groups and the BW, dry matter intake (DMI), average daily gain (ADG), and feed conversion ratio (FCR) were measured. Results The in vitro experiment showed that the IVDMD, IVCPD, and VFA production of the Rice TMR were higher than those of the Corn TMR (p0.05) in the final BW, ADG, DMI, and FCR between the two TMRs. Conclusion The use of rice grains instead of corn grains did not exhibit any negative effects on the rumen fermentation or growth performance, thereby rice grains with a DM of less than 20% could be used as a starch source in the diet of growing steers.

The aim of this study was to investigate the effects of 3-nitrooxypropanol (NOP) on gas production, rumen fermentation, and animal performances depending on animal type using a meta-analysis approach. A database consisted of data from 14 studies, 18 experiments and 55 treatments. The supplementation of NOP linearly decreased methane (CH4) emissions [g/kg dry matter intake (DMI)] regardless of animal type and length of experimental period (beef, p < 0.0001, R2 = 0.797; dairy, p = 0.0003, R2 = 0.916; and long term, p < 0.0001, R2 = 0.910). The total volatile fatty acids (VFA) concentration and the proportion of acetate, based on beef cattle database, were significantly decreased with increasing NOP supplementation (p = 0.0015, R2 = 0.804 and p = 0.0003, R2 = 0.918), whereas other individual VFAs was increased. Based on the dairy database, increasing levels of NOP supplementation linearly decreased proportion of acetate (p = 0.0284, R2 = 0.769) and increased that of valerate (p = 0.0340, R2 = 0.522), regardless of significant change on other individual VFAs. In animal performances, the DMI, from beef cattle database, tended to decrease when the levels of NOP supplementation increased (p = 0.0574, R2 = 0.170), whereas there was no significant change on DMI from dairy cattle database. The NOP supplementation tended to decrease milk yield (p = 0.0606, R2 = 0.381) and increase milk fat and milk protein (p = 0.0861, R2 = 0.321, p = 0.0838, R2 = 0.322). NOP is a viable candidate as a feed additive because of its CH4 mitigation effects, regardless of animal type and experiment period, without adverse effects on animal performances.

Objective Reducing roughage feeding without negatively affecting rumen health is of interest in ruminant nutrition. We investigated the effects of roughage sources and concentrate types on growth performance, ruminal fermentation, and blood metabolite levels in growing cattle. Methods In this 24-week trial, 24 Hanwoo cattle (224±24.7 kg) were fed similar nitrous and energy levels of total mixed ration formulated using two kinds of roughage (timothy hay and ryegrass straw) and two types of concentrate mixes (high starch [HS] and high fiber [HF]). The treatments were arranged in a 2×2 factorial, consisting of 32% timothy–68% HS, 24% timothy–76% HF, 24% ryegrass–76% HS, and 17% ryegrass–83% HF. Daily feed intakes were measured. Every four weeks, blood were sampled, and body weight was measured before morning feeding. Every eight weeks, rumen fluid was collected using a stomach tube over five consecutive days. Results The mean dry matter intake (7.33 kg) and average daily gain (1,033 g) did not differ among treatments. However, significant interactions between roughage source and concentrate type were observed for the rumen and blood parameters (p

Objective Among stress responses, the unfolded protein response (UPR) is a well-known mechanism related to endoplasmic reticulum (ER) stress. ER stress is induced by a variety of external and environmental factors such as starvation, ischemia, hypoxia, oxidative stress, and heat stress. Inositol requiring enzyme 1α (IRE1α)-X-box protein 1 (XBP1) is the most conserved pathway involved in the UPR and is the main component that mediates IRE1α signalling to downstream ER-associated degradation (ERAD)- or UPR-related genes. XBP1 is a transcription factor synthesised via a novel mechanism called ‘frame switch splicing’, and this process has not yet been studied in the horse XBP1 gene. Therefore, the aim of this study was to confirm the frame switch splicing of horse XBP1 and characterise its dynamics using Thoroughbred muscle cells exposed to heat stress. Methods Primary horse muscle cells were used to investigate heat stress-induced frame switch splicing of horse XBP1. Frame switch splicing was confirmed by sequencing analysis. XBP1 amino acid sequences and promoter sequences of various species were aligned to confirm the sequence homology and to find conserved cis-acting elements, respectively. The expression of the potential XBP1 downstream genes were analysed by quantitative real-time polymerase chain reaction. Results We confirmed that splicing of horse XBP1 mRNA was affected by the duration of thermal stress. Twenty-six nucleotides in the mRNA of XBP1 were deleted after heat stress. The protein sequence and the cis-regulatory elements on the promoter of horse XBP1 are highly conserved among the mammals. Induction of putative downstream genes of horse XBP1 was dependent on the duration of heat stress. We confirmed that both the mechanisms of XBP1 frame switch splicing and various binding elements found in downstream gene promoters are highly evolutionarily conserved. Conclusion The frame switch splicing of horse XBP1 and its dynamics were highly conserved among species. These results facilitate studies of ER-stress in horse.

Objective This study was conducted to evaluate whether the co-injection of antioxidants together with foot-and-mouth disease (FMD) vaccination has the potential to attenuate the negative effects caused by vaccination in Holstein finishing steers. Methods A total of 36 finishing Holstein steers (body weight [BW]: 608±45.6 kg, 17 months old) were randomly allocated to one of three treatments: i) control (CON, only FMD vaccination without any co-injection), ii) co-injection of commercial non-steroidal anti-inflammatory drugs (NSAID) with FMD vaccination at a ratio of 10:1 (NSAID vol/FMD vaccine vol) as a positive control (PCON), iii) co-injection of commercial mixture of vitamin E and selenium with FMD vaccination (VITESEL) (1 mL of FMD vaccine+1 mL of antioxidants per 90 kg of BW). Changes in growth performance and blood parameters because of treatments were determined. Results No significant difference in BW, average daily gain, and dry matter intake of the steers was observed among the treatments. The FMD vaccination significantly increased white blood cells (WBC), neutrophils, platelets, and mean platelet volume (p

Objective The objective of this study was to investigate the effects of essential oil mixture (EOM) supplementation on rumen fermentation characteristics and microbial changes in an in vitro. Methods Three experimental treatments were used: control (CON, no additive), EOM 0.1 (supplementation of 1 g EOM/kg of substrate), and EOM 0.2 (supplementation of 2 g EOM/kg of substrate). An in vitro fermentation experiment was carried out using strained rumen fluid for 12 and 24 h incubation periods. At each time point, in vitro dry matter digestibility (IVDMD), neutral detergent fiber digestibility (IVNDFD), pH, ammonia nitrogen (NH3-N), and volatile fatty acid (VFA) concentrations, and relative microbial diversity were estimated. Results After 24 h incubation, treatments involving EOM supplementation led to significantly higher IVDMD (treatments and quadratic effect; p = 0.019 and 0.008) and IVNDFD (linear effect; p = 0.068) than did the CON treatment. The EOM 0.2 supplementation group had the highest NH3-N concentration (treatments; p = 0.032). Both EOM supplementations did not affect total VFA concentration and the proportion of individual VFAs; however, total VFA tended to increase in EOM supplementation groups, after 12 h incubation (linear; p = 0.071). Relative protozoa abundance significantly increased following EOM supplementation (treatments, p

Objective To investigate changes in rumen fermentation characteristics and bacterial community by a sudden change to a high concentrate diet (HC) in Korean domestic ruminants. Methods Major Korean domestic ruminants (each of four Hanwoo cows; 545.5±33.6 kg, Holstein cows; 516.3±42.7 kg, and Korean native goats; 19.1±1.4 kg) were used in this experiment. They were housed individually and were fed ad libitum with a same TMR (800 g/kg timothy hay and 200 g/kg concentrate mix) twice daily. After two-week feeding, only the concentrate mix was offered for one week in order to induce rapid rumen acidosis. The rumen fluid was collected from each animals twice (on week 2 and week 3) at 2 h after morning feeding using an oral stomach tube. Each collected rumen fluid was analyzed for pH, volatile fatty acid (VFA), and NH3-N. In addition, differences in microbial community among ruminant species and between normal and an acidosis condition were assessed using two culture-independent 16S polymerase chain reaction (PCR)-based techniques (terminal restriction fragment length polymorphism and quantitative real-time PCR). Results The HC decreased ruminal pH and altered relative concentrations of ruminal VFA (p

The objective of this study was to evaluate the effects of dietary supplementation with red ginseng byproduct (RGB) on rumen fermentation, growth performance, blood metabolites, and mRNA expression of heat shock proteins (HSP) in fattening Hanwoo steers under heat stress. Two experimental total mixed rations (TMR) were prepared: (1) a TMR meeting the requirement of fattening beef having an average daily gain (ADG) 0.8 kg/day (CON) and (2) a TMR that included 2% RGB on a dry matter (DM) basis (GINSENG). In vitro rumen fermentation and in vivo growth experiments were conducted using two experimental diets. A total of 22 Hanwoo steers were distributed to two treatments (CON vs. GINSENG) in a completely randomized block design according to body weight (BW). The experiment was conducted during the summer season for five weeks. The final BW, ADG, DM intake, and feed conversion ratio did not differ between treatments in the growth trial. In the mRNA expression results, only HSP 90 showed an increasing tendency in the GINSENG group. The use of 2%DM RGB did not improve the growth performance or alleviate heat stress in fattening Hanwoo steers during the summer season.

Objective This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results The maximum activity of recombinant Cel5A (rCel5A) was observed at 50°C and pH 4.0. The enzyme was constant at the temperature range of 20°C to 40°C but also, at the pH range of 3 to 9. The metal ions including Ca2+, K+, Ni2+, Mg2+, and Fe2+ increased the endoglucanase activity but the addition of Mn2+, Cu2+, and Zn2+ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and 45.66 μmol/min/mg. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was 96.69 (s−1) and 6.88 (mL/mg/s), respectively. Conclusion Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

We evaluated whether olive leaves (OLs) are effective as feed additives and supplements for ruminants and the potential methane reduction effects during in vitro fermentation. Two Hanwoo cows (460 ± 20 kg) equipped with cannula were fed Timothy hay and corn-based feed 3% of the body weight at a ratio of 6:4 (8:30 a.m. and 5:00 p.m.). Ruminal fluid from the cows was collected and mixed before morning feeding. In vitro batch fermentation was monitored after 12 and 24 h of incubation at 39 °C, and OLs were used as supplements to achieve the concentration of 5% in the basal diet. At 12 h of fermentation, methane production decreased in the 5% OLs group compared to that in the control group, but not at 24 h. The proportion of cellulose-degrading bacteria, Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens, tended to increase in the 5% OLs group at 12 h. The amount of ammonia produced was the same as the polymerase chain reaction result for Prevotella ruminicola. At 12 h, the proportion of Prevotella ruminicola was significantly higher in the 5% OLs group. OLs may be used incorporated with protein byproducts or other methane-reducing agents in animal feed.

Objective The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture (143.8±10.5 to 159.2±14.8) was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT) groups (31.4±8.3 to 33.4±11.1). After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p

The objective of this study was to evaluate the effects of dietary supplementation with bacteriophage and β-mannanase on health and growth performance in calves. Thirty-six pre-weaning male Holstein calves were randomly allocated to one of four dietary treatments with a 2 × 2 factorial arrangement: no supplementation, 0.1% β-mannanase, 0.1% bacteriophage, and both 0.1% bacteriophage and 0.1% β-mannanase supplementation in a starter on a dry matter basis. The experiment lasted from 2 weeks before weaning to 8 weeks after weaning. Twenty-two calves survived to the end of the experiment. No interaction was observed between the two different feed additives. The bacteriophage supplementation tended to increase the odds ratio of survival (p = 0.09). The number of Escherichia coli in feces significantly decreased by bacteriophage supplementation one week after weaning. β-mannanase supplementation increased the concentrate intake (p < 0.01) and tended to increase the final BW (p = 0.08). Analysis of repeated measures indicated β-mannanase supplementation increased weekly body weight gain (p = 0.018). We conclude that bacteriophage supplementation may have a positive effect on calf survival rate, while β-mannanase supplementation may increase the growth rate and starter intake by calves just before and after weaning.

Streptococcus bovis (S. bovis) is one of the critical initiators of acute acidosis in ruminants. Therefore, we aimed to develop and characterize the endolysin LyJH307, which can lyse ruminal S. bovis. We tested the bactericidal activity of recombinant LyJH307 against S. bovis JB1 under a range of pH, temperature, NaCl, and metal ion concentrations. In silico analyses showed that LyJH307 has a modular design with a distinct, enzymatically active domain of the NLPC/P60 superfamily at the N-terminal and a cell wall binding domain of the Zoocin A target recognition domain (Zoocin A_TRD) superfamily at the C-terminal. The lytic activity of LyJH307 against S. bovis JB1 was the highest at pH 5.5, and relatively higher under acidic, than under alkaline conditions. LyJH307 activity was also the highest at 39 °C, but was maintained between 25°C and 55°C. LyJH307 bactericidal action was retained under 0-500 mM NaCl. While the activity of LyJH307 significantly decreased on treatment with ethylenediaminetetraacetic acid (EDTA), it was only restored with supplementation of 10 mM Ca2+. Analyses of antimicrobial spectra showed that LyJH307 lysed Lancefield groups D (S. bovis group and Enterococcus faecalis) and H (S. sanguinis) bacteria. Thus, LyJH307 might help to prevent acute ruminal acidosis.

Much research on animal feed has focused on finding alternative feed ingredients that can replace conventional ones (e.g., grains and beans) to reduce feed costs. The objective of this study was to evaluate the economic, as well as nutritional value of spent coffee grounds (SCG) and Japanese mugwort (Artemisia princeps) residues (APR) as alternative feed ingredients for ruminants. We also investigated whether pre-fermentation using Lactobacillus spp. was a feasible way to increase the feed value of these by-products. Chemical analyses and an in vitro study were conducted for SCG, APR, and their pre-fermented forms. All the experimental diets for in vitro ruminal fermentation were formulated to contain a similar composition of crude protein, neutral detergent fiber and total digestible nutrients at 1x maintenance feed intake based on the dairy National Research Council (NRC). The control diet was composed of ryegrass, corn, soybean meal, whereas the treatments consisted of SCG, SCG fermented with Lactobacillus spp. (FSCG), APR, and its fermented form (FAPR). The treatment diets replaced 100 g/kg dry matter (DM) of the feed ingredients in the control. Costs were lower for the all treatments, except FAPR, than that of the control. After 24-h incubation, the NDF digestibility of the diets containing SCG and its fermented form were significantly lower than those of the other diets (P < 0.01); pre-fermentation tended to increase NDF digestibility (P = 0.07), especially for APR. Supplementation of SCG significantly decreased total gas production (ml/g DM) after 24-h fermentation in comparison with the control (P < 0.05); however, there were no significant differences between the control and the SCG or the APR diets in total gas production, as expressed per Korean Won (KRW). Diets supplemented with SCG or FSCG tended to have a higher total volatile fatty acid (VFA) concentration, expressed as per KRW, compared with the control (P = 0.06). Conversely, the fermentation process of SCG and APR significantly decreased total gas production and VFA production as expressed per KRW (P < 0.05). Because of their nutrient composition and relatively lower cost, we concluded that SCG and APR could be used as alternative feed sources, replacing conventional feed ingredients. However, pre-fermentation of agricultural by-products, such as SCG and APR, may be inappropriate for improving their nutritive considering the increase in production costs.